Methods and devices for detecting hemolysis

Abstract

A sensor assembly enables hemolysis detection by a diagnostic analyzer. The sensor assembly can be included in a diagnostic cartridge that can be coupled to the diagnostic analyzer and can deliver a sample fluid to the sensor assembly. The sensor assembly includes a primary oxygen sensor configured to provide a first measurement and a modified oxygen sensor including an oxidant configured to oxidize hemoglobin iron to methemoglobin and configured to provide a second measurement. A hemolysis detection module of the diagnostic analyzer provides the level of hemolysis in the sample based on the difference between the first measurement and the second measurement. Numerous other diagnostic analyzers, diagnostic cartridges, and detection methods are provided.

Description

[Technical Field]

[0001] This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 63 / 482,123, filed January 30, 2023. The entire contents of the above-referenced patent applications are expressly incorporated herein by reference.

[0002] This application relates to diagnostic testing methods and devices, and more particularly to methods and devices that enable detection of hemolysis levels in blood samples. [Background technology]

[0003] Hemolysis is the destruction of red blood cells, which in some cases can result from preanalytical causes associated with blood collection or sample handling. For example, hemolysis in blood samples can be caused by using the wrong needle size, improper tube mixing, incorrectly filling the tube, using too much suction, applying a tourniquet for too long, prolonged storage, extreme temperatures, processing delays, and / or other disturbances in blood sample collection or sample handling.

[0004] Similarly, hemolysis of a blood sample can be caused by certain diseases or conditions, such as hemolytic anemia, autoimmune diseases, bone marrow failure, and inherited blood disorders such as sickle cell disease or thalassemia.

[0005] Hemolysis has traditionally been detected by manual visual inspection of a blood sample by a technician after separating the plasma (or serum) portion (i.e., by centrifugation) and comparing the color of the plasma (or serum) to a colored hemolysis chart. The chart shows the color of the separated sample associated with an increased concentration of free (extracellular) hemoglobin contained in the plasma (or serum). Thus, a technician can determine the hemolysis index based on the visual color of the separated plasma (or serum).

[0006] Hemolysis due to improper or incorrect handling during specimen collection is a potentially undesirable prerequisite that can affect the accuracy of results and the reliability of blood gas testing. For example, the impact of in vitro hemolysis on measured potassium concentrations is well known. In such cases, reported potassium concentrations may be clinically inaccurate, the magnitude of which depends on the degree of hemolysis. Many other analytes may also be affected by the interfering effects of in vitro hemolysis on biological assays. For example, when free (extracellular) hemoglobin is present in a blood sample, its properties can interfere with certain types of diagnostic tests, such as those involving optical measurement techniques. In particular, free extracellular hemoglobin present in a sample can interfere with certain assays due to its absorption properties. Therefore, identifying samples containing hemolysis is desirable for laboratory or point-of-care testing, as it allows for the possibility of raising suspicion and / or redrawing certain results. Furthermore, automated testing is desirable to expedite the hemolysis detection process while minimizing the subjectivity of manual visual hemolysis determinations made by medical professionals. Eliminating the centrifugation process for testing for hemolysis would also be beneficial. Summary of the Invention [Problem to be solved by the invention]

[0007] Therefore, devices and methods that can enable automated hemolysis detection and improved speed, while minimizing or eliminating the judgments and inconsistencies associated with manual visual inspection, are desirable. [Means for solving the problem]

[0008] In some embodiments provided herein, a diagnostic sensor assembly is provided. The diagnostic sensor assembly can be implemented in a diagnostic cartridge configured to connect to a diagnostic analyzer to determine the level of hemolysis in a sample. The diagnostic sensor assembly includes a sample inlet configured to receive a sample, a sample passageway extending from the sample inlet, a primary oxygen sensor configured to contact the sample along the sample passageway, and a modified oxygen sensor configured to contact the sample along the sample passageway, the modified oxygen sensor being configured to convert hemoglobin to methemoglobin (Fe 2+ From Fe 3+ and an oxidizing agent configured to oxidize, i.e., change the heme iron configuration from a ferrous state to a ferric state. From the signals obtained from the main oxygen sensor and the modified oxygen sensor, a quantification of the level of hemolysis in the sample can be obtained.

[0009] In some embodiments provided herein, a diagnostic analyzer includes an analyzer body including a cartridge receiver and an electrical connector; a diagnostic cartridge receivable in the cartridge receiver, the cartridge body configured to couple to the cartridge receiver, the cartridge body including electrical contacts connectable to the electrical connector when the cartridge body is received in the cartridge receiver; a sample inlet configured to receive a sample; a sample passage extending into the cartridge body from the sample inlet and configured to receive the sample; a primary oxygen sensor configured to contact the sample in the sample passage and to provide a first measurement of the sample; and a primary oxygen sensor configured to contact the sample in the sample passage and to convert hemoglobin to methemoglobin (Fe). 2+ From Fe 3+ a diagnostic cartridge including an oxidant configured to oxidize a sample and a modified oxygen sensor configured to provide a second measurement value of the sample; and a controller coupled to the electrical connector and configured to receive the first and second measurements of the sample and configured to provide a level of hemolysis in the sample based on the first and second measurements.

[0010] In some embodiments provided herein, a method for determining hemolysis of a sample is provided, the method comprising coupling a diagnostic cartridge to a cartridge receiver of a diagnostic analyzer, the diagnostic cartridge including a sample inlet configured to receive a sample, a sample passageway extending from the sample inlet into a cartridge body, a primary oxygen sensor configured to contact the sample in the sample passageway, and a modified oxygen sensor configured to contact the sample in the sample passageway, the modified oxygen sensor detecting the conversion of hemoglobin to methemoglobin (Fe 2+ From Fe 3+ to) oxidize the sample; passing the sample through a sample passage to contact the primary oxygen sensor and the modified oxygen sensor; obtaining a first measurement of the sample from the primary oxygen sensor; obtaining a second measurement of the sample from the modified oxygen sensor; and providing a level of hemolysis in the sample based on the first measurement and the second measurement.

[0011] Other features and aspects of the present disclosure will become more fully apparent from the following detailed description, claims, and accompanying drawings.

[0012] The drawings described below are for illustrative purposes only and are not necessarily drawn to scale. The drawings are not intended to limit the scope of the present disclosure in any way. [Brief explanation of the drawings]

[0013] [Figure 1] 1 is a perspective view of an example of a diagnostic analyzer (e.g., for point-of-care deployment) including a diagnostic cartridge (including a diagnostic sensor assembly) receivable in a cartridge receiving portion according to embodiments provided herein. [Figure 2] FIG. 1 is a bottom view of an example diagnostic cartridge containing a sensor assembly configured to provide measurements for use in hemolysis level detection, according to embodiments provided herein. [Figure 3A]3A-3A is a partial cross-sectional view of an example diagnostic cartridge taken along section line 3A-3A in FIG. 2, illustrating an exemplary structure of a sensor assembly in which an oxidant is included in an outer membrane layer that covers an internal electrolyte chamber. The membrane layer may be homogeneous or heterogeneous, according to embodiments provided herein. [Figure 3B] 3B is a partial cross-sectional view of another example of a diagnostic cartridge, as in FIG. 3A, in which an oxidizer is intermixed with a membrane layer, which may be homogeneous or heterogeneous according to embodiments provided herein. [Figure 3C] FIG. 2 is a partial cross-sectional view of an example of a modified oxygen sensor in which an oxidant is mixed into the membrane, according to embodiments provided herein. [Figure 3D] FIG. 1 is a schematic top view of an example diagnostic cartridge including main and modified oxygen sensors, as well as a reference sensor and other additional sensors, and a common ground, according to embodiments provided herein. [Figure 3E] FIG. 2 is a partial cross-sectional view of an example of a modified oxygen sensor in which an oxidizer is mixed into the outermost layer of a multilayer film, according to embodiments provided herein. [Figure 4] FIG. 1 is a schematic diagram of an example of a diagnostic analyzer including a diagnostic cartridge containing an acceptable sensor assembly, showing its sensor components and connections to a controller of the diagnostic analyzer, according to embodiments provided herein. [Figure 5] FIG. 10 is a bottom view of another example of a diagnostic cartridge containing a sensor assembly configured to provide measurements used in hemolysis level detection, the sample path of which includes multiple branches or legs that can separate from a main path, according to embodiments provided herein. [Figure 6] 6 is a cross-sectional view of a first passage portion that can be separated from the main passage of an exemplary diagnostic cartridge taken along section line 6-6 of FIG. 5 according to embodiments provided herein. [Figure 7] 7 is a cross-sectional view of a second passage portion that can be separated from the main passage of an exemplary diagnostic cartridge taken along section 7-7 of FIG. 5 according to embodiments provided herein. [Figure 8] 1 is a flow diagram of an exemplary method for detecting hemolysis using a diagnostic sensor assembly and a diagnostic analyzer, according to an embodiment of the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0014] Individuals with male, female, or other gender identification are included within the term, regardless of grammatical use of the term.

