Optical electrode analyte detection system and methods of use

Abstract

The present disclosure provides, in part, devices and compositions comprising an optochemical sensor (optode) and optode spin disk for detecting one or more analytes in a biological sample, and methods of using the same.

Description

OPTICAL ELECTRODE ANALYTE DETECTION SYSTEM AND METHODS OF USEFIELD

[0001] The present disclosure relates to, in part, devices and compositions comprising an optochemical sensor (optode) for detecting one or more analytes in a biological sample and methods of using the same.CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of and priority to U.S. Provisional Application No. 63 / 528,225, filed July 21, 2023, the entire contents of which are incorporated by reference herein.BACKGROUND

[0003] Ionophore-based optochemical sensors (IBOS), also referred to as “optodes,” are devices which have a chemical sensor that can interact with an analyte to initiate an optically accessible change in the sensing material. Typically, this can be accomplished with an indicator dye that changes its spectral properties (e.g., absorbance and / or fluorescence) upon contact with and binding of an analyte from a sample. A salient example of such a sensor is a pH indicator, where the pH of a sample influences the protonation degree of the indicator dye, which determines the color change of the sensor. Another example is an optical oxygen sensor, where the luminescence intensity and decay time are influenced by the excited-state quenching with an analyte. Indirect sensing schema can also be used, where a receptor element does not change its optical properties upon contact with an analyte, but rather a reporter sensor initiates the optical change.

[0004] Compared with their electrochemical counterparts, optochemical sensors share important advantages regarding their applications and sensor formats. Optical sensors are typically easy to miniaturize and cost less to manufacture. They also allow signal readout without physical contact ( / .e., remote sensing) and enable researchers using optical imaging techniques to obtain spatially resolved analyte patterns. For IBOS in particular, ion imaging enables sophisticated microelectrode arrays to be replaced with planar optodes or particle suspensions to achieve two-and three-dimensional analyte concentration maps.

[0005] IBOS are typically based on a shared operating principle: selective extraction of the target analyte into a lipophilic liquid membrane introduces either i) coextraction of an oppositely charged reference ion or ii) release of a reference ion with the same charge. Accordingly, coextraction is most often found for anion sensors, and ion exchange is more prominent for cation-selective IBOS. Lipophilic ionic sites in theform of ion-exchanger salts are often added to compensate for the charges of the active compounds to obey the charge balance condition inside the sensing phase.

[0006] Although optochemical sensors have been successfully achieved for oxygen and pH detection, few successful results have been published using optode devices for other ions, or combinations of ions. There are several reasons for the observed dearth of optodes. For example, there is a lack of available water- soluble chromogenic and fluorogenic indicators for various ions. Consequently, for imaging purposes, a dissolved indicator approach is often not feasible for some ions or for some sample types. Moreover, ion indicators suffer from modest selectivity and flexibility in terms of their dynamic range, where reliable signal over small physiological ranges is difficult to achieve.

[0007] There remains a need for optodes that can transmit reversible optical signal with a dynamic range that appropriately reflects analyte sensing over physiological concentration ranges for multiple ions. Additionally, there is an unmet need for implementing optodes into devices that permit higher throughput and more accessible analysis of biological samples.SUMMARY

[0008] Accordingly, the present disclosure provides, in part, compositions and methods of use involving an optochemical sensor (optode) for detecting one or more analytes in a biological sample comprising a matrix comprising one or more polymers and one or more plasticizers, where the matrix has disposed therein one or more chromoionophores, one or more ionophores, one or more ion exchangers, and where the optode is configured to generate at least one optical signal as a function of the detection of the one or more analytes by the one or more chromoionophores and the one or more analytes by the one or more ionophores.

[0009] In embodiments, the one or more polymers comprises one or more of polyvinyl chloride (PVC), polyethylene terephthalate (PET), sulfonated polyether ether ketone (SPEEK), cellulose acetate, and cellulose diacetate. In embodiments, the one or more polymers comprises polyvinyl chloride (PVC).

[0010] In embodiments, the one or more plasticizers comprises one or more of dioctyl sebacate (DOS), ortho-nitrophenyloctylether (NPOE), bis(1 -butylpentyl) adipate (BBPA), dioctyl phthalate (DEHP / DOP), dioctyl terephthalate (DEHT / DOPT), trioctyl trimellitate (TOTM), trioctyl phosphate (TOP), tritolyl phosphate (TCP), triphenyl phosphate (TPhP), 2-nitrophenyl dodecyl ether (NPDE), 2-nitrophenyl tetradecyl ether (NPTE), polycaprolactone (PCL), phthalate ester, dibutyl sebacate, citrate ester, triacetin, adipate, azelate, benzoate, ortho-phthalate, terephthalate, and trimellitate. In embodiments, the one or more plasticizerscomprises dioctyl sebacate (DOS), ortho-nitrophenyloctylether (NPOE), bis(1 -butylpentyl) adipate (BBPA), dioctyl phthalate (DEHP / DOP), dioctyl terephthalate (DEHT / DOPT), and trioctyl trimellitate (TOTM).

[0011] In embodiments, the one or more chromoionophores is configured to detect one or more monovalent cations or anions from the biological sample.

[0012] In embodiments, the one or more monovalent cations comprises H+. In embodiments, the one or more chromoionophores is configured to detect H+at a concentration range of at least about pH 6.8 to at least about pH 9. In embodiments, the one or more chromoionophores comprises one or more of nitron-8- hydroxyquinoline-5-sulfonate, 4,4'-bis[(N,N-dialkylamino)methyl]-azobenzene, azobenzene, o-cresolphtalein octadecyl ester, Cresol Red, Phenol Red, 1-(2-Pyridylazo)-2-naphthol (PAN), o-Cresolphthalein, alpha- Naphtholphthalein, Phenolphthalein, Thymol Blue, m-Cresol Purple, Turmeric (Curcumin), 4-[(2,6-dibromo- 4-nitrophenyl)azo]-2-octadecyloxy-1 -naphthol, octyl bis(2,4-dinitrophenyl)acetate, 4-nonadecylpyridine (ETH 1907), 5-Octadecanoyloxy-2-(4-nitrophenylazo)phenol (ETH 2412), octadecyl 2-(4- dipropylaminophenylazo)benzoate (ETH 2418), ETH 2430, 9-dimethylamino-5-[4-(16-butyl-2,14-dioxo-3,15- dioxaeicosyl)phenylimino]benzo[a]phenoxazine (ETH 2439), ETH 2458, ETH 2462, ETH 2468, ETH 2479, ETH 3531 , 3-octadecanoylimino-7-(diethylamino)-1,2-benzophenoxazine (ETH 5294), ETH 5348, 9- (diethylamino)-5-[(2-octyldecyl)imino]benzo[a]phenoxazine (ETH 5350), ETH 5355, 9-Dimethylamino-5-[4- (15-butyl-1,13-dioxo-2,14-dioxanonadecyl)phenylimino]benzo[a]phenoxazine(ETH 5418), 3',3",5',5"-Tetrabromophenolphthaleinethyl ester (TBPE), 1-Hydroxy-4-[4-(2- hydroxyethylsulfonyl)phenylazo]naphthalene-2-sulfonic acid potassium salt (GJM-541), ETH 7058, ETH 7061, and ETH 7075.

[0013] In embodiments, at least one of the one or more ionophores is configured to detect at least one monovalent cation. In embodiments, the one or more monovalent cations comprises one or more of potassium (K+) and sodium (Na+).

[0014] In embodiments, the one or more ionophores is configured to detect sodium (Na+) at a concentration range of at least about 80 mM to at least about 180 mM. In embodiments, the one or more ionophores is configured to detect potassium (K+) at a concentration range of at least about 2.0 mM to at least about 10 mM.

[0015] In embodiments, the one or more ionophores is configured to detect sodium (Na+) and comprises one or more of gramicidin A, laidlomycin, lasalocid, maduramicin, monensin, narasin, nigericin, nonactin, nystatin, salinomycin, valinomycin, dicyclohexano-18-crown-6, calix[4]arenecrown-4, potassiumtetrakis(p-chlorophenyl)borate (KTpCIPB), ortho-nitrophenyloctylether, sodium tetrakis(p- chlorophenyl)borate (NaTFPB), Sodium tetrakis[3,5-bis(trifluoromethyl)phenyl]borate (NaBARF), Potassium tetrakis[3,5-bis(trifluoromethyl)phenyl]borate (KTFPB), bis-2-ethylhexylsebacate, 2,7-di-tert-butyl-9,9- dimethyl-4,5-xanthenediamine, tridodecylmethylammonium chloride (TDDMACI), 1 ,8-naphthalenediol, bis(1 - butylpentyl)adipate (BBPA), crown ether ionophores, bis[(12-crown-4)methyl] dodecylmethylmalonate, di(o- methoxy)-stilbeno-24-crown-8, 15-octadecyloxymethyl-15- methy 1-16-crown-5, dibenzo-16-crown-5 nitrophenol, 4-tert-butylcalix[4]arene-tetraacetic acid tetraethyl ester, Sodium ionophore IV (Cas #: 172883-29-3), N,N'-dibenzyl-N,N'-diphenyl-1,2-phenylenedioxydiacetamide (ETH 157), N,N',N"-triheptyl-N,N',N"- trimethyl-4,4',4"-propylidynetris(3-oxabutyramide) (ETH 227), N,N,N',N'-tetracyclohexyl-1 ,2- phenylenedioxydiacetamide (ETH 2120), and ETH 4120.

[0016] In embodiments, the one or more ionophores is configured to detect potassium (K+) and comprises one or more of nonactin, monactin, valinomycin, nigericin, hydroxylated polyvinyl chloride (PVC- OH), dicyclohexano-18-crown-6, dibenzo-18-crown-6, benzo- 15-crown-5, dibenzo-30-crown-10, dinaphtho-30-crown-10, 4-nitrobenzo-18-crown-6, 4'-picrylamino-5'-nitrobenzo-18-crown-6, 2, 3-naphtho-15-crown-5, (hexanoyloxymethyl)benzo-l 5-crown-5, octadecanoyloxy-methyl-15-crown-5, benzo-15-crown-5 ether, bis[(benzo-15-crown-5)-4'-ylmethyl] pimelate, and 2-dodecyl-2-methyl-1,3-propanediyl bis[N-[5'-nitro(benzo- 15-crown-5)-4'-yl]carbamate] (BME-44).

[0017] In embodiments, at least one of the one or more ionophores is configured to detecta monovalent anion from the biological sample. In embodiments, the one or more monovalent anions comprises chloride (CI-). In embodiments, the one or more ionophores is configured to detect chloride (CI-) at a concentration range of at least about 60 mM to at least about 140 mM.

[0018] In embodiments, the one or more ionophores is configured to detect chloride (CI-) and comprises one or more of triphenyltin chloride, tributyltin chloride, trioctyltin chloride, macrocyclic tetramercury, dodecylbenzylmercury(ll) chloride, tricyclohexyltin chloride, triphenylgermanium chloride, phenylmercuric chloride, hafnocene dichloride, chloro(octaethylporphyrinato)indium-(lll), meso-Tetraphenylporphyrin manganese(lll)-chloride, 4,5-Bis-[N'-(butyl)thioureido]-2,7-di-tert-butyl-9,9-dimethylxanthene, ETH 2412, 4,5- Dimethyl-3,6-dioctyloxy-o-phenylene-bis(mercurytrifluoroacetate) (ETH 9009), and 3,6-didodecyloxy-4,5- dimethyl-o-phenylene-bis(mercury chloride) (ETH 9033).

[0019] In embodiments, the one or more ionophores and / or one or more chromoionophores comprises a lipophilic indicator dye.

[0020] In embodiments, the one or more ion exchangers comprises one or more of potassium tetrakis(p- chlorophenyl)borate (KTpCIPB), sodium tetrakis(p-chlorophenyl)borate (NaTFPB), Sodium tetrakis[3,5- bis(trifluoromethyl)phenyl]borate (NaBARF), Potassium tetrakis[3,5-bis(trifluoromethyl)phenyl]borate (KTFPB), Tridodecylmethylammonium chloride (TDDMACI). In embodiments, the one or more ion exchangers comprises a photo-regenerative ion-exchange ionophore configured to extract one or more analyte cations or anions and release the one or more analyte cations or anions as a function of exposure to one or more wavelengths of electromagnetic radiation from about 280 nm to about 1000 nm.

[0021] In embodiments, the optode is configured to provide the at least one optical signal as a function of detection of one or more of H+ (pH), potassium (K+), sodium (Na+), and chloride (CI-) within a range of a human physiological concentration.

[0022] In embodiments, the optode is configured to provide at least one optical signal as a function of H+ concentration (pH), optionally where the optical signal is calibrated to provide an optical signal as a function of the range of the human physiological concentration of at least about pH 6.8 to at least about pH 9.

[0023] In embodiments, the optode is configured to provide at least one optical signal as a function of potassium (K+) concentration, optionally where the optical signal is calibrated to provide an optical signal as a function of the range of the human physiological concentration of at least about 2.0 mM to at least about 10 mM.

[0024] In embodiments, the optode is configured to provide at least one optical signal as a function of sodium (Na+) concentration, optionally where the optical signal is calibrated to provide an optical signal as a function of the range of the human physiological concentration of at least about 80 mM to at least about 180 mM.

[0025] In embodiments, the optode is configured to provide at least one optical signal as a function of chloride (CI-) concentration, optionally where the optical signal is calibrated to provide an optical signal as a function of the range of the human physiological concentration of at least about 60 mM to at least about 140 mM.

[0026] In embodiments, the optode is not substantially selective for one or more ions of calcium, magnesium, lithium, rubidium, cesium, ammonium, beryllium, strontium, barium, carbon dioxide, carbonate, bicarbonate, oxygen, thiocyanate, nitrate, nitrite, hydroxide, hydrogen sulfide, phosphate, sulfide, sulfite,sulfate, perchlorate, arsenic, iodide, and transition metals, optionally including antimony, molybdenum, iron, lead, copper, zinc, chromium, silver, selenium, mercury, uranium, thallium, samarium, and cadmium.

[0027] In embodiments, the biological sample comprises one or more of blood, sera, plasma, bone marrow, lymph, saliva, sputum, mucus, respiratory or nasal secretion, oropharyngeal swab, nasopharyngeal swab, oral swab, ductal lavage, bronchoalveolar lavage, cerebrospinal fluid, skin swab, vaginal swab, gastric juice, ascites, peritoneal fluid, pleural fluid, gynecological fluids, pus, perspiration, tears, urine, and stool. In embodiments, the biological sample comprises blood.

[0028] In embodiments, the optode is reusable. In embodiments, the optode is configured to be washed with an inorganic solvent and detect one or more analytes from at least a second biological sample with less than about 0.01% loss in sensitivity, less than about 0.05% loss in sensitivity, less than about 0.1% loss in sensitivity, less than about 0.2% loss in sensitivity, less than about 0.3% loss in sensitivity, less than about 0.4% loss in sensitivity, less than about 0.5% loss in sensitivity, less than about 0.6% loss in sensitivity, less than about 0.7% loss in sensitivity, less than about 0.8% loss in sensitivity, less than about 0.9% loss in sensitivity, less than about 1 .0% loss in sensitivity, less than about 1 .5% loss in sensitivity, or less than about 2.0% loss in sensitivity.

[0029] In embodiments, the inorganic solvent is one or more of deionized water, phosphate-buffered saline (PBS), sodium chloride (NaCI), sodium hydroxide, tris base, HEPES, MOPS, PIPES, urea, guanidine hydrochloride, and detergent.

[0030] In embodiments, the at least one optical signal is detectable in a range of at least about 300 nm to at least about 800 nm. In embodiments, the optode is configured to generate at least one optical signal for detecting each analyte. In embodiments, the at least one optical signal comprises one or more of a fluorescence signal, colorimetric signal, luminescent signal, absorption signal, and reflectance signal.

[0031] In embodiments, the at least one optical signal comprises a signal detectable at about 340 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the at least one optical signal comprises a signal detectable at about 405 nm, optionally for detecting one or more of H+, K+, Na+, and Cl- . In embodiments, the at least one optical signal comprises a signal detectable at about 505 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the at least one optical signal comprises a signal detectable at about 545 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the at least one optical signal comprises a signal detectable at about 660 nm, optionally fordetecting one or more of H+, K+, Na+, and CI-. In embodiments, the at least one optical signal comprises a signal detectable at about 720 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

[0032] In embodiments, the matrix is flexible. In embodiments, the matrix is configured to be applied to a surface, optionally where the matrix has a uniform thickness of about or at least about 1 pm, about or at least about 5 pm, about or at least about 10 pm, about or at least about 50 pm, about or at least about 100 pm, about or at least about 500 pm, about or at least about 1 .0 mm, about or at least about 1 .5 mm, about or at least about 2.0 mm, about or at least about 2.5 mm, about or at least about 3.0 mm, about or at least about 3.5 mm, about or at least about 4.0 mm, or about or at least about 5.0 mm.

[0033] In embodiments, the surface is transparent to at least a portion of an electromagnetic spectrum from at least about 280 nm to at least about 1000 nm.

[0034] The present disclosure provides, in part, an optochemical sensor (optode) for detecting one or more analytes from a biological sample comprising a disk comprising one or more wells disposed within the disk, where the one or more wells comprises an optode, as described herein, is disposed on a surface within the well, an inlet port for providing the biological sample into the well, at least a portion of an outer surface of the well that is transparent to at least a portion of an electromagnetic spectrum from at least about 280 nm to at least about 1000 nm, where the disk and the one or more wells are configured to be spun and withstand a centrifugal force, and where the optode within the well is configured to generate at least one optical signal in the range of at least about 300 nm to at least about 800 nm as a function of binding one or more analytes in the biological sample.

[0035] In embodiments, the optode comprises a matrix comprising polyvinyl chloride (PVC) dioctyl sebacate (DOS), ortho-nitrophenyloctylether (NPOE), bis(1 -butylpentyl) adipate (BBPA), dioctyl phthalate (DEHP / DOP), dioctyl terephthalate (DEHT / DOPT), trioctyl trimellitate (TOTM), or a mixture thereof, and the matrix has disposed therein a chromoionophore that is proton-selective and configured to detect H+ concentration (pH) in a range of at least about pH 6.8 to at least about pH 9, a first ionophore that is potassium-selective and configured to detect potassium (K+) concentration in a range of at least about 2.0 mM to at least about 10 mM, a second ionophore that is sodium-selective and configured to detect sodium (Na+) concentration in a range of at least about 80 mM to at least about 180 mM, a third ionophore that is chloride-selective and configured to detect chloride (CI-) concentration in a range of at least about 60 mM to at least about 140 mM, and at least one ion exchanger.

[0036] In embodiments, the optode is not substantially selective for one or more ions of calcium, magnesium, lithium, rubidium, cesium, ammonium, beryllium, strontium, barium, carbon dioxide, carbonate, bicarbonate, oxygen, thiocyanate, nitrate, nitrite, hydroxide, hydrogen sulfide, phosphate, sulfide, sulfite, sulfate, perchlorate, arsenic, iodide, and transition metals, optionally including antimony, molybdenum, iron, lead, copper, zinc, chromium, silver, selenium, mercury, uranium, thallium, samarium, and cadmium.

[0037] In embodiments, the biological sample comprises one or more of blood, sera, plasma, bone marrow, lymph, saliva, sputum, mucus, respiratory or nasal secretion, oropharyngeal swab, nasopharyngeal swab, oral swab, ductal lavage, bronchoalveolar lavage, cerebrospinal fluid, skin swab, vaginal swab, gastric juice, ascites, peritoneal fluid, pleural fluid, gynecological fluids, pus, perspiration, tears, urine, and stool. In embodiments, the biological sample comprises blood.

[0038] In embodiments, the optode spin disk, or one or more layers thereof, is composed of plastic, polymethyl methacrylate, carbon fiber, and / or metal, optionally wherein the metal is aluminum and / or titanium. In embodiments, the disk further comprises a substrate to hold the optode in place, optionally wherein the disk comprises one or more of a structural support layer, inlet layer, channel layer, and capping layer, and further optionally wherein the optode in held place on a capping layer. In embodiments, the substrate and / or capping layer comprises one or more of polyethylene terephthalate (PET), polyethylene terephthalate glycol (PETG), polyurethane, polyethylene, polypropylene, and glass. In embodiments, the substrate is chemically resistant to one or more solvents, optionally including, cyclohexanone, N,N- dimethylformamide (DMF), acetonitrile, butanone, and dichloromethane.

[0039] In embodiments, the disk comprises at least about 1 well, at least about 2 wells; at least about 3 wells, at least about 4 wells, at least about 5 wells, at least about 6 wells, at least about 7 wells, at least about 8 wells, at least about 9 wells, at least about 10 wells, at least about 12 wells, at least about 14 wells, at least about 16 wells, at least about 18 wells, at least about 20 wells, at least about 22 wells, at least about 24 wells, at least about 26 wells, at least about 28 wells, at least about 30 wells, at least about 35 wells, or at least about 40 wells.

[0040] In embodiments, the disk is configured to be spun at a rate of at least about 100 revolutions per minute (RPM), at least about 200 RPM, at least about 300 RPM, at least about 400 RPM, at least about 500 RPM, at least about 600 RPM, at least about 700 RPM, at least about 800 RPM, at least about 900 RPM, at least about 1000 RPM, at least about 1200 RPM, at least about 1400 RPM, at least about 1600 RPM, at least about 1800 RPM, at least about 2000 RPM, at least about 2200 RPM, at least about 2400 RPM, at leastabout 2600 RPM, at least about 2800 RPM, at least about 2900 RPM, at least about 3000 RPM, at least about 3500 RPM, at least about 4000 RPM, at least about 4500 RPM, or at least about 5000 RPM.

