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

A diagnostic kit and potassium ion technology are applied in the field of medical examination and determination, which can solve the problems that a biochemical analyzer cannot measure potassium ion and sodium ion content at the same time, cannot be popularