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Process for determining contect of potassium ion by enzyme method and kit for diagnosing potassium ion thereof

A diagnostic kit and potassium ion technology, applied in the field of medical testing and determination, can solve the problems that cannot be popularized and applied, and the biochemical analyzer cannot simultaneously measure potassium ion and sodium ion content, etc.

Inactive Publication Date: 2006-06-14
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because there are holes in the macrocyclic structure, the oxygen atoms in the molecule have unshared electron pairs that can be combined with metal ions. According to the size of the holes, metal ions of different diameters can be selectively combined to achieve the purpose of measuring the ion concentration (determination Blood potassium, generally using common crown ether anionic dyes for colorimetric quantification); atomic spectrophotometry can also be used to detect K in serum + 、Na + , but the operation is complicated, the error is large, and it is not as simple as the flame photometry
[0007] The method for measuring potassium ion content by enzymatic method has good consistency with flame photometer method or ion selective electrode method. This method has strong anti-interference and good stability, but it cannot simultaneously measure potassium ion and The respective content of sodium ions makes this method unable to be popularized and applied.

Method used

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  • Process for determining contect of potassium ion by enzyme method and kit for diagnosing potassium ion thereof
  • Process for determining contect of potassium ion by enzyme method and kit for diagnosing potassium ion thereof
  • Process for determining contect of potassium ion by enzyme method and kit for diagnosing potassium ion thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment one (single agent)

[0057] Prepare the potassium ion diagnostic kit according to the following ingredients and dosage:

[0058] Glycine buffer 80mmol / l,

[0059] Adenosine diphosphate 3mmol / l,

[0060] Phosphoenolpyruvate 2mmol / l,

[0061] Glucose 5mmol / l,

[0062] thio-NADH 1mmol / l,

[0063] Pyruvate kinase 6000U / l,

[0064] Hexokinase 10000U / l,

[0065] Glucose phosphate dehydrogenase 10000U / l,

[0066] Ethylene glycol 50% (accounting for the total volume of the reagent).

[0067] Set on the automatic biochemical analyzer (Hitachi-7080): temperature 37°C, reaction time 10 minutes, test main wavelength 340nm, test secondary wavelength 405nm or more, the volume ratio of the measured potassium ion sample to the reagent is 1:25, The reaction direction is positive reaction (absorbance rises, the same below).

[0068] Add the sample and the prepared single agent, and the two are automatically mixed inside the analyzer to detect and record the increase o...

Embodiment 2

[0069] Embodiment two (two doses)

[0070] Prepare the potassium ion diagnostic kit according to the following ingredients and dosage:

[0071] Reagent I——

[0072] Imidazole ~ hydrochloric acid buffer 100mmol / l,

[0073] Adenosine diphosphate 5mmol / l,

[0074] Phosphoenolpyruvate 3mmol / l,

[0075] Glucose 12mmol / l,

[0076] NADPH 2mmol / l,

[0077] Hexokinase 15000U / l,

[0078] Glucose phosphate dehydrogenase 15000U / l,

[0079] Glycerol 50% (accounting for the total volume of reagent I);

[0080] Reagent II——

[0081] Imidazole ~ hydrochloric acid buffer 100mmol / l,

[0082] Pyruvate kinase 9000U / l,

[0083] Ethylene glycol 50% (accounting for the total volume of reagent II).

[0084] Set on the automatic biochemical analyzer (Hitachi-7080): temperature 30°C, reaction time 15 minutes, test main wavelength 340nm, test secondary wavelength 405nm or more, the ratio between the total volume of the measured potassium ion sample and reagent I and reagent II The ratio is 1...

Embodiment 3

[0086] Embodiment three (three doses)

[0087] Prepare the potassium ion diagnostic kit according to the following ingredients and dosage:

[0088] Reagent I——

[0089] Triethanolamine buffer 120mmol / l,

[0090] Adenosine diphosphate 7mmol / l,

[0091] Phosphoenolpyruvate 4mmol / l,

[0092] Glucose 20mmol / l,

[0093] thio-NADPH 3mmol / l,

[0094] Propylene glycol 20mmol / l;

[0095] Reagent II——

[0096] Triethanolamine buffer 120mmol / l,

[0097] Hexokinase 20000U / l,

[0098] Glucose phosphate dehydrogenase 20000U / l,

[0099] Propylene glycol 50% (accounting for the total volume of reagent II);

[0100] Reagent III——

[0101] Triethanolamine buffer 120mmol / l,

[0102] Pyruvate kinase 12000U / l,

[0103] Ethylene glycol 50% (accounting for the total volume of reagent III).

[0104] Set on the automatic biochemical analyzer (Hitachi-7080): temperature 25°C, reaction time 20 minutes, test main wavelength 340nm, test secondary wavelength 405nm or more, the tested potassium ...

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Abstract

The invention relates to potassium ion content measuring method by enzyme method and potassium ion diagnosis kit. It is utilized potassium ion to activate pyruvate kinase to make adenosine diphosphate react with phosphoric acid enol pyruvic acid to generate adenosine triphosphate, couple hexokinase glucose phosphate dehydrogenase to react, and reduce oxidized coenzyme to reduced coenzyme, and quantitatively response potassium ion content by measuring main wavelength 340nm absorbance vertical speed. The method is not influence for sodium ion in sample, and not polluted by internal and foreign matter. And it has high specificity, precise testing result, and good accuracy. The diagnosis kit is formed to double or three reagents to reduce cross influence, keep reagent stability to be convenient for storing for long time. The method can realize fast testing by common ultraviolet / visible light analyzer or semi / full automatic biochemical analyzer. Thus its testing cost is low; and it is convenient for popularizing and applying in industry.

Description

technical field [0001] The invention relates to a method for measuring potassium ion content in samples such as serum and plasma by enzymatic method, and a potassium ion diagnostic kit prepared by applying the method, which belongs to the technical field of medical inspection and determination. Background technique [0002] The content of potassium ions in serum (referred to as "serum potassium") has very important clinical significance. Under normal circumstances, there is a reasonable reference value range for blood potassium in the human body. Hypokalemia occurs when serum potassium falls below the lower reference limit due to renal tubular disease, hyperaldosteronism, hyperinsulinemia, metabolic alkalosis, or the use of insulin for diabetic ketosis, use of diuretics Potassium ions are transferred to the inside of cells during the treatment of certain diseases; when blood potassium is higher than the upper limit of the reference value, it may be caused by glomerular disea...

Claims

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Application Information

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IPC IPC(8): G01N21/75G01N21/31
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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