Potassium ion concentration measuring reagent, measuring method and kit

Abstract

The invention discloses a potassium ion concentration measuring reagent. The measuring reagent is a single agent, comprising: a buffer, a coenzyme, an acetaldehyde dehydrogenase, a reaction substrate, a surfactant, a protective agent and a stabilizer; or the measuring reagent is a double agent reagent composed of a reagent 1 and a reagent 2, wherein the reagent 1 comprises: a buffer, an acetaldehyde dehydrogenase, a reaction substrate, a surfactant, a protective agent, a stabilizer and a diaphorase or a coenzyme; and the reagent 2 comprises: a buffer, a coenzyme and a diaphorase, a surfactant, a protective agent and a chromogen substrate; the buffer is a Tris-HCl buffer, and the pH value of the Tris-HCl buffer is 7-8.5; the coenzyme is one of NAD+ or NADP+; the reaction substrate is an aldehyde compound; the chromogen substrate is an NBT substrate or an INT substrate; the surfactant is a Triton series or Tween series surfactant; the protective agent is a mixed solution of one or more of BSA, sucrose and trehalose; and the stabilizer is a mixed solution of one or more of polyols; and a potassium ion concentration measuring method and a corresponding kit are also disclosed.

Description

Technical Field

[0001] The present invention relates to the technical field of potassium ion detection, and in particular to a potassium ion concentration measuring reagent, a measuring method and a kit. Background Art

[0002] Potassium is an important metal ion for maintaining cellular physiological activities in the body. It plays an important role in maintaining acid-base balance, participating in protein and sugar metabolism, and maintaining normal excitability of the myocardium and neuromuscular system.

[0003] Under normal circumstances, the concentration of serum potassium is between 3.5 and 5.5 mmol / L, with an average of 4.2 mmol / L. When serum potassium is >5.5 mmol / L, it is called hyperkalemia. The main causes of hyperkalemia include acute renal failure, potassium excretion disorders in uremia, adrenocortical insufficiency, shock, tissue crush injury, hypoaldosteronism, severe hemolysis, oral or injected potassium-containing solution, etc. When serum potassium is <3.5 mmol / L, it is called hypokalemia. There are many causes of hypokalemia, mainly adrenocortical hyperfunction, severe vomiting, diarrhea, taking diuretics and insulin, barium salt poisoning, metabolic alkalosis, low potassium diet, periodic paralysis, etc. Hyperkalemia and hypokalemia caused by potassium metabolism disorders, if not corrected in time, can lead to corresponding disorders in the physiological functions of various organ systems, especially the cardiovascular system and nervous system, and the body's material metabolism, and in severe cases, death.

[0004] The human body needs about 2-3g of potassium per day, which mainly comes from potassium salts in food. 90% of the daily potassium intake is excreted through urine, 10% is excreted through feces, and very little potassium is lost through the skin. The kidneys can balance potassium according to the needs of the body, and have the functions of potassium conservation and potassium excretion to maintain water, electrolyte and acid-base balance. Urine potassium measurement can reflect the condition of kidney disease. If the potassium content increases, it is predicted that adrenal cortical hyperfunction, renal tubular acidosis, diabetic ketoacidosis and taking diuretics may occur; if the potassium content decreases, it is predicted that adrenal cortical insufficiency, renal failure, and renal tubular potassium excretion disorders may occur. Taking certain drugs can also cause changes in urine potassium levels. For example, taking potassium-excreting diuretics (such as ethacrynic acid, furosemide, etc.), adrenocortical hormones (such as prednisone, etc.), amphotericin B, gentamicin, etc., can increase urine potassium. Taking spironolactone, triamterene, captopril, etc. can reduce urine potassium.

[0005] Currently, the main methods for potassium ion determination include flame photometry, ion selective electrode method, enzyme method, etc.

