Potassium ion detection reagent, detection method and use thereof

Abstract

The present invention relates to a potassium ion detection reagent comprising sodium tetraphenylborate, a buffer, a thickener, and a surfactant. It also relates to a kit containing the reagent, a method for detecting potassium ions, and uses of the reagent. Furthermore, the present invention relates to the use of the potassium ion detection reagent in assessing alkaline cleaning solution residues in medical devices, as well as a method for assessing the residual rate of alkaline cleaning solution in medical devices. The present invention overcomes the poor stability of sodium tetraphenylborate and improves the analytical sensitivity of the potassium ion detection reagent.

Description

Technical Field

[0001] The present invention relates to the field of potassium ion detection, and in particular to the detection of residual levels of alkaline cleaning solutions in the medical field. Background Art

[0002] Potassium ion determination methods include flame photometry, sodium tetraphenylborate turbidimetry, atomic absorption spectrometry, dry chemical methods, chromogenic ion carrier colorimetry, ion-selective electrode (ISE), and enzyme kinetics. In the sodium tetraphenylborate turbidimetry method, sodium tetraphenylborate, in a slightly alkaline medium and with the presence of a stabilizer, reacts with potassium ions to form a stable, low-solubility white substance called potassium tetraphenylborate. Its turbidity is proportional to the potassium ion concentration. This relationship allows for the detection of potassium ion concentration by measuring the optical signal. This method boasts high sensitivity and has been successfully applied to the determination of potassium ion content in soil equipment, pharmaceuticals, and other applications.

[0003] However, the main drawback of the sodium tetraphenylborate turbidimetric method is that its aqueous solution is extremely unstable, typically only stable for two hours after preparation, requiring it to be prepared immediately before use. Furthermore, due to its poor stability, the concentration of sodium tetraphenylborate in solution cannot exceed 30g / L. These drawbacks limit the detection effectiveness (e.g., sensitivity) and application range of the sodium tetraphenylborate turbidimetric method.

[0004] Therefore, in the field of potassium ion detection, there is a strong demand to increase the concentration and stability of sodium tetraphenylborate solution in the solution. Summary of the Invention

[0005] To overcome the above problems, the inventors studied the potassium ion detection method and found that adding a surfactant to the detection reagent based on the sodium tetraphenylborate turbidimetry method can effectively improve the stability of the sodium tetraphenylborate solution, increase its sensitivity, and expand its application range.

[0006] Therefore, in a first aspect, the present invention provides a potassium ion detection reagent based on sodium tetraphenylborate turbidimetry, comprising sodium tetraphenylborate, a buffer, a thickener and a surfactant.

[0007] The addition of a surfactant to the reagent of the present invention allows the detection reagent containing sodium tetraphenylborate to remain stable at room temperature for more than six months, making the potassium ion detection reagent based on the sodium tetraphenylborate method easy to store and use, thereby reducing usage costs and facilitating its promotion. Furthermore, the improved reagent of the present invention also optimizes the analytical sensitivity when detecting potassium ions.

[0008] In the present invention, sodium tetraphenylborate can also be called sodium tetraphenylborate, sodium tetraphenylborate, sodium tetraphenylborate, sodium tetraphenylborate, and sodium tetraphenylborate. 24 H 20 BNa, substance with CAS registration number 143-66-8.

[0009] In some embodiments, sodium tetraphenylborate in the reagent of the present invention may be present at a concentration of 30 g / L or greater.

[0010] Due to the poor stability of sodium tetraphenylborate itself, its concentration in aqueous solutions usually cannot be higher than 30g / L. However, in the improved reagent of the present invention, sodium tetraphenylborate can be present at a concentration of more than 30g / L, thus giving sodium tetraphenylborate turbidimetry the potential for application in detection scenarios requiring higher sensitivity.

[0011] In a specific embodiment, sodium tetraphenylborate in the reagent of the present invention may be present in a concentration of about 30 to about 60 g / L.