[0015] In some embodiments, a diagnostic sensor assembly is provided. The diagnostic sensor assembly can be implemented in a diagnostic cartridge (e.g., a card-like member) that includes a cartridge body that houses a passageway and multiple sensors. In some embodiments, the diagnostic cartridge can be, for example, a single-use cartridge. The diagnostic cartridge can be received (e.g., inserted or otherwise coupled) by a diagnostic analyzer configured to provide blood analysis, particularly hemolysis level detection (quantification) of a blood sample. The diagnostic analyzer can be, for example, a handheld or benchtop diagnostic analyzer. A blood sample can be provided to the diagnostic sensor assembly of the diagnostic cartridge. The blood sample can be, for example, whole blood. In some cases, the sample can be serum or plasma, which may contain hemolysis. The volume of whole blood used by the test can be very small, such as 100 μL or less.

[0016] When the sensor assembly is included in a diagnostic cartridge, a cartridge body configured to interface with a diagnostic analyzer can be provided in any suitable configuration to provide measurement signals from the various sensors of the diagnostic cartridge. The cartridge body can include a diagnostic sensor assembly having a sample inlet configured to receive a sample and a sample passageway extending from the sample inlet into, for example, the cartridge body. The sample (e.g., whole blood) can be provided (e.g., injected) through the sample inlet into the sample passageway, such as by use of a syringe, pump mechanism, or other suitable sample delivery device.

[0017] The diagnostic sensor assembly of the diagnostic cartridge further includes a plurality of sensors. In the embodiments described herein, the sensor assembly includes two oxygen sensors. In particular, the sensor assembly includes a primary oxygen sensor configured to contact the sample along the sample passageway and a modified oxygen sensor also configured to contact the sample along the sample passageway. The modified oxygen sensor differs from the primary oxygen sensor in that it includes (i.e., contains or is accompanied by) an oxidant, which converts extracellular hemoglobin in the sample to methemoglobin (Fe ). 2+ From Fe 3+ The modified oxygen sensor is configured to oxidize extracellular hemoglobin (hemoglobin) before reaching the adjacent oxygen sensor. In particular, in some embodiments, the modified oxygen sensor includes an oxidant, which can be provided within the membrane of the modified oxygen sensor. In other embodiments, the modified oxygen sensor includes an oxidant such that the oxidant is located in the sample passageway, not in the membrane, but adjacent (e.g., upstream) to the oxygen sensor, thereby enabling the sample to flow over the oxidant before flowing over the oxygen sensor. In this way, extracellular hemoglobin is oxidized before reaching the adjacent oxygen sensor. This embodiment therefore involves measuring a "corrected" oxygen level in the sample, and thus the term modified oxygen sensor also refers to the combination of the upstream oxidant and the adjacent oxygen sensor. The amount of sample modification will depend on the degree and presence of hemolysis in the sample due to the upstream reaction of the sample with the oxidant provided within the passageway.

[0018] In each embodiment, a quantification of the hemolysis level of a sample can be obtained based on signals generated by the primary and modified oxygen sensors. These signals represent local O measurements in the vicinity of the primary and modified sensors of the diagnostic sensor assembly, respectively. These signals can be provided to a controller of a diagnostic analyzer for processing and hemolysis level determination.

[0019] The presence of the oxidant causes oxygen to be liberated in the vicinity of the modified oxygen sensor when free (extracellular) hemoglobin is present in the sample, thereby providing a higher level of local oxygen for sensing by the modified oxygen sensor compared to a main oxygen sensor without the oxidant. The oxidant is configured to contact the sample. The difference in oxygen signals between the main oxygen sensor and the modified oxygen sensor can be correlated to the hemoglobin level present in the sample, thus providing a detection (quantification) of the hemolysis level in the sample. The hemolysis level can be processed to provide a hemolysis index. This index can range, for example, from zero to a maximum value, and can be displayed to the operator of the diagnostic analyzer or otherwise electronically communicated within a hospital information system. Calibration can be performed before (or even after) performing hemolysis detection to ensure that the diagnostic sensor assembly provides appropriate results.

[0020] In another embodiment, a diagnostic analyzer configured to provide detection (quantification) of hemolysis in a blood sample is provided. The diagnostic analyzer has an analyzer body including a cartridge receiver and an electrical connector. The electrical connector is configured to form an electrical connection between a controller and a diagnostic sensor assembly configured in a diagnostic cartridge receivable in the cartridge receiver. The cartridge body is configured to be received by the cartridge receiver, and the cartridge body includes electrical contacts configured to couple to the electrical connector when the cartridge body is received by the cartridge receiver. The diagnostic sensor assembly in the cartridge body includes a sample inlet and a sample passageway configured to receive the sample.

[0021] The primary oxygen sensor is configured to contact a sample provided in the sample passage and to provide a first measurement of the sample (a signal correlating to the amount of oxygen in the vicinity of the primary oxygen sensor). Similarly, the modified oxygen sensor is configured to contact a sample provided in the sample passage and an oxidant (of the modified oxygen sensor) oxidizes extracellular hemoglobin to methemoglobin (Fe 2+ From Fe3+ The modified oxygen sensor is therefore configured to provide a second measurement (a signal that correlates to the amount of oxygen in the sample in the vicinity of the modified oxygen sensor).

[0022] The controller of the diagnostic analyzer is electrically coupled to the electrical connector (of the analyzer body). The electrical connector can include multiple conductive paths for connecting to the multiple electrical contacts (of the sensor assembly) and carrying sensor information from each of the sensors of the diagnostic sensor assembly. In particular, the controller is configured to receive the first measurement and the second measurement via the connection between the electrical connector and the electrical contacts of the diagnostic sensor assembly.

[0023] The controller is further configured to provide a hemolysis level measurement (quantification) based on the first measurement and the second measurement. In particular, this processing is performed by the controller's processor, and the controller's hemolysis detection module is configured to execute a difference detection routine. The difference between the first measurement and the second measurement obtained by subtraction correlates to the degree of hemolysis present in the sample. In particular, an elevated second measurement compared to the first measurement quantitatively correlates to the degree of hemolysis in the sample (index level), and a higher index level indicates a greater amount of hemolysis in the sample.

[0024] In yet another embodiment, a method for detecting hemolysis in a sample (e.g., whole blood, plasma, or serum) is provided. The method includes coupling a diagnostic sensor assembly to a diagnostic analyzer. The sensor assembly can be implemented as part of a diagnostic cartridge that is connectable to, or configured to connect to, a cartridge receptacle of the diagnostic analyzer. The sensor assembly of the diagnostic cartridge can include a sample inlet configured to receive a sample, a sample passageway extending from the sample inlet (e.g., into the cartridge body), a primary oxygen sensor configured to contact the sample in the sample passageway, and a modified oxygen sensor configured to contact the sample in the sample passageway, the modified oxygen sensor converting extracellular hemoglobin in the sample to methemoglobin (Fe2+ From Fe 3+ The method further includes passing a sample through a sample passageway into contact with the primary oxygen sensor and the modified oxygen sensor, and obtaining a first measurement of the sample from the primary oxygen sensor and a second measurement of the sample from the modified oxygen sensor. These measurements may include signals correlating to localized oxygen readings. The first measurement correlates to a first partial pressure measurement of oxygen in the sample near the primary oxygen sensor. The second measurement correlates to a second partial pressure measurement of oxygen in the sample near the modified oxygen sensor. According to this method, a level of hemolysis is determined based on the first and second measurements, which quantify the degree (index level) of hemolysis contained in the sample.

[0025] These and other embodiments of the present disclosure will be described in detail herein with reference to FIGS. 1 to 8 herein.