[0041] In embodiments, the disk is configured to withstand a centrifugal force of at least about 100 x g, at least about 200 x g, at least about 300 x g, at least about 400 x g, at least about 500 x g, at least about 600 x g, at least about 700 x g, at least about 800 x g, at least about 900 x g, at least about 1000 x g, at least about 1200 x g, at least about 1400 x g, at least about 1600 x g, at least about 1800 x g, at least about 2000 x g, at least about 2200 x g, at least about 2400 x g, at least about 2600 x g, at least about 2800 x g, at least about 3000 x g, at least about 3500 x g, at least about 4000 x g, at least about 4500 x g, or at least about 5000 x g.

[0042] In embodiments, the well comprises a total well volume of at least about 5 pL, at least about 10 pL, at least about 15 pL, at least about 20 pL, at least about 25 pL, at least about 30 pL, at least about 35 pL, at least about 40 pL, at least about 45 pL, at least about 50 pL, at least about 55 pL, at least about 60 pL, at least about 65 pL, at least about 70 pL, at least about 75 pL, at least about 80 pL, at least about 85 pL, at least about 90 pL, at least about 95 pL, at least about 100 pL, at least about 110 pL, at least about 120 pL, at least about 130 pL, at least about 140 pL, at least about 150 pL, at least about 160 pL, at least about 170 pL, at least about 180 pL, at least about 190 pL, or at least about 200 pL. In embodiments, the well comprises a well width of at least about 1 mm, at least about 2 mm, at least about 3 mm, at least about 4 mm, at least about 5 mm, at least about 6 mm, at least about 7 mm, at least about 8 mm, at least about 9 mm, or at least about 10 mm.

[0043] In embodiments, the optode disposed on the surface within the well comprises a surface area of at least about 1 mm2, at least about 2 mm2, at least about 3 mm2, at least about 4 mm2, at least about 5 mm2, at least about 6 mm2, at least about 7 mm2, at least about 8 mm2, at least about 9 mm2, at least about 10 mm2, at least about 15 mm2, at least about 20 mm2, at least about 25 mm2, at least about 50 mm2, at least about 100 mm2, at least about 200 mm2, at least about 300 mm2, at least about 400 mm2, at least about 500 mm2, and optionally where the optode has a uniform thickness of about or at least about 1 pm, about or at least about 5 pm, about or at least about 10 pm, about or at least about 50 pm, about or at least about 100 pm, about or at least about 500 pm, about or at least about 1 .0 mm, about or at least about 1 .5 mm, about or at least about 2.0 mm, about or at least about 2.5 mm, about or at least about 3.0 mm, about or at least about 3.5 mm, about or at least about 4.0 mm, or about or at least about 5.0 mm.

[0044] In embodiments, the well comprises a overflow well, optionally where the overflow well has a total volume of at least about 3 pL, at least about 5 pL, at least about 10 pL, at least about 15 pL, at least about 20 pL, at least about 25 pL, at least about 30 pL, at least about 35 pL, at least about 40 pL, at least about 45 pL, at least about 50 pL, at least about 55 pL, at least about 60 pL, at least about 65 pL, at least about 70 pL, at least about 75 pL, at least about 80 pL, at least about 85 pL, at least about 90 pL, at least about 95 pL, or at least about 100 pL.

[0045] In embodiments, the inlet port comprises a diameter of at least about 0.5 mm, at least about 0.6 mm, at least about 0.7 mm, at least about 0.8 mm, at least about 0.9 mm, at least about 1 mm, at least about 1.5 mm, or at least about 2 mm. In embodiments, the well comprises an outlet port, optionally where the outlet port comprises a diameter of at least about 0.5 mm, at least about 0.6 mm, at least about 0.7 mm, at least about 0.8 mm, at least about 0.9 mm, at least about 1 mm, at least about 1.5 mm, or at least about 2 mm.

[0046] In embodiments, the well comprises a covermask, optionally where the covermask comprises a diameter of at least about of at least about 0.5 mm, at least about 1 mm, at least about 1 .5 mm, or at least about 2 mm, or at least about 2.5 mm, or at least about 3.0 mm, or at least about 3.5 mm, or at least about 4 mm, or at least about 4.5 mm, or at least about 5 mm. In embodiments, the portion of the outer surface of the well that is transparent is comprised of one or more of glass, quartz, plastic, polystyrene, polyethylene terephthalate, polycarbonate, polymethyl methacrylate (acrylic), polyethylene, polyurethane, polypropylene, thermoplastic elastomers (TPE), acrylonitrile butadiene styrene (ABS), epoxy, cyclic olefin copolymer (COC), cyclic olefin polymer (COP), poly dimethyl siloxane (PDMS), poly ether ester ketone (PEEK), polyetherimide (ULTEM), and nylon.

[0047] In embodiments, the optode is configured to be washed with a solvent, originally where the solvent comprises one or more of deionized water, phosphate-buffered saline (PBS), sodium chloride (NaCI), sodium hydroxide, tris base, HEPES, MOPS, PIPES, urea, guanidine hydrochloride, and detergent.

[0048] In embodiments, the optode spin disk has a fluid circuit embedded within the disk that fluidically connects to the one or more wells by a channel or tubing. In embodiments, the fluid circuit fluidically connects the one or more wells to one or more pH fix chambers, pH priming circuits, bead mixing chambers, overflow chambers, separator chambers, siphons, and air volume ballasts. In embodiments, the one or more pH fix chambers are configured to fix the ionic content of the biological sample to prevent deviation in concentrations of the one or more analytes during an optode sensing phase. In embodiments, t the fluid circuit fluidicallyconnects the one or more pH fix chambers to one or more separator chambers and / or siphons. In embodiments, the one or more separator chambers is configured to separate cells, cell debris, and / or plasma from the biological sample during and / or after a centrifugal force is applied to the disk. In embodiments, the fluid circuit fluidically connects the one or more separator chambers to one or more sample reservoirs and / or siphons. In embodiments, the one or more sample reservoirs is configured to hold the biological sample and comprises a volume of about or at least about 10 pL, about or at least about 20 pL, about or at least about 50 pL, about or at least about 0.1 mL, about or at least about 0.2 mL, about or at least about 0.3 mL, about or at least about 0.4 mL, about or at least about 0.5 mL, about or at least about 0.6 mL, about or at least about 0.7 mL, about or at least about 0.8 mL, about or at least about 0.9 mL, or about or at least about 1 .0 mL.

[0049] In embodiments, the one or more wells comprise the inlet port and an outlet port and the one or more wells are configured to be connected to a microfluidic platform. In embodiments, the microfluidic platform is in fluid connection with the inlet port and the outlet port of the one or more wells. In embodiments, the microfluidic platform comprises at least one value, a section of a channel or tubing, and a fluid pumping mechanism.

[0050] In aspects, the present disclosure includes, in embodiments, a method for detecting one or more analytes in a biological sample comprising providing the optochemical sensor (optode), as described herein, dispensing one or more aliquots of the biological sample into one or more wells of the optode, applying a centrifugal force to the optode, detecting at least one optical signal from the one or more wells in the range of at least about 300 nm to at least about 800 nm as a function of binding to one or more analytes in the biological sample.

[0051] In embodiments, the biological sample comprises one or more of blood, sera, plasma, bone marrow, lymph, saliva, sputum, mucus, respiratory or nasal secretion, oropharyngeal swab, nasopharyngeal swab, oral swab, ductal lavage, bronchoalveolar lavage, cerebrospinal fluid, skin swab, vaginal swab, gastric juice, ascites, peritoneal fluid, pleural fluid, gynecological fluids, pus, perspiration, tears, urine, and stool. In embodiments, the biological sample comprises blood.

[0052] In embodiments, methods herein comprise dispensing one or more aliquots of the biological sample into one or more sample reservoirs on the optode, optionally where the one or more aliquots comprises a volume of a total volume of at least about 3 pL, at least about 5 pL, at least about 10 pL, at least about 15 pL, at least about 20 pL, at least about 25 pL, at least about 30 pL, at least about 35 pL, at leastabout 40 pL, at least about 45 pL, at least about 50 pL, at least about 55 pL, at least about 60 pL, at least about 65 pL, at least about 70 pL, at least about 75 pL, at least about 80 pL, at least about 85 pL, at least about 90 pL, at least about 95 pL, at least about 100 pL, at least about 110 pL, at least about 120 pL, at least about 130 pL, at least about 140 pL, at least about 150 pL, at least about 160 pL, at least about 170 pL, at least about 180 pL, at least about 190 pL, at least about 0.2 mL, about or at least about 0.3 mL, about or at least about 0.4 mL, about or at least about 0.5 mL, about or at least about 0.6 mL, about or at least about 0.7 mL, about or at least about 0.8 mL, about or at least about 0.9 mL, or about or at least about 1 .0 mL.

[0053] In embodiments, applying the centrifugal force comprises spinning the optode at a rate of at least about 100 RPM, at least about 200 RPM, at least about 300 RPM, at least about 400 RPM, at least about 500 RPM, at least about 600 RPM, at least about 700 RPM, at least about 800 RPM, at least about 900 RPM, at least about 1000 RPM, at least about 1200 RPM, at least about 1400 RPM, at least about 1600 RPM, at least about 1800 RPM, at least about 2000 RPM, at least about 2200 RPM, at least about 2400 RPM, at least about 2600 RPM, at least about 2800 RPM, at least about 2900 RPM, at least about 3000 RPM, at least about 3500 RPM, at least about 4000 RPM, at least about 4500 RPM, or at least about 5000 RPM. In embodiments, applying the centrifugal force comprises a centrifugal force of at least about 100 x g, at least about 200 x g, at least about 300 x g, at least about 400 x g, at least about 500 x g, at least about 600 x g, at least about 700 x g, at least about 800 x g, at least about 900 x g, at least about 1000 x g, at least about 1200 x g, at least about 1400 x g, at least about 1600 x g, at least about 1800 x g, at least about 2000 x g, at least about 2200 x g, at least about 2400 x g, at least about 2600 x g, at least about 2800 x g, at least about 3000 x g, at least about 3500 x g, at least about 4000 x g, at least about 4500 x g, or at least about 5000 x g-

[0054] In embodiments, methods herein further comprise separating cells and / or cell debris from the biological sample during and / or after the centrifugal force is applied to the optode.

[0055] In embodiments, methods herein further comprise fixing the ionic content of the biological sample to prevent deviation in concentrations of the one or more analytes during the detecting.

[0056] In embodiments, detecting the at least one optical signal comprises measuring one or more of a fluorescence signal, colorimetric signal, luminescent signal, absorption signal, and reflectance signal. In embodiments, measuring comprises using one or more of a sensor monitor, absorbance spectrometer, reflectometric photometer, LED photometer, fluorimeter, and luminometer.

[0057] In embodiments, the at least one optical signal comprises a signal detectable at about 340 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the at least one optical signal comprises a signal detectable at about 405 nm, optionally for detecting one or more of H+, K+, Na+, and Cl- . In embodiments, the at least one optical signal comprises a signal detectable at about 505 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the at least one optical signal comprises a signal detectable at about 545 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the at least one optical signal comprises a signal detectable at about 660 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the at least one optical signal comprises a signal detectable at about 720 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

[0058] In embodiments, the detecting is not substantially selective for one or more ions of calcium, magnesium, lithium, rubidium, cesium, ammonium, beryllium, strontium, barium, carbon dioxide, carbonate, bicarbonate, oxygen, thiocyanate, nitrate, nitrite, hydroxide, hydrogen sulfide, phosphate, sulfide, sulfite, sulfate, perchlorate, arsenic, iodide, and transition metals, optionally including antimony, molybdenum, iron, lead, copper, zinc, chromium, silver, selenium, mercury, uranium, thallium, samarium, and cadmium.

[0059] In embodiments, the method further comprises preparing the optode with one or more organic solvents comprising cyclohexanone, N,N-dimethylformamide (DMF), acetonitrile, butanone, and dichloromethane.

[0060] In embodiments, the method further comprises washing the one or more wells with an inorganic solvent, optionally where the inorganic solvent comprise one or more of deionized water, phosphate-buffered saline (PBS), sodium chloride (NaCI), sodium hydroxide, tris base, HEPES, MOPS, PIPES, urea, guanidine hydrochloride, and detergent.

[0061] In embodiments, dispensing the one or more aliquots of the biological sample and / or washing the one or more wells comprises using a microfluidic platform, where the microfluidic platform is in fluid connection with the one or more wells.

[0062] In embodiments, the method comprises a reusable optode which includes dispensing at least a second biological sample into the one or more wells of the optode. In embodiments, the method comprises detecting at least one optical signal from the one or more wells after dispensing at least a second biological sample, and where the detection does not result in a substantially significant reduction in sensitivity. In embodiments, the substantially significant reduction in sensitivity is less than about 0.01% loss in sensitivity, less than about 0.05% loss in sensitivity, less than about 0.1% loss in sensitivity, less than about 0.2% lossin sensitivity, less than about 0.3% loss in sensitivity, less than about 0.4% loss in sensitivity, less than about 0.5% loss in sensitivity, less than about 0.6% loss in sensitivity, less than about 0.7% loss in sensitivity, less than about 0.8% loss in sensitivity, less than about 0.9% loss in sensitivity, less than about 1.0% loss in sensitivity, less than about 1.5% loss in sensitivity, or less than about 2.0% loss in sensitivity.

[0063] In embodiments, the method comprises two or more cycles of said washing, dispensing, and detecting one or more analytes from an additional biological sample.

[0064] In aspects, the present disclosure provides, in embodiments, a method of diagnosing one or more medical conditions and / or disorders by performing the methods described herein and diagnosing the one or more medical conditions and / or disorders as a function of detecting one or more of pH (H+), sodium (Na+) concentration, potassium (K+) concentration, and chloride (CI-) concentration. In embodiments, the one or more medical conditions and / or disorders comprises a medical condition and / or disorder associated with a blood concentration of one or more of H+, Na+, K+, and CI-.

[0065] In embodiments, the one or more medical conditions and / or disorders comprises one or more of acidosis, alkalosis, hypokalemia, hyperkalemia, hyponatremia, hypernatremia, hyperchloremia, hypochloremia, Cushing syndrome, Barter syndrome, kidney disease, heart failure, heart disease, albuminuria, hyperaldosteronism, ketoacidosis, lactic acidosis, renal tubular acidosis, diabetes, dehydration, hyperglycemia, hyperparathyroidism, effects of a prescribed therapeutic agent, including effects of diuretics (e.g., hydrochlorothiazide, furosemide, torsemide, and indapamide) and / or effects carbonic anhydrase inhibitors (e.g., glaucoma treatment), ethylene glycol poisoning, methanol poisoning, salicylate toxicity (e.g., aspirin overdose), ureteral diversion, syndrome of inappropriate antidiuretic hormone secretion (SIADH), and cancer (e.g., endocrine neoplasia, kidney cancer, liver cancer).

[0066] In embodiments, methods herein further comprise prescribing one or more therapeutic agents as a function of the detecting. In embodiments, the one or more therapeutic agents comprises a therapeutic agent associated with a blood concentration of one or more of H+, Na+, K+, and CI-. In embodiments, the one or more therapeutic agents comprises one or more of electrolyte supplementation (e.g., potassium salt, chloride salt, sodium salt), sodium zirconium cyclosilicate (LOKELMA), diuretics (e.g., loop diuretics, amiloride triamterene), hypertonic or hypotonic saline, vasopressin receptor antagonists, demeclocycline therapy, bicarbonate therapy, hemodialysis, continuous renal replacement therapy (CRRT), alkalinizing agents, mineralocorticoids, vitamin D supplementation, carbonic anhydrase inhibitors, acetazolamide, insulin, ketoconazole, mitotane (LYSODREN), metyrapone (METOPIRONE), Mifepristone (KORLYM,MIFEPREX), angiotensin-converting enzyme (ACE) inhibitors, angiotensin receptor blockade (ARB), aldosterone antagonists, spironolactone, indomethacin, ibuprofen, non-steroidal anti-inflammatory drugs (NSAIDs), growth hormone (GH), dapagliflozin (FARXIGA), blood thinners, vasodilators, enalapril (VASOTEC, EPANED), lisinopril (ZESTRIL, QBRELIS), captopril, and alcohol dehydrogenase enzyme (ADH) antagonists (e.g., Fomepizole).DESCRIPTION OF THE DRAWINGS

[0067] Fig. 1 depicts an illustrative, non-limiting diagrammatic representation of an analyte-specific optode. The optode changes one or more spectral properties as a function of changes in ion concentration for its cognate ion.

[0068] Figs. 2A-2E depict illustrative, non-limiting schematic representations of ion-exchange and coextraction processes. Fig. 2A shows triggered upon increase of the analyte ion concentration, where R- indicates a negatively charged ion exchanger. Calcium ionophore molecules (L) stabilize the calcium inside the lipophilic liquid / polymeric film (sensorfilm). Electroneutrality is maintained by expelling an equivalent number of protons, which triggers a detectable change in the color of the reporter dye (Ind). Fig. 2B shows an illustrative schematic of an optode with plasticized PVC used for ion-exchange with sodium (Na) and potassium (K). Fig. 2C shows an illustrative schematic of an optode with plasticized PVC used for ionexchange and co-extraction with chloride (Cl). Fig. 2D shows an illustrative schematic of a cation-selective optode with ionophores and chromophores embedded in a polymer-based membrane, where the ionophore captures the cation of interest (e.g., sodium, (Na+) or potassium (K+)) which alters the charge across the membrane. This in turn triggers a chromophore to lose a proton (H+) to balance charge neutrality, resulting in a measurable color change used to calculate the concentration of the electrolyte in the sample. Fig. 2E shows an illustrative schematic of an anion-selective optode with ionophores and chromophores embedded in a polymer-based membrane, where the ionophore captures the anion of interest (e.g., chloride (Cl-)) which alters the charge across the membrane. This in turn triggers a chromophore to lose a proton (H+) to balance charge neutrality, resulting in a measurable color change used to calculate the concentration of the electrolyte in the sample.

[0069] Fig. 3 depicts an illustrative, non-limiting schematic of three detection principles: (a) equilibrium based, (b) dynamic change, and (c) exhaustive exchange. Abbreviations: c, concentration; Ind, lipophilic pH indicator; L, ionophore; MD, any cation; S, signal.

[0070] Fig. 4 depicts an illustrative, non-limiting graphical representation of a theoretical response equation of an optode sensor.

[0071] Fig. 5 depicts an illustrative, non-limiting graphical representation of a theoretical prediction of an optode response for potassium, showing predicted exponential optode signal change in the physiological range for potassium (K) isotopes (top), and color changes in optode in the visual spectrum as a function of ionophore and chromoionophore ratio (bottom).

[0072] Figs. 6A-6B depict a graphical representation of a measurement procedure for K+, pH, Na+, and CI-. The analysis method for protonation degree: 1 - a = Aprot / (Aprot + Adeprot), where the analysis done at 405 nm, 505 nm, 660 nm, and 720 nm over the spectra of 400 nm to 1000 nm. Fig. 6A shows the calibration analysis where 720 nm absorbance is used as a baseline for subtracting both protonation peak (660 nm) and deprotonation peak (505 nm); top curve is 4 mM buffer; bottom curve is normalized value. Fig. 6B shows a more accurate method for plasma analysis, where a 2-point calibration using 720 nm as the baseline for the protonation peak and 405 nm absorbance is used as a baseline for the deprotonation peak; top curve and bottom curve are two replicates of normalized curves.

[0073] Fig. 7 depicts a diagrammatic representation of an optode membrane fabrication.

[0074] Fig. 8 depicts a graphical representation of a potassium-selective optode signal. Response was demonstrated across a human blood physiological range (2 mM to 8 mM), where calibration, linearity of optical signal, and precision between replicates was demonstrated in buffer and blood plasma samples. Sample volume was reduced from about 50 pL to about 8 pL.

[0075] Figs. 9A-9C depict a graphical representation of pH, Na+, and Cl- detection from buffer and / or blood samples. Analysis method for protonation degree: 1 - a = Aprot / (Aprot + Adeprot), where analysis was performed at 505 nm, 660 nm, and 720 nm. Fig. 9A depicts pH detection over the physiological range of pH 6.8 to pH 9. Fig. 9B depicts sodium ion detection over the physiological concentration range of 80 mM to 180 mM. Fig. 9C depicts chloride ion detection over the physiological concentration range of 0 mM to 140 mM.

[0076] Fig. 10 depicts a diagrammatic representation of an optochemical sensor (optode) spin disc.

[0077] Fig. 11 depicts an illustrative, non-limiting diagrammatic representation of an exemplary fluid circuit-embedded optochemical sensor (optode) spin disc.

[0078] Fig. 12 depicts an illustrative, non-limiting diagrammatic representation of an exemplary optode well.

[0079] Fig. 13 depicts an illustrative, non-limiting diagrammatic representation of sample flow direction from near the center of the optode spin disk, outward to the optode wells due to the force of centrifugation.

[0080] Fig. 14 depicts an illustrative, non-limiting diagrammatic representation of an optode spin disk construction, with top and bottom structural supports, an inlet layer (e.g., .5 mm thickness, comprised of polyethylene terephthalate (PET)), a channel layer (e.g., comprised of PET and two layers of pressure sensitive adhesive (PSA)), and a capping layer (e.g., composed of PET with optodes cast thereon).

[0081] Fig. 15 depicts an illustrative, non-limiting diagrammatic representation of a fluid channel of an optode spin disk for plasma, where 1) shows a reservoir for loading and directing plasma, 2) shows a bead mixing chamber, 3) shows a pH well, 4) represents sample wells where optodes are located, and 5) shows an overflow chamber.

[0082] Fig. 16 depicts an illustrative, non-limiting diagrammatic representation of a fluid channel of an optode spin disk for whole blood, where 1) shows a sample loading chamber, 2) shows a plasma separator chamber, 3) shows a chamber for plasma overflow, 4) shows a bead mixing chamber, 5) shows a pH priming circuit, and 6) depicts the optode wells.