[0012] In the present invention, "buffer" refers to a solution used to keep the pH of the reagent relatively stable in a potassium ion detection reagent based on the sodium tetraphenylborate method. The present invention has no particular restrictions on the type of buffer, and any common buffer used in the sodium tetraphenylborate turbidimetric method can be used. Exemplary buffers can be Tris buffer, phosphate buffer, HEPES buffer, TAPS buffer, Mops buffer, but the present invention is not limited thereto. Those skilled in the art can appropriately adjust the pH or buffer concentration of the buffer according to the desired pKa. Typically, such a buffer can have a concentration of about 50mM to about 300mM and have a pH value range of about 7.5 to about 9.5.

[0013] In some embodiments, the buffer is selected from one or more of Tris buffer, phosphate buffer, HEPES buffer, TAPS buffer and Mops buffer.

[0014] In some embodiments, the buffer concentration of the buffer is about 50 mM to about 300 mM.

[0015] In some embodiments, the pH of the buffer is in the range of about 7.5 to about 9.5.

[0016] In the present invention, "thickener" refers to a substance that increases the viscosity of the reagent. In the sodium tetraphenylborate turbidimetry, the use of a thickener can maintain sodium tetraphenylborate and the product potassium tetraphenylborate in a uniform state, thereby avoiding stratification. The present invention has no particular limitation on the type of thickener, and any common thickener used in the sodium tetraphenylborate turbidimetry can be used. Exemplary thickeners can be glycerol and dextran, but the present invention is not limited thereto. Those skilled in the art can determine the appropriate amount of thickener to be added, as long as sodium tetraphenylborate and the product potassium tetraphenylborate can be kept in a uniform state. Typically, the thickener can be present in a concentration of about 5 g / L to about 50 g / L.

[0017] In some embodiments, the thickener is selected from one or more of glycerol and dextran.

[0018] In some embodiments, the concentration of the thickener is 5 to 50 g / L.

[0019] In a specific embodiment, the surfactant is selected from one or more of Tween, such as Tween-20, Tween-80; TX, such as TX-100, TX-405; and Brij, such as Brij35, Brij98, and Brij L23.

[0020] In a preferred embodiment, the surfactant is Tween-80.

[0021] As demonstrated in the examples below, when Tween-80 is selected as the surfactant, the reagent and the corresponding detection method of the present invention can further achieve better analytical sensitivity.

[0022] In some embodiments, the concentration of the surfactant of the present invention is about 0.5 to about 25 g / L, such as 1 g / L, 2 g / L, 3 g / L, 4 g / L, 5 g / L, 6 g / L, 7 g / L, 8 g / L, 9 g / L, 10 g / L, 11 g / L, 12 g / L, 13 g / L, 14 g / L, 15 g / L, 16 g / L, 17 g / L, 18 g / L, 19 g / L, 20 g / L, 21 g / L, 22 g / L, 23 g / L, or 24 g / L. In specific embodiments, the concentration of the surfactant of the present invention is about 1 to about 20 g / L.

[0023] In a preferred embodiment, the concentration of the surfactant of the present invention is about 4 to about 12 g / L. In another preferred embodiment, the concentration of the surfactant of the present invention is about 5 to about 10 g / L.

[0024] As demonstrated in the examples below, when the concentration of the surfactant is selected from about 4 to about 12 g / L or about 5 to about 10 g / L, the reagent of the present invention and the corresponding detection method have better stability and high sensitivity.

[0025] Alternatively, the reagent of the present invention may further include other components for sodium tetraphenylborate turbidimetry. Exemplary other components may be preservatives for extending the shelf life of the reagent, such as sodium azide, the proclin series, p-hydroxybenzoic acid, benzoic acid, or a combination thereof. The reagent of the present invention can be used to detect the presence and / or level of potassium ions.

[0026] In a second aspect, the present invention provides a kit comprising the reagents defined in the first aspect of the present invention.

[0027] Optionally, the kit of the present invention may further include other reagents and / or equipment for sodium tetraphenylborate turbidimetry. Exemplary other reagents may include reagents for sample pretreatment, for example, reagents for treating soil samples may include an acidic solution (such as HCl) and an alkaline solution (NaOH). Exemplary other equipment may include colorimetric tubes, etc.