[0026] Referring to FIG. 1 , a perspective view of an example diagnostic analyzer 100 is shown in accordance with embodiments provided herein. The diagnostic analyzer 100 includes an analyzer body 102 configured to accommodate various user interfaces, such as user-controlled haptics (e.g., buttons, switches, touchscreens, etc.). In the depicted embodiment, the analyzer body 102 of the diagnostic analyzer 100 can include a computing device 104. In some embodiments, the computing device 104 can be removably attached to a device mount 102M on the base 102B of the diagnostic analyzer 100. The computing device 104 can be a handheld computing device, such as a personal digital assistant (PDA), tablet, or other similar computing device. In some diagnostic analyzers, the processing and memory capabilities of the computing device 104 can be housed within the analyzer body 102 rather than in a detachable / detachable form of the computing device 104.

[0027] The diagnostic analyzer 100 includes a controller 105, which in this embodiment is configured to include a first controller 105C1, which can be part of the base 102B of the analyzer body 102, and a second controller 105C2, which can be part of or integrated with the computing device 104. The first controller 105C1 and the second controller 105C2 are in electronic communication with each other and can perform different functions. In this embodiment, the computing device 104 can include a display 104D that enables visual display of user input, operational information, test results, and other information. In some embodiments, the display 104D can be tiltable about a pivot axis 102A. For example, a device mount 102M on the base 102B can receive the computing device 104 and can be pivotable about the pivot axis 102A at a location 102L to allow for adjustment of the viewing angle.

[0028] The display 104D can be a touchscreen having a user interface that, together with one or more haptics (e.g., buttons, switches, or other user control devices), allows an operator to control the operation of the diagnostic analyzer 100, observe measurement results from performed diagnostic tests, and / or perform other auxiliary functions. The first controller 105C1 can include electronics that enable communication with the diagnostic sensor assembly 103 implemented within the diagnostic cartridge 106, including signal conditioning (including filtering, analog-to-digital conversion, and / or possibly amplification) of various sensor signals received from the various sensors of the diagnostic sensor assembly 103 of the diagnostic cartridge 106, as described in more detail herein. The first controller 105C1 can further include electronics that enable providing an amperometric or potentiometric input to the diagnostic sensor assembly 103 as a baseline input for carrying a signal correlating to the sensor measurement signal.

[0029] The second controller 105C2 can be operable to perform processing and can include a hemolysis detection module 430 (FIG. 4), which is a software module containing a difference-finding routine. The software module containing the difference-finding routine can be stored in the memory 431 of the diagnostic analyzer 100 and can be executable on the processor 432. The hemolysis detection module 430 receives sensor readings (signals) from the various oxygen sensors described herein, compares the values, and obtains a difference between them. From this difference, the hemolysis detection module 430 can subsequently detect the level of hemolysis contained in the sample 111. Signals from any additional sensors in the diagnostic sensor assembly 103 can also be received and processed. In some embodiments, test results and other information can be transmitted to the hospital information system (HIS) 101.

[0030] Referring again to FIG. 1 , the diagnostic analyzer 100 can include a cartridge receiver 102P, which can be a port, opening, or other suitable coupling feature configured to receive and couple to the diagnostic cartridge 106. The diagnostic cartridge 106 can be coupled or connected (e.g., inserted) to the cartridge receiver 102P, thereby forming an electrical connection between the diagnostic sensor assembly 103 and the controller 105 and enabling processing. As shown, the cartridge receiver 102P can include a slot sized to accept the diagnostic cartridge 106. The diagnostic cartridge 106 can resemble a playing card, thin compared to its width and length. For example, the diagnostic cartridge 106 can have a length of approximately 85 mm, a width of approximately 55 mm, and a thickness of approximately 1.2 mm. However, the diagnostic cartridge 106 can also include other dimensions and / or shapes.

[0031] As shown in FIG. 1, the diagnostic sensor assembly 103 can be configured as part of a cartridge body 108 of a diagnostic cartridge 106 configured to connect to a diagnostic analyzer 100. The diagnostic sensor assembly 103 includes a sample inlet 109 configured to receive a sample 111 to be tested, which can be a port, an opening, a receiving element, or the like. The sample inlet 109 can be provided in a top layer 108T of the cartridge body 108, as shown in FIG. 1. The sample inlet 109 can include a circular or other shaped opening that provides a port configured to receive the sample 111. In some embodiments, the sample inlet 109 can have a width or diameter dimension of about 4 mm to about 8 mm, although other diameters, dimensions, or shapes can also be used.

[0032] The sample 111 can be whole blood, and the sample inlet 109 can be configured to allow a syringe or other suitable pump or delivery device to be sealingly coupled to the sample inlet 109 to receive the sample 111 and inject and flow the sample 111 into the sample passage 210 (shown in FIG. 2). The cartridge body 108 can be made from multiple layers of material bonded together to form the sample passage 210. For example, the cartridge body 108 can include a bottom layer 208B (FIG. 2), a top layer 108T (FIG. 1), and optionally, a middle layer 308I (FIGS. 3A-3C and 3E), which can be coupled and sealed together by any suitable means, such as using adhesives, mechanical interlocks, combinations thereof, etc. Portions of the sample passage 210 can be formed by the interaction of mating portions of the top layer 108T, the bottom layer 208B, and, if present, the middle layer 308I. In some embodiments, the top layer 108T, the bottom layer 208B, and the middle layer 308I can be formed from the same type of material. For example, the material can be a plastic such as polypropylene. Alternatively, different materials (e.g., different types of plastic, paper, foil, and / or laminates thereof) can be used for the middle layer 108I, the top layer 108T, and the bottom layer 208B. In some embodiments, the top layer 108T and / or the bottom layer 208B can be a clear (transparent or translucent) material to allow visual observation of the flow of the sample 111 therein. Other structures for forming the sample passage 210 can also be used, such as other two-piece, three-piece, or other multi-piece designs.

[0033] Referring again to FIG. 2 , a bottom view of the diagnostic cartridge 106 including the diagnostic sensor assembly 103 is shown. In the depicted embodiment, the sample path 210 can include a first portion 210A extending from the sample inlet 109 to a second portion 210B. The second portion 210B can include a sensor array 212 comprised of multiple sensors, including at least two oxygen sensors. In some embodiments, the sample path 210 can extend from the sample inlet 109 into the cartridge body 108. The second portion 210B can have different dimensions compared to the first portion 210A. For example, the second portion 210B can be wider to accommodate the dimensions of the various sensors 214, 216, 217 housed therein. Thus, in some embodiments, the second portion 210B can resemble a chamber. A waste passage 219 may be connected to the downstream end of the second portion 210B, which includes a conduit or passage configured to receive the sample effluent after the sample 111 contacts an additional sensor or the last sensor or component in the sensor array 212, such as ground 217.

[0034] The sample passage 210 may be, for example, 12,500 μm 2 ~0.8mm 2 In some embodiments, the width-to-height ratio W:H of the sample passage 210 can be about 5:1 or greater. The height H is the dimension across the sample passage 210 as shown in FIG. 3A, and the width W is the dimension across the sample passage 210 as shown in FIG. 2. The width W can be about 250 μm to about 2 mm, and the height H can be about 50 μm to about 400 μm. The length L along the sample passage 210, extending from the sample inlet 109 to the beginning of the waste passage 219, can be about 1.25 mm to about 100 mm, or greater. Other relationships between the length L, width W, and / or height H can be used, and other suitable length L, height H, and / or width W dimensions can be used.

[0035] More specifically, the sensor array 212 of the diagnostic sensor assembly 103 includes a main oxygen sensor 214 configured to contact the sample 111 along the sample path 210 and a modified oxygen sensor 216, 316 also configured to contact the sample 111 along the sample path 210. As shown in the depicted embodiment of FIGS. 2-3C and 3E, either of the oxygen sensors 214, 216, or 316 can be provided in the second portion 210B. The modified oxygen sensor 216, 316 can be located downstream (to the left as shown in FIG. 2) of the main oxygen sensor 214 so that the oxidant 324 ( FIGS. 3A-3C and 3E ) associated with the modified oxygen sensor 216, 316 does not alter the sample 111 exposed to the main oxygen sensor 214. Other additional sensors and / or grounds 217 can be provided between the main oxygen sensor 214 and the modified oxygen sensor 216, 316 or can be otherwise disposed within the sensor array 212.