[0083] Fig. 17 depicts an illustrative, non-limiting diagrammatic representation of a fluid channel plasma separator within an optode spin disk, where 1) shows a sample loading chamber, 2) shows an air volume ballast and siphon to prevent backflow, 3) shows a chamber for blood cells to be removed, 4) shows a clean plasma reservoir, and 5) shows a priming siphon.

[0084] Fig. 18 depicts an illustrative, non-limiting diagrammatic representation of a fluid channel plasma separator within an optode spin disk, where 1) shows bead chambers, 2) shows a pH aliquot, 3) shows an air volume ballast and siphon to prevent backflow, and 4) shows an optional pH well with an optode located therein to measure pH. While circles denote Lyo beads, and the darker circle in “4” represents the pH optode.

[0085] Fig. 19 depicts an illustrative, non-limiting diagrammatic representation of a fluid channel design within an optode spin disk for minimizing sample volume. The white circle near the middle of the fluid circuit represents a 1x Lyo bead, circles near the bottom are optode apertures located within the well.

[0086] Fig. 20 depicts a graphical representation of difference in spectra signal intensity at 505 nm and 660 nm for varying concentrations of NaCI (shown is signal for 0.3 mM to 100 mM NaCI).

[0087] Fig. 21 depicts imaging of optodes cast within a capping layer (e.g., resembling the fluid flow depicted in Fig. 13), with visible color differences in the optodes between those on the left and right, detecting small differences in buffer potassium ion concentration between 6.7 mM (left) and 2.7 mM (right).

[0088] Figs. 22A and 22B depicts graphical representation of data analysis for determining ion concentration from optode readings using spectra signal intensity at 405 nm, 505 nm, 545 nm, 660 nm, and 720 nm. Fig. 22A shows an analysis using a 50% maxima analysis. Fig. 22B shows an analysis without a 50% maxima analysis. Points placed in the top graph of both Fig. 22A and Fig. 22B illustrate peak locations identified in each wavelength (e.g., 405 nm, 505 nm, 545 nm, 660 nm, and 720 nm).

[0089] Figs. 23A and 23B depicts a non-limiting illustrative and graphical representation, respectively, of optode centration analysis for reading optode signal from a spin disk. Fig. 23A shows that an optode with a diameter (0) of 2.2 mm may not be centered in the well having a 3 mm diameter, where the beam path for reading the optode in the well having a diameter of 1.6 mm passing through the well’s center may not pass through the center of the optode. Fig. 23B shows an analysis for correcting for optode position within the well for reading the spin disk, e.g., with representative spectra signal intensity at 505 nm (bottom curve), 660 nm (topmost curve), and 720 nm shown (middle curve), and the optode true center depicted by the black arrows.

[0090] Fig. 24 depicts a graphical representation of data analysis for determining ion concentration from optode readings using spectra signal intensity at 505 nm (topmost curve), 660 nm (bottom curve), and 720 nm (middle curve).

[0091] Fig. 25 depicts a graphical representation of calibration curves of optode signal which demonstrates sensitive ion detection within a physiological range, pH 7.3 (topmost curve), pH 7.5 (middle curve), pH 7.6 (bottom curve).DETAILED DESCRIPTION

[0092] The present disclosure provides, in part, compositions of optochemical sensors, referred to herein as “optodes,” which can be integrated into a device for analyzing the analyte content of biological samples. In embodiments, the optode is an ion-selective matrix which is composed of plasticized polyvinyl chloride ( / .e., PVC + dioctyl sebacate (DOS)) impregnated with distinct ionophores (e.g., one to detect K+, one to detect Na+, and one to detect CI-), a proton chromoionophore ( / .e., to detect H+ (pH)), and an ionexchanger (e.g., a lipophilic anionic additive). In embodiments, the optode can be cast onto a surface of one or more sample wells of a spin-on device, which can be a plate or disk. This plate or disk, in embodiments,can be composed of plastic, carbon fiber, or metal (e.g., aluminum or titanium), having one or more bores for screwing onto or otherwise securing the plate or disk onto a motor shaft for rotation, like a centrifuge rotor onto a centrifuge. The optode spin disk, in embodiments, is constructed to withstand centrifugal forces in the range of at least about 100 x g to at least about 5000 x g, where the biological sample can be “incubated” during a brief spin period (e.g., on the order of minutes). In embodiments, the optode matrix can generate one or more optical signals as a function of detecting one or more analytes (e.g., H+, K+, Na+, and / or CI-), where the detection is calibrated to measure in the appropriate physiological range.

[0093] In embodiments, the optode spin disk can also include a fluid circuit embedded therein which has one or more sample reservoirs, a separating chamber, e.g., to remove cells and debris from the biological sample, a sample fixing chamber, e.g., to fix the pH of the biological sample.

[0094] Additionally, in embodiments, each well of the optode spin disk can have an inlet port and / or an outlet port. In embodiments, the inlet / outlet ports can be used for connecting microfluidic devices, such as valves, tubing, and fluid propulsion (e.g., peristaltic pump). The spin disk, in embodiments, can have biological sample (e.g., blood, sera, plasma, etc.) pumped through a microfluidic device into each individual well. In embodiments, the wells can be disconnected after dispensing the sample, and then centrifugation and detection can be performed.

[0095] In embodiments, described herein, in part, are methods of using the optode spin disk to evaluate the presence of and / or concentration of one or more analytes in a biological sample. In embodiments, the method is an assay that is performed by dispensing a biological sample into one or more wells of the optode spin disk, applying a centrifugal force to the optode spin disk as an incubation step, and then detecting the change in one or more optical signals (e.g., by analyzing the wells for the presence of a change in optical property at about 300 nm to about 800 nm). In non-limiting embodiments, the biological sample is blood, and the assay is performed by dispensing the blood into one or more sample reservoirs of an optode spin disk, applying a centrifugal force to the optode spin disk, separating whole cells and debris from the blood sample, fixing the pH of the blood sample, dispensing the cell-depleted, pH-fixed blood sample into one or more wells, each well having one or more optodes (e.g., a sodium-selective optode, a potassium-selective optode, a chloride-selective optode, and a pH-selective optode) disposed therein, and then measuring a change in one or more optical signals from the one or more optodes as a function of binding and detecting its cognate ion in the blood sample.Optochemical Sensors (Optodes)

[0096] In aspects, provided herein, in embodiments, are optochemical sensors (optodes) for detecting one or more analytes in a biological sample, where the optode includes a matrix with one or more polymers and one or more plasticizers, where the matrix has disposed therein one or more chromoionophores, one or more ionophores, one or more ion exchangers, and where the optode is configured to generate at least one optical signal as a function of the detection of one or more analytes by the one or more chromoionophores and the detection of one or more analytes by the one or more ionophores. In embodiments, each optode can exhibit colorimetric changes as a function of concentration of its cognate ion in a sample (e.g., as illustrated in Fig. 1).

[0097] The one or more polymers, in embodiments, can include polyvinyl chloride (PVC), polyethylene terephthalate (PET), sulfonated polyether ether ketone (SPEEK), polysiloxane, poly-bisphenol A carbonate, cellulose acetate, and / or cellulose diacetate. In embodiments where cellulose diacetate is used, for example where the optode is to be affixed to a surface, a thin film such as CLARIFOIL (e.g., about 14 pm to about 520 pm thick cellulose diacetate film) can be used as the polymer for the optode matrix. In embodiments, the optode matrix can include a mixture of about 60% polysiloxane and about 40% poly-bisphenol A carbonate. In embodiments, the optode matrix includes polyvinyl chloride (PVC). In embodiments, the optode matrix includes plasticized PVC.

[0098] The one or more plasticizers, in embodiments, include dioctyl sebacate (DOS, also referred to as Bis(2-ethylhexyl) sebacate), ortho-nitrophenyloctylether (NPOE), bis(1 -butylpentyl) adipate (BBPA), dioctyl phthalate (DEHP / DOP, also referred to as bis(2-ethylhexyl) phthalate, diethylhexyl phthalate), dioctyl terephthalate (DEHT / DOPT, also referred to as bis(2-ethylhexyl) terephthalate, di(ethylhexyl) terephthalate), trioctyl trimellitate (TOTM), trioctyl phosphate (TOP, also referred to as tris(2-ethylhexyl) phosphate), tritolyl phosphate (TCP, also referred to as tricresyl phosphate), triphenyl phosphate (TPhP), 2-nitrophenyl dodecyl ether (NPDE), 2-nitrophenyl tetradecyl ether (NPTE), polycaprolactone (PCL), phthalate ester, dibutyl sebacate, citrate ester, triacetin, adipate, azelate, benzoate, ortho-phthalate, terephthalate, and / or trimellitate. In embodiments, the one or more plasticizers includes dioctyl sebacate (DOS). In embodiments, the optode matrix includes PVC plasticized with DOS, NPOE, or a mixture of DOS and NPOE.

[0099] In embodiments, the optode matrix possess one or more features that aid its practical application, including the ability to adhere to a variety of substrates / surfaces (e.g., glass, quarts, plastic, etc.), the ability to bind, immobilize, or otherwise impregnate a variety of ionophores, chromoionophores, lipophilicadditives, etc., and resistance to washing / rinsing with organic and / or inorganic solvents for reuse without an appreciable loss in sensitivity.

[0100] The one or more chromoionophores, in embodiments, include one or more proton (H+) sensors. In embodiments, the chromoionophore can be any molecule, such as a lipophilic indicator dye, that is able to be embedded into the optode matrix that is selective for protons, where the molecule can change spectral properties depending on the protonation degree ( / .e., [H+], H+ activity, and / or pH).

[0101] In embodiments, the one or more proton-selective chromoionophores in the matrix includes one or more of nitron-8-hydroxyquinoline-5-sulfonate, 4,4'-bis[(N,N-dialkylamino)methyl]-azobenzene, azobenzene, o-cresolphtalein octadecyl ester, Cresol Red, Phenol Red, 1-(2-Pyridylazo)-2-naphthol (PAN), o-Cresolphthalein, alpha-Naphtholphthalein, Phenolphthalein, Thymol Blue, m-Cresol Purple, Turmeric (Curcumin), 4-[(2,6-dibromo-4-nitrophenyl)azo]-2-octadecyloxy-1 -naphthol, octyl bis(2,4- dinitrophenyl)acetate, 4-nonadecylpyridine (ETH 1907), 5-Octadecanoyloxy-2-(4-nitrophenylazo)phenol (ETH 2412), octadecyl 2-(4-dipropylaminophenylazo)benzoate (ETH 2418), ETH 2430, 9-dimethylamino-5- [4-(16-butyl-2,14-dioxo-3,15-dioxaeicosyl)phenylimino]benzo[a]phenoxazine (ETH 2439), ETH 2458, ETH 2462, ETH 2468, ETH 2479, ETH 3531, 3-octadecanoylimino-7-(diethylamino)-1,2-benzophenoxazine (ETH 5294), ETH 5348, 9-(diethylamino)-5-[(2-octyldecyl)imino]benzo[a]phenoxazine (ETH 5350), ETH 5355, 9- Dimethylamino-5-[4-(15-butyl-1,13-dioxo-2,14-dioxanonadecyl)phenylimino]benzo[a]phenoxazine(ETH 5418), 3',3",5',5"-Tetrabromophenolphthaleinethyl ester (TBPE), 1-Hydroxy-4-[4-(2- hydroxyethylsulfonyl)phenylazo]naphthalene-2-sulfonic acid potassium salt (GJM-541), ETH 7058, ETH 7061, and ETH 7075. In embodiments, the one or more proton-selective chromoionophores includes a derivative of one or more molecule described herein, as well as Nile Blue derivatives, fluorescein derivatives, hydroxy coumarins, anthraquinone derivatives, hydroxypyridinecarboxamide derivatives, substituted diphenylamines (e.g., resembling Takagi reagent), crown ether ionophores, calixarene ionophores, and / or acridine derivatives.

[0102] The one or more ionophores, in embodiments, can selectively bind a cation or anion analyte ion. In embodiments, the ionophore can include a fluoroionophore (fluorescence emission spectrum changes) or chromogenic ionophore (absorbance changes). In embodiments, one or more ionophores in the matrix includes at least one cation-selective sensor and at least one anion-selective sensor. In embodiments, one or more ionophores in the matrix includes at least two cation-selective sensors and at least one anionselective sensor.

[0103] In embodiments, at least one of the one or more ionophores is configured to detect at least one monovalent cation. In embodiments, the one or more monovalent cations comprises one or more of potassium (K+) and sodium (Na+).

[0104] In embodiments, the one or more ionophores is configured to detect sodium (Na+) at a concentration range of at least about 80 mM to at least about 180 mM. In embodiments, the one or more ionophores is configured to detect sodium (Na+) at a concentration range of about or at least about 80 mM to about or at least about 90 mM, about or at least about 90 mM to about or at least about 100 mM, about or at least about 100 mM to about or at least about 110 mM, about or at least about 110 mM to about or at least about 120 mM, about or at least about 120 mM to about or at least about 130 mM, about or at least about 130 mM to about or at least about 140 mM, about or at least about 140 mM to about or at least about 150 mM, about or at least about 150 mM to about or at least about 160 mM, or about or at least about 170 mM to about or at least about 180 mM.

[0105] In embodiments, the one or more ionophores is configured to detect potassium (K+) at a concentration range of at least about 2.0 mM to at least about 10 mM. In embodiments, the one or more ionophores is configured to detect potassium (K+) at a concentration range of about or at least about 2 mM to about or at least about 3 mM, about or at least about 3 mM to about or at least about 4 mM, about or at least about 4 mM to about or at least about 5 mM, about or at least about 5 mM to about or at least about 6 mM, about or at least about 6 mM to about or at least about 7 mM, about or at least about 7 mM to about or at least about 8 mM, about or at least about 8 mM to about or at least about 9 mM, or about or at least about 9 mM to about or at least about 10 mM.

[0106] In embodiments, the optode includes one or more ionophores that are sodium (Na+)-selective. In embodiment the one or more Na+-selective ionophores (Na+ sensors) includes one or more of the one or more ionophores is configured to detect sodium (Na+) and comprises one or more of gramicidin A, laidlomycin, lasalocid, maduramicin, monensin, narasin, nigericin, nonactin, nystatin, salinomycin, valinomycin, dicyclohexano-18-crown-6, calix[4]arenecrown-4, potassium tetrakis(p-chlorophenyl)borate (KTpCIPB), ortho-nitrophenyloctylether, sodium tetrakis(p-chlorophenyl)borate (NaTFPB), Sodium tetrakis[3,5-bis(trifluoromethyl)phenyl]borate (NaBARF), Potassium tetrakis[3,5- bis(trifluoromethyl)phenyl]borate (KTFPB), bis-2-ethylhexylsebacate, 2,7-di-tert-butyl-9,9-dimethyl-4,5- xanthenediamine, tridodecylmethylammonium chloride (TDDMACI), 1 ,8-naphthalenediol, bis(1- butylpentyl)adipate (BBPA), crown ether ionophores, bis[(12-crown-4)methyl] dodecylmethylmalonate, di(o-methoxy)-stilbeno-24-crown-8, 15-octadecyloxymethyl-15- methy 1-16-crown-5, dibenzo-16-crown-5 nitrophenol, 4-tert-butylcalix[4]arene-tetraacetic acid tetraethyl ester, Sodium ionophore IV (Cas #: 172883- 29-3), N,N'-dibenzyl-N,N'-diphenyl-1,2-phenylenedioxydiacetamide (ETH 157), N,N',N"-triheptyl-N,N',N"- trimethyl-4,4',4"-propylidynetris(3-oxabutyramide) (ETH 227), N,N,N',N'-tetracyclohexyl-1 ,2- phenylenedioxydiacetamide (ETH 2120), and ETH 4120. In embodiments, the one or more sodium-selective ionophores includes a derivative of one or more molecule described herein, as well as noncyclic di- and triamides, hexabutyl isomerides, crown ether ionophores (e.g., bis(15-crown-5), bis(12-crown-4), dibenzo- 16-crown-5), malonates, calixarene ionophores (e.g., calix[4]arene derivatives, such as tetraamides and tetraesters), and hemispherand ionophores.

[0107] In embodiments, the optode includes one or more ionophores that are potassium (K+)-selective. In embodiment the one or more K -^selective ionophores (K+ sensors) includes one or more of nonactin, monactin, valinomycin, nigericin, hydroxylated polyvinyl chloride (PVC-OH), dicyclohexano-18-crown-6, dibenzo-18-crown-6, benzo-15-crown-5, dibenzo-30-crown-10, dinaphtho-30-crown-10, 4-nitrobenzo-18- crown-6, 4'-picrylamino-5'-nitrobenzo-18-crown-6, 2, 3-naphtho-15-crown-5, (hexanoy loxy methy l)benzo-15- crown-5, octadecanoyloxy-methyl-15-crown-5, benzo-15-crown-5 ether, bis[(benzo-15-crown-5)-4'-ylmethyl] pimelate, and 2-dodecyl-2-methyl-1 ,3-propanediyl bis[N-[5'-nitro(benzo-15-crown-5)-4'-yl]carbamate] (BME- 44). In embodiments, the one or more potassium-selective ionophores includes a derivative of one or more molecule described herein, as well as derivatives of antibiotics, crown ionophores and bis-crown ionophores, hemispherand ionophores, calixarene ionophores, KRYPTOFIX 222, Takegi reagent, and alkyl-acridine orange.

[0108] In embodiments, at least one of the one or more ionophores is configured to detecta monovalent anion from the biological sample. In embodiments, the one or more monovalent anions comprises chloride (CI-). In embodiments, the one or more ionophores is configured to detect chloride (CI-) at a concentration range of at least about 60 mM to at least about 140 mM. In embodiments, the one or more ionophores is configured to detect chloride (CI-) at a concentration range of about or at least about 60 mM to about or at least about 70 mM, about or at least about 70 mM to about or at least about 80 mM, about or at least about 80 mM to about or at least about 90 mM, about or at least about 90 mM to about or at least about 100 mM, about or at least about 100 mM to about or at least about 110 mM, about or at least about 110 mM to about or at least about 120 mM, about or at least about 120 mM to about or at least about 130 mM, or about or at least about 130 mM to about or at least about 140 mM.

[0109] In embodiments, the optode includes one or more ionophores that are chloride (Cl-)-selective. In embodiment the one or more (Cl-)-selective ionophores (Cl- sensors) includes one or more of triphenyltin chloride, tributyltin chloride, trioctyltin chloride, macrocyclic tetramercury, dodecylbenzylmercury(ll) chloride, tricyclohexyltin chloride, triphenylgermanium chloride, phenylmercuric chloride, hafnocene dichloride, chloro(octaethylporphyrinato)indium-(lll), meso-Tetraphenylporphyrin manganese(lll)-chloride, 4,5-Bis-[N'- (buty l)th iou reido]-2, 7-d i-tert-buty I-9, 9-di methy Ixanth ene, ETH 2412, 4, 5- Di methy I-3, 6-d iocty loxy-o- phenylene-bis(mercurytrifluoroacetate) (ETH 9009), and 3,6-didodecyloxy-4,5-dimethyl-o-phenylene- bis(mercury chloride) (ETH 9033). In embodiments, the one or more chloride-selective ionophores includes a derivative of one or more molecule described herein, as well as derivatives of tetraalkylammonium chloride, CAPRIQUAT salts, octaethyl derivatives, and organomercury compounds.

[0110] In embodiments, the one or more ionophores and / or one or more chromoionophores comprises a lipophilic indicator dye.

[0111] In embodiments, the optode includes one or more ion exchangers, which refers to molecules that possess one or more ionic sites (e.g., a lipophilic ionic site) which function as an ion-exchanger salt that is added into the optode matrix to compensate for changes in charge of the matrix and sample to obey the charge balance condition during the sensing phase. In embodiments, the one or more ion exchangers comprises one or more of potassium tetrakis(p-chlorophenyl)borate (KTpCIPB), sodium tetrakis(p- chlorophenyl)borate (NaTFPB), Sodium tetrakis[3,5-bis(trifluoromethyl)phenyl]borate (NaBARF), Potassium tetrakis[3,5-bis(trifluoromethyl)phenyl]borate (KTFPB), Tridodecylmethylammonium chloride (TDDMACI). In embodiments, the one or more ion exchangers comprises a photo-regenerative ion-exchange ionophore configured to extract one or more analyte cations or anions and release the one or more analyte cations or anions as a function of exposure to one or more wavelengths of electromagnetic radiation from about 280 nm to about 1000 nm. In embodiments, ion exchangers are molecules that act as proton carriers ( / .e., lipophilic pH indicator) which change the protonation degree of the optode matrix surface and / or biological sample. As analytes bind the sensor surface, in embodiments, the ion exchanger can release protons back into the liquid sample as an analyte balance during the sensing phase.

[0112] During selective extraction of the target analyte (e.g., into a lipophilic membrane), the one or more “ion exchangers,” as used herein in embodiments, function to introduce either i) coextraction of an oppositely charged reference ion, or ii) release of a reference ion with the same charge. In embodiments, the ion exchanger functions as an analyte balance, such that as analytes are detected, e.g., becoming bound bythe optode matrix, the ion exchanger can help balance the charge between the matrix and the biological sample during the sensing phase (e.g., as shown in Figs. 2A-2E and 3). In embodiments, the reference ions are protons. In embodiments, coextraction is used for anion-selective sensors, and release of a reference ion is used for cation-selective sensors.