[0028] The kit of the present invention can be used to detect the presence and / or level of potassium ions.

[0029] In a third aspect, the present invention provides a method for detecting potassium ions (based on sodium tetraphenylborate turbidimetry), the method comprising the following steps:

[0030] Mix the reagent with the sample and allow it to react for a period of time; and determine the potassium ion content.

[0031] The reagent contains sodium tetraphenylborate, a buffer solution, a thickener and a surfactant.

[0032] In some embodiments, the method further comprises a step of pre-treating the sample before the mixing step.

[0033] In a specific embodiment, the pretreatment step may include, for example, dissolving a solid sample and removing potential interference from other components in the sample. For example, when testing a soil sample, the soil is dissolved with 1 mM HCl, stirred, and centrifuged to obtain the supernatant. The supernatant is then treated with 1% 1 M NaOH to dissolve the supernatant, and then centrifuged to obtain the supernatant for analysis. The pretreatment step may also include dissolving potassium in the sample. For example, since the sample may adhere to the surface of an object in a non-solubility form, the object may be rinsed before testing to dissolve any potassium present. However, the present invention is not limited thereto.

[0034] The present invention has no particular limitations on the reaction time and temperature between the reagent and the sample. Conventional sodium tetraphenylborate turbidimetry can be used, as long as the sodium tetraphenylborate reacts sufficiently with the potassium ions to form stable potassium tetraphenylborate. An exemplary reaction time is 5 minutes, and an exemplary reaction temperature is 37°C. However, the present invention is not limited thereto.

[0035] In the present invention, a sample refers to an analyte that may contain potassium ions. Exemplary samples may be selected from soil, blood, serum, reagents containing potassium ions or their residues, waste liquids, pharmaceuticals, and fertilizers. Preferably, the reagents containing potassium ions or their residues are alkaline detergents or their residues.

[0036] As explained in the description of the samples targeted by the detection method, the potassium ion detection reagent of the present invention is applicable to a variety of different application scenarios, for example, it can be used to detect potassium ions in soil samples or in clinical blood samples. In addition, during the research process, the inventors surprisingly discovered that the residual level of alkaline cleaning solution after cleaning medical equipment can be assessed by detecting potassium ion content. Therefore, in additional aspects, the present invention further expands the scope of application of potassium ion detection reagents.

[0037] Specifically, in the field of biochemical analysis, samples from complexity, such as serum, blood plasma, urine, pleural effusion, cerebrospinal fluid, etc., are often tested by medical equipment (such as fully automatic medical equipment), and reagents with different compositions are also used during testing. After the test, these samples and reagents are likely to remain in at least some parts of the medical equipment, so it is usually necessary to use alkaline cleaning fluid to clean these parts. However, while cleaning, alkaline cleaning agents may also cause secondary contamination, such as residual potassium ions that may affect the potassium ion detection of ISE, and residual metal chelating agents that may affect the determination of some metal ions such as Ca, Cu and Zn, etc. Therefore, the residual rate of alkaline cleaning agents is an important evaluation index of medical equipment cleaning efficiency. For example, for biochemical analyzers, the maximum allowable residual rate of reaction cups and stirring rods is 200ppm, and the maximum allowable residual rate of reagent needles and sample needles is 100ppm.

[0038] Currently, the standard method for evaluating cross-contamination is Orange G (YYT 0654-2017 Pharmaceutical Industry Standard of the People's Republic of China - Fully Automatic Biochemical Analyzer). This method uses the specific absorption peak of Orange G for detection. However, the standard solution is an aqueous solution with properties (such as surface tension and viscosity) that differ greatly from those of the detection reagent and alkaline cleaning solution. As a result, the Orange G residual rate assessment result is lower than the actual value, and therefore cannot simulate the actual cleaning effect of the equipment. In other words, there is still a lack of effective methods for evaluating the residual rate of alkaline cleaning solutions.

[0039] To address this issue, in a fourth aspect, the present invention provides a use of a potassium ion detection reagent in evaluating alkaline cleaning solution residues in medical equipment.