[0036] The modified oxygen sensor 216, 316 includes an oxidant 324 (FIGS. 3A-3C and 3E) configured to oxidize extracellular (free) hemoglobin to methemoglobin. In particular, the oxidant 324 converts extracellular hemoglobin iron to the ferrous state (Fe +2 ) to the ferric state (Fe +3 ) In various depicted embodiments, partial cross-sectional views are shown in Figures 3A-3C and 3E.

[0037] 3A, the oxidant 324 may be embodied as part of the membrane 316M. The membrane 316M may be formed from any semi-permeable, wettable material, such as a polymeric material. For example, the polymeric material may be a polyurethane-based material, a polyacrylate-based material, copolymers thereof, or the like. The sensor chamber 316C, at least partially formed by the membrane 316M of the modified oxygen sensor 216, may include an electrolyte 316E disposed within the sensor chamber 316C.

[0038] The electrolyte 316E can be any solid-state proton-conducting polymer, such as a mixture of Nafion™ and polyvinylpyrrolidone, which can be mixed in a 4:1 ratio. In some cases, the electrolyte 316E can be a hydrogel, for example, including poly-N-vinylpyrrolidone K90 (PNPV) and 2,6-bis(4-azidobenzylidene)-4-methylcyclohexanone. Other suitable liquid or gel electrolytes suitable for such pump-type and Clark-type oxygen sensors can also be used.

[0039] The oxidizing agent 324 can be mixed into the polymer material to form the film 316M. The oxidizing agent 324 can be homogeneously present in the film 316M, or optionally can be present in a graded manner, i.e., a concentration gradient, such that a higher concentration of the oxidizing agent 324 is provided on the surface of the film 316M adjacent the sample 111 located in the second portion 210B of the sample passage 210. For example, a graded change can be achieved by depositing layers of the same polymer material with varying levels of the oxidizing agent 324, with the highest concentration provided in the outermost layer adjacent the sample 111.

[0040] In principle, film 316M can be formed from any suitable process, such as deposition involving a vaporizable liquid, including the desired film base material and including oxidizer 324. In such deposition, the liquid mixture can be dispensed from a tip. Optionally, film 316M can be formed from spin coating, dip coating, screen printing, spray coating, etc.

[0041] In the embodiment of Figures 3B and 3C, the oxidant 324 can be mixed into a matrix of a heterogeneous membrane 316M, as shown in the modified oxygen sensor 316. In this heterogeneous embodiment of Figures 3B and 3C, the membrane 316M is provided in direct contact with the sample 111 and is positioned between the sample 111 and one or more electrodes 225. For purposes of illustration, the one or more electrodes 225 are shown as a single electrode (e.g., a working electrode). The reference electrode can be located at a different location. However, it should be understood that in some embodiments, the one or more electrodes 225 can also consist of a working electrode and a counter electrode, and / or a reference electrode. Any conventional electrode structure or arrangement can be used.

[0042] The membrane 316M can include a hydrophobic polymer blended with a hydrophilic component. For example, the membrane 316M can include a heterogeneous membrane composition having a hydrophilic electrolyte-containing compartment and a hydrophobic compartment that accommodates gas (e.g., O) and water vapor transport. The hydrophobic compartment can include a polymer.

[0043] Exemplary polymeric materials for the membrane 316M can include polysiloxane, polyorganophosphazene, poly-1-trimethyl-silyl-1-propyne, poly-4-methyl-2-pentyne, and mixtures thereof. The hydrophilic component of the admixture can include, for example, hydrophilic polymers such as polyvinyl alcohol (PVA), polyacrylate polymers such as hydroxymethacrylate, polyacrylamide, polysaccharides, cellulosic polymers, and / or gelatin. The hydrophilic component can further include some or all of an emulsifier, a hydrophilic polymer adhesive, an electrolyte salt, a viscosity modifier, and other optional dissolved components. Other optional components of the hydrophilic compartment can include one or more components such as a crosslinker, a catalyst, a redox agent, a buffer, and / or a surfactant that can be incorporated into the membrane 316M upon formation.

[0044] The gas diffusion coefficients of the various phases should differ by 10 or more, 50 or more, or even 100 or more, with the hydrophobic component being significantly higher. In one method, the membrane 316M and oxidizer 324 can be deposited, for example, in a well formed in the bottom layer 208B. The bottom layer 208B can be an electrically insulating material, such as an epoxy layer, a polymer composite, or other suitable electrically insulating material. The membrane-forming solution can be dried to form the membrane 316M, including the heterogeneous membrane. Further discussion of the structure and materials of such conventional heterogeneous sensors can be found in U.S. Pat. No. 7,094,330.

[0045] 3E, the sensor 316 can include a membrane 316M containing multiple layers. For example, the base layer 316B can be any oxygen-permeable, non-wetting material, such as a polyethylene or polytetrafluoroethylene (PTFE) material, silicone, paraffin wax, or the like. The top layer 316T is configured to contact the sample 111 and can be made from a wetting material, such as a polyurethane-based material, a polyacrylate-based material, or copolymers thereof. The wetting material comprising the top layer 316T includes an oxidizing agent 324.

[0046] The wettable material of the membrane 316M of the embodiment of Figure 3A and the top layer 316T of the embodiment of Figure 3E is hydrophilic and can include a contact angle of greater than 0 degrees and less than or equal to 90 degrees. In some embodiments, the surface of the wettable material can have a contact angle of less than 45 degrees, or even less than 30 degrees. In some embodiments, the surface of the membrane 316M and top layer 316T of the wettable material can be modified to further enhance its wettability and availability to the oxidizing agent 324.

[0047] For example, the top surface of film 316M (FIG. 3A) and the top surface of top layer 316T (FIG. 3E) can be treated to enhance wettability. For example, the top surface can be plasma treated, i.e., treated with ionized gases and / or radicals, for a time sufficient to obtain a smaller contact angle (e.g., 10 seconds to 5 minutes). Optionally, the top surface can be treated with ultraviolet ozone (UVO) for a suitable time to obtain a smaller contact angle (e.g., about 5 minutes). Other suitable treatment methods for enhancing wettability, as well as combinations of the above-described contact angle-reducing treatments, can also be used.

[0048] Wettability measurements can be measured using the sessile drop method with an optical tensiometer. In some embodiments, the outer portion of top layer 316T adjacent sample 111 can be provided with a higher concentration of oxidant 324 to maximize the amount of oxidant 324 available to oxidize the extracellular hemoglobin contained in sample 111.

[0049] In all embodiments described herein, the oxidant 324 included in the modified oxygen sensor 316 may include, for example, potassium ferricyanide. In some cases, the oxidant 324 may convert, for example, extracellular hemoglobin in the sample 111 to methemoglobin (Fe 2+ From Fe 3+ The oxidizing agent 324 may include any chemical compound from other known classes of oxidizing agents, such as organic or inorganic nitrites, aromatic amines, or quinones, that function to cause the oxidation of extracellular hemoglobin in the sample 111 to methemoglobin (Fe 2+ From Fe 3+The oxidant 324 may be provided in an amount effective to cause sufficient oxidation of hemoglobin (to oxygen). The goal is to have a sufficient concentration of oxidant 324 available to oxidize hemoglobin so as to provide a sufficient differential between the readings sensed by the primary oxygen sensor 214 and the modified oxygen sensors 216, 316. In particular, the oxidant 324 used should not cause interference with the readings of one or more other sensors along the sample path 210, 310. In some embodiments, such as those shown in FIGS. 3B-3C, one or more additional thin layers of membrane 316M may be formed adjacent to one or more electrodes 225; these thin layers may be of the same material as membrane 316M but may not contain oxidant 324, thus minimizing deleterious redox-type interactions at one or more electrodes 225.

[0050] In any of the above-described embodiments, oxidizing agent 324 can be provided to, i.e., mixed with, film 316M (FIGS. 3A-3C and 3E). In some embodiments, oxidizing agent 324 can be provided in a suitable amount, i.e., weight percent (wt%), from about 0.01 wt % to about 25 wt %, or even from about 0.5 wt % to about 5.0 wt %, based on the total weight of film 316M including oxidizing agent 324.

[0051] As can be seen, the oxidant 324 in the membrane 316M converts extracellular hemoglobin to methemoglobin (Fe 2+ From Fe 3+ to the primary oxygen sensor 214, which in turn locally liberates bound oxygen from the extracellular hemoglobin in the sample 111. In response, the liberated / generated oxygen becomes detectable by the modified oxygen sensor 216, 316, and thus, as further described herein, a quantification of hemolysis in the sample 111 can be obtained specifically (e.g., as a difference between readings from the primary oxygen sensor 214 and the modified oxygen sensor 216, 316) when free (extracellular) hemoglobin is present in the sample 111.