[0006] 1. The principle of flame photometry is: when the serum sample is transformed into fine mist by the atomization device and sent into the flame for combustion, K+ emits a spectrum of special wavelengths after obtaining energy, and the light passes through the filter and is received by the photodetector. By measuring the radiated light energy through the photoelectric system and calibrating with the known high and low values ​​of each analyte, the potassium content can be obtained. Flame photometry is widely used in clinical practice due to its small amount of blood, simple operation, accurate and reliable results, good specificity and low cost in determining serum potassium, especially in primary medical units. Its disadvantage is that the sensitivity of flame photometry is affected by the gas pressure, the pressure of the combustion-supporting gas, and the sample injection speed during the detection process. The sample must be fully mixed after adding distilled water, otherwise the analyzed sample will have a certain deviation from the actual value. In addition, the biggest drawback of flame photometry is that it uses propane and other fuels, which brings safety hazards to the laboratory.

[0007] 2. The ion selective electrode method is a quantitative analysis method based on measuring the potential of the battery. The activity of K+ causes the ion selective electrode to produce different potentials, and its activity is proportional to the ion concentration. Therefore, the ion selective electrode method can quantitatively analyze the potassium ion concentration in the specimen, and is highly selective and sensitive. This method has the characteristics of small specimen usage, fast detection, stability, accuracy and reliability, and is more suitable for small clinical specimens and emergency use. However, the ion selective electrode method requires continuous calibration, and the electrodes need to be replaced every six months, and the maintenance cost is relatively high. When testing together with other biochemical projects, the samples also need to be subsampled, which increases the workload. The analyzed data must be re-summarized and then entered into the report form, which is prone to human errors.

[0008] 3. Enzymatic method At present, it mainly refers to pyruvate kinase method, the principle of which is that phosphoenolpyruvate (PEP) and adenosine diphosphate (ADP) generate pyruvate and adenosine triphosphate (ATP) under the catalysis of potassium-dependent pyruvate kinase (PK). Then, under the catalysis of lactate dehydrogenase, the generated pyruvate reacts with NADH to generate lactate and NAD. The consumption of NADH in the reaction is proportional to the potassium ion concentration in the sample. Therefore, the potassium ion content can be calculated by monitoring the rate of decrease of absorbance at 340nm. Enzymatic determination of K+ makes large-scale fully automatic determination possible. At the same time, this method has the advantages of simple operation, wide linear range, low contamination rate, good correlation with ion-selective electrodes, accuracy, rapidity, good repeatability, and suitability for the detection of a large number of specimens. Generally, serum potassium detection can be completed on a biochemical analyzer, which can fully meet clinical needs and has good clinical application value. However, the stability of its reagents is not ideal, and it is expensive, and it also has high requirements for water quality. The enzymatic method itself is easily interfered by temperature, pH value of the matrix, jaundice, lipid turbidity, etc., especially high bilirubin and chylomicronemia, which affect the enzymatic determination. Summary of the invention

[0009] The purpose of the present invention is to overcome the defects of the prior art and to provide a potassium ion concentration measuring reagent, a measuring method and a kit for the instability of liquid reagents in the related art. The measuring method is a new enzyme colorimetric method, and potassium ion detection is performed using potassium-activated acetaldehyde dehydrogenase. By adding substances such as an activator and an enzyme stabilizer, the sensitivity and accuracy of the reaction can be quickly improved while ensuring the stability of the reagent. At the same time, the measuring reagent and the measuring method can not only be used for detection under ultraviolet light (340nm), but also for detection under visible light (405nm-680nm) by adding different substrates in a targeted manner, which greatly increases the promotion and application of the reagent and the method.

[0010] The first aspect of the present invention is to provide a potassium ion concentration measuring reagent, wherein the measuring reagent is a single agent, comprising: a buffer, a coenzyme, acetaldehyde dehydrogenase, a reaction substrate, a surfactant, a protective agent and a stabilizer; or the measuring reagent is a double agent reagent consisting of a reagent 1 and a reagent 2, wherein:

[0011] The reagent 1 comprises: a buffer, acetaldehyde dehydrogenase, a reaction substrate, a surfactant, a protective agent, a stabilizer and diaphorase or a coenzyme;

[0012] The reagent 2 includes: a buffer, a coenzyme and a diaphorase, a surfactant, a protective agent and a chromogen substrate; if the measuring reagent is a single dose, the measuring reagent also includes: a diaphorase and / or a chromogen substrate.

[0013] Preferably, the buffer is Tris-HCl buffer, and the pH value of the Tris-HCl buffer is 7-8.5.

[0014] Preferably, the coenzyme is one of NAD+ or NADP+.

[0015] Preferably, the reaction substrate is an aldehyde compound.

[0016] Preferably, the chromogen substrate is an NBT substrate or an INT substrate.

[0017] Preferably, the surfactant is a Triton series or Tween series surfactant.

[0018] Preferably, the protective agent is a mixed solution of one or more of BSA, sucrose and trehalose; and the stabilizer is a mixed solution of one or more of polyols.

[0019] The second aspect of the present invention is to provide a method for measuring potassium ion concentration, which is implemented based on the potassium ion concentration measuring reagent described in the first aspect, comprising:

[0020] S1, setting basic parameters on the fully automatic biochemical analyzer, wherein the basic parameters are measurement temperature, reaction time between the measured sample and the potassium ion concentration measuring reagent, measurement main wavelength, measurement sub-wavelength, volume ratio of the measured potassium ion sample to the reagent, reaction direction, delay time and detection time;

[0021] S2, adding the potassium ion concentration measuring reagent to the sample to be tested according to the volume ratio of the sample to the reagent, and waiting for the time corresponding to the reaction time; during the reaction, the acetaldehyde dehydrogenase activated by potassium ions converts the reaction substrate into the corresponding acid compound by catalyzing the dehydrogenation reaction of the reaction substrate, and at the same time reduces the coenzyme to a reduced coenzyme, and the generated reduced coenzyme is proportional to the concentration of the potassium ions;

[0022] S3, within the detection time, measuring the absorbance change of the reduced coenzyme at the measurement main wavelength and / or the measurement sub-wavelength, and calculating the concentration of potassium ions in the sample under test based on the absorbance change.

[0023] Preferably, the method further comprises:

[0024] S4, based on the addition of different chromogenic substrates, a reaction color change occurs within the visible light range, and the concentration range of potassium ions in the sample being tested is reflected based on the color change.

[0025] The third aspect of the present invention is to provide a kit for measuring potassium ion concentration, comprising:

[0026] A reaction device and a potassium ion concentration measuring reagent according to the first aspect, wherein the reaction device has at least one reaction hole for detecting potassium ion concentration, and the reaction hole is used to add a sample to be tested to implement the potassium ion concentration measuring method according to the second aspect.

[0027] Beneficial effects of the measuring reagent, measuring method and kit of the present invention:

[0028] (1) Compared with the pyruvate kinase method, the measuring reagent of the present invention has good stability and a positive reaction. The acetaldehyde dehydrogenase activated by potassium ions catalyzes the dehydrogenation reaction of acetaldehyde to convert acetaldehyde into acetic acid. At the same time, the coenzyme can be reduced to a reduced coenzyme. The generated reduced coenzyme is proportional to the potassium ion concentration. The potassium ion concentration can be measured by measuring the absorbance change of the reduced coenzyme at 340 nm. The measuring process is simple and easy to operate.

[0029] (2) If different chromogenic substrates are added to the entire reaction system during the measurement process, the reaction color change can occur within the visible light range, which facilitates the promotion and application of reagents and measurement methods and improves the practicality of the invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the related technologies, the drawings required for use in the specific embodiments or the related technical descriptions will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0031] Figure 1 The present invention is a flow chart of a method for measuring potassium ion concentration according to an embodiment of the present invention. DETAILED DESCRIPTION

[0032] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0033] In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, the terms "first", "second", and "third" are used for descriptive purposes only, and cannot be understood as indicating or implying relative importance.

[0034] In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances. Embodiment 1

[0035] This embodiment provides a potassium ion concentration measuring reagent, which is a single agent and includes: a buffer, a coenzyme, acetaldehyde dehydrogenase, a reaction substrate, a surfactant, a protective agent, and a stabilizer;

[0036] As a preferred embodiment, the measuring reagent further comprises: diaphorase. Diaphorase, as a hydrolase, can catalyze dehydrogenation reactions. Diaphorase is a light brown freeze-dried crystal or a fluorescent yellow-green powder, and has the following properties:

[0037] (1) Composition: Diaphorase is mainly composed of protein and is secreted by glycosyl collagenase in the human body.

[0038] (2) Reaction type: Diaphorase is a hydrolase that breaks hyaluronic acid molecules into small fragments through hydrolysis.

[0039] (3) Catalytic activity: Diaphorase can catalyze the cleavage of the ester bond between the sugar group and water molecules in hyaluronic acid, thereby degrading the hyaluronic acid molecules.

[0040] (4) Substrate specificity: Diaphorase mainly acts on hyaluronic acid, but can also act on some polysaccharides with similar structures.

[0041] As a preferred embodiment, the measuring reagent further comprises: a chromogen substrate. Embodiment 2

[0042] This embodiment provides a potassium ion concentration measuring reagent, which is a two-dose reagent consisting of reagent 1 and reagent 2, wherein:

[0043] The reagent 1 comprises: a buffer, acetaldehyde dehydrogenase, a reaction substrate, a surfactant, a protective agent, a stabilizer and diaphorase or a coenzyme;

[0044] The reagent 2 comprises: a buffer, a coenzyme and a diaphorase, a surfactant, a protective agent, and a chromogen substrate;

[0045] That is, the positions of the coenzyme and the diaphorase in the reagent 1 or the reagent 2 are not limited.

[0046] As a preferred embodiment, the buffer is Tris-HCl buffer, and the pH value of the Tris-HCl buffer is 7-8.5.

[0047] As a preferred embodiment, the coenzyme is one of NAD+ and NADP+.

[0048] As a preferred embodiment, the reaction substrate is an aldehyde compound.

[0049] In this embodiment, the reaction substrate is acetaldehyde, n-butyraldehyde, cinnamaldehyde or benzaldehyde.

[0050] As a preferred embodiment, the chromogen substrate is an NBT substrate or an INT substrate.

[0051] As a preferred embodiment, the surfactant is a Triton series or Tween series surfactant.

[0052] As a preferred embodiment, the protective agent is a mixed solution of one or more of BSA, sucrose and trehalose.

[0053] As a preferred embodiment, the stabilizer is a mixed solution of one or more polyols.

[0054] In this embodiment, the polyol is ethylene glycol or propylene glycol. Of course, those skilled in the art should know that it may also be other appropriate polyols, all of which are within the protection scope of the present invention. Embodiment 3

[0055] like Figure 1 As shown, this embodiment provides a method for measuring potassium ion concentration, which is implemented based on the potassium ion concentration measuring reagent of Example 1 and Example 2, including:

[0056] S1, setting basic parameters on the fully automatic biochemical analyzer, wherein the basic parameters are measurement temperature, reaction time between the measured sample and the potassium ion concentration measuring reagent, measurement main wavelength, measurement sub-wavelength, volume ratio of the measured potassium ion sample to the reagent, reaction direction, delay time and detection time;

[0057] In this embodiment, the basic parameters are that the measurement temperature is 37° C., the reaction time between the sample to be measured and the potassium ion concentration measurement reagent is 6 minutes, the main wavelength is 340nm-605nm, the secondary wavelength is 412nm-700nm, the volume ratio of the sample to be measured and the reagent is 20:200, the reaction direction is a positive reaction (ascending reaction), the delay time is about 1 minute, and the detection time is about 5 minutes;