[0040] By measuring the potassium ion content remaining in the reaction cup and the stirring rod, the residual level of the alkaline cleaning solution can be indirectly reflected, thereby providing an accurate and effective optional method for evaluating the residual rate of potassium-containing alkaline cleaning solution.

[0041] In a preferred embodiment, the potassium ion detection reagent includes a second reagent, wherein the second reagent contains sodium tetraphenylborate, a buffer, a thickener, and a surfactant. The second reagent of the present invention can be used to determine the potassium ion content in the residual alkaline cleaning solution based on the sodium tetraphenylborate turbidimetric method.

[0042] In a more preferred embodiment, the potassium ion detection reagent further includes a first reagent comprising a buffer, a thickener, and a surfactant. The first reagent of the present invention can be used to rinse out the alkaline cleaning solution remaining in the medical device, thereby obtaining a sample containing the alkaline cleaning solution to be evaluated. At the same time, the first reagent can also be used to better dissolve the sample and play an anti-interference role, such as through timed turbidity detection, to eliminate interference in the sample.

[0043] The definitions and descriptions of sodium tetraphenylborate, buffer, thickener and surfactant in the reagent of the present invention are also applicable to the corresponding components in the first reagent and the second reagent of the present invention.

[0044] It is understood that the buffer, thickener and / or surfactant commonly contained in the first reagent and the second reagent of the present invention can be of the same or different types, such as the thickener in the first reagent is glycerol, while the thickener in the second reagent is dextran; and / or the same or different parameters, such as the buffer in the first reagent is 100mM Tris buffer, while the buffer in the second reagent is 150mM Tris buffer.

[0045] In a preferred embodiment, the first reagent and the second reagent of the present invention use the same buffer, thickener and surfactant.

[0046] In the present invention, an alkaline cleaning solution refers to a potassium-containing alkaline cleaning solution commonly used in cleaning medical equipment. Those skilled in the art will appreciate the specific types of these alkaline cleaning solutions. Exemplary alkaline cleaning solutions include those containing potassium hydroxide, such as Alkaline Cleaning Solution CD80 (Mindray) or Alkaline Cleaning Solution CX (Beckman).

[0047] In a fifth aspect, the present invention provides a method for evaluating the residual rate of alkaline cleaning solution in medical equipment, comprising the following steps:

[0048] treating the medical device with a first reagent of the present invention to collect a sample to be evaluated;

[0049] adding the second reagent of the present invention to the collected sample, mixing and reacting for a period of time;

[0050] Determine the potassium ion content in the sample after the reaction; and

[0051] The residual rate of the alkaline cleaning solution is obtained based on the obtained potassium ion content.

[0052] In the present invention, the first reagent can be used to treat one or more locations of the medical device, depending on actual needs. For example, when evaluating the residual alkaline cleaning solution at the reaction cup and / or stirring rod, the aforementioned location of the first reagent treatment device is used to obtain the residual rate level at that location. For another example, when evaluating the residual alkaline cleaning solution at the reagent needle and / or sample needle, the aforementioned location of the first reagent treatment device is used to obtain the residual rate level at that location.

[0053] In some embodiments, the methods of the present invention may further include determining whether the residual level is within a maximum allowable residual level. For example, when evaluating samples from a cuvette and / or a stirrer, the residual level may be further determined to be greater than the corresponding maximum allowable residual level (200 ppm). Furthermore, if the residual level exceeds the corresponding maximum allowable residual level, the methods of the present invention may further include removing the alkaline cleaning solution residue from the medical device.

[0054] The present invention has no particular limitations on the reaction time and conditions between the second reagent and the sample. Conventional sodium tetraphenylborate turbidimetry can be used, as long as the sodium tetraphenylborate reacts sufficiently with the potassium ions to form stable potassium tetraphenylborate. An exemplary reaction time is 5 minutes, and an exemplary reaction temperature is 37°C. However, the present invention is not limited thereto.