[0052] The primary oxygen sensor 214 is configured to provide a first measurement that correlates to the partial pressure of oxygen in the sample 111 in the vicinity of the primary oxygen sensor 214. Similarly, the modified oxygen sensors 216, 316 are configured to provide a second measurement that correlates to the partial pressure of oxygen in the sample 111 in the vicinity of the modified oxygen sensors 216, 316. In a sample 111 that has free hemoglobin, excess oxygen is liberated by an oxidation reaction between extracellular hemoglobin in the sample 111 and an oxidant 324, which can then be sensed locally by the modified oxygen sensors 216, 316. A differential measurement can then be obtained that is compared to the reading from the primary oxygen sensor 214.

[0053] Referring again to Figures 3C and 3E, an example of a modified oxygen sensor 316 having a solid-state integrated chip structure is shown. The modified oxygen sensor 316 may be provided within a diagnostic cartridge 306, as shown in Figure 3D, and in use may be connected to the inlet 351 and outlet 352 of the diagnostic analyzer 100 (Figure 1), or may be otherwise connected to the sample passage 210 and waste passage 219. The modified oxygen sensor 316 of Figures 3C and 3E may be included within a diagnostic cartridge such as that shown in Figure 2.

[0054] 3D, inlet 351 delivers sample 111 to sensor array 212, which includes primary oxygen sensor 214 and modified sensor (MO2) 316. Diagnostic cartridge 306 may further include one or more additional sensors and / or ground 217. The additional sensors may be configured to measure other analytes and / or conditions, such as Cl-, Mg++, Na+, K+, pCO2, Ca++, glucose, lactate, creatinine, etc. Additionally or alternatively, other additional sensors 217 configured to sense other analytes (e.g., BUN, Hct, and / or TCO2) and / or conditions (e.g., pH) may also be included.

[0055] The diagnostic cartridge 306 may further include one or more reference sensors 354 configured to provide a reference signal. Optionally, the reference signal may be obtained inside the diagnostic analyzer 100 or elsewhere along the sample path 210 or 310. The diagnostic cartridge 306 may further include a common ground in any suitable location connectable to the controller 105. The placement of the sensors may be other than that shown. However, the modified oxygen sensor 316 may be located downstream of the main oxygen sensor 214. Furthermore, the modified oxygen sensor 316 and the main oxygen sensor 214 may be separated by one or more additional sensors 217 or a suitable space so that the main oxygen sensor 214 is not affected by excess oxygen liberated near the modified oxygen sensor 316.

[0056] The sensors 214, 316, 217 and one or more reference sensors 354, as well as ground (if used), can be electrically coupled to a detection system of the diagnostic analyzer 100 (FIGS. 1 and 4), which can include any suitable electronics to enable the potential difference (or current difference) between the primary oxygen sensor 214 and the modified oxygen sensors 216, 316 to be read as a measurable signal. Again, the construction of the detection system and reference sensor 354 is well known and will not be further described herein. For example, the reference system and reference sensor 354 can be of the type used in the epoc® blood analysis system available from Siemens Medical Solutions, or as shown in FIG. 3D. However, the reference sensor 354 can also be located elsewhere along the sample path 210, 310 besides the second portion 210B, 310B.

[0057] More specifically, and with further reference to FIGS. 3C and 3E, the modified oxygen sensor 316 can include a cartridge body 308 including an oxygen-permeable membrane 316M coupled to the bottom layer 208B or the like. The walls 356 of the diagnostic cartridge 306 and the sensors (e.g., the primary oxygen sensor 214, the modified oxygen sensor 316, the reference sensor 354 (see FIGS. 3D, 3E, and 4), and any additional sensors 217) form the second portion 310B that receives the sample 111. The membrane 316M can be formed from a thin polymer sheet that is selective for O. The membrane 316M can have, for example, a diameter or maximum dimension of about 200 μm to about 1,700 μm and a thickness of about 10 μm to 200 μm. Other suitable diameters or maximum dimensions and / or thicknesses can also be used. The chemical composition of the membrane 316M that renders it selective for O can be as described above herein or any other suitably oxygen-permeable material.

[0058] The modified oxygen sensor 316, the reference sensor 354, and any additional sensors and / or grounds 317 can include one or more electrodes 225, and in some embodiments, the one or more electrodes 225 can be located adjacent to the membrane 316M. The one or more electrodes 225 can include working, counter, and / or reference electrodes and can be made of any suitable structure, such as conductive traces, masked deposits, etc. Conductors can extend from each of the one or more electrodes 225, the reference sensor 354, and any other additional sensors and / or grounds 317 to corresponding electrical contacts (e.g., 218 shown in FIGS. 2 and 4 ), which can be provided on the cartridge body 108, 308 (e.g., at the bottom thereof) of the diagnostic cartridge 106, 306, and the electrical contacts 218 can be interconnected to the diagnostic analyzer 100 when the diagnostic cartridge 106, 306 is coupled to the diagnostic analyzer 100.

[0059] For example, one or more of the electrodes 225 can include silver (AG) elements, which can be coated with, for example, a silver chloride (AGCl) coating. In some cases, the one or more of the electrodes 225 can include, for example, gold or platinum, or some combination of the above. Other suitably conductive materials or combinations of conductive materials can also be used. The connection between the one or more electrodes 225 and the electrical contacts 218 on the cartridge body 108, 308 of the diagnostic cartridge 306 can be any suitable conductive material and can be a trace or conductor formed in any suitable configuration, such as by printing, deposition, or other known conductive conduit or trace formation methods.

[0060] In some embodiments, the sensor array 212, 312 can include one or more additional sensors 217 other than the primary oxygen sensor 214 and the correction oxygen sensor 216, 316. For example, the other sensors 217 can be configured to sense other analytes and / or conditions described above. In some embodiments, the number of additional sensors 217 can be one or more, five or more, or even ten or more. The total number of sensors in the sensor array 212, 312, including the primary oxygen sensor 214 and the correction oxygen sensor 216, 316, can range from two to fifteen, for example. As shown in FIGS. 3A, 3B, and 3D, the one or more additional sensors 217 can be located between the primary oxygen sensor 214 and the correction oxygen sensor 216, 316 in the second portion 210B, 310B.

[0061] 2, the cartridge body 108 of the diagnostic cartridge 106 can include a control portion 220 that can be used to supply a calibration fluid to the sample passage 210 for calibrating the operation of the diagnostic sensor assembly 103 of the diagnostic cartridge 106. Such control portions 220 are known to those skilled in the art and will not be described further herein.

[0062] 1-4, in the depicted embodiment, operation of the diagnostic analyzer 100 may involve inserting the diagnostic cartridge 106, 306 into the cartridge receiver 102P and engaging the diagnostic sensor assembly 103, 303 of the diagnostic cartridge 106, 306 with the controller 105 via an electrical connection between the diagnostic sensor assembly 103, 303 and the controller 105. The sample inlet 109 receives a sample 111 (e.g., contained in a syringe or other delivery means) and directs the sample 111 into the sample passage 210, 310 for delivery to the second portion 210B, 310B containing the main oxygen sensor 214 and the modified oxygen sensor 216, 316, as well as to any other additional sensors 217 that may be included in the sample passage 210, 310. When the sample 111 contacts the oxygen sensors 214, 216, 316, signals correlating to the oxygen levels sensed by the oxygen sensors 214, 216, 316 are provided to the controller 105. The controller 105 can then perform processing of the various received sensor signals to generate a sensed level of the sensed analyte, condition, or component (e.g., O2) for each sensor 214, 216, 316, and 217. From the sensed signals from the primary oxygen sensor 214 and the modified oxygen sensors 216, 316, the controller 105 can operate to detect the level of hemolysis in the sample 111.

[0063] Waste sample fluid after passing through second portion 210B, 310B can flow into waste passage 219 for storage. In some embodiments, waste passage 219 can be formed in cartridge body 108 of diagnostic cartridge 106, 306, for example, as a trench, groove, or similar structure, and can have a serpentine or other non-linear shape. Waste passage 219 should be long enough to hold waste fluid (e.g., sample 111 after passing through second portion 210B, 310B, and optionally control fluid from a calibration operation in control portion 220).