[0113] In embodiments, the signal is dependent on the concentration, where the activity ratio between target ions and protons, and the resultant cross sensitivity to sample pH can include buffering the samples during measurement or simultaneous pH measurement with an independent sensor. In embodiments, the ion exchanger is a molecule that is added to the matrix to maintain charge neutrality and assist with ion diffusion / transport in the sample as the proton-selective chromoionophore detects H+ concentration (pH) (e.g., as illustrated in Figs. 2A-2E). This is done, in embodiments, so that the equilibrium does not change sufficiently to re-extract protons from the chromoionophore, leading to inaccurate detection. In embodiments, pH dependency can also be exploited to tune the sensitivity toward the target ion (e.g., K+, Na+, or Cl- selectivity) of the one or more ionophores.

[0114] In embodiments, the optode is configured to provide the at least one optical signal as a function of detection of one or more of H+ (pH), potassium (K+), sodium (Na+), and chloride (CI-) within a range of a human physiological concentration. For example, a theoretical optode response curves for detecting and providing an optical signal as a function of potassium in a physiological concentration range (e.g., 2-5 mM) is shown and described in Figs. 4-5.

[0115] In embodiments, the optode is configured to provide at least one optical signal as a function of proton (H+) concentration ( / .e., pH). In embodiments, the optical signal is calibrated to provide an optical signal as a function of pH in the range of a human physiological concentration of at least about pH 6.8 ( / .e., approximately 1.585 x 104mM) to at least about pH 9 ( / .e., approximately 1.0 x 106mM). In embodiments, the optode is configured to provide at least one optical signal as a function of H+ concentration (pH), where the optical signal is detectable in a range of at least about 300 nm to at least about 800 nm. In embodiments, the optode is configured to provide an optical signal with a sensitivity of about or at least about 0.01 pH units, or about or at least about 1.0 x 1011mM [H+], where a difference in optical signal intensity or amplitude can be observed with changes of about or at least about 0.01 pH units, about or at least about 0.05 pH units, about or at least about 0.1 pH units, or about or at least about 0.2 pH units. In embodiments, the optode matrix is configured such that the optical signal is calibrated to provide a minimal optical signal (or optical signal change) at about pH 6.8 and a maximal optical signal (or optical signal change) at about pH 9.

[0116] In embodiments, the optode is configured to provide at least one optical signal as a function of potassium (K+) concentration. In embodiments, the optical signal is calibrated to provide an optical signal as a function of potassium ion concentration in the range of a human physiological concentration of at least about 2.0 mM to at least about 10 mM. In embodiments, the optode is configured to provide at least one optical signal as a function of potassium ion concentration, where the optical signal is detectable in a range of at least about 300 nm to at least about 800 nm. In embodiments, the optode is configured to provide an optical signal with a sensitivity of about or at least about 0.01 mM [K+], where a difference in optical signal intensity or amplitude can be observed with changes of about or at least about 0.01 mM potassium, about or at least about 0.05 mM potassium, about or at least about 0.1 mM potassium, about or at least about 0.2 mM potassium, about or at least about 0.5 mM potassium, about or at least about 1 .0 mM potassium, or about or at least about 2.0 mM potassium. In embodiments, the optode matrix is configured such that the optical signal is calibrated to provide a minimal optical signal (or optical signal change) at about 2.0 mM [K+] and a maximal optical signal (or optical signal change) at about 10 mM [K+],

[0117] In embodiments, the optode is configured to provide at least one optical signal as a function of sodium (Na+) concentration. In embodiments, the optical signal is calibrated to provide an optical signal as a function of sodium ion concentration in the range of a human physiological concentration of at least about 80 mM to at least about 180 mM. In embodiments, the optode is configured to provide at least one optical signal as a function of sodium ion concentration, where the optical signal is detectable in a range of at least about 300 nm to at least about 800 nm. In embodiments, the optode is configured to provide an optical signal with a sensitivity of about or at least about 0.01 mM [Na+], where a difference in optical signal intensity or amplitude can be observed with changes of about or at least about 0.01 mM sodium, about or at least about 0.05 mM sodium, about or at least about 0.1 mM sodium, about or at least about 0.2 mM sodium, about or at least about 0.5 mM sodium, about or at least about 1 .0 mM sodium, about or at least about 2.0 mM sodium, or about or at least about 5.0 mM sodium. In embodiments, the optode matrix is configured such that the optical signal is calibrated to provide a minimal optical signal (or optical signal change) at about 80 mM [Na+] and a maximal optical signal (or optical signal change) at about 180 mM [Na+],

[0118] In embodiments, the optode is configured to provide at least one optical signal as a function of chloride (CI-) concentration. In embodiments, the optical signal is calibrated to provide an optical signal as a function of chloride ion concentration in the range of a human physiological concentration of at least about 60 mM to at least about 140 mM. In embodiments, the optode is configured to provide at least one optical signal as a function of chloride ion concentration, where the optical signal is detectable in a range of at leastabout 300 nm to at least about 800 nm. In embodiments, the optode is configured to provide an optical signal with a sensitivity of about or at least about 0.01 mM [CI-], where a difference in optical signal intensity or amplitude can be observed with changes of about or at least about 0.01 mM chloride, about or at least about 0.05 mM chloride, about or at least about 0.1 mM chloride, about or at least about 0.2 mM chloride, about or at least about 0.5 mM chloride, about or at least about 1.0 mM chloride, about or at least about 2.0 mM chloride, or about or at least about 5.0 mM chloride. In embodiments, the optode matrix is configured such that the optical signal is calibrated to provide a minimal optical signal (or optical signal change) at about 60 mM [CI-] and a maximal optical signal (or optical signal change) at about 140 mM [Cl-].

[0119] In embodiments, the sensitivity and tuning of the optode specificity, e.g., to maximize signal change within a particular range of ion concentration, can be achieved by altering the concentrations of the one or more chromoionophores, ionophores, and / or ion exchangers. In embodiments, different concentrations mixed into each matrix for a specific measurement range and sensitivity can be done to reduce cross sensitivity, for example to pH.

[0120] In embodiments, the optical signal change as a function of ion concentration relates to a change in the optical signal intensity or amplitude concomitant with an increase or decrease in ion concentration. In embodiments, the optode is manufactured such that the optical signal intensity or amplitude is calibrated to provide a sensitivity that allows a spectrum of signal change in intensity or amplitude according to detecting each ion within a physiological range, for instance in non-limiting examples, of human blood. This can be achieved, in embodiments, by controlling the selection and concentration of chromoionophore, ionophore, and / or ion exchanger that is embedded within the solvent-accessible surface in the optode matrix.

[0121] In embodiments, at least one optical signal (e.g., as spectra) is detectable in a range of at least about 300 nm to at least about 800 nm (e.g., as shown in Fig. 6A-6B). The optical signal for ion concentration(s) described herein, in embodiments, can be detected in a range of at least about 300 nm to at least about 320 nm, a range of at least about 320 nm to at least about 340 nm, a range of at least about 340 nm to at least about 360 nm, a range of at least about 360 nm to at least about 380 nm, a range of at least about 380 nm to at least about 400 nm, at least about 400 nm to at least about 420 nm, a range of at least about 420 nm to at least about 440 nm, a range of at least about 440 nm to at least about 460 nm, a range of at least about 460 nm to at least about 480 nm, a range of at least about 480 nm to at least about 500 nm, a range of at least about 500 nm to at least about 520 nm, a range of at least about 520 nm to at least about 540 nm, a range of at least about 540 nm to at least about 560 nm, a range of at least about 560nm to at least about 580 nm, a range of at least about 580 nm to at least about 600 nm, a range of at least about 600 nm to at least about 620 nm, a range of at least about 620 nm to at least about 640 nm, a range of at least about 640 nm to at least about 660 nm, a range of at least about 660 nm to at least about 680 nm, a range of at least about 680 nm to at least about 700 nm, a range of at least about 700 nm to at least about 720 nm, a range of at least about 720 nm to at least about 740 nm, a range of at least about 740 nm to at least about 760 nm, a range of at least about 760 nm to at least about 780 nm, a range of at least about 780 nm to at least about 800 nm, including values and ranges therein.

[0122] In embodiments, the optode is configured to generate at least one optical signal for detecting each analyte. In embodiments, the at least one optical signal comprises one or more of a fluorescence signal, colorimetric signal, luminescent signal, absorption signal, and reflectance signal.

[0123] In embodiments, the optode is configured to generate a spectra (e.g., a single signal) from the detection of multiple analytes, which can be read / analyzed to provide a distinct concentration measure for each individual analyte detected in a biological sample. In embodiments, the optode spectra is within the near-UV, VIS, and / or near-IR range of wavelengths. In embodiments, the spectra includes at least one optical signal detectable at about 340 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the spectra includes at least one optical signal detectable at about 405 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the spectra includes at least one optical signal detectable at about 505 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the spectra includes at least one optical signal detectable at about 545 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the spectra includes at least one optical signal detectable at about 660 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the spectra includes at least one optical signal detectable at about 720 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the optode can detect each of H+, K+, Na+, and CI-, providing four distinct optical signals from a single sample.

[0124] In embodiments, the optode is not substantially selective for one or more contaminating or non- selective ions, including calcium, magnesium, lithium, rubidium, cesium, ammonium, beryllium, strontium, barium, carbon dioxide, carbonate, bicarbonate, oxygen, thiocyanate, nitrate, nitrite, hydroxide, hydrogen sulfide, phosphate, sulfide, sulfite, sulfate, perchlorate, arsenic, iodide, and metals and transition metals, such as antimony, molybdenum, iron, lead, copper, zinc, chromium, silver, selenium, mercury, uranium, thallium, samarium, and cadmium. In embodiments, “substantially selective,” with respect to detectingcontaminating ions, or non-specific ion binding, refers to a negligible impact on optical signal due to the presence of a naturally-occurring concentration within a biological sample. In embodiments, the negligible impact can refer to a change in an optical signal of about or less than about 0.01 %, about or less than about 0.05%, about or less than about 0.1%, about or less than about 0.15%, about or less than about 0.2%, about or less than about 0.5%, about or less than about 1.0%, about or less than about 1.5%, about or less than about 2.0%, or about or less than about 5.0%, where the change in signal is due to the presence of a containing ion versus a control reading of a sample analyte without the containing ion.

[0125] In non-limiting embodiments, the optode is not substantially selective for one or more naturally- occurring concentrations of ions in human blood, which can be present in about or at least about 5000 pg / L (e.g., for zinc), about or at least about 1500 pg / L (e.g., for copper), about or at least about 200 pg / L (e.g., for selenium), about or at least about 100 pg / L (e.g., for barium), about or at least about 1 pg / L to about 50 pg / L (e.g., for antimony, arsenic, cesium, chromium, manganese, and nickel), less than about 1 pg / dL to about 1 pg / L (e.g., for cadmium, cobalt, and lead).

[0126] In embodiments, the optode measures one or more analytes from a biological sample. In embodiments, the biological sample comprises a bodily fluid, liquid, or homogenized tissue or sample. In embodiments, the biological sample is an animal sample or human sample. In embodiments, non-limiting examples of biological samples include blood, sera, plasma, bone marrow, lymph, saliva, sputum, mucus, respiratory or nasal secretion, oropharyngeal swab, nasopharyngeal swab, oral swab, ductal lavage, bronchoalveolar lavage, cerebrospinal fluid, skin swab, vaginal swab, gastric juice, ascites, peritoneal fluid, pleural fluid, gynecological fluids, pus, perspiration, tears, urine, and stool. In embodiments, the biological sample can include obtaining a swab (e.g., skin swabs, vaginal swabs, oral swabs, nasal swabs, etc.), obtaining a washing or lavages, such as ductal lavages or bronchoalveolar lavages, aspirates, scrapings, specimens (e.g., bone marrow specimens, tissue biopsy specimens, surgical specimens, etc.), among other body fluids, secretions, and / or excretions, and the like. In embodiments, the biological sample is a homogenized tissue or biopsy sample, cell-containing body fluid, free floating nucleic acid sample, or a resuspended sample containing biological solids, such as the case with feces or gastrointestinal contents. In embodiments, the biological sample is a dilution or serial dilution of a biological sample. In embodiments, the biological sample is blood.

[0127] In embodiments, the optode is reusable, where the optode matrix can be washed, rinsed, or otherwise cleared of ions and take multiple measures of analytes. In embodiments, the optode is configuredto be washed with an inorganic solvent and detect one or more analytes from at least a second biological sample with less than about 0.01% loss in sensitivity, less than about 0.05% loss in sensitivity, less than about 0.1 % loss in sensitivity, less than about 0.2% loss in sensitivity, less than about 0.3% loss in sensitivity, less than about 0.4% loss in sensitivity, less than about 0.5% loss in sensitivity, less than about 0.6% loss in sensitivity, less than about 0.7% loss in sensitivity, less than about 0.8% loss in sensitivity, less than about 0.9% loss in sensitivity, less than about 1.0% loss in sensitivity, less than about 1.5% loss in sensitivity, or less than about 2.0% loss in sensitivity. In embodiments, the inorganic solvent is one or more of deionized water, phosphate-buffered saline (PBS), sodium chloride (NaCI), sodium hydroxide, tris base, HEPES, MOPS, PIPES, urea, acetic acid, guanidine hydrochloride, and detergent.

[0128] In embodiments, the optode matrix is flexible, pliable, or can otherwise be manipulated. In embodiments, the optode matrix is configured to be applied to a surface (e.g., as shown cast onto a PET substrate in Fig. 7). In embodiments, the surface is transparent to at least a portion of an electromagnetic spectrum from at least about 280 nm to at least about 1000 nm.Optochemical Sensor (Optode) Spin Disk

[0129] In aspects, provided in embodiments herein, is an optochemical sensor (optode) for detecting one or more analytes from a biological sample including a disk or plate that includes one or more wells disposed therein, where the one or more wells includes an optode, as described herein, (e.g., often referred to herein as an “optode spin disk”) is disposed on a surface within the well, an inlet port for providing the biological sample into the well, and at least a portion of an outer surface of the well that is transparent to at least a portion of an electromagnetic (EM) spectrum from at least about 280 nm to at least about 1000 nm, where the disk and the one or more wells are configured to be spun and withstand a centrifugal force, and where the optode within the well is configured to generate at least one optical signal in the range of at least about 300 nm to at least about 800 nm as a function of binding one or more analytes in the biological sample.

[0130] In embodiments, the optode spin disk includes a matrix comprising polyvinyl chloride (PVC) and dioctyl sebacate (DOS), ortho-nitrophenyloctylether (NPOE), or a mixture of DOC and NPOE. In embodiments, the optode spin disk matrix has disposed therein a chromoionophore that is proton-selective and configured to detect H+ concentration (pH) in a range of at least about pH 7 to at least about pH 8, a first ionophore that is potassium-selective and configured to detect potassium (K+) concentration in a range of at least about 2.0 mM to at least about 10 mM, a second ionophore that is sodium-selective and configured to detect sodium (Na+) concentration in a range of at least about 80 mM to at least about 180 mM, a thirdionophore that is chloride-selective and configured to detect chloride (CI-) concentration in a range of at least about 60 mM to at least about 140 mM, and at least one ion exchanger.

[0131] In embodiments, the optode spin disk includes an optode that is configured to generate at least one optical signal detectable in the range of at least about 300 nm to at least about 800 nm as a function of the detection of H+, K+, Na+, and Cl- in the biological sample (e.g., as shown in Figs. 8 and 9A-9D). In embodiments, the at least one optical signal comprises one or more of a fluorescence signal, colorimetric signal, luminescent signal, absorption signal, and reflectance signal.

[0132] In embodiments, the at least one optical signal comprises a signal detectable at about 340 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the at least one optical signal comprises a signal detectable at about 405 nm, optionally for detecting one or more of H+, K+, Na+, and Cl- . In embodiments, the at least one optical signal comprises a signal detectable at about 505 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the at least one optical signal comprises a signal detectable at about 545 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the at least one optical signal comprises a signal detectable at about 660 nm, optionally for detecting one or more of H+, K+, Na+, and CI-. In embodiments, the at least one optical signal comprises a signal detectable at about 720 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

[0133] In embodiments, the optode spin disk is not substantially selective for one or more contaminating or non-selective ions, as described herein. In embodiments, the optode spin disk can detect one or more analytes from a biological sample, as described herein. In embodiments, the optode spin disk biological sample is blood.

[0134] In embodiments, the optode spin disk is composed of plastic, polymethyl methacrylate, carbon fiber, and / or metal. In embodiments, the metal is aluminum and / or titanium. In embodiments, the optode spin disk can be composed of any material that is used for manufacturing centrifugal rotors, or any other materials used for parts with are spun or withstand centrifugal force, for example as used in laboratory devices and equipment.

[0135] In embodiments, the disk further comprises a substrate to hold the optode in place. In embodiments, the substrate is located (e.g., disposed on) the surface of the one or more wells where the optode is disposed. In embodiments, the substrate is a uniform, thin layer (e.g., having a thickness of about or at least about 1 pm, to about or at least about 5.0 mm) which holds the optode in place on a surface. In embodiments, the optode and / or substrate meant to embed or hold the optode can be prepared using oneor more of tetrahydrofuran (THF), cyclohexanone, N,N-dimethylformamide (DMF), acetonitrile, butanone, and dichloromethane. In embodiments, the substrate is composed of one or more of polyethylene terephthalate (PET), polyethylene terephthalate glycol (PETG), polyurethane, polyethylene, polypropylene, and glass. In embodiments, the substrate and / or optode embedded therein is chemically resistant to one or more solvents, optionally including cyclohexanone, N,N-dimethylformamide (DMF), acetonitrile, butanone, and dichloromethane. In embodiments, the substrate is chemically resistant to one or more solvents, including cyclohexanone, N,N-dimethylformamide (DMF), acetonitrile, butanone, and dichloromethane. In embodiments, the “chemically resistant” nature, as used herein refers to the ability of the substrate and / or optode to not substantially interact and / or react with one or more solvents, e.g., one or more organic solvents used to cast the optode onto a surface and / or one or more inorganic solvents used to wash the optode.

[0136] In embodiments, the optode spin disk includes at least about 1 well, at least about 2 wells; at least about 3 wells, at least about 4 wells, at least about 5 wells, at least about 6 wells, at least about 7 wells, at least about 8 wells, at least about 9 wells, at least about 10 wells, at least about 12 wells, at least about 14 wells, at least about 16 wells, at least about 18 wells, at least about 20 wells, at least about 22 wells, at least about 24 wells, at least about 26 wells, at least about 28 wells, at least about 30 wells, at least about 35 wells, or at least about 40 wells. In embodiments, the optode spin disk includes an even number of wells, where each well contains a symmetrically opposed well as a counterbalance, e.g., as done centrifuge rotors.

[0137] In embodiments, the optode spin disk is configured to be spun at a rate of at least about 100 revolutions per minute (RPM), at least about 200 RPM, at least about 300 RPM, at least about 400 RPM, at least about 500 RPM, at least about 600 RPM, at least about 700 RPM, at least about 800 RPM, at least about 900 RPM, at least about 1000 RPM, at least about 1200 RPM, at least about 1400 RPM, at least about 1600 RPM, at least about 1800 RPM, at least about 2000 RPM, at least about 2200 RPM, at least about 2400 RPM, at least about 2600 RPM, at least about 2800 RPM, at least about 2900 RPM, at least about 3000 RPM, at least about 3500 RPM, at least about 4000 RPM, at least about 4500 RPM, or at least about 5000 RPM.

[0138] In embodiments, the optode spin disk is configured to withstand a centrifugal force of at least about 100 x g, at least about 200 x g, at least about 300 x g, at least about 400 x g, at least about 500 x g, at least about 600 x g, at least about 700 x g, at least about 800 x g, at least about 900 x g, at least about 1000 x g, at least about 1200 x g, at least about 1400 x g, at least about 1600 x g, at least about 1800 x g, at least about 2000 x g, at least about 2200 x g, at least about 2400 x g, at least about 2600 x g, at least about 2800x g, at least about 3000 x g, at least about 3500 x g, at least about 4000 x g, at least about 4500 x g, or at least about 5000 x g. In embodiments, the optode spin disk being “configured to withstand,” with reference to a centrifugal force or spinning relates to the ability of the optode spin disk to withstand an applied force without appreciable degradation of analyte binding or optical signal of the optode matrix, appreciable degradation of adherence of the optode matrix to the surface of the well, or leaking, delamination, imbalance and vibration, or other catastrophic failure.

[0139] In embodiments, the optode spin disk well includes a total well volume of at least about 3 pL, at least about 5 pL, at least about 10 pL, at least about 15 pL, at least about 20 pL, at least about 25 pL, at least about 30 pL, at least about 35 pL, at least about 40 pL, at least about 45 pL, at least about 50 pL, at least about 55 pL, at least about 60 pL, at least about 65 pL, at least about 70 pL, at least about 75 pL, at least about 80 pL, at least about 85 pL, at least about 90 pL, at least about 95 pL, at least about 100 pL, at least about 110 pL, at least about 120 pL, at least about 130 pL, at least about 140 pL, at least about 150 pL, at least about 160 pL, at least about 170 pL, at least about 180 pL, at least about 190 pL, or at least about 200 pL. In embodiments, the optode spin disk well can accommodate any volume or sample weight that typical benchtop or standing laboratory centrifuges can accommodate, for example, about or at least about 1 mL to about or at least about 50 mL (e.g., for capable sample tubes).

[0140] In embodiments, the optode spin disk well includes a well width of at least about 1 mm, at least about 2 mm, at least about 3 mm, at least about 4 mm, at least about 5 mm, at least about 6 mm, at least about 7 mm, at least about 8 mm, at least about 9 mm, or at least about 10 mm. In embodiments, the optode spin disk well has a well width that can accommodate a pipette tip, for example, a 10 pL pipette tip, a 200 pL pipette tip, a 1000 pL pipette tip, or a serological pipette tip / bulb (e.g., 5 mL, 10 mL, 25 mL, or 50 mL).