[0055] It will be appreciated that those skilled in the art can determine the residual rate of the corresponding alkaline cleaning solution based on the specific type and content of the potassium component in the alkaline cleaning solution. For example, when the alkaline cleaning solution used contains potassium hydroxide (alkaline cleaning solution CD80), the alkaline cleaning solution is first diluted to obtain a multi-point calibration standard (ppm) to plot a standard curve. The value subsequently measured is the residual rate of the alkaline cleaning solution.

[0056] In a sixth aspect, the present invention further provides a kit for determining residual alkaline cleaning solution, comprising the first reagent and the second reagent of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] Figure 1 The standard curve for detecting potassium ion concentration using the reagents of Experiment 2 is shown;

[0058] Figure 2 The standard curve for detecting potassium ion concentration using the reagent of Experiment 6 is shown;

[0059] Figure 3 The standard curve for detecting potassium ion concentration using the reagent of Experiment 9 is shown;

[0060] Figure 4 The standard curve for detecting the residual alkaline cleaning solution using the reagent of Experiment 2 is shown. DETAILED DESCRIPTION

[0061] The following is a clear and complete description of the technical methods in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0062] Preparation of the first reagent

[0063] Take the preparation of 1L as an example:

[0064] 1) In a 1 L beaker, weigh 900 ml of deionized water, add 24.228 g of Tris base, and stir for at least 10 minutes to ensure complete dissolution. Adjust the pH (at 25°C) to 8.0 ± 0.1 with 3 M hydrochloric acid.

[0065] 2) Weigh a certain amount of surfactant and add it to the above solution, stirring for more than 10 minutes;

[0066] 3) Weigh 10 g of dextran (molecular weight 60K) and add it to the above solution. Stir for more than 10 minutes and finally adjust the volume to 1 L.

[0067] Preparation of the second reagent

[0068] 1) In a 1 L beaker, weigh 900 ml of deionized water, add 24.228 g of Tris base, and stir for at least 10 minutes to ensure complete dissolution. Adjust the pH (at 25°C) to 8.0 ± 0.1 with 3 M hydrochloric acid.

[0069] 2) Weigh a certain amount of surfactant and add it to the above solution, stirring for more than 10 minutes;

[0070] 3) Weigh 10 g of dextran (molecular weight 60K) and add it to the above solution, stirring for more than 10 minutes;

[0071] 4) Weigh 40 g of sodium tetraphenylborate and add it to the above solution. Stir for 30 minutes until dissolved, and finally adjust the volume to 1 L.

[0072] 5) Filter with a 0.45 μm filter membrane to remove a small amount of particles.

[0073] Example 1

[0074] The first and second reagents were prepared according to the methods of “Preparation of the first reagent” and “Preparation of the second reagent” above, wherein the type of surfactant was Tween 80, and the amounts used were shown in Table 1 below.

[0075] Table 1

[0076]

[0077]

[0078] Example 2

[0079] One day after preparation, the second reagent in the comparative example showed obvious white precipitate visually, while no precipitate was observed in Experiments 1-4, which added a surfactant. To further analyze the long-term storage stability of Experiments 1-4, a 37°C thermal acceleration test was conducted: the reagents in Experiments 1-4 were stored at 37°C and tested in a biochemical analyzer on days 2, 4, 6, 8, and 10 with 3 μmol / L and 10 μmol / L KCl solutions. The relative deviation from day 0 was calculated, and the results are shown in Table 2 below.

[0080] Table 2

[0081]

[0082] The results showed that under the conditions of thermal acceleration at 37°C, the reagents in experiments 1 to 4 were able to remain stable for 2 days, 4 days, 6 days, 8 days and 10 days, and the relative deviation was within 10%.