[0064] Referring again to Figures 3A-3C, the diagnostic cartridge 106, 306 can include one or more electrodes 225 for each sensor 214, 216, 217, 316, and 354, each electrically connected to an electrical contact 218. While only one electrical contact 218 is shown in Figure 2 for simplicity, it should be understood that each of the sensors 214, 216, 217, 316, and 354 has one or more electrical contacts dedicated to providing a signal, such as a voltage or current. Changes in potential or current from sensor operation can then be detected. If the sensor type is designed for potentiometry, a baseline voltage can be provided, and changes in that baseline voltage can be detected. For example, a low voltage of 1 mV to 500 mV, or even 1 mV to 50 mV, can be provided as a baseline voltage in the measurement circuit. Similarly, if the sensor's measurement system is amperometric, a baseline current can be provided, and changes therefrom can be detected. For example, a low current of 1 nA to 500 nA or even 1 nA to 50 nA can be provided as a baseline current in the measurement circuit. Other suitable voltage or current baseline values ​​can also be used. Any conventional circuitry that enables voltage and / or current measurements can also be used.

[0065] As shown in FIG. 4 , for example, one or more electrodes of the main oxygen sensor 214, the modified oxygen sensor 216, 316, the additional sensor or ground 217, and the reference sensor 354 can each have a conductive path extending to the controller 105. For example, in some embodiments, a conductive path can be provided to or from a corresponding electrical contact 218 configured on the surface of the cartridge body 108, 308 of the diagnostic cartridge 106, 306. For example, as shown in FIG. 4 , the conductive path can include a connection to each of the electrical contacts 218A, 218B (only a few of which are labeled). The electrical contacts 218 are contacted by mating contacts 221 (e.g., mating contacts 221A, 221B, only a few of which are labeled) of the electrical connector 458, which connect to the controller 105. Similarly, the ground 217 (if used) can be connected to contact 218G, which is contacted by mating contact 221G of the electrical connector 458. These signals from the main oxygen sensor 214 and the modified oxygen sensors 216, 316, as well as the optional additional sensor 217 and the reference sensor 354, may subsequently be received by the controller 105. An electrical connection between the electrical contact 218 and the mating contact 221 may be formed when the diagnostic cartridge 106, 306 is coupled to the cartridge receiver 102P.

[0066] Referring again to FIG. 4, the diagnostic analyzer 100 can also include a hemolysis detection module 430 within the controller 105. The hemolysis detection module 430 can be stored as programmed code within the memory 431 and executed by the processor 432. The processor 432 can control the operation of the sensors 214, 216, 316, 354, 217, the memory 431, and / or the display 104D. Signals from the various sensors 214, 216, 217 can be manipulated to provide values ​​correlating to the analyte being sensed. In the case of the primary oxygen sensor 214 and the modified oxygen sensors 216, 316, the signals therefrom are manipulated to derive values ​​correlating to the oxygen level. From these signal values, the hemolysis detection module 430 can determine the level of hemolysis in the sample 111 based on the difference between them. This difference can be calculated by difference-finding program code. The greater the difference found via the difference finding program of the hemolysis detection module 430, the greater the oxygen liberated as a result of the oxidation reaction with the oxidant 324 generated in the vicinity of the modified oxygen sensor 216, 316, and such difference correlates to the level of hemolysis (extracellular hemoglobin) present in the sample 111. The greater the difference determined or calculated via the difference finding program, the greater the level or amount of hemolysis present in the sample 111.

[0067] The processor 432 may be any suitable computational resource, such as, but not limited to, a microprocessor, a microcontroller, an embedded microcontroller, a digital signal processor (DSP), or a field programmable gate array (FPGA) configured to be implemented as a microcontroller.

[0068] The memory 431 may be any suitable type of memory, such as, but not limited to, one or more of volatile memory, non-volatile memory, or a combination thereof. Volatile memory may include, but is not limited to, static random access memory (SRAM) or dynamic random access memory (DRAM). Non-volatile memory may include, but is not limited to, electrically programmable read-only memory (EPROM), electrically erasable programmable read-only memory (EEPROM), flash memory, etc. The memory 431 may have a plurality of instructions stored therein that, when executed by the processor 432, cause the processor 432 to perform various operations specified by one or more of the stored instructions, including a difference detection program.

[0069] The user interface can include one or more display screens (e.g., display 104D). The user interface can be controlled by processor 432, and the functionality of the user interface can be implemented at least in part by computer-executable instructions (e.g., program code or software) stored in memory 431 and / or executed by processor 432 of diagnostic analyzer 100. In some embodiments, processor 432 can receive measurement results from oxygen sensors 214, 216, 316, and from additional sensors 217 if one or more other additional sensors are included, such as reference sensor 354, process the measurement results to generate calculations, and present the calculations and / or other information, such as patient information, via display 104D of the user interface. For example, user interface and display 104D can be configured to present one or more measurement and / or calculation results of hemolysis and optionally other analyte and / or condition measurements to a user of diagnostic analyzer 100. Such results can likewise be communicated to HIS 101.

[0070] FIG. 5 illustrates an alternative embodiment of a diagnostic sensor assembly 503 that can be included in a diagnostic cartridge 506. This embodiment includes a sample passage 510 similar to the previously described embodiment, but the sample passage 510 includes at least two branches or tributaries, such as a first passage 510B and a second passage 510C. In some embodiments, the first passage 510B and the second passage 510C can optionally split off from a primary passage 510A extending from the sample inlet 109. In this embodiment, the main oxygen sensor 214 can be located in the first passage 510B, and the modified oxygen sensor 516 can be located in the second passage 510C. For example, the main oxygen sensor 214 can be located in the first passage 510B split off from the primary passage 510A, and the modified oxygen sensor 516 can be located in the second passage 510C split off from the primary passage 510A. For example, one or more optional additional sensors or grounds 217 can be provided in one or both of the first and second passages 510B, 510C of the same type as discussed above. Alternatively, or in addition, other sensor types can be included. Similarly, in each of the embodiments of FIG. 5 described herein, a reference sensor can be included anywhere along one or both of the first and second passages 510B, 510C, or as part of the oxygen sensors 214, 214B. Again, for simplicity, the electrical contacts 518 are shown as a single circle, but each of the main oxygen sensor 214, oxygen sensor 214B, additional sensors and / or grounds 217, and reference sensor 354 can have their own electrical contacts. A diagnostic cartridge 506 containing the diagnostic sensor assembly 503 can be connected to the diagnostic analyzer 100 in a manner similar to that shown in FIG. 4.

[0071] Advantageously, because the sample passage 510 is bifurcated and the two oxygen sensors 214, 516 are not located in series within the same single sample passage, there is a reduced or eliminated risk that the oxidant 524 associated with the modified oxygen sensor 516 will alter the sample exposed to the primary oxygen sensor 214. As previously mentioned, the diagnostic cartridge 506 may include a control portion 220 and a waste passage 219 extending from each of the first passage 510B and the second passage 510C.

[0072] This embodiment of the modified oxygen sensor 516 is configured such that the sample 111 flows over the oxidant 524 to convert extracellular hemoglobin to the ferrous state (Fe +2 ) to the ferric state (Fe +3 ), which then liberates oxygen, which can be sensed by oxygen sensor 214B. Oxygen sensor 214B may be identical to main oxygen sensor 214 located in first passage 510B and may be of conventional construction. As shown, modified oxygen sensor 516 is composed of or includes oxidant 524 and oxygen sensor 214B, which may be conventional like main oxygen sensor 214. Oxidant 524 may be disposed and located upstream of oxygen sensor 214B.

[0073] In an alternative embodiment, the modified oxygen sensor 516 can differ from the main oxygen sensor 214 in the same manner as described above with respect to the modified oxygen sensor 216, 316 in which the membrane 316M includes the oxidant 324. This alternative embodiment differs from the embodiment of FIG. 2 in that the modified oxygen sensor 516 can be located in a different branch of the sample passage 510 (e.g., the second passage 510C), and therefore the first and second measurements can be taken in any order. For example, in this alternative embodiment of FIG. 5, the first and second measurements can be taken simultaneously (or in any other order), depending on the location of the oxygen sensors in the respective passages. The use of a bifurcated passage can serve to reduce or minimize the risk that the modified oxygen sensor 516 will alter the sample exposed to the main oxygen sensor 214 and to remove the requirement that the sample must flow past the main sensor before flowing past the modified sensor.