[0141] In embodiments, the optode of the spin disk is disposed on a surface within the well that includes a surface area of at least about 1 mm2, at least about 2 mm2, at least about 3 mm2, at least about 4 mm2, at least about 5 mm2, at least about 6 mm2, at least about 7 mm2, at least about 8 mm2, at least about 9 mm2, at least about 10 mm2, at least about 15 mm2, at least about 20 mm2, at least about 25 mm2, at least about 50 mm2, at least about 100 mm2, at least about 200 mm2, at least about 300 mm2, at least about 400 mm2, at least about 500 mm2. In embodiments, the optode of the spin disk is disposed on a surface within the well that is equivalent to the surface area sufficient to obtain an optical reading of, for example, a single well of a 384-well plate, a single well of a 96-well plate, a single well of a 12-well plate, a single well of a 6-well plate, a cuvette, and the like.

[0142] In embodiments, the optode of the spin disk has a thin, uniform thickness. In embodiments, the optode matrix is cast onto a surface, where the optode matrix thickness is uniform having a thickness of about or at least about 1 pm, about or at least about 5 pm, about or at least about 10 pm, about or at least about 50 pm, about or at least about 100 pm, about or at least about 500 pm, about or at least about 1 .0 mm, about or at least about 1 .5 mm, about or at least about 2.0 mm, about or at least about 2.5 mm, about or at least about 3.0 mm, about or at least about 3.5 mm, about or at least about 4.0 mm, or about or at least about 5.0 mm. In embodiments, the optode is deposited in small volumes onto a surface substrate to allow a uniform thickness to form. This surface substrate, in embodiments, can be a surface of the optode or a “capping substrate,” which is a distinct piece that is composed of a material having resistance to the solvents that the optode molecules are dissolved in (e.g., THF, CHx), and is a material that can be affixed to the spin disk plate after the optode is cast.

[0143] In embodiments, uniform thickness is achieved on the capping substrate by surface treating the material (e.g., plasma charging, adding a hydrophobic layer, adding a hydrophilic layer) so that the optode matrix layer forms uniformly. In embodiments, the optode thickness contributes to the theoretical modeling (e.g., as described in Figs. 4 and 6), which demonstrates that the background subtraction and protonation state are necessary to relax the dependence on thickness, and / or reduce sensitivity to thickness. In embodiments, the optode is formed on the surface of a capping substrate (which is generally a different material than the optode spin disk plate) where small volume droplets (e.g., 50-200 nL) which have the optode molecules dissolved are deposited onto the surface.

[0144] To ensure proper thickness, in embodiments, the optode is deposited in small volumes onto the surface, for example at about or at least about 10 nL, about or at least about 20 nL, about or at least about 50 nL, about or at least about 100 nL, about or at least about 150 nL, about or at least about 200 nL, about or at least about 500 nL, about or at least about 1 pL, about or at least about 2 pL, about or at least about 4 pL, about or at least about 6 pL, about or at least about 8 pL, about or at least about 10 pL, about or at least about 20 pL, about or at least about 30 pL, about or at least about 40 pL, about or at least about 50 pL, about or at least about 100 pL, about or at least about 150 pL, or about or at least about 200 pL.

[0145] Once the optode matrix is formed on the capping substrate, in embodiments, it can be affixed to the optode spin disk plate, e.g., by adhesive, solvent bonding, or thermal bonding. In embodiments, this layer is then laminated to be set in place to withstand centrifugal force.

[0146] In embodiments, the disk one or more layers (e.g., as shown in Fig. 14), comprising one or more of a structural support layer, inlet layer, channel layer, and capping layer, e.g., where the optode(s) are held place on a capping layer, sandwiched between the channel layer and a structural support layer. In embodiments, the fluid channels are formed by the inlet layer and top support layer sandwiching the channel layer between the capping layer holding the optodes. In embodiments, this creates a fluid-tight series of channels embedded within the disk to accommodate volumes of biological sample, which travel throughout the fluidic channel during rotation of the optode spin disk. In embodiments, one or more of the inlet layer, channel layer, and capping layer are composed of PET and / or pressure sensitive adhesive (PSA).

[0147] In embodiments, the optode spin disk well includes an overflow well ( / .e., “dead volume”), where the overflow well has a total volume of at least about 5 pL, at least about 10 pL, at least about 15 pL, at least about 20 pL, at least about 25 pL, at least about 30 pL, at least about 35 pL, at least about 40 pL, at least about 45 pL, at least about 50 pL, at least about 55 pL, at least about 60 pL, at least about 65 pL, at least about 70 pL, at least about 75 pL, at least about 80 pL, at least about 85 pL, at least about 90 pL, at least about 95 pL, or at least about 100 pL. In embodiments, the overflow well prevents loss or sample and / or spilling during a centrifugation process. In embodiments, the overflow well helps to account for inaccuracies in pipetting biological samples, for example where differences in viscosities between samples can lead to inaccuracies in pipetting small volumes (e.g., accuracy can suffer when working with whole blood).

[0148] In embodiments, the optode spin disk well includes an inlet port that includes a diameter of at least about 0.5 mm, at least about 0.6 mm, at least about 0.7 mm, at least about 0.8 mm, at least about 0.9 mm, at least about 1 mm, at least about 1.5 mm, or at least about 2 mm. In embodiments, the optode spin disk well has an inlet port width that can accommodate a pipette tip, for example, a 10 pL pipette tip, a 200 pL pipette tip, a 1000 pL pipette tip, or a serological pipette tip / bulb (e.g., 5 mL, 10 mL, 25 mL, or 50 mL). In embodiments, the optode spin disk well has an inlet port width that can accommodate tubing, coupling, and / or valves for fluidic communication of a microfluidic device for aliquoting the biological sample into one or more wells of the spin disk.

[0149] In embodiments, the optode spin disk well includes an outlet port for venting air pressure. In embodiments the outlet port includes a diameter of at least about 0.5 mm, at least about 0.6 mm, at least about 0.7 mm, at least about 0.8 mm, at least about 0.9 mm, at least about 1 mm, at least about 1 .5 mm, or at least about 2 mm. In embodiments, the optode spin disk well has an outlet port width suitable for tubing, coupling, and / or valves for fluidic communication of a fluid handling or microfluidic device for draining,aspirating, removing, or otherwise manipulating the biological sample from the one or more wells of the spin disk. In embodiments, the optode spin disk well has an outlet port width that can accommodate a pipette tip, for example, a 10 pL pipette tip, a 200 pL pipette tip, a 1000 pL pipette tip, or a serological pipette tip / bulb (e.g., 5 mL, 10 mL, 25 mL, or 50 mL) to extract or otherwise recover the biological sample.

[0150] In embodiments, the optode spin disk well includes at least a portion of an outer surface that is transparent to at least a portion of the EM spectrum, including the visible spectrum, from at least about 280 nm to at least about 1000 nm. In embodiments, this outer portion is transparent to at least a portion of the EM spectrum to enable analyzing or measuring the analyte detection in each well. In embodiments, the portion of the outer surface of the well that is transparent to at least a portion of the EM spectrum is comprised of one or more of glass, quartz, plastic, polystyrene, polyethylene terephthalate (PET), polycarbonate, polymethyl methacrylate (acrylic), polyethylene, polyurethane, polypropylene, thermoplastic elastomers (TPE), acrylonitrile butadiene styrene (ABS), epoxy, cyclic olefin copolymer (COC), cyclic olefin polymer (COP), poly dimethyl siloxane (PDMS), polyether ester ketone (PEEK), polyetherimide (ULTEM), and nylon. In embodiments, the optode spin disk itself can be composed of the same material as the well. In embodiments, the optode spin disk material has a thickness that is equivalent to the thickness of the well, i.e., the well is flush in line with the optode spin disk surface.

[0151] In embodiments, the optode spin disk is reusable, where the optode matrix disposed within each well, the well surface material, and / or the optode disk material can be washed, rinsed, or otherwise cleared of ions and / or biological sample to take multiple measurements. In embodiments, the optode spin disk is configured to be washed with an inorganic solvent and detect one or more analytes from at least a second biological sample with less than about 0.01 % loss in sensitivity, less than about 0.05% loss in sensitivity, less than about 0.1% loss in sensitivity, less than about 0.2% loss in sensitivity, less than about 0.3% loss in sensitivity, less than about 0.4% loss in sensitivity, less than about 0.5% loss in sensitivity, less than about 0.6% loss in sensitivity, less than about 0.7% loss in sensitivity, less than about 0.8% loss in sensitivity, less than about 0.9% loss in sensitivity, less than about 1.0% loss in sensitivity, less than about 1.5% loss in sensitivity, or less than about 2.0% loss in sensitivity between measurements. In embodiments, the optode is configured to be washed with a solvent. In embodiments the solvent is one or more of deionized water, phosphate-buffered saline (PBS), sodium chloride (NaCI), sodium hydroxide, tris base, HEPES, MOPS, PIPES, urea, guanidine hydrochloride, and detergent.

[0152] In embodiments, the optode spin disk is useful for high-throughput screening of biological samples. For example, in embodiments, the optode spin disk can have 40 or more wells which can be loaded with biological samples, incubated, and analyzed for ion detection in about 30 seconds to about 20 minutes, where each optode spin disk well can be washed, reloaded with sample, and again analyzed for two or more cycles of detection, repeated indefinitely. In embodiments, this enables high-throughput screening of biological samples, with about or at least about 3 rounds of analyte detection to at least about 120 rounds of analyte detection, which can equate to about or at least about 120 samples per hour to about or at least about 4800 samples per hour. Thus, the optode spin disk described herein and the associated methods of use, in embodiments, represent a throughput advantage to traditional optode analyte detection.

[0153] In reference to Fig. 10, an exemplary embodiment of an optochemical sensor (optode) spin disk 1000 is illustrated. In embodiments, the optode spin disk 1000 can be a circular disk or plate which can have disposed within the disk or plate one or more bores 1002 for attachment to an implement for spinning the disk. In embodiments, the bore 1002 can be threaded, for instance in non-limiting examples, like for screwing onto a rotor head (e.g., like a centrifuge rotor), or can be threadless in a manner that fits onto a motor shaft, where one or more additional implements (e.g., a nut) fastens or affixes the optode spin disk 1000 onto a motor shaft. In embodiments, the optode spin disk 1000 can be composed of a material configured to withstand a centrifugal force ( / .e., spinning) in the range of at least about 50 x g to at least about 5,000 x g, such as carbon fiber, plastic (e.g., polymethacrylate), or metal (e.g., titanium or aluminum). In embodiments, the optode spin disk 1000 can have disposed therein one or more wells 1004. In embodiments, each well 1004 can have a total well volume for holding a liquid sample that is at least about 5 pL to at least about 200 pL, a well width that is at least about 1 mm to at least about 10 mm, an overflow well volume that is at least about 5 pL to at least about 50 pL. In embodiments, each well 1004 can also have a covermask 1006. In some embodiments, the covermask 1006 is optional. The covermask 1006, in embodiments, comprises an aperture that more accurately defines the region where the optode is read optically. In embodiments, a covermask is optically opaque except for the aperture areas. In some embodiments, the entire covermask region is substantially optically transparent. In embodiments, each well 1004 can have immobilized therein - on at least a portion of one surface - an optode matrix 1008, as described herein. In embodiments, the covermask 1006 can cover at least a portion of the well that is read for analyte detection (e.g., by a light source, LED, laser, camera, fluorimeter, luminometer, etc.). In embodiments, this portion of the well 1004 within the optode spin disk 1000 plate can be transparent to at least a portion of the EM spectrum between 280 nm and 1000 nm such that emission, reflectance, etc., can be analyzed (e.g., read via laser detection).In embodiments, the covermask 1006 portion (or aperture therein) of the well 1004 can be read at 505 nm, 660 nm, and / or 720 nm. In embodiments, each well 1004 can have an inlet port 1010 for pipetting liquid sample into the well 1004. In embodiments, the inlet port 1010 can be at last about 0.5 mm in diameter to at least about 2.0 mm in diameter. In embodiments, each well 1004 can also have an outlet port 1012, where the outlet port 1012 can be at least about 0.5 mm to at least about 2 mm in diameter.

[0154] In embodiments, the optode spin disc can have a fluid circuit embedded within the disk plate (e.g., as shown in Figs. 11 and 13-19). In embodiments, the fluid circuit can have several portions, each having a different function for analyzing the biological sample. In embodiments, the biological sample is fed through the fluid circuit embedded on the optode spin disk via centrifugal force. In embodiments, the optode spin disk is configured to be spun in at least two different directions, for example, where applying a first direction of centrifugal force (e.g., spinning in one direction) can force the biological sample through a first portion of the fluid circuit, and applying a second direction of centrifugal force (e.g., spinning in the opposite direction) can force the biological sample through a next portion of the fluid circuit. In embodiments, the optode is configured to be spun in two diametrically opposed directions, to force the biological sample through the fluid circuit.

[0155] In embodiments, the optode spin disk has a fluid circuit embedded within the disk that fluidically connects to the one or more wells by a channel or tubing (e.g., as shown in Figs. 11 and 13-19). In embodiments, the one or more pH fix chambers are configured to fix the ionic content of the biological sample to prevent deviation in concentrations of the one or more analytes during an optode sensing phase. In embodiments, the fluid circuit fluidically connects the one or more wells to one or more pH fix chambers. In embodiments, the “pH fix chamber” is also referred to as “pH well” (e.g., as in Figs. 15 and 18), which is a fluid chamber located within the optode disk for combining sample with one or more soluble pH optode (which is deposited as a solid on the capping layer) and containing the resulting mixed solution after dissolution. In embodiments, the fluid circuit comprises a pH priming circuit (e.g., as shown in Fig. 16), which is a circuit used to measure a dedicated volume of sample for mixing with the pH optode and is used to control the introduction of the sample into the pH well by changing disc RPM / centrifugal force. In embodiments, the pH priming circuit is unique to the pH optode due to the optode its solubility in the sample, where it uses its own metering technique in comparison to one or more other optodes because the dye may escape the chamber. In embodiments, the fluid circuit comprises a pH aliquot, which is a chamber that stores a sample volume to be aliquoted for pH optode mixing before the pH priming circuit (e.g., shown as a “U” shaped chamber in Fig. 18).

[0156] In embodiments, the fluid circuit fluidically connects the one or more pH fix chambers to one or more separator chambers and / or siphons. In embodiments, the terms “siphon” and “priming siphon” are used interchangeably (e.g., as in Figs. 17 and 18). In embodiments, the siphon and / or priming siphon function to prevent liquid from moving from a first chamber to a second chamber until disc RPM / centrifugal force is reduced enough for pneumatic pressure inside a chamber (e.g., sample separation chamber) to overcome centrifugal force, where once the liquid reaches the crest of the chamber, it siphons out to the next chamber. In embodiments, the one or more separator chambers and / or siphons is configured to separate cells, cell debris, and / or plasma from the biological sample during and / or after a centrifugal force is applied to the spin disk, embodiments, the one or more separator chambers and / or siphons is configured to prevent backflow of sample. In embodiments, the fluid circuit fluidically connects the one or more separator chambers to one or more sample reservoirs and / or siphons. In embodiments, the one or more sample reservoirs is configured to hold the biological sample and comprises a volume of about or at least about or at least about 10 pL, about or at least about 20 pL, about or at least about 50 pL, about or at least about 0.1 mL, about or at least about 0.2 mL, about or at least about 0.3 mL, about or at least about 0.4 mL, about or at least about 0.5 mL, about or at least about 0.6 mL, about or at least about 0.7 mL, about or at least about 0.8 mL, about or at least about 0.9 mL, or about or at least about 1 .0 mL.

[0157] For example, in reference to Fig. 11, an exemplary embodiment of a fluid circuit-embedded optode spin disk 1100 for analyzing blood is illustrated. In embodiments, the fluid circuit-embedded optode spin disk 1100 has at least one sample reservoir 1102 for receiving blood. In embodiments, the sample reservoir 1102 is configured to hold a volume of about or at least about 10 pL, about or at least about 20 pL, about or at least about 50 pL, about or at least about 0.1 mL, about or at least about 0.2 mL, about or at least about 0.3 mL, about or at least about 0.4 mL, about or at least about 0.5 mL, about or at least about 0.6 mL, about or at least about 0.7 mL, about or at least about 0.8 mL, about or at least about 0.9 mL, or about or at least about 1.0 mL. In embodiments, the fluid circuit-embedded optode spin disk 1100 can have multiple sample reservoirs 1102 for accommodating several different samples (e.g., a dilution series of a single sample, or multiple, distinct samples). In embodiments, the fluid circuit-embedded optode spin disk 1100 can have 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more sample reservoirs. In embodiments, the sample reservoir 1102 is located nearer the center of the spin disk, such that as centrifugal force is applied, the biological sample is pulled outwards through the fluid circuit towards the edge of the disk.

[0158] In embodiments, the fluid circuit-embedded optode spin disk 1100 has at least one separator chamber 1104 for each sample reservoir 1102. In embodiments, blood can be moved via centrifugal forcefrom the sample reservoir 1102 into the separator chamber 1104. In embodiments, blood is forced into the separator chamber 1104 such that as the spin disk is spun, larger components of the blood are separated, for example, whole cells (e.g., red blood cells, white blood cells, and platelets) are separated from the plasma. In embodiments, the separator chamber 1104 can remove about or at least about 10% of whole cells, about or at least about 20% of whole cells, about or at least about 30% of whole cells, about or at least about 40% of whole cells, about or at least about 50% of whole cells, about or at least about 60% of whole cells, about or at least about 70% of whole cells, about or at least about 80% of whole cells, about or at least about 90% of whole cells, about or at least about 95% of whole cells, about or at least about 99% of whole cells, for example as measured by total cell count and / or cell mass in comparison to the total cell count and / or cell mass in the original sample disposed in the sample reservoir 1102.

[0159] In embodiments, the fluid circuit-embedded optode spin disk 1100 has at least one pH fix chamber 1106 for each sample reservoir 1102. In embodiments, blood can be moved via centrifugal force from the separator chamber 1104 into the pH fix chamber 1106. In embodiments, blood is forced into the pH fix chamber 1104 such that the blood analyte content (e.g., H+ concentration, pH) can be “fixed” so there are no significant deviations in blood pH during the sensing phase in each of the optode wells. In embodiments, no significant deviations in blood pH refer to a change of less than about 0.01 pH (e.g., less than about a change of 1.0 x 1011mM [H+]), less than about 0.02 pH, less than about 0.03 pH, less than about 0.04 pH, less than about 0.05 pH, less than about 0.1 pH, or less than a deviation of about 0.2 pH from the original blood sample disposed in the sample reservoir 1102. In embodiments, the pH fix chamber 1104 can have one or more components for binding ions, as described herein.

[0160] In embodiments, the fluid circuit-embedded optode spin disk 1100 has one or more wells having disposed therein one or more optodes 1108 for each sample reservoir 1102. In embodiments, the one or more optodes 1108 includes a sodium (Na+)-selective optode, a potassium (K+)-selective optode, a chloride (Cl-)-selective optode, and a pH (H+)-selective optode. In embodiments, separated plasma extracted from blood can be moved via pneumatic ballast pressure to force liquid from the pH fix chamber 1106 over a siphon into the next chamber with the one or more optodes 1108.

[0161] In reference to Fig. 12, an exemplary embodiment of an optode spin disk well 1200 is illustrated. In embodiments, the well 1200 has disposed on at least a portion of one surface an optode matrix 1204. In embodiments, the optode matrix 1204 is first cast onto a substrate material 1202 (e.g., a “capping substrate” made of polyethylene terephthalate (PET)) which can be affixed to the spin disk surface, covering the well1200, using an adhesive layer 1206 flanking the edges of the well 1200. In embodiments, the substrate material 1202 is resistant to organic solvents that dissolve optode molecules, such as THF and hydrocarbon (CHx) solvents. In embodiments, the substrate material 1202 surface can be modified by a specific coating to allow the application of a thin, uniform thickness optode matrix. In embodiments, this provides a liquid-tight seal around the edge of the well 1200, providing an encircled cavity, or liquid channel 1208 within. In embodiments, the substrate material 1202 is transparent to at least a portion of the EM spectrum. In embodiments, the material of the well 1200 on the opposite side is transparent to at least a portion of the EM spectrum. In embodiments, the substrate material 1202 can be further sealed by thin lamination so that the well can withstand centrifugal force.

[0162] In embodiments, the optode spin disc can be configured to be single component of a multicomponent system. In embodiments, the multi-component system includes a microfluidic platform and / or device that is in fluid communication with the optode spin disc. In embodiments, the one or more wells of the optode spin disk can include an inlet port and an outlet port where the inlet port and / or outlet port are configured to be connected to a microfluidic platform. In embodiments, the microfluidic platform and / or device is in fluid connection with the inlet port and the outlet port of the one or more wells. In embodiments, the microfluidic platform and / or device includes at least one valve, a section of tubing, and a fluid pumping mechanism (e.g., peristaltic pump).

[0163] In embodiments, the multi-component system includes one or more detectors and / or sensor monitors for reading the signal output from the one or more optodes of the spin disk, such as a charge-couple device (CCD) imaging device, reflectometric photometer, light emitting diode (LED) photometer, fluorometer, luminometer, absorbance plate reader (e.g., absorption spectroscopy), and the like. In embodiments, the multi-component system includes a SPECTRAMAX plate reader (Molecular Devices Corp.), a SERALYZER reflectometric photometer (Miles Laboratories, Inc.), a luminescence measuring oxygen sensor (LUMOS) system, or a GLOMAX luminometer (Promega). In embodiments, the multi-component system can include a computing device, such as a desktop computer, laptop, tablet, or smartphone.Methods of Assaying Analytes from Biological Samples

[0164] The present disclosure provides, in embodiments, methods for detecting one or more analytes in a biological sample using an optochemical sensor (optode), as described herein, by dispensing one or more aliquots of the biological sample into one or more wells of the optode, applying a centrifugal force to the optode, and detecting at least one optical signal (e.g., spectra) from the one or more wells in the rangeof at least about 300 nm to at least about 800 nm as a function of binding to one or more analytes in the biological sample.