[0083] Example 3

[0084] The analytical sensitivity of the first and second reagents in experiments 1 to 4 was tested for potassium ion content. Similarly, the sensitivity test of the second reagent in the comparative example could not be performed due to obvious precipitation. The specific steps are as follows:

[0085] 1) Prepare KCl calibrants Cal 1 to 9 (1.0 μmol / L, 2.0 μmol / L, 3.0 μmol / L, 5 μmol / L, 10 μmol / L, 20 μmol / L, 40 μmol / L, 80 μmol / L, and 100 μmol / L);

[0086] 2) Set the parameters of the fully automatic biochemical analyzer (Mindray, BS800) to 605 nm as the dominant wavelength, the volume ratio of the first reagent, the second reagent, and the sample = 150 μl:150 μl:2 μl, the endpoint method, the reaction time of 5 min, and select the potassium ion detection item;

[0087] 3) Based on the measured reactivity, a standard curve for determining the potassium ion concentration of the standard substance was drawn ( Figure 1 , taking Experiment 2 as an example), the standard of analytical sensitivity is that the relative deviation between the lowest concentration that can be detected and the true concentration does not exceed 10%.

[0088] The experimental results show that when the reagents in Experiments 1 to 4 are used for determination, the analytical sensitivity of potassium ions can reach 5.1μmol / L, 1.4μmol / L, 2.1μmol / L and 3.1μmol / L respectively.

[0089] Example 4

[0090] The first and second reagents were prepared according to the methods of “Preparation of the first reagent” and “Preparation of the second reagent” above, wherein the type of surfactant was TX-100, and the amounts used were shown in Table 3 below.

[0091] Table 3

[0092]

[0093] One day after preparation, no precipitation was observed in the second reagents in Experiments 5 to 8. The above experiments were subjected to a 37°C thermal acceleration test according to the method in Example 2. The results are shown in Table 4 below.

[0094] Table 4

[0095]

[0096] As shown in Table 4, under the condition of thermal acceleration at 37°C, the second reagent in Experiments 5 to 8 can remain stable for 2 days, 4 days, 6 days, 8 days, and 10 days, respectively, and the relative deviation is within 10%.

[0097] Next, according to the method in Example 3, the analytical sensitivity of the reagents in Experiments 5 to 8 in detecting potassium ion content was investigated ( Figure 2 , taking Experiment 6 as an example). The results show that when using the reagents in Experiments 5 to 8, the analytical sensitivity of potassium ions can reach 4.6 μmol / L, 2.0 μmol / L, 3.0 μmol / L, and 3.8 μmol / L, respectively.

[0098] Example 5

[0099] Experiment 9 was prepared according to the above-mentioned methods of “Preparation of the first reagent” and “Preparation of the second reagent”, wherein the surfactant in the first reagent was 5 g / L TX-100, and the surfactant in the second reagent was 5 g / L Brij L23.

[0100] The stability and analytical sensitivity of Experiment 9 were tested according to the methods in Examples 2 and 3. The results showed that under the condition of thermal acceleration at 37°C, the reagent in Experiment 9 could remain stable for 8 days with a relative deviation within 10%. When the reagent in Experiment 9 was used, the analytical sensitivity of potassium ions was 1.2 μmol / L ( Figure 3 ).

[0101] Example 6

[0102] Use the reagents of Experiment 2 to determine the alkaline cleaning solution residue in the biochemical analyzer. The specific steps are as follows:

[0103] 1) Use a fully automatic biochemical analyzer (Mindray, BS800) and the accompanying alkaline cleaning solution to perform the preset enhanced cleaning program;

[0104] 2) Set the biochemical analyzer parameters, the detection wavelength is 605nm, the volume ratio of the first reagent, the second reagent and the sample is 150μl:150μl:2μl, the endpoint method, the reaction time is 5min. Select the potassium ion detection project, dilute the cleaning solution to a concentration of 5×10 -6 As (10ppm) calibration. In a similar manner, multi-point calibration products of 2.5ppm, 10ppm, 20ppm, 40ppm and 100ppm were prepared, and the standard curve was obtained ( Figure 4 );

[0105] 3) Evaluation of cleaning residues: Select the potassium ion test item, add the first reagent and the second reagent, and test a full circle of reaction cups. The test result of each reaction cup is Ai.

[0106] Example 7

[0107] The sample carryover contamination rate detection method in the industry standard "YYT 0654-2017 Fully Automatic Biochemical Analyzer" was used, and the residual rate of the fully automatic analyzer (Mindray, BS800) was measured using the same alkaline cleaning solution and cleaning process as in Example 6.