[0074] In another embodiment, the oxidant 524 can be provided upstream of the oxygen sensor 214B in the second passage 510C and also in the membrane of the oxygen sensor 214B. The amount at each location varies from hemoglobin to methemoglobin (Fe 2+ From Fe 3+ The temperature can be adjusted to achieve maximum oxidation of the

[0075] 6 and 7 show partial cross-sectional views taken along section lines 6-6 and 7-7, respectively, in FIG. 5. The diagnostic cartridge 506 can be constructed from multiple layers bonded together as described above. The first and second passages 510B and 510C can each include an oxygen sensor 214, 214B, both of which can be conventional oxygen sensors, similar to the main oxygen sensor 214 described above. The second passage 510C can include an oxidizer 524. The oxidizer 524 can be located upstream from the oxygen sensor 514B. The oxidizer 524 can be provided in any suitable form, such as a segmented cylinder or other shaped member, and can be sprayed and dried onto the passage portions of one or both of the top and bottom layers 108T and 208B, or even onto the middle layer 308I thereof.

[0076] In this embodiment, the oxidizing agent 524 may or may not include an adhesive material, such as a soluble aqueous polymeric material, such as polyvinyl alcohol or polyacrylic acid. The oxidizing agent 524 may be mixed with the soluble aqueous polymeric material. The soluble aqueous polymeric material may be soluble upon contact with the sample 111. The adhesive material may optionally be non-soluble (e.g., cross-linked), but may be sufficiently wettable or porous to allow sufficient interaction with the sample 111 to facilitate oxidation of any extracellular hemoglobin present. If the adhesive is not soluble, this may allow for repeated use conditions as opposed to single-use applications. In this configuration, the oxidizing agent 524 may also be potassium ferricyanide. However, other suitable oxidizing agent materials capable of sufficiently oxidizing hemoglobin may also be used.

[0077] The oxidizing agent 524 can be provided at about 0.01 to 99% by weight, based on the total weight of the adhesive and oxidizing agent 524. It is desirable to have a very small amount of adhesive relative to the oxidizing agent 524. For example, some adhesives can be used at about 1% to about 3% by weight, simply as a means to "hold" the oxidizing agent 524 in place until dissolved by the sample 111. The thickness and length of the cylinder or other shaped member containing the oxidizing agent 524 can be of a length and thickness or dimension sufficient to effect a suitable measurable change in oxygen level in the vicinity of the oxygen sensor 214B in the presence of free extracellular hemoglobin, compared to the oxygen level at the primary oxygen sensor 214.

[0078] In some embodiments, the oxidizing agent 524 can be provided in one of the wells in the bottom layer 208B located upstream from the oxygen sensor 214B, such as the well containing the membrane 316M in the modified oxygen sensor 316 of Figure 3B. However, in this embodiment, the well need not contain the electrode 225. The oxidizing agent 524 can be mixed into a membrane formed in the well, or can be provided in some other form, such as a form that is dissolvable by the sample, as described above.

[0079] 8 shows a flow diagram of an example of a method 800 for determining hemolysis of a sample 111 using a diagnostic sensor assembly 103, 303, 503 and a diagnostic analyzer 100 according to an embodiment of the present disclosure. The method 800 begins at block 802 by coupling a diagnostic cartridge 106, 306, 506 to a cartridge receptacle 102P of the diagnostic analyzer 100. For example, the diagnostic cartridge 106, 306, 506 can be coupled (e.g., inserted) into the cartridge receptacle 102P of the analyzer body 102, thereby forming an electrical connection between the diagnostic sensor assembly 103, 303, 503 of the diagnostic cartridge 106, 306, 506 and the controller 105.

[0080] In the depicted embodiment, the diagnostic sensor assembly 103, 303, 503 of the diagnostic cartridge 106, 306, 506 includes a sample inlet 109 configured to receive a sample 111, a sample passage 210, 310, 510 extending from the sample inlet 109 into the cartridge body 108, 308, 508, a main oxygen sensor 214 configured to contact the sample 111 in the sample passage 210, 310, 510, and a modified oxygen sensor 216, 316, 516 configured to contact the sample 111 in the sample passage 210, 310, 510, wherein the modified oxygen sensor 216, 316, 516 includes an oxidant 224, 324, 524 configured to oxidize extracellular hemoglobin iron to methemoglobin as described above. In some embodiments, before using the diagnostic analyzer 100, the display 104D and / or user interface may prompt the user to connect a diagnostic cartridge 106, 306, 506, enter user identification information, scan a name tag or other bar code, enter a password or otherwise authenticate the user's identity, and / or provide a sample 111 to the sample inlet 109.

[0081] After the cartridge body 108, 308, 508 is coupled to the cartridge receiver 102P of the diagnostic analyzer 100, at block 804, the method 800 can further include passing the sample 111 through the sample passage 210, 310, 510 and into contact with the main oxygen sensor 214 and the modified oxygen sensor 216, 316, 516. The sample 111 also contacts additional sensors or ground 217 and reference sensor 534 located along the sample passage 210, 310, 510. For example, a syringe or other sample delivery device can be used to interface with the sample inlet 109 of the cartridge body 108, 308, 508. The syringe or other device can pass the sample 111 into the sample inlet 109, through the sample passage 210, 310, 510, and into the waste passage 219.

[0082] The method 800 further includes, at block 806, obtaining a first measurement of the sample 111 from the primary oxygen sensor 214, and, at block 808, obtaining a second measurement of the sample 111 from the modified oxygen sensor 216, 316, 516. At block 810, the method 800 includes providing a level of hemolysis in the sample based on the first measurement and the second measurement. Such information can be collected by the controller 105, and the hemolysis level can be displayed on a user interface (e.g., display 104D) for communication to a user of the diagnostic analyzer 100. Additionally, any other test results and other information, as well as the hemolysis level, can be transmitted to the hospital information system (HIS) 101 via a wired connection or wirelessly. For example, WIFI or wired LAN communication can be used.

[0083] In some embodiments, the sample 111 used for testing can be a volume of 100 μL or less, or even 50 μL or less, although other sample 111 volumes can be used. As will be appreciated, the diagnostic sensor assembly 103, 303, 503 embodied in a diagnostic cartridge 106, 306, 506 can be used in a diagnostic analyzer 100 to perform multiple oxygen measurements to detect the level of hemolysis in the sample 111. It will also be appreciated that the diagnostic sensor assembly 103, 303, 503 can be included in a diagnostic analyzer without being embodied in a diagnostic cartridge, and can include a cleaning system in communication with the diagnostic sensor assembly 103, 303, 503 for reuse of the diagnostic sensor assembly 103, 303, 503.

[0084] In some embodiments, only the sensor array 212, 312 can be included as a disposable cartridge that can be coupled to a diagnostic analyzer, which itself can include the sample inlets and passageways, waste passageways, control portions, and a washing system that enables washing of the sensor array 212, 312 after use. Thus, the diagnostic cartridge can include inlets (such as inlet 351) and outlets (such as outlet 352) that sealingly connect to the passageways and waste passageways of the diagnostic analyzer when the cartridge is coupled to the analyzer. Thus, in this case, the diagnostic cartridge includes the primary and modified oxygen sensors described herein, as well as electrical contacts, such that the cartridge is detachable / removable from the diagnostic analyzer after multiple uses.