[0165] In embodiments, methods of assaying analytes from biological samples include dispensing one or more blood samples into one or more sample reservoirs of an optode spin disk, applying a centrifugal force to the optode spin disk, separating whole cells from the blood sample, fixing the pH of the blood sample, dispensing the cell-depleted, pH-fixed blood sample into one or more wells, each well having one or more optodes (e.g., a sodium-selective optode, a potassium-selective optode, a chloride-selective optode, and a pH-selective optode) disposed therein, and then measuring a change in one or more optical signals emitted from the one or more optodes as a function of binding and detecting its cognate ion in the blood sample.

[0166] Methods herein, in embodiments, can include assaying the analyte concentration of one or more biological samples. In embodiments, the biological sample comprises one or more of blood, sera, plasma, bone marrow, lymph, saliva, sputum, mucus, respiratory or nasal secretion, oropharyngeal swab, nasopharyngeal swab, oral swab, ductal lavage, bronchoalveolar lavage, cerebrospinal fluid, skin swab, vaginal swab, gastric juice, ascites, peritoneal fluid, pleural fluid, gynecological fluids, pus, perspiration, tears, urine, and stool. In embodiments, the biological sample is blood, and the analyte is one or more ions found in blood.

[0167] In embodiments, methods herein include dispensing one or more aliquots of the biological sample where the aliquots have a total volume of at least about 3 pL, at least about 5 pL, at least about 10 pL, at least about 15 pL, at least about 20 pL, at least about 25 pL, at least about 30 pL, at least about 35 pL, at least about 40 pL, at least about 45 pL, at least about 50 pL, at least about 55 pL, at least about 60 pL, at least about 65 pL, at least about 70 pL, at least about 75 pL, at least about 80 pL, at least about 85 pL, at least about 90 pL, at least about 95 pL, at least about 100 pL, at least about 110 pL, at least about 120 pL, at least about 130 pL, at least about 140 pL, at least about 150 pL, at least about 160 pL, at least about 170 pL, at least about 180 pL, at least about 190 pL, or at least about 200 pL. Methods herein, in embodiments, can include diluting the biological sample prior to aliquoting, where the diluent can include one or more of deionized water, phosphate-buffered saline (PBS), tris base, HEPES, MOPS, PIPES, among other solutions commonly used in biochemistry. In embodiments, the diluent does not substantially alter the concentrations of one or more of H+, K+, Na+, or Cl- ions. In embodiments, methods herein account for any changes in analyte concentration from the addition of diluent.

[0168] In embodiments, methods herein include applying the centrifugal force by spinning the optode spin disk at a rate of at least about 100 RPM, at least about 200 RPM, at least about 300 RPM, at least about 400 RPM, at least about 500 RPM, at least about 600 RPM, at least about 700 RPM, at least about 800 RPM, at least about 900 RPM, at least about 1000 RPM, at least about 1200 RPM, at least about 1400 RPM, at least about 1600 RPM, at least about 1800 RPM, at least about 2000 RPM, at least about 2200 RPM, at least about 2400 RPM, at least about 2600 RPM, at least about 2800 RPM, at least about 2900 RPM, at least about 3000 RPM, at least about 3500 RPM, at least about 4000 RPM, at least about 4500 RPM, or at least about 5000 RPM. In embodiments, methods herein include applying a centrifugal force of at least about 100 x g, at least about 200 x g, at least about 300 x g, at least about 400 x g, at least about 500 x g, at least about 600 x g, at least about 700 x g, at least about 800 x g, at least about 900 x g, at least about 1000 x g, at least about 1200 x g, at least about 1400 x g, at least about 1600 x g, at least about 1800 x g, at least about 2000 x g, at least about 2200 x g, at least about 2400 x g, at least about 2600 x g, at least about 2800 x g, at least about 3000 x g, at least about 3500 x g, at least about 4000 x g, at least about 4500 x g, or at least about 5000 x g.

[0169] In embodiments, methods herein include dispensing one or more aliquots of the biological sample into one or more sample reservoirs on the optode spin disk. In embodiments, the one or more aliquots can include a volume (e.g., of blood) that is about or at least about 10 pL, about or at least about 20 pL, about or at least about 50 pL, about or at least about 0.1 mL, about or at least about 0.2 mL, about or at least about 0.3 mL, about or at least about 0.4 mL, about or at least about 0.5 mL, about or at least about 0.6 mL, about or at least about 0.7 mL, about or at least about 0.8 mL, about or at least about 0.9 mL, or about or at least about 1.0 mL.

[0170] In embodiments, methods herein include separating cells and / or cell debris from the biological sample during and / or after the centrifugal force is applied to the optode spin disk, e.g., such as when using an optode spin disk with a separator chamber.

[0171] In embodiments, methods herein include fixing the ionic content of the biological sample to prevent deviation in concentrations of the one or more analytes during the detecting, for example so that the concentrations of H+, Na+, K+, and / or Cl- do not substantially change during the sensing phase of optode binding.

[0172] In embodiments, methods herein include detecting at least one optical signal by measuring one or more of absorption signal (e.g., absorption spectroscopy), fluorescence signal, colorimetric signal,luminescent signal, and reflectance signal. In embodiments, the measuring can include using one or more of charge-couple device (CCD) imaging device, light emitting diode (LED) light source, reflectometric photometer, LED photometer, fluorometer, luminometer, or a absorption spectroscopy plate reader. In embodiments, the measuring can include using one or more of a SPECTRAMAX plate reader (Molecular Devices Corp.), a SERALYZER reflectometric photometer (Miles Laboratories, Inc.), a luminescence measuring oxygen sensor (LUMOS) system, or a GLOMAX luminometer (Promega).

[0173] Methods herein include, in embodiments, measuring at least one optical signal detectable at about 340 nm for detecting one or more of H+, K+, Na+, and CI-. Methods herein include, in embodiments, measuring at least one optical signal detectable at about 405 nm for detecting one or more of H+, K+, Na+, and CI-. Methods herein include, in embodiments, measuring at least one optical signal detectable at about 505 nm for detecting one or more of H+, K+, Na+, and CI-. Methods herein include, in embodiments, measuring at least one optical signal detectable at about 545 nm for detecting one or more of H+, K+, Na+, and CI-. Methods herein include, in embodiments, measuring at least one optical signal detectable at about 660 nm for detecting one or more of H+, K+, Na+, and CI-. Methods herein include, in embodiments, measuring at least one optical signal detectable at about 720 nm for detecting one or more of H+, K+, Na+, and CI-. In embodiments, methods herein include assaying analyte content of one or more biological samples at any wavelength, or range of wavelengths, as described herein.

[0174] Methods herein, in embodiments, can assay analyte content without substantially detecting ( / .e., an assay that is not substantially selective for) one or more of calcium, magnesium, lithium, rubidium, cesium, ammonium, beryllium, strontium, barium, carbon dioxide, carbonate, bicarbonate, oxygen, thiocyanate, nitrate, nitrite, hydroxide, hydrogen sulfide, phosphate, sulfide, sulfite, sulfate, perchlorate, arsenic, iodide, and metals and transition metals, such as antimony, molybdenum, iron, lead, copper, zinc, chromium, silver, selenium, mercury, uranium, thallium, samarium, and cadmium. In embodiments, “substantially selective,” with respect to detecting contaminating ions, or non-specific ion binding, refers to a negligible impact on optical signal due to the presence of a naturally-occurring concentration within a biological sample. In embodiments, the negligible impact can refer to a change in an optical signal of about or less than about 0.01%, about or less than about 0.05%, about or less than about 0.1%, about or less than about 0.15%, about or less than about 0.2%, about or less than about 0.5%, about or less than about 1 .0%, about or less than about 1.5%, about or less than about 2.0%, or about or less than about 5.0%, where the change in signal is due to the presence of a containing ion versus a control reading of a sample analyte without the containing ion.

[0175] In embodiments, methods herein are not substantially selective for one or more naturally- occurring concentrations of ions in human blood, which can be present in about or at least about 0.5 mg / dL to about or at least about 10 mg / dL (e.g., for calcium, magnesium, phosphate), about or at least about 20 mM / L to about or at least about 35 mM / L (e.g., for bicarbonate), about or at least about 5000 pg / L (e.g., for zinc), about or at least about 1500 pg / L (e.g., for copper), about or at least about 200 pg / L (e.g., for selenium), about or at least about 100 pg / L (e.g., for barium), about or at least about 1 pg / L to about 50 pg / L (e.g., for antimony, arsenic, cesium, chromium, manganese, and nickel), less than about 1 pg / dL to about 1 pg / L (e.g., for cadmium, cobalt, and lead). In embodiments, methods herein are specifically selective for K+, Na+, CI-.

[0176] In embodiments, methods herein include preparing the optode with one or more organic solvents comprising tetrahydrofuran (THF), cyclohexanone, N,N-dimethylformamide (DMF), acetonitrile, butanone, and dichloromethane. In embodiments, the optode is prepared prior to aliquoting the biological sample into the one or more wells, for example, between uses.

[0177] In embodiments, methods herein include washing and / or rinsing the one or more wells with an inorganic or aqueous solvent. In embodiments, the solvent comprises one or more of deionized water, phosphate-buffered saline (PBS), sodium chloride (NaCI), sodium hydroxide, tris base, HEPES, MOPS, PIPES, urea, guanidine hydrochloride, and detergent. In embodiments, the optode is single-use (e.g., disposable) and does not require washing.

[0178] In embodiments, methods herein include dispensing the one or more aliquots of the biological sample and / or washing the one or more wells using a pipette, liquid handling device, or microfluidic platform. In embodiments, methods herein include using the optode spin disk in conjunction with a liquid handling device or microfluidic platform which is in fluid connection with the one or more wells, e.g., for high-throughput or automated sampling.

[0179] In embodiment, methods herein include detecting at least one optical signal (e.g., spectra) from the one or more wells after dispensing at least a second biological sample, where the detection does not result in or experience a substantially significant reduction in sensitivity. In embodiments, the substantially significant reduction in sensitivity is less than about 0.01% loss in sensitivity, less than about 0.05% loss in sensitivity, less than about 0.1% loss in sensitivity, less than about 0.2% loss in sensitivity, less than about 0.3% loss in sensitivity, less than about 0.4% loss in sensitivity, less than about 0.5% loss in sensitivity, less than about 0.6% loss in sensitivity, less than about 0.7% loss in sensitivity, less than about 0.8% loss in sensitivity, less than about 0.9% loss in sensitivity, less than about 1.0% loss in sensitivity, less than about1 .5% loss in sensitivity, or less than about 2.0% loss in sensitivity. In embodiments, measuring the sensitivity, including any losses thereof, and / or calibrating of sample concentration ( / .e., to make dilutions), include measuring the optical signal from the optode, e.g., as described herein.

[0180] In embodiments, methods herein include two or more cycles of said washing, dispensing, and detecting one or more analytes from an additional biological sample. In embodiments, the two or more cycles can be a part of high-throughput assaying of analyte content in the biological sample.

[0181] In embodiments, methods herein include performing repetitive assaying, including dispensing at least a second biological sample into the one or more wells of the optode spin disk. In embodiments, the methods herein are useful for high-throughput screening of biological samples. For example, in embodiments, the optode spin disk can have 40 or more wells which can be loaded with biological samples, incubated, and analyzed for ion detection in about 30 seconds to about 20 minutes, where each optode spin disk well can be washed, reloaded with sample, and again analyzed for two or more cycles of detection, repeated indefinitely. In embodiments, this enables methods of high-throughput screening of biological samples, with about or at least about 120 rounds to about or at least about 3 rounds of analyte detection, which can equate to about or at least about 4800 samples to about or at least about 120 samples per hour. Thus, the optode spin disk described herein and the associated methods of use, in embodiments, represent a throughput advantage to traditional optode analyte detection.

[0182] In embodiments, a non-limiting assay using the optode spin disk can include using a pipette to dispense a small volume of approximately 25 pL of a liquid biological sample (e.g., blood, sera, or plasma) into one or more wells, and spinning the optode spin disk at approximately 2,500 RPM for about 2 min as an incubation period (e.g., sensing phase for the optode). Next, in embodiments, each well can be rotated in front of a detector to be read, for example, where the sample is analyzed at 720 nm in triplicate, at 660 nm in triplicate, and 505 nm in triplicate. In embodiments, the amplitude or intensity of emission, reflectance, etc., at each wavelength can be indicative of the concentration of each analyte in the sample (e.g., K+, Na+, and CI-), with the optode configured and / or calibrated to be to be sensitive within the range of a human physiological range for the biological sample.Methods of Diagnosis and / or Treatment

[0183] Methods herein, in embodiments, include performing the methods as described herein and transforming the information from the analyte content into diagnosing one or more medical conditions and / or disorders as a function of the analyte detecting. In embodiments, the one or more medical conditions and / ordisorders comprises a medical condition and / or disorder associated with a blood concentration of one or more of H+, Na+, K+, and CI-.

[0184] In embodiments, methods herein include diagnosing one or more medical conditions and / or disorders according to the detection of analyte content in a biological sample. In embodiments, the one or more medical conditions and / or disorders can include one or more of acidosis, alkalosis, hypokalemia, hyperkalemia, hyponatremia, hypernatremia, hyperchloremia, hypochloremia, Cushing syndrome, Barter syndrome, kidney disease, heart failure, heart disease, albuminuria, hyperaldosteronism, ketoacidosis, lactic acidosis, renal tubular acidosis, diabetes, dehydration, hyperglycemia, hyperparathyroidism, effects of a prescribed therapeutic agent, including effects of diuretics (e.g., hydrochlorothiazide, furosemide, torsemide, and indapamide) and / or effects carbonic anhydrase inhibitors (e.g., glaucoma treatment), ethylene glycol poisoning, methanol poisoning, salicylate toxicity (e.g., aspirin overdose), ureteral diversion, syndrome of inappropriate antidiuretic hormone secretion (SIADH), and cancer (e.g., endocrine neoplasia, kidney cancer, liver cancer).

[0185] In embodiments, methods herein include determining adherence to one or more therapeutic agents and / or medications which have an effect on the blood concentration of one or more of H+, Na+, K+, and CI-. For instance, in non-limiting embodiments, a heart disease therapeutic, metabolic disorder therapeutic, lifestyle disease therapeutic, and the like, can have an effect on one or more of H+, Na+, K+, and Cl- ion concentration in the blood, where the methods herein can determine the degree of adherence to the medication according to the changes in concentration of the subject taking the medication.

[0186] In embodiments, methods herein include prescribing one or more therapeutic agents as a function of the detecting of analyte content in a biological sample. In embodiments, the one or more therapeutic agents include a therapeutic agent associated with increasing, decreasing, or otherwise modulating a blood concentration of one or more of H+, Na+, K+, and CI-.

[0187] In embodiments, the one or more therapeutic agents comprises one or more of electrolyte supplementation (e.g., potassium salt, chloride salt, sodium salt), sodium zirconium cyclosilicate (LOKELMA), diuretics (e.g., loop diuretics, amiloride triamterene), hypertonic or hypotonic saline, vasopressin receptor antagonists, demeclocycline therapy, bicarbonate therapy, hemodialysis, continuous renal replacement therapy (CRRT), alkalinizing agents, mineralocorticoids, vitamin D supplementation, carbonic anhydrase inhibitors, acetazolamide, insulin, ketoconazole, mitotane (LYSODREN), metyrapone (METOPIRONE), Mifepristone (KORLYM, MIFEPREX), angiotensin-converting enzyme (ACE) inhibitors, angiotensin receptorblockade (ARB), aldosterone antagonists, spironolactone, indomethacin, ibuprofen, non-steroidal antiinflammatory drugs (NSAIDs), growth hormone (GH), dapagliflozin (FARXIGA), blood thinners, vasodilators, enalapril (VASOTEC, EPANED), lisinopril (ZESTRIL, QBRELIS), captopril, and alcohol dehydrogenase enzyme (ADH) antagonists (e.g., Fomepizole).Kits

[0188] The present disclosure provides, in embodiments, kits including optodes, optode spin disk(s), and / or additional components for carrying out methods and assays described herein. In embodiments, the components of the present disclosure are assembled into a kit.

[0189] In embodiments, the kit comprises various samples of formulated optodes, for examples vials, tubes, containers, and / or ampules, which contain about or at least about a single sensor to about or at least about 1000+ sensors. In embodiments, the vials, tubes, containers, and / or ampules containing optode sensors (e.g., the sensing molecule components alone, or formulated into a matrix) can be resealable. In embodiments, the optode sensors can have a shelf life that allows them to be stored and used over time. Optode sensors herein, in embodiments, can be stably stored and effective for analyte detection for about or at least about 1 day, about or at least about 1 week, about or at least about 1 month, about or at least about 2 months, about or at least about 4 months, about or at least about 6 months, about or at least about 1 year, about or at least about 2 years, about or at least about 5 years, or about or at least about 10 years. In embodiments, optode sensors can be stored at a range of temperatures, including at about or at least about -80°C, about or at least about -20°C, about or at least about 0°C, about or at least about 4°C, about or at least about 25°C, about or at least about 37°C.

[0190] The kits described herein, in embodiments, can include one or more solutions for performing methods described herein, for example solutions suitable for washing / rinsing the optode or optode disk wells without damaging the components of their sensitivity. In embodiments, the kit can include one or more suitable diluents, e.g., such as deionized water or PBS for diluting biological samples.

[0191] The kits described herein, in embodiments, can include one or more implements for isolating, handling, dissolving, homogenizing, or otherwise preparing a biological sample for use with the optode and / or optode spin disk, including for example, a syringe and / or needle, a pipette and / or pipette tips, vials, tubes, and the like.

[0192] The kits described herein, in embodiments, can include one or more containers housing components for performing the methods described herein and optionally instructions for use. Any of the kits described herein, in embodiments, can further include components needed for performing the manufacturing methods described herein, e.g., for generating the optode spin disk. Each component of the kit, where applicable, can be provided in liquid, suspension, or fluid form, for example, where the components can be admixed and cast onto a surface, depending on the desired matrix thickness, surface area, and surface type to be applied to. In embodiments, some of the components are reconstitutable or otherwise processible, for example, by the addition of a suitable solvent or other species, which may or may not be provided with the kit. Alternatively, in embodiments, each component of the kit can be pre-assembled and ready for use, for example, where the optode is pre-cast into the wells of the spin disk, and the spin disk is sealed and primed for use.

[0193] In embodiments, the kits may optionally include instructions and / or promotion for use of the components provided. As used herein, "instructions" can define a component of instruction and / or promotion, and typically involve written instructions on or associated with packaging of the disclosure. Instructions also can include any oral or electronic instructions provided in any manner such that a user will clearly recognize that the instructions are to be associated with the kit, for example, audiovisual (e.g., videotape, DVD, etc.), Internet, and / or web-based communications, etc. As used herein, "promoted" includes all methods of doing business including methods of education, engineering instruction, scientific inquiry, discovery or development, academic research, manufacturing, chemical, cosmetic, and pharmaceutical industry activity including sales, and any advertising or other promotional activity including written, oral, and electronic communication of any form, associated with the disclosure. Additionally, the kits may include other components depending on the specific application, as described herein.

[0194] The kits may have a variety of forms, such as a blister pouch, a shrink-wrapped pouch, a vacuum sealable pouch, a sealable thermoformed tray, or a similar pouch or tray form, with the accessories loosely packed within the pouch, one or more tubes, containers, a box, or a bag.

[0195] Without further elaboration, it is believed that one skilled in the art can, based on the above description, utilize the present disclosure to its fullest extent. The following specific embodiments are, therefore, to be construed as merely illustrative, and not limiting of the remainder of the disclosure in any way whatsoever.EXAMPLESExample 1: Manufacturing an Optode Spin Disk to Detect pH, Na+, K+, and Cl- in Blood.

[0196] Whole blood (plasma + cells) are deposited into a holding chamber, and by using centrifugal force, the blood is pushed through a channel or tubing into a separation chamber, where the plasma is separated from whole cells (RBC / WBC), and then further pushed through a channel or tubing into a Lyo bead chamber to fix the pH and ionic content so that changes in analyte concentration do not occur during measurement, where the pH-fixed plasma is then distributed into a plurality of wells (at least 4) which individually measure pH, Na+, K+, and Cl- content.

[0197] The optode formulation for each individual analyte is concentration controlled for sensitivity in the physiological range, as well as for thickness. For uniform thickness, the capping substrate is constructed that it is: 1) made from material that is resistance to organic solvents (e.g., THF, CHx) which dissolve the chromo / ionophore molecules, and 2) surface treated (e.g., plasma charged, hydrophobic layer, hydrophilic layer) so that the layer forms uniformly. Uniform, thin thickness is achieved which, as summarized in the equation(s) and theoretical modeling presented herein, illustrate that the background subtraction and protonation state relax the dependence on thickness, and / or reduce sensitivity to thickness.

[0198] The optode is formed on the surface of a capping substrate (which is generally a different material than the optode spin disk plate) where small volume (e.g., 50-200 nL) droplets which have the optode molecules dissolved therein are deposited onto the surface. Once the optode matrix is formed on the capping substrate, it is affixed to the optode spin disk plate by adhesive. This layer is then laminated to be set in place to withstand centrifugal force.