[0058] S2, adding the potassium ion concentration measuring reagent to the sample to be tested according to the volume ratio of the sample to the reagent, and waiting for the time corresponding to the reaction time; during the reaction, the acetaldehyde dehydrogenase activated by potassium ions converts the reaction substrate into the corresponding acid compound by catalyzing the dehydrogenation reaction of the reaction substrate, and at the same time reduces the coenzyme to a reduced coenzyme, and the generated reduced coenzyme is proportional to the concentration of the potassium ions;

[0059] S3, measuring the absorbance change (ΔA / min) of the reduced coenzyme at the measurement main wavelength and / or the measurement sub-wavelength within the detection time, and calculating the potassium ion concentration of the measured sample based on the absorbance change.

[0060] As a preferred embodiment, the method further comprises:

[0061] S4, based on the addition of different chromogenic substrates, a reaction color change occurs within the visible light range, and the concentration range of potassium ions in the sample being tested is reflected based on the color change.

[0062] Specifically, the concentration range is characterized as high concentration, medium concentration or low concentration. Embodiment 4

[0063] This embodiment provides a kit for measuring potassium ion concentration, comprising:

[0064] A reaction device and a potassium ion concentration measuring reagent according to Example 1 or Example 2, wherein the reaction device has at least one reaction hole for detecting potassium ion concentration, and the reaction hole is used to add a sample to be tested to implement the potassium ion concentration measuring method described in Example 3.

[0065] Application Examples

[0066] Experiments show that considering the accuracy of the test results and the economic efficiency of the preparation cost, there is no

[0067] Regardless of whether it is a single dose or a double dose, the potassium ion determination kit of this application embodiment with the following composition relationship is more ideal:

[0068]

[0069] The potassium ion determination kit of this application embodiment can be a single dose, including: buffer, coenzyme, acetaldehyde dehydrogenase, reaction substrate, surfactant, protective agent, stabilizer, diaphorase, and chromogen substrate.

[0070] The above single-dose reagent can also be prepared into the following double-dose reagent:

[0071] Reagent 1: buffer, acetaldehyde dehydrogenase, reaction substrate, surfactant, protective agent, stabilizer, diaphorase.

[0072] Reagent 2: buffer, coenzyme, surfactant, protective agent, chromogen substrate.

[0073] The position of the coenzyme or diaphorase in reagent 1 or reagent 2 may not be limited.

[0074] The buffer is Tris-HCL buffer, and the pH value of the buffer is 7-8.5.

[0075] The coenzyme may be NAD + or NADP + One of a kind.

[0076] The reaction substrate can be aldehyde compounds such as acetaldehyde, n-butyraldehyde, cinnamaldehyde, benzaldehyde, etc.

[0077] The chromogen substrate may be an NBT substrate or an INT substrate, or may not be added.

[0078] The surfactant is a Triton series or Tween series surfactant.

[0079] The protective agent is one or more mixed solutions of BSA, sucrose and trehalose.

[0080] The stabilizer is a mixed solution of one or more polyols such as ethylene glycol and glycerol.

[0081] The present invention will be further described below in conjunction with implementation examples.

[0082] Application Example 1

[0083] The potassium ion determination reagent of this embodiment is a single reagent, comprising:

[0084] Table 1

[0085]

[0086] According to Table 1 above, prepare the potassium ion determination single reagent. First prepare the Tris-HCL solution with pH 8.5, then weigh each component separately and add it to the buffer solution. Mix thoroughly and set aside.

[0087] The following settings were made on the fully automatic biochemical analyzer: temperature 37°C, reaction time 6 minutes, main test wavelength 340nm, secondary test wavelength 412nm, volume ratio of potassium ion sample to reagent 20:200, reaction direction positive reaction (ascending reaction), delay time about 1 minute, and detection time about 5 minutes. By calculating the absorbance change (△A / min), the concentration of potassium ions in the sample to be tested can be calculated.