[0085] The following is a list of non-limiting exemplary embodiments disclosed herein. 1. A diagnostic sensor assembly comprising: a sample inlet configured to receive a sample; a sample passageway extending from the sample inlet; a primary oxygen sensor configured to contact the sample along the sample passage; and a modified oxygen sensor configured to contact the sample along the sample path, the modified oxygen sensor including an oxidant configured to oxidize hemoglobin to methemoglobin, from which a quantification of hemolysis of the sample can be obtained. 2. The diagnostic sensor assembly of exemplary embodiment 1, wherein the diagnostic sensor assembly is configured as part of a cartridge body of a diagnostic cartridge configured to connect to a diagnostic analyzer. 3. The diagnostic sensor assembly of any one of the preceding exemplary embodiments, wherein the primary oxygen sensor is configured to provide a first measurement that correlates to the partial pressure of oxygen in the sample. 4. The diagnostic sensor assembly of any one of the preceding exemplary embodiments, wherein the oxidizer of the modified oxygen sensor comprises potassium ferricyanide. 5. Oxidizing agents convert hemoglobin to the ferrous state (Fe +2 ) to the ferric state (Fe +3) to oxidize the sensor to the desired concentration. 6. The diagnostic sensor assembly of any one of the preceding exemplary embodiments, wherein the modified oxygen sensor is configured to provide a second measurement value that correlates to the partial pressure of oxygen in the sample. 7. The diagnostic sensor assembly of any one of the preceding exemplary embodiments, wherein the oxidant is provided within a membrane of the modified oxygen sensor. 8. The oxidant in the modified oxygen sensor is designed to convert extracellular hemoglobin to the ferrous state (Fe +2 ) to the ferric state (Fe +3 20. The diagnostic sensor assembly of claim 19, wherein the sample passage is provided at a location within the sample passage that enables oxidation of the sample to . 9. The diagnostic sensor assembly of any one of the preceding exemplary embodiments, wherein the main oxygen sensor is provided in a first passage split from the primary passage, and the correction oxygen sensor is provided in a second passage split from the primary passage. 10. A diagnostic analyzer comprising: an analyzer body including a cartridge receiving portion and an electrical connector; A diagnostic cartridge receivable in a cartridge receiving portion, a cartridge body configured to couple to the cartridge receiving portion, the cartridge body comprising: an electrical contact connectable to the electrical connector when the cartridge body is received in the cartridge receiving portion; a sample inlet configured to receive the sample; a sample passageway extending from the sample inlet into the cartridge body and configured to receive the sample; a primary oxygen sensor configured to contact the sample in the sample passage and configured to provide a first measurement of the sample; a diagnostic cartridge configured to contact the sample in the sample passage, the diagnostic cartridge including an oxidant configured to oxidize hemoglobin to methemoglobin, and a modified oxygen sensor configured to provide a second measurement of the sample; a controller coupled to the electrical connector and configured to receive a first measurement and a second measurement of the sample, and configured to provide a level of hemolysis in the sample based on the first measurement and the second measurement. 11. The diagnostic analyzer of exemplary embodiment 10, wherein the first measurement correlates to a first partial pressure measurement of oxygen in the sample. 12. The diagnostic analyzer of any one of the preceding exemplary embodiments, wherein the second measurement correlates to a second partial pressure measurement of oxygen in the sample. 13. The diagnostic analyzer of any one of the preceding exemplary embodiments, wherein an elevated second measurement compared to the first measurement quantitatively correlates to the level of hemolysis in the sample. 14. The diagnostic analyzer of any one of the preceding exemplary embodiments, wherein the controller is configured to determine a difference between the first measurement and the second measurement. 15. The diagnostic analyzer of any one of the preceding exemplary embodiments, wherein the difference quantitatively correlates to the level of hemolysis in the sample. 16. A method for determining hemolysis in a sample, comprising: coupling a diagnostic cartridge to a cartridge receiver of a diagnostic analyzer, the diagnostic cartridge including a sample inlet configured to receive a sample, a sample passageway extending from the sample inlet into the cartridge body, a primary oxygen sensor configured to contact the sample in the sample passageway, and a modified oxygen sensor configured to contact the sample in the sample passageway, the modified oxygen sensor including an oxidant configured to oxidize extracellular hemoglobin to methemoglobin; passing the sample through the sample passageway and contacting the primary oxygen sensor and the modified oxygen sensor; obtaining a first measurement of the sample from a primary oxygen sensor; obtaining a second measurement of the sample from the modified oxygen sensor; and providing a level of hemolysis in the sample based on the first measurement and the second measurement.

[0086] The foregoing description discloses only exemplary embodiments of the present disclosure. Modifications of the above-disclosed diagnostic cartridge and diagnostic analyzer, and method for detecting hemolysis, which fall within the scope of the present invention, will be readily apparent to those skilled in the art. It is therefore to be understood that other embodiments, as defined by the appended claims and their equivalents, may also fall within the scope of the present invention.

Claims

1. 1. A diagnostic sensor assembly comprising: a sample inlet configured to receive a sample; a sample passageway extending from the sample inlet; a primary oxygen sensor configured to contact the sample along the sample passage; and a modified oxygen sensor configured to contact the sample along the sample path, the modified oxygen sensor including an oxidant configured to oxidize hemoglobin to methemoglobin, from which a quantification of hemolysis of the sample can be obtained.

2. The diagnostic sensor assembly of claim 1 , wherein the diagnostic sensor assembly is configured as part of a cartridge body of a diagnostic cartridge configured to connect to a diagnostic analyzer.

3. The diagnostic sensor assembly of claim 1 , wherein the primary oxygen sensor is configured to provide a first measurement that correlates to a partial pressure of oxygen in the sample.

4. 10. The diagnostic sensor assembly of claim 1, wherein the oxidizer of the modified oxygen sensor comprises potassium ferricyanide.

5. The oxidizing agent converts hemoglobin to the ferrous state (Fe ＋２ ) to the ferric state (Fe ＋３ 10. The diagnostic sensor assembly of claim 1, wherein the diagnostic sensor assembly is configured to oxidize the oxidized cations to cations of the ...

6. The diagnostic sensor assembly of claim 1 , wherein the modified oxygen sensor is configured to provide a second measurement that correlates to the partial pressure of oxygen in the sample.

7. The diagnostic sensor assembly of claim 1 , wherein the oxidizer is provided within a membrane of the modified oxygen sensor.

8. The oxidant in the modified oxygen sensor converts extracellular hemoglobin to the ferrous state (Fe ＋２ ) to the ferric state (Fe ＋３ 10. The diagnostic sensor assembly of claim 1, wherein the sample path is provided at a location effective to oxidize the sample to the desired ion concentration.

9. 9. The diagnostic sensor assembly of claim 8, wherein the main oxygen sensor is provided in a first passageway split from the primary passageway and the correction oxygen sensor is provided in a second passageway split from the primary passageway.

10. 1. A diagnostic analyzer comprising: an analyzer body including a cartridge receiving portion and an electrical connector; A diagnostic cartridge receivable in a cartridge receiving portion, comprising: a cartridge body configured to couple to a cartridge receiver, the cartridge body comprising: an electrical contact connectable to the electrical connector when the cartridge body is received in the cartridge receiving portion; a sample inlet configured to receive a sample; a sample passageway extending from the sample inlet into the cartridge body and configured to receive the sample; a primary oxygen sensor configured to contact the sample in the sample passage and to provide a first measurement of the sample; a diagnostic cartridge configured to contact the sample in the sample passage, the diagnostic cartridge including an oxidant configured to oxidize hemoglobin to methemoglobin, and a modified oxygen sensor configured to provide a second measurement of the sample; a controller coupled to the electrical connector and configured to receive a first measurement and a second measurement of the sample, and configured to provide a level of hemolysis in the sample based on the first measurement and the second measurement.

11. 11. The diagnostic analyzer of claim 10, wherein the first measurement correlates to a first measurement of partial pressure of oxygen in the sample.

12. 11. The diagnostic analyzer of claim 10, wherein the second measurement correlates to a second partial pressure measurement of oxygen in the sample.

13. 13. The diagnostic analyzer of claim 12, wherein an elevated second measurement compared to the first measurement quantitatively correlates to the level of hemolysis in the sample.

14. The diagnostic analyzer of claim 10 , wherein the controller is configured to determine a difference between the first measurement and the second measurement.

15. 15. The diagnostic analyzer of claim 14, wherein the difference quantitatively correlates to the level of hemolysis in the sample.

16. 1. A method for determining hemolysis in a sample, comprising: coupling a diagnostic cartridge to a cartridge receiver of a diagnostic analyzer, the diagnostic cartridge including a sample inlet configured to receive a sample, a sample passageway extending from the sample inlet into a cartridge body, a primary oxygen sensor configured to contact the sample in the sample passageway, and a modified oxygen sensor configured to contact the sample in the sample passageway, the modified oxygen sensor including an oxidant configured to oxidize extracellular hemoglobin to methemoglobin; passing a sample through a sample passageway to contact the primary oxygen sensor and the correction oxygen sensor; obtaining a first measurement of the sample from a primary oxygen sensor; obtaining a second measurement of the sample from the modified oxygen sensor; and providing a level of hemolysis in the sample based on the first measurement and the second measurement.

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