[0108] Among them, the steps of the industry standard method are:

[0109] 1) Dissolve an appropriate amount of Orange G in human serum to prepare an Orange G stock solution with an absorbance of 200 at 340 nm.

[0110] 2) Dilute the Orange G stock solution accurately 200-fold and measure the absorbance of the dilution relative to deionized water at 340 nm on a photometer. Repeat the measurement 20 times and calculate the average of the 20 absorbances. Multiply this average by the dilution factor to obtain the theoretical absorbance, A, of the Orange G stock solution.

[0111] 3) Using deionized water as the reagent, Orange G stock solution and deionized water as the samples, with the sample addition amount being the maximum sample amount specified for the analyzer, the absorbance of the samples at the end of the reaction was measured on the analyzer in the order of stock solution, stock solution, stock solution, deionized water, deionized water, and deionized water. Perform a total of five sets of measurements.

[0112] 4) In each group of measurements, the absorbance of the fourth sample is Ai4, and the absorbance of the sixth sample is Ai6, where i is the serial number of the measurement group;

[0113] 5) Calculate the carryover rate according to the following formula and take the maximum carryover rate as the result.

[0114]

[0115] Where V S V is the volume of sample added; R is the volume of reagent added.

[0116] Example 8

[0117] The tests of Examples 6 and 7 were performed in parallel a total of 159 times, and the results are summarized in Table 5 below.

[0118] Table 5

[0119]

[0120]

[0121]

[0122] As shown in Table 5, the average cleaning residue rate measured by the present invention's detection method is 48.867 ppm, while the average cleaning residue rate measured by the industry standard method is 10 ppm. The average residual rate after the cleaning process of the Mindray BS-800 biochemical analyzer with the accompanying alkaline cleaning solution is known to be around 50 ppm.

[0123] It can be seen that the current industry standard detection method (i.e., the Orange G method in Example 7) cannot reflect the true cleaning efficiency, while the detection method of the present invention more truly reflects the actual residual situation.

[0124] Example 9

[0125] In order to verify the ability of the second reagent of the present invention to detect potassium ion-containing samples, the second reagent in Experiment 2 was further used to measure the potassium ion content of a soil standard (the potassium ion content of which has been calibrated by the China Metrology Bureau to be 120 mg / kg). The specific steps are as follows:

[0126] 1) Dissolve 10 g of soil standard in 1000 ml of 1 mM HCl and stir for 30 minutes. Centrifuge and remove the supernatant, which is then dissolved in 10% 1 M NaOH. Treat at 50-90°C for 10 minutes. Cool and centrifuge, then remove the supernatant as the test sample (concentration: 2.80 mmol / L).

[0127] 2) Calibration with 10 μmol / L potassium chloride;

[0128] 3) Set the biochemical analyzer parameters: the dominant wavelength is 605 nm, the volume ratio of the first reagent, the second reagent, and the sample is 150 μl:150 μl:2 μl, the endpoint method is used, and the reaction time is 5 minutes. Select the potassium ion test and set the sample dilution parameter to 100x.

[0129] The test results showed that the potassium ion concentration was 2.68mmol / L.

[0130] According to the formula "potassium ion concentration × 39 × 1.1 × volume / soil mass", the potassium ion content of the soil sample is 115 mg / kg, with a relative deviation of 4.2%, which shows that the reagent of the present invention can accurately measure the potassium ion content in the soil sample.

[0131] The above description is only a preferred embodiment of the present invention and does not limit the patent scope of the present invention. All equivalent structural transformations made by using the contents of the present description and drawings under the inventive concept of the present invention, or direct / indirect application in other related technical fields are included in the patent protection scope of the present invention.