[0088] Application Example 2

[0089] The potassium ion determination reagent of this embodiment is a single reagent, comprising:

[0090] Table 2

[0091]

[0092] Prepare the potassium ion determination reagent according to Table 2 above. First prepare the Tris-HCL solution with pH 8.5, then weigh each component separately and add it to the buffer solution. Mix thoroughly and set aside.

[0093] The following settings were made on the fully automatic biochemical analyzer: temperature 37°C, reaction time 6 minutes, test main wavelength 605nm, test secondary wavelength 700nm, volume ratio of potassium ion sample to reagent 20:200, reaction direction positive reaction (ascending reaction), delay time about 1 minute, and detection time about 5 minutes. By calculating the absorbance change (△A / min), the concentration of potassium ions in the sample to be tested can be calculated.

[0094] Application Example 3

[0095] The potassium ion determination reagent of this embodiment is a double reagent, comprising:

[0096] Table 3-1

[0097] Reagent 1

[0098]

[0099] Table 3-2

[0100] Reagent 2

[0101]

[0102] Prepare potassium ion determination reagent 1 and reagent 2 according to Table 3-1 and Table 3-2 above, respectively. First prepare pH 8.5 Tris-HCL solution, then weigh each component and add it to the buffer solution, mix thoroughly and set aside.

[0103] The following settings were made on the fully automatic biochemical analyzer: temperature 37°C, reaction time 6 minutes, main test wavelength 605nm, secondary test wavelength 700nm, the volume ratio of the potassium ion sample to be tested, reagent 1 and reagent 2 was 10:100:100, the reaction direction was positive reaction (ascending reaction), the delay time was about 1 minute, and the detection time was about 5 minutes. By calculating the absorbance change (△A / min), the concentration of potassium ions in the sample to be tested can be calculated.

[0104] Application Example 4

[0105] The potassium ion determination reagent of this embodiment is a double reagent, comprising:

[0106] Table 4-1

[0107] Reagent 1

[0108]

[0109] Table 4-2

[0110] Reagent 2

[0111]

[0112] Prepare potassium ion determination reagent 1 and reagent 2 according to Table 4-1 and Table 4-2 above, respectively. First prepare pH 8.5 Tris-HCL solution, then weigh each component and add it to the buffer solution, mix thoroughly and set aside.

[0113] The following settings were made on the fully automatic biochemical analyzer: temperature 37°C, reaction time 6 minutes, main test wavelength 505nm, secondary test wavelength 700nm, the volume ratio of the potassium ion sample to be tested, reagent 1 and reagent 2 was 10:100:100, the reaction direction was positive reaction (ascending reaction), the delay time was about 1 minute, and the detection time was about 5 minutes. By calculating the absorbance change (△A / min), the concentration of potassium ions in the sample to be tested can be calculated.

[0114] Through experimental verification, other measuring methods described in the above invention contents can achieve the purpose of the present invention. Since the measuring steps and other conditions are similar to the above embodiments, no other examples are given.

[0115] Experiments have proved that the determination method of the present invention can completely obtain the required determination results through general biochemical analysis instruments: the absorbance change of the blank reagent (△A / min) ≤ 0.015; the absorbance time response curve should be an upward curve until the end point; the reagent can measure the effective (R ≥ 0.99) linear range of up to 1-50mmol / L; the accuracy of the reagent test, its relative deviation does not exceed ± 5%; the coefficient of variation (CV) of the precision (repeatability) of the reagent test ≤ 5%. The present invention has good repeatability, good accuracy, and a wide linear range, which is sufficient for easy promotion and application.

[0116] The experimental results of the blank limit test, repeatability test, linearity test and accuracy test of the test data of the third embodiment are shown in Table 5-1, Table 5-2, Table 5-3 and Table 5-4 respectively; wherein the limit of blank (LOB) refers to the maximum detection result observed for the blank sample under the specified possible conditions.