[0199] Numerous formulations with varying chromoionophore / ionophore concentrations and ratios are tested for each analyte to arrive at optodes which are sensitive in the physiological range of the desired blood analyte, while reducing cross-reactivity to other ions.Example 2: Optode spectra signal intensity analysis for determining ion concentration in physiological ranges

[0200] Buffer solutions of sodium chloride (NaCI) at 0.01 mM to 100 mM ionic concentration were produced and subjected to an optode at ambient temperature. Absorbance spectra from 400 nm to 800 nm was measured, with intensities at 505 nm and 660 nm particularly measured. As shown in Fig. 20, decreasing concentration of NaCI is observed as a decrease in signal intensity at 660 nm with a concomitant increase in signal intensity at 505 nm.

[0201] As shown in Fig. 21, color change optodes cast within a capping layer as illustrated in Fig. 14 (e.g., resembling the fluid flow channel depicted in Fig. 13), demonstrated color differences in the visible EM range as a function of potassium ion concentration [K+] change in the physiological range (e.g., in comparison from left 6.7 mM to right 2.7 mM).

[0202] This data demonstrated, inter alia, that observable spectra differences within the range of 400 nm to 800 nm are detectable in the physiological range for bodily fluid ions using an optode.

[0203] Spectra analysis was then performed with an optode spin disk to determine if spectra from a series of optode wells is resolvable as a function of the disk spinning, and to locate the physical center of the optical mask aperture and optode. As shown in Fig. 22A, an algorithm for single wavelength peaks as a function of time (e.g., also as a function of optode disk rotation) is illustrated. First, derivatives were calculated from each individual peak in the spectra, e.g., as shown in the illustrative function at the bottom in Fig. 22A. The derivatives are used to mark the maxima and minima (e.g., first derivative) and / or to determine the inflection points of the function (e.g., second derivative). The index peaks are determined from “blank wells” that flank a series of optode well spectra, for example as shown in the graphical representation, e.g., denoted as “2” in the graph at the top with the spheroids marking the index wells as visual markers. This confirmed the ability to resolve the pattern of optode spectra and determined the index marks for rotations, where index to index is a single rotation through a plurality of optodes (or subset of optodes). For each peak, the crossover point was determined with 50% maxima, e.g., denoted as “3” in the bottom function. For each peak, the centroid was taken between the start and end, which was used to average the peak height within a signal region. This reading was used to calculate the optical density (OD) relative to the refence well. Fig. 22B similarly shows a series of calculations which are maxima-independent, yielding similar results.

[0204] To account for optodes not being homogeneous or not being located in exactly the same location relative to each individual well (e.g., as shown in Figs. 23A-23B), optode centration analysis was performed. As shown in Figs. 23A-23B, the center position is robust to a small amount of misalignment, where differences in spectra signal intensity were resolved such that the optode true center was determined from the spectra during rotation.

[0205] Signals were processed to determine ion concentration, e.g., with representative spectra shown in Fig. 24. The following equation was sued with representative spectra intensity at 505 nm, 660 nm, and 720 nm:Color signal protonation = ,ODS0Snm- 0D72Qnm) / 0D^nm+ OD660nm- 2 X 0D720nm)

[0206] The average intensity over each center peak was calculated, then each average intensity over multiple rotations was averaged, and the optical density (OD) at each wavelength was calculated relative to the plasma blank, and the “color signal protonation” was calculated. The concentration was then obtained using this value relative to a calibration curve.

[0207] As shown in Fig. 25, calibration curves over a physiological concentration range and pH range is illustrated for the spin disk optode. Calibration curves were calculated for each of the blood ions, sodium, chloride, and potassium as follows:Concentation [Na, Cl, or pH] = A x (protonation) + BConcentration [7C] = B X protonationA

[0208] Sodium [Na+], chloride [CI-], and pH are calculated using linear fits, where potassium [K] is a power fit.

[0209] pH error term can be accounted for as linear offset to protonation (prot-C*pH) for small deviations about nominal pH, for example:[ ] = ((prot — C x pH) / BA[Na, Cl, pH] = prot — C x pH) — B) / A

[0210] These calibration curves were built for the purpose of calibrating blood plasma samples at different pH levels because plasma samples exhibit different pH. pH optodes were used to obtain the pH value of the plasma samples. Calibration curves were calculated at different pH levels and the plasma sample ions were evaluated.

[0211] These data demonstrated, inter alia, that the spin disk optode was useable to determine ion concentrations of sodium [Na+], potassium [K+], chloride [CI-], and pH within the physiological range present in bodily fluids, such as blood and plasma. Further, spectral analysis can account for variations in optode spin disk rotation reads, as well as optode placement within the well and pH drift in samples during readings.DEFINITIONS

[0212] The following definitions are used in connection with the disclosure disclosed herein. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of skill in the art to which this disclosure belongs.

[0213] As used herein, “a,” “an,” or “the” can mean one or more than one.

[0214] Further, the term “about” when used in connection with a referenced numeric indication means the referenced numeric indication plus or minus up to 10% of that referenced numeric indication. For example, the language “about 50” covers the range of 45 to 55.

[0215] As referred to herein, all compositional percentages are by weight of the total composition, unless otherwise specified. As used herein, the word “include,” and its variants, is intended to be non-limiting, such that recitation of items in a list is not to the exclusion of other like items that may also be useful in the compositions and methods of this technology. Similarly, the terms “can” and “may” and their variants are intended to be non-limiting, such that recitation that an embodiment can or may comprise certain elements or features does not exclude other embodiments of the present technology that do not contain those elements or features.

[0216] Although the open-ended term “comprising,” as a synonym of terms such as including, containing, or having, is used herein to describe and claim the disclosure, the present disclosure, or embodiments thereof, may alternatively be described using alternative terms such as “consisting of” or “consisting essentially of.”

[0217] In embodiments, as used herein, the words “preferred” and “preferably” refer to embodiments of the technology that afford certain benefits, under certain circumstances. However, other embodiments may also be preferred, under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful, and is not intended to exclude other embodiments from the scope of the technology.EQUIVALENTS

[0218] While the disclosure has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the disclosure following, in general, the principles of the disclosure and including such departures from the present disclosure as come within known or customary practice within the art to which the disclosure pertains and as may be applied to the essential features hereinbefore set forth and as follows in the scope of the appended claims.

[0219] Those skilled in the art will recognize, or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific embodiments described specifically herein. Such equivalents are intended to be encompassed in the scope of the following claims.INCORPORATION BY REFERENCE

[0220] All patents and publications referenced herein are hereby incorporated by reference in their entireties.

[0221] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present disclosure is not entitled to antedate such publication by virtue of prior disclosure.

[0222] As used herein, all headings are simply for organization and are not intended to limit the disclosure in any manner. The content of any individual section may be equally applicable to all sections.

Claims

CLAIMSWhat is claimed is:

1. An optochemical sensor (optode) for detecting one or more analytes in a biological sample comprising: a matrix comprising one or more polymers and one or more plasticizers, wherein the matrix has disposed therein: i) one or more chromoionophores; ii) one or more ionophores; and ill) one or more ion exchangers, wherein the optode is configured to generate at least one optical signal as a function of the detection of the one or more analytes by the one or more chromoionophores and the one or more ionophores.

2. The optode of claim 1 , wherein the one or more polymers comprises one or more of polyvinyl chloride (PVC), polyethylene terephthalate (PET), sulfonated polyether ether ketone (SPEEK), cellulose acetate, and cellulose diacetate.

3. The optode of claim 1 or 2, wherein the one or more polymers comprises polyvinyl chloride (PVC).

4. The optode of any one of claims 1-3, wherein the one or more plasticizers comprises one or more of dioctyl sebacate (DOS), ortho-nitrophenyloctylether (NPOE), bis(1 -butylpentyl) adipate (BBPA), dioctyl phthalate (DEHP / DOP), dioctyl terephthalate (DEHT / DOPT), trioctyl trimellitate (TOTM), trioctyl phosphate (TOP), tritolyl phosphate (TCP), triphenyl phosphate (TPhP), 2-nitrophenyl dodecyl ether (NPDE), 2-nitrophenyl tetradecyl ether (NPTE), polycaprolactone (PCL), phthalate ester, dibutyl sebacate, citrate ester, triacetin, adipate, azelate, benzoate, ortho-phthalate, terephthalate, and trimellitate.

5. The optode of any one of the preceding claims, wherein the one or more plasticizers comprises dioctyl sebacate (DOS), ortho-nitrophenyloctylether (NPOE), bis(1 -butylpentyl) adipate (BBPA), dioctyl phthalate (DEHP / DOP), dioctyl terephthalate (DEHT / DOPT), and trioctyl trimellitate (TOTM).

6. The optode of any one of the preceding claims, wherein the one or more chromoionophores is configured to detect one or more monovalent cations or anions from the biological sample.

7. The optode of claim 6, wherein the one or more monovalent cations comprises H+.

8. The optode of claim 6 or 7, wherein the one or more chromoionophores is configured to detect H+ at a concentration range of at least about pH 6.8 to at least about pH 9.

9. The optode of any one of claims 6-8, wherein the one or more chromoionophores comprises one or more of nitron-8-hydroxyquinoline-5-sulfonate, 4,4'-bis[(N,N-dialkylamino)methyl]-azobenzene, azobenzene, o-cresolphtalein octadecyl ester, Cresol Red, Phenol Red, 1-(2-Pyridylazo)-2-naphthol (PAN), o-Cresolphthalein, alpha-Naphtholphthalein, Phenolphthalein, Thymol Blue, m-Cresol Purple, Turmeric (Curcumin), 4-[(2,6-dibromo-4-nitrophenyl)azo]-2-octadecyloxy-1-naphthol, octyl bis(2,4-dinitrophenyl)acetate, 4-nonadecylpyridine (ETH 1907), 5-Octadecanoyloxy-2-(4- nitrophenylazo)phenol (ETH 2412), octadecyl 2-(4-dipropylaminophenylazo)benzoate (ETH 2418), ETH 2430, 9-dimethylamino-5-[4-(16-butyl-2,14-dioxo-3,15- dioxaeicosyl)phenylimino]benzo[a]phenoxazine (ETH 2439), ETH 2458, ETH 2462, ETH 2468, ETH 2479, ETH 3531, 3-octadecanoylimino-7-(diethylamino)-1 ,2-benzophenoxazine (ETH 5294), ETH 5348, 9-(diethylamino)-5-[(2-octyldecyl)imino]benzo[a]phenoxazine (ETH 5350), ETH 5355, 9- Dimethylamino-5-[4-(15-buty I- 1 , 13-dioxo-2, 14- dioxanonadecyl)phenylimino]benzo[a]phenoxazine(ETH 5418), 3',3",5',5"-Tetrabromophenolphthaleinethyl ester (TBPE), 1-Hydroxy-4-[4-(2- hydroxyethylsulfonyl)phenylazo]naphthalene-2-sulfonic acid potassium salt (GJM-541), ETH 7058, ETH 7061, and ETH 7075.

10. The optode of any one of the preceding claims, wherein at least one of the one or more ionophores is configured to detect at least one monovalent cation.

11. The optode of claim 10, wherein the one or more monovalent cations comprises one or more of potassium (K+) and sodium (Na+).

12. The optode of claim 10 or 11, wherein the one or more ionophores is configured to detect sodium (Na+) at a concentration range of at least about 80 mM to at least about 180 mM.

13. The optode of claim 10, wherein the one or more ionophores is configured to detect potassium (K+) at a concentration range of at least about 2.0 mM to at least about 10 mM.

14. The optode of any one of claims 10-12, wherein the one or more ionophores is configured to detect sodium (Na+) and comprises one or more of gramicidin A, laidlomycin, lasalocid, maduramicin, monensin, narasin, nigericin, nonactin, nystatin, salinomycin, valinomycin, dicyclohexano-18-crown-6, calix[4]arenecrown-4, potassium tetrakis(p-chlorophenyl)borate (KTpCIPB), orthonitrophenyloctylether, sodium tetrakis(p-chlorophenyl)borate (NaTFPB), Sodium tetrakis[3,5- bis(trifluoromethyl)phenyl]borate (NaBARF), Potassium tetrakis[3,5- bis(trifluoromethyl)phenyl]borate (KTFPB), bis-2-ethylhexylsebacate, 2,7-di-tert-butyl-9,9-dimethyl- 4,5-xanthenediamine, tridodecylmethylammonium chloride (TDDMACI), 1 ,8-naphthalenediol, bis(1- butylpentyl)adipate (BBPA), crown ether ionophores, bis[(12-crown-4)methyl] dodecylmethylmalonate, di(o-methoxy)-stilbeno-24-crown-8, 15-octadecyloxymethyl-15-methy I- 16- crown-5, dibenzo-16-crown-5 nitrophenol, 4-tert-butylcalix[4]arene-tetraacetic acid tetraethyl ester, Sodium ionophore IV (Cas #: 172883-29-3), N,N'-dibenzyl-N,N'-diphenyl-1 ,2- phenylenedioxydiacetamide (ETH 157), N,N',N"-triheptyl-N,N',N"-trimethyl-4,4',4"-propylidynetris(3- oxabutyramide) (ETH 227), N,N,N',N'-tetracyclohexyl-1 ,2-phenylenedioxydiacetamide (ETH 2120), and ETH 4120.

15. The optode of any one of claims 10-11 or 13, wherein the one or more ionophores is configured to detect potassium (K+) and comprises one or more of nonactin, monactin, valinomycin, nigericin, hydroxylated polyvinyl chloride (PVC-OH), dicyclohexano-18-crown-6, dibenzo-18-crown-6, benzo- 15-crown-5, dibenzo-30-crown-10, dinaphtho-30-crown-10, 4-nitrobenzo-18-crown-6, 4'- picrylamino-5'-nitrobenzo-18-crown-6, 2, 3-naphtho-15-crown-5, (hexanoyloxymethyl)benzo-l 5- crown-5, octadecanoyloxy-methyl-15-crown-5, benzo-15-crown-5 ether, bis[(benzo-15-crown-5)-4'- ylmethyl] pimelate, and 2-dodecyl-2-methyl-1,3-propanediyl bis[N-[5'-nitro(benzo-15-crown-5)-4'- yl]carbamate] (BME-44).

16. The optode of any one of claims 1 -9, wherein at least one of the one or more ionophores is configured to detect a monovalent anion from the biological sample.

17. The optode of claim 16, wherein the one or more monovalent anions comprises chloride (CI-).

18. The optode of claim 16 or 17, wherein the one or more ionophores is configured to detect chloride (CI-) at a concentration range of at least about 60 mM to at least about 140 mM.

19. The optode of any one of claims 16-18, wherein the one or more ionophores is configured to detect chloride (CI-) and comprises one or more of triphenyltin chloride, tributyltin chloride, trioctyltin chloride, macrocyclic tetramercury, dodecylbenzylmercury(ll) chloride, tricyclohexyltin chloride, triphenylgermanium chloride, phenylmercuric chloride, hafnocene dichloride, chloro(octaethylporphyrinato)indium-(lll), meso-Tetraphenylporphyrin manganese(lll)-chloride, 4,5-Bis-[N'-(butyl)thioureido]-2,7-di-tert-butyl-9,9-dimethylxanthene, ETH 2412, 4,5-Dimethyl-3,6- dioctyloxy-o-phenylene-bis(mercurytrifluoroacetate) (ETH 9009), and 3,6-didodecyloxy-4,5- dimethyl-o-phenylene-bis(mercury chloride) (ETH 9033).

20. The optode of any one of the preceding claims, wherein the one or more ionophores and / or one or more chromoionophores comprises a lipophilic indicator dye.21 . The optode of any one of the preceding claims, wherein the one or more ion exchangers comprises one or more of potassium tetrakis(p-chlorophenyl)borate (KTpCIPB), sodium tetrakis(p- chlorophenyl)borate (NaTFPB), Sodium tetrakis[3,5-bis(trifluoromethyl)phenyl]borate (NaBARF), Potassium tetrakis[3,5-bis(trifluoromethyl)phenyl]borate (KTFPB), Tridodecylmethylammonium chloride (TDDMACI).

22. The optode of any one of the preceding claims, wherein the one or more ion exchangers comprises a photo-regenerative ion-exchange ionophore configured to extract one or more analyte cations or anions and release the one or more analyte cations or anions as a function of exposure to one or more wavelengths of electromagnetic radiation from about 280 nm to about 1000 nm.

23. The optode of any one of the preceding claims, wherein the optode is configured to provide the at least one optical signal as a function of detection of one or more of H+ (pH), potassium (K+), sodium (Na+), and chloride (CI-) within a range of a human physiological concentration.

24. The optode of claim 23, wherein the optode is configured to provide at least one optical signal as a function of H+ concentration (pH), optionally wherein the optical signal is calibrated to provide an optical signal as a function of the range of the human physiological concentration of at least about pH 6.8 to at least about pH 9.

25. The optode of claim 23, wherein the optode is configured to provide at least one optical signal as a function of potassium (K+) concentration, optionally wherein the optical signal is calibrated to provide an optical signal as a function of the range of the human physiological concentration of at least about 2.0 mM to at least about 10 mM.

26. The optode of claim 23, wherein the optode is configured to provide at least one optical signal as a function of sodium (Na+) concentration, optionally wherein the optical signal is calibrated to provide an optical signal as a function of the range of the human physiological concentration of at least about 80 mM to at least about 180 mM.

27. The optode of claim 23, wherein the optode is configured to provide at least one optical signal as a function of chloride (CI-) concentration, optionally wherein the optical signal is calibrated to provide an optical signal as a function of the range of the human physiological concentration of at least about 60 mM to at least about 140 mM.

28. The optode of any one of the preceding claims, wherein the optode is not substantially selective for one or more ions of calcium, magnesium, lithium, rubidium, cesium, ammonium, beryllium, strontium, barium, carbon dioxide, carbonate, bicarbonate, oxygen, thiocyanate, nitrate, nitrite, hydroxide, hydrogen sulfide, phosphate, sulfide, sulfite, sulfate, perchlorate, arsenic, iodide, and transition metals, optionally including antimony, molybdenum, iron, lead, copper, zinc, chromium, silver, selenium, mercury, uranium, thallium, samarium, and cadmium.

29. The optode of any one of the preceding claims, wherein the biological sample comprises one or more of blood, sera, plasma, bone marrow, lymph, saliva, sputum, mucus, respiratory or nasal secretion, oropharyngeal swab, nasopharyngeal swab, oral swab, ductal lavage, bronchoalveolar lavage, cerebrospinal fluid, skin swab, vaginal swab, gastric juice, ascites, peritoneal fluid, pleural fluid, gynecological fluids, pus, perspiration, tears, urine, and stool.

30. The optode of claim 29, wherein the biological sample comprises blood.31 . The optode of any one of the preceding claims, wherein the optode is reusable.

32. The optode of any one of the preceding claims, wherein the optode is configured to be washed with an inorganic solvent and detect one or more analytes from at least a second biological sample with less than about 0.01% loss in sensitivity, less than about 0.05% loss in sensitivity, less than about 0.1% loss in sensitivity, less than about 0.2% loss in sensitivity, less than about 0.3% loss in sensitivity, less than about 0.4% loss in sensitivity, less than about 0.5% loss in sensitivity, less than about 0.6% loss in sensitivity, less than about 0.7% loss in sensitivity, less than about 0.8% loss in sensitivity, less than about 0.9% loss in sensitivity, less than about 1 .0% loss in sensitivity, less than about 1 .5% loss in sensitivity, or less than about 2.0% loss in sensitivity.

33. The optode of claim 32, wherein the inorganic solvent is one or more of deionized water, phosphate- buffered saline (PBS), sodium chloride (NaCI), sodium hydroxide, tris base, HEPES, MOPS, PIPES, urea, guanidine hydrochloride, and detergent.

34. The optode of any one of the preceding claims, wherein the at least one optical signal is detectable in a range of at least about 300 nm to at least about 800 nm.

35. The optode of any one of the preceding claims, wherein the optode is configured to generate at least one optical signal for detecting each analyte.

36. The optode of any one of the preceding claims, wherein the at least one optical signal comprises one or more of a fluorescence signal, colorimetric signal, luminescent signal, absorption signal, and reflectance signal.

37. The optode of any one of claims 34-36, wherein the at least one optical signal comprises a signal detectable at about 340 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

38. The optode of any one of claims 34-36, wherein the at least one optical signal comprises a signal detectable at about 405 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

39. The optode of any one of claims 34-36, wherein the at least one optical signal comprises a signal detectable at about 505 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

40. The optode of any one of claims 34-36, wherein the at least one optical signal comprises a signal detectable at about 545 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

41. The optode of any one of claims 34-36, wherein the at least one optical signal comprises a signal detectable at about 660 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

42. The optode of any one of claims 34-36, wherein the at least one optical signal comprises a signal detectable at about 720 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

43. The optode of any one of the preceding claims, wherein the matrix is flexible.

44. The optode of any one of the preceding claims, wherein the matrix is configured to be applied to a surface, optionally wherein the matrix has a uniform thickness of about or at least about 1 pm, about or at least about 5 pm, about or at least about 10 pm, about or at least about 50 pm, about or at least about 100 pm, about or at least about 500 pm, about or at least about 1.0 mm, about or at least about 1.5 mm, about or at least about 2.0 mm, about or at least about 2.5 mm, about or at least about 3.0 mm, about or at least about 3.5 mm, about or at least about 4.0 mm, or about or at least about 5.0 mm.

45. The optode of claim 44, wherein the surface is transparent to at least a portion of an electromagnetic spectrum from at least about 280 nm to at least about 1000 nm.

46. An optochemical sensor (optode) for detecting one or more analytes from a biological sample comprising: a disk comprising one or more wells disposed within the disk, wherein the one or more wells comprises: an optode of any one of claims 1-44 disposed on a surface within the well; an inlet port for providing the biological sample into the well; and at least a portion of an outer surface of the well that is transparent to at least a portion of an electromagnetic spectrum from at least about 280 nm to at least about 1000 nm; and wherein the disk and the one or more wells are configured to be spun and withstand a centrifugal force; and wherein the optode within the well is configured to generate at least one optical signal in the range of at least about 300 nm to at least about 800 nm as a function of binding one or more analytes in the biological sample.