Claims

1. A reagent for detecting potassium ions, characterized in that, Including sodium tetraphenylborate, buffer, thickener and surfactant, Wherein, the surfactant is selected from one or more of Tween, TX and Brij, and the concentration of the surfactant is 0.5-25 g / L; The buffer is selected from one or more of Tris buffer, phosphate buffer, HEPES buffer, TAPS buffer and Mops buffer, and the concentration of the buffer is 50-300 mM and the pH is 7.5-9.5; and The concentration of the sodium tetraphenylborate is greater than or equal to 30 g / L.

2. The reagent according to claim 1, characterized in that The invention comprises a first reagent containing a buffer, a thickener and a surfactant; and a second reagent containing sodium tetraphenylborate, a buffer, a thickener and a surfactant.

3. The reagent according to claim 1, characterized in that The surfactant is Tween-80.

4. The reagent according to any one of claims 1 to 3, characterized in that The concentration of the surfactant is 4-12 g / L.

5. The reagent according to any one of claims 1 to 3, characterized in that The thickener is selected from one or more of glycerol and dextran.

6. The reagent according to any one of claims 1 to 3, characterized in that The concentration of the thickener is 5-50 g / L.

7. The reagent according to any one of claims 1 to 3, characterized in that The concentration of sodium tetraphenylborate is 30-60 g / L.

8. A kit for detecting potassium ions, characterized in that The method comprises the reagent according to any one of claims 1 to 7.

9. A method for detecting potassium ions, characterized in that, The following steps are involved: mixing the reagent according to any one of claims 1 to 7 with a sample and reacting them for a period of time; as well as The potassium ion content was measured based on the suspended matter.

10. The method according to claim 9, characterized in that The step of mixing the reagent according to any one of claims 1 to 7 with the sample and reacting the mixture for a period of time comprises the following steps: Mixing a first reagent with a sample to obtain a mixture of the first reagent and the sample; wherein the first reagent contains a buffer, a thickener, and a surfactant; and A second reagent is mixed with the mixture and reacted for a period of time; wherein the second reagent contains sodium tetraphenylborate, a buffer, a thickener and a surfactant.

11. The method according to any one of claims 9 to 10, characterized in that Before the mixing step, a sample pretreatment step is also included.

12. The method according to any one of claims 9 to 10, characterized in that The sample is selected from soil, blood, serum, reagents containing potassium ions or their residues, waste liquid, medicines and fertilizers.

13. The method according to claim 12, characterized in that The potassium ion-containing reagent or its residue is an alkaline cleaning solution or its residue.

14. A method for evaluating the residual rate of alkaline cleaning solution in medical equipment, characterized in that: The following steps are involved: treating the medical device with a first reagent to collect a sample to be evaluated; mixing a second reagent with the collected sample and reacting for a period of time; Determine the potassium ion content in the sample; as well as The residual rate of the alkaline cleaning solution is obtained according to the obtained potassium ion content. wherein the first reagent contains a buffer, a thickener, and a surfactant; and the second reagent contains sodium tetraphenylborate, a buffer, a thickener, and a surfactant, In which, in the second reagent, the surfactant is selected from one or more of Tween, TX and Brij, and the concentration of the surfactant is 0.5~25 g / L; the buffer is selected from one or more of Tris buffer, phosphate buffer, HEPES buffer, TAPS buffer and Mops buffer, and the concentration of the buffer is 50~300 mM, and the pH is 7.5~9.5; and the concentration of sodium tetraphenylborate is greater than or equal to 30 g / L.

15. The method according to claim 14, characterized in that The first reagent is used to treat one or more of the following locations of the medical device: a reaction cup, a stirring rod, a reagent needle, and a sample needle.

16. The use of potassium ion detection reagents in evaluating alkaline cleaning solution residues in medical equipment, including: The potassium ion detection reagent includes sodium tetraphenylborate, a buffer, a thickener and a surfactant. Wherein, the surfactant is selected from one or more of Tween, TX and Brij, and the concentration of the surfactant is 0.5-25 g / L; The buffer is selected from one or more of Tris buffer, phosphate buffer, HEPES buffer, TAPS buffer and Mops buffer, and the concentration of the buffer is 50-300 mM and the pH is 7.5-9.5; and The concentration of the sodium tetraphenylborate is greater than or equal to 30 g / L.

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