[0117] Table 5-1

[0118]

[0119] Table 5-2

[0120]

[0121] Table 5-3

[0122]

[0123] Table 5-4

[0124] .

[0125] Through the description of the above implementation methods, those skilled in the art can clearly understand that the above embodiments can be implemented by software, or by means of software plus necessary general hardware platforms. Based on such understanding, the technical solutions of the above embodiments can be embodied in the form of software products, which can be stored in a non-volatile storage medium (which can be a CD-ROM, a USB flash drive, a mobile hard disk, etc.), including several instructions for enabling a computer device (which can be a personal computer, a server, or a network device, etc.) to execute the methods described in the various embodiments of the present invention.

[0126] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A potassium ion concentration measuring reagent, characterized in that, The measuring reagent is a single agent, comprising: a buffer, a coenzyme, acetaldehyde dehydrogenase, a reaction substrate, a surfactant, a protective agent and a stabilizer; or the measuring reagent is a double agent reagent consisting of reagent 1 and reagent 2, wherein: The reagent 1 comprises: a buffer, acetaldehyde dehydrogenase, a reaction substrate, a surfactant, a protective agent, a stabilizer and diaphorase or a coenzyme; The reagent 2 comprises: a buffer, a coenzyme and a diaphorase, a surfactant, a protective agent and a chromogen substrate; When the measuring reagent is a single agent, the measuring reagent further comprises: diaphorase and / or chromogen substrate; The buffer is a Tris-HCl buffer, and the pH value of the Tris-HCl buffer is 7-8.5; The coenzyme is one of NAD+ or NADP+; The reaction substrate is an aldehyde compound; The chromogen substrate is an NBT substrate or an INT substrate; The surfactant is a Triton series or Tween series surfactant; The protective agent is a mixed solution of one or more of BSA, sucrose and trehalose; the stabilizer is a mixed solution of one or more of polyols; For the single-dose or double-dose reagent, the following composition relationship is met: The buffer is 10-500mmol / L; the coenzyme is 0.5-10mmol / L; the acetaldehyde dehydrogenase is 0.5-10mmol / L; the reaction substrate is 2-50mmol / L; the surfactant is 0.1-1%; the protective agent is 0.5-5%, and the stabilizer is 1-15%; the diaphorase is 2-40KU / L, and the chromogen substrate is 0.1-5mmol / L.

2. A method for measuring potassium ion concentration, characterized in that, The potassium ion concentration measuring reagent according to claim 1 is implemented, comprising: S1, setting basic parameters on the fully automatic biochemical analyzer, wherein the basic parameters are measurement temperature, reaction time between the measured sample and the potassium ion concentration measuring reagent, measurement main wavelength, measurement sub-wavelength, volume ratio of the measured potassium ion sample to the reagent, reaction direction, delay time and detection time; S2, adding the potassium ion concentration measuring reagent to the sample to be tested according to the volume ratio of the sample to the reagent, and waiting for the time corresponding to the reaction time; during the reaction, the acetaldehyde dehydrogenase activated by potassium ions converts the reaction substrate into the corresponding acid compound by catalyzing the dehydrogenation reaction of the reaction substrate, and at the same time reduces the coenzyme to a reduced coenzyme, and the generated reduced coenzyme is proportional to the concentration of the potassium ions; S3, measuring the absorbance change of the reduced coenzyme at the measurement main wavelength and / or the measurement sub-wavelength within the detection time, and calculating the concentration of potassium ions of the measured sample based on the absorbance change; S4, based on the addition of different chromogenic substrates, a reaction color change occurs within the visible light range, and the concentration range of potassium ions in the sample being tested is reflected based on the color change.

3. A kit for measuring potassium ion concentration, comprising: A reaction device and a potassium ion concentration measuring reagent as claimed in claim 1, wherein the reaction device has at least one reaction hole for detecting potassium ion concentration, and the reaction hole is used to add a sample to be tested to implement the potassium ion concentration measuring method as claimed in claim 2.

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