47. The optode of claim 46, wherein the optode comprises a matrix comprising polyvinyl chloride (PVC) dioctyl sebacate (DOS), ortho-nitrophenyloctylether (NPOE), bis(1 -butylpentyl) adipate (BBPA), dioctyl phthalate (DEHP / DOP), dioctyl terephthalate (DEHT / DOPT), trioctyl trimellitate (TOTM), or a mixture thereof, wherein the matrix has disposed therein: a chromoionophore that is proton-selective and configured to detect H+ concentration (pH) in a range of at least about pH 6.8 to at least about pH 9; a first ionophore that is potassium-selective and configured to detect potassium (K+) concentration in a range of at least about 2.0 mM to at least about 10 mM; a second ionophore that is sodium-selective and configured to detect sodium (Na+) concentration in a range of at least about 80 mM to at least about 180 mM; a third ionophore that is chloride-selective and configured to detect chloride (CI-) concentration in a range of at least about 60 mM to at least about 140 mM; and ill) at least one ion exchanger.

48. The optode of claim 46 or 47, wherein the optode is not substantially selective for one or more ions of calcium, magnesium, lithium, rubidium, cesium, ammonium, beryllium, strontium, barium, carbon dioxide, carbonate, bicarbonate, oxygen, thiocyanate, nitrate, nitrite, hydroxide, hydrogen sulfide, phosphate, sulfide, sulfite, sulfate, perchlorate, arsenic, iodide, and transition metals, optionally including antimony, molybdenum, iron, lead, copper, zinc, chromium, silver, selenium, mercury, uranium, thallium, samarium, and cadmium.

49. The optode of any one of claims 46-48, wherein the biological sample comprises one or more of blood, sera, plasma, bone marrow, lymph, saliva, sputum, mucus, respiratory or nasal secretion, oropharyngeal swab, nasopharyngeal swab, oral swab, ductal lavage, bronchoalveolar lavage, cerebrospinal fluid, skin swab, vaginal swab, gastric juice, ascites, peritoneal fluid, pleural fluid, gynecological fluids, pus, perspiration, tears, urine, and stool.

50. The optode of claim 49, wherein the biological sample comprises blood.51 . The optode of any one of claims 46-50, wherein the disk, or one or more layers thereof, is composed of plastic, polymethyl methacrylate, carbon fiber, and / or metal, optionally wherein the metal is aluminum and / or titanium.

52. The optode of any one of claims 46-51, wherein the disk further comprises a substrate to hold the optode in place, optionally wherein the disk comprises one or more of a structural support layer, inlet layer, channel layer, and capping layer, and further optionally wherein the optode in held place on a capping layer.

53. The optode of claim 52, wherein one or more of the structural support layer, inlet layer, channel layer, and capping layer comprises one or more of polyethylene terephthalate (PET), pressure sensitive adhesive (PSA), polyethylene terephthalate glycol (PETG), polyurethane, polyethylene, polypropylene, and glass.

54. The optode of claim 52 or 53, wherein the substrate is chemically resistant to one or more solvents, optionally including cyclohexanone, N,N-dimethylformamide (DMF), acetonitrile, butanone, and dichloromethane.

55. The optode of any one of claims 46-54, wherein the disk comprises at least about 1 well, at least about 2 wells; at least about 3 wells, at least about 4 wells, at least about 5 wells, at least about 6 wells, at least about 7 wells, at least about 8 wells, at least about 9 wells, at least about 10 wells, atleast about 12 wells, at least about 14 wells, at least about 16 wells, at least about 18 wells, at least about 20 wells, at least about 22 wells, at least about 24 wells, at least about 26 wells, at least about 28 wells, at least about 30 wells, at least about 35 wells, or at least about 40 wells.

56. The optode of any one of claims 46-55, wherein the disk is configured to be spun at a rate of at least about 100 revolutions per minute (RPM), at least about 200 RPM, at least about 300 RPM, at least about 400 RPM, at least about 500 RPM, at least about 600 RPM, at least about 700 RPM, at least about 800 RPM, at least about 900 RPM, at least about 1000 RPM, at least about 1200 RPM, at least about 1400 RPM, at least about 1600 RPM, at least about 1800 RPM, at least about 2000 RPM, at least about 2200 RPM, at least about 2400 RPM, at least about 2600 RPM, at least about 2800 RPM, at least about 2900 RPM, at least about 3000 RPM, at least about 3500 RPM, at least about 4000 RPM, at least about 4500 RPM, or at least about 5000 RPM.

57. The optode of any one of claims 46-56, wherein the disk is configured to withstand a centrifugal force of at least about 100 x g, at least about 200 x g, at least about 300 x g, at least about 400 x g, at least about 500 x g, at least about 600 x g, at least about 700 x g, at least about 800 x g, at least about 900 x g, at least about 1000 x g, at least about 1200 x g, at least about 1400 x g, at least about1600 x g, at least about 1800 x g, at least about 2000 x g, at least about 2200 x g, at least about2400 x g, at least about 2600 x g, at least about 2800 x g, at least about 3000 x g, at least about3500 x g, at least about 4000 x g, at least about 4500 x g, or at least about 5000 x g.

58. The optode of any one of claims 46-57, wherein the well comprises a total well volume of at least about 3 pL, at least about 5 pL, at least about 10 pL, at least about 15 pL, at least about 20 pL, at least about 25 pL, at least about 30 pL, at least about 35 pL, at least about 40 pL, at least about 45 pL, at least about 50 pL, at least about 55 pL, at least about 60 pL, at least about 65 pL, at least about 70 pL, at least about 75 pL, at least about 80 pL, at least about 85 pL, at least about 90 pL, at least about 95 pL, at least about 100 pL, at least about 110 pL, at least about 120 pL, at least about 130 pL, at least about 140 pL, at least about 150 pL, at least about 160 pL, at least about 170 pL, at least about 180 pL, at least about 190 pL, or at least about 200 pL.

59. The optode of any one of claims 46-58, wherein the well comprises a well width of at least about 1 mm, at least about 2 mm, at least about 3 mm, at least about 4 mm, at least about 5 mm, at least about 6 mm, at least about 7 mm, at least about 8 mm, at least about 9 mm, or at least about 10 mm.

60. The optode of any one of claims 46-59, wherein the optode disposed on the surface within the well comprises a surface area of at least about 1 mm2, at least about 2 mm2, at least about 3 mm2, at least about 4 mm2, at least about 5 mm2, at least about 6 mm2, at least about 7 mm2, at least about 8 mm2, at least about 9 mm2, at least about 10 mm2, at least about 15 mm2, at least about 20 mm2, at least about 25 mm2, at least about 50 mm2, at least about 100 mm2, at least about 200 mm2, at least about 300 mm2, at least about 400 mm2, at least about 500 mm2, and optionally wherein the optode has a uniform thickness of about or at least about 1 pm, about or at least about 5 pm, about or at least about 10 pm, about or at least about 50 pm, about or at least about 100 pm, about or at least about 500 pm, about or at least about 1 .0 mm, about or at least about 1 .5 mm, about or at least about 2.0 mm, about or at least about 2.5 mm, about or at least about 3.0 mm, about or at least about 3.5 mm, about or at least about 4.0 mm, or about or at least about 5.0 mm.

61. The optode of any one of claims 46-60, wherein the well comprises a overflow well, optionally wherein the overflow well has a total volume of at least about 3 pL, at least about 5 pL, at least about 10 pL, at least about 15 pL, at least about 20 pL, at least about 25 pL, at least about 30 pL, at least about 35 pL, at least about 40 pL, at least about 45 pL, at least about 50 pL, at least about 55 pL, at least about 60 pL, at least about 65 pL, at least about 70 pL, at least about 75 pL, at least about 80 pL, at least about 85 pL, at least about 90 pL, at least about 95 pL, or at least about 100 pL.

62. The optode of any one of claims 46-61 , wherein the inlet port comprises a diameter of at least about 0.5 mm, at least about 0.6 mm, at least about 0.7 mm, at least about 0.8 mm, at least about 0.9 mm, at least about 1 mm, at least about 1 .5 mm, or at least about 2 mm.

63. The optode of any one of claims 46-62, wherein the well comprises an outlet port, optionally wherein the outlet port comprises a diameter of at least about 0.5 mm, at least about 0.6 mm, at least about 0.7 mm, at least about 0.8 mm, at least about 0.9 mm, at least about 1 mm, at least about 1.5 mm, or at least about 2 mm.

64. The optode of any one of claims 46-63, wherein the well comprises a covermask, optionally wherein the covermask comprises a diameter of at least about of at least about 0.5 mm, at least about 1 mm, at least about 1 .5 mm, or at least about 2 mm, or at least about 2.5 mm, or at least about 3.0 mm, or at least about 3.5 mm, or at least about 4 mm, or at least about 4.5 mm, or at least about 5 mm.

65. The optode of any one of claims 46-64, wherein the portion of the outer surface of the well that is transparent is comprised of one or more of glass, quartz, plastic, polystyrene, polyethyleneterephthalate, polycarbonate, polymethyl methacrylate (acrylic), polyethylene, polyurethane, polypropylene, thermoplastic elastomers (TPE), acrylonitrile butadiene styrene (ABS), epoxy, cyclic olefin copolymer (COC), cyclic olefin polymer (COP), poly dimethyl siloxane (PDMS), poly ether ester ketone (PEEK), polyetherimide (ULTEM), and nylon.

66. The optode of any one of claims 46-65, wherein the optode is configured to be washed with a solvent, optionally wherein the solvent comprises one or more of deionized water, phosphate-buffered saline (PBS), sodium chloride (NaCI), sodium hydroxide, tris base, HEPES, MOPS, PIPES, urea, guanidine hydrochloride, and detergent.

67. The optode of any one of claims 46-66, wherein the optode has a fluid circuit embedded within the disk that fluidically connects to the one or more wells by a channel or tubing.

68. The optode of claim 67, wherein the fluid circuit fluidically connects the one or more wells to one or more pH fix chambers, pH priming circuits, bead mixing chambers, overflow chambers, separator chambers, siphons, and air volume ballasts.

69. The optode of claim 68, wherein the one or more pH fix chambers are configured to fix the ionic content of the biological sample to prevent deviation in concentrations of the one or more analytes during an optode sensing phase.

70. The optode of any one of claims 67-69, wherein the fluid circuit fluidically connects the one or more pH fix chambers to one or more separator chambers and / or siphons.

71. The optode of any one of claims 67-71, wherein the one or more separator chambers is configured to separate cells, cell debris, and / or plasma from the biological sample during and / or after a centrifugal force is applied to the disk.

72. The optode of any one of claims 67-71 , wherein the fluid circuit fluidically connects the one or more separator chambers to one or more sample reservoirs and / or siphons.

73. The optode of claim 72, wherein the one or more sample reservoirs is configured to hold the biological sample and comprises a volume of about or at least about 10 pL, about or at least about 20 pL, about or at least about 50 pL, about or at least about 0.1 mL, about or at least about 0.2 mL, about or at least about 0.3 mL, about or at least about 0.4 mL, about or at least about 0.5 mL, about or at least about 0.6 mL, about or at least about 0.7 mL, about or at least about 0.8 mL, about or at least about 0.9 mL, or about or at least about 1 .0 mL.

74. The optode of any one of claims 46-73, wherein the one or more wells comprise the inlet port and an outlet port and the one or more wells are configured to be connected to a liquid handling device or microfluidic platform.

75. The optode of claim 74, wherein the liquid handling device or microfluidic platform is in fluid connection with the inlet port and the outlet port of the one or more wells.

76. The optode of claim 74 or 75, wherein the microfluidic platform comprises at least one value, a section of tubing, and a fluid pumping mechanism.

77. A method for detecting one or more analytes in a biological sample comprising: providing the optochemical sensor (optode) of any one of claims 45-75; dispensing one or more aliquots of the biological sample into one or more wells of the optode; applying a centrifugal force to the optode; and detecting at least one optical signal from the one or more wells in the range of at least about 300 nm to at least about 800 nm as a function of binding to one or more analytes in the biological sample.

78. The method of claim 77, wherein the biological sample comprises one or more of blood, sera, plasma, bone marrow, lymph, saliva, sputum, mucus, respiratory or nasal secretion, oropharyngeal swab, nasopharyngeal swab, oral swab, ductal lavage, bronchoalveolar lavage, cerebrospinal fluid, skin swab, vaginal swab, gastric juice, ascites, peritoneal fluid, pleural fluid, gynecological fluids, pus, perspiration, tears, urine, and stool.

79. The method of claim 78, wherein the biological sample comprises blood.

80. The method of any one of claims 77-79, further comprising dispensing one or more aliquots of the biological sample into one or more sample reservoirs on the optode, optionally wherein the one or more aliquots comprises a volume of a total volume of at least about 3 pL, at least about 5 pL, at least about 10 pL, at least about 15 pL, at least about 20 pL, at least about 25 pL, at least about 30 pL, at least about 35 pL, at least about 40 pL, at least about 45 pL, at least about 50 pL, at least about 55 pL, at least about 60 pL, at least about 65 pL, at least about 70 pL, at least about 75 pL, at least about 80 pL, at least about 85 pL, at least about 90 pL, at least about 95 pL, at least about 100 p L, at least about 110 p L, at least about 120 p L, at least about 130 p L, at least about 140 p L, at least about 150 pL, at least about 160 pL, at least about 170 pL, at least about 180 pL, at least about 190L, at least about 0.2 mL, about or at least about 0.3 mL, about or at least about 0.4 mL, about or at least about 0.5 mL, about or at least about 0.6 mL, about or at least about 0.7 mL, about or at least about 0.8 mL, about or at least about 0.9 mL, or about or at least about 1 .0 mL.

81. The method of any one of claims 77-80, wherein applying the centrifugal force comprises spinning the optode at a rate of at least about 100 RPM, at least about 200 RPM, at least about 300 RPM, at least about 400 RPM, at least about 500 RPM, at least about 600 RPM, at least about 700 RPM, at least about 800 RPM, at least about 900 RPM, at least about 1000 RPM, at least about 1200 RPM, at least about 1400 RPM, at least about 1600 RPM, at least about 1800 RPM, at least about 2000 RPM, at least about 2200 RPM, at least about 2400 RPM, at least about 2600 RPM, at least about 2800 RPM, at least about 2900 RPM, at least about 3000 RPM, at least about 3500 RPM, at least about 4000 RPM, at least about 4500 RPM, or at least about 5000 RPM.

82. The method of any one of claims 77-81 , wherein applying the centrifugal force comprises a centrifugal force of at least about 100 x g, at least about 200 x g, at least about 300 x g, at least about 400 x g, at least about 500 x g, at least about 600 x g, at least about 700 x g, at least about 800 x g, at least about 900 x g, at least about 1000 x g, at least about 1200 x g, at least about 1400 x g, at least about 1600 x g, at least about 1800 x g, at least about 2000 x g, at least about 2200 x g, at least about 2400 x g, at least about 2600 x g, at least about 2800 x g, at least about 3000 x g, at least about 3500 x g, at least about 4000 x g, at least about 4500 x g, or at least about 5000 x g.

83. The method of any one of claims 77-82, further comprising separating cells and / or cell debris from the biological sample during and / or after the centrifugal force is applied to the optode.

84. The method of any one of claims 77-83, further comprising fixing the ionic content of the biological sample to prevent deviation in concentrations of the one or more analytes during the detecting.

85. The method of any one of claims 77-84, wherein detecting the at least one optical signal comprises measuring one or more of a fluorescence signal, colorimetric signal, luminescent signal, absorption signal, and reflectance signal.

86. The method of claim 85, wherein measuring comprises using one or more of a sensor monitor, absorbance spectrometer, reflectometric photometer, LED photometer, fluorimeter, and luminometer.

87. The method of any one of claims 77-86, wherein the at least one optical signal comprises a signal detectable at about 340 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

88. The method of any one of claims 77-86, wherein the at least one optical signal comprises a signal detectable at about 405 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

89. The method of any one of claims 77-86, wherein the at least one optical signal comprises a signal detectable at about 505 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

90. The method of any one of claims 77-86, wherein the at least one optical signal comprises a signal detectable at about 545 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

91. The method of any one of claims 77-86, wherein the at least one optical signal comprises a signal detectable at about 660 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

92. The method of any one of claims 77-86, wherein the at least one optical signal comprises a signal detectable at about 720 nm, optionally for detecting one or more of H+, K+, Na+, and CI-.

93. The method of any one of claims 77-92, wherein the detecting is not substantially selective for one or more ions of calcium, magnesium, lithium, rubidium, cesium, ammonium, beryllium, strontium, barium, carbon dioxide, carbonate, bicarbonate, oxygen, thiocyanate, nitrate, nitrite, hydroxide, hydrogen sulfide, phosphate, sulfide, sulfite, sulfate, perchlorate, arsenic, iodide, and transition metals, optionally including antimony, molybdenum, iron, lead, copper, zinc, chromium, silver, selenium, mercury, uranium, thallium, samarium, and cadmium.

94. The method of any one of claims 77-93, further comprising preparing the optode with one or more organic solvents comprising tetrahydrofuran (THF), cyclohexanone, N,N-dimethylformamide (DMF), acetonitrile, butanone, and dichloromethane.

95. The method of any one of claims 77-94, further comprising washing the one or more wells with an inorganic solvent, optionally wherein the inorganic solvent comprise one or more of deionized water, phosphate-buffered saline (PBS), sodium chloride (NaCI), sodium hydroxide, tris base, HEPES, MOPS, PIPES, urea, guanidine hydrochloride, and detergent.

96. The method of any one of claims 77-95, wherein dispensing the one or more aliquots of the biological sample and / or washing the one or more wells comprises using a microfluidic platform, wherein the microfluidic platform is in fluid connection with the one or more wells.

97. The method of any one of claims 77-96, comprising dispensing at least a second biological sample into the one or more wells of the optode.

98. The method of claim 97, comprising detecting at least one optical signal from the one or more wells after dispensing at least a second biological sample, and wherein the detection does not result in a substantially significant reduction in sensitivity.

99. The method of claim 98, wherein the substantially significant reduction in sensitivity is less than about 0.01% loss in sensitivity, less than about 0.05% loss in sensitivity, less than about 0.1% loss in sensitivity, less than about 0.2% loss in sensitivity, less than about 0.3% loss in sensitivity, less than about 0.4% loss in sensitivity, less than about 0.5% loss in sensitivity, less than about 0.6% loss in sensitivity, less than about 0.7% loss in sensitivity, less than about 0.8% loss in sensitivity, less than about 0.9% loss in sensitivity, less than about 1.0% loss in sensitivity, less than about 1.5% loss in sensitivity, or less than about 2.0% loss in sensitivity.

100. The method of any one of claims 95-99, comprising two or more cycles of said washing, dispensing, and detecting one or more analytes from an additional biological sample.

101. A method of diagnosing one or more medical conditions and / or disorders comprising performing the method of any one of claims 77-100 and diagnosing the one or more medical conditions and / or disorders as a function of detecting one or more of pH (H+), sodium (Na+) concentration, potassium (K+) concentration, and chloride (CI-) concentration.

102. The method of claim 101, wherein the one or more medical conditions and / or disorders comprises a medical condition and / or disorder associated with a blood concentration of one or more of H+, Na+, K+, and CI-.

103. The method of claim 101 or 102, wherein the one or more medical conditions and / or disorders comprises one or more of acidosis, alkalosis, hypokalemia, hyperkalemia, hyponatremia, hypernatremia, hyperchloremia, hypochloremia, Cushing syndrome, Barter syndrome, kidney disease, heart failure, heart disease, albuminuria, hyperaldosteronism, ketoacidosis, lactic acidosis, renal tubular acidosis, diabetes, dehydration, hyperglycemia, hyperparathyroidism, effects of a prescribed therapeutic agent, including effects of diuretics (hydrochlorothiazide, furosemide, torsemide, and indapamide) and / or effects carbonic anhydrase inhibitors (glaucoma treatment), ethylene glycol poisoning, methanol poisoning, salicylate toxicity (aspirin overdose), ureteraldiversion, syndrome of inappropriate antidiuretic hormone secretion (SIADH), and cancer (endocrine neoplasia, kidney cancer, liver cancer).

104. The method of any one of claims 101-103, further comprising prescribing one or more therapeutic agents as a function of the detecting.

105. The method of claim 104, wherein the one or more therapeutic agents comprises a therapeutic agent associated with a blood concentration of one or more of H+, Na+, K+, and CI-.

106. The method of claim 104 or 105, wherein the one or more therapeutic agents comprises one or more of electrolyte supplementation (potassium salt, chloride salt, sodium salt), sodium zirconium cyclosilicate (LOKELMA), diuretics (loop diuretics, amiloride triamterene), hypertonic or hypotonic saline, vasopressin receptor antagonists, demeclocycline therapy, bicarbonate therapy, hemodialysis, continuous renal replacement therapy (CRRT), alkalinizing agents, mineralocorticoids, vitamin D supplementation, carbonic anhydrase inhibitors, acetazolamide, insulin, ketoconazole, mitotane (LYSODREN), metyrapone (METOPIRONE), mifepristone (KORLYM, MIFEPREX), angiotensin-converting enzyme (ACE) inhibitors, angiotensin receptor blockade (ARB), aldosterone antagonists, spironolactone, indomethacin, ibuprofen, non-steroidal anti-inflammatory drugs (NSAIDs), growth hormone (GH), dapagliflozin (FARXIGA), blood thinners, vasodilators, enalapril (VASOTEC, EPANED), lisinopril (ZESTRIL, QBRELIS), captopril, and alcohol dehydrogenase enzyme (ADH) antagonists (Fomepizole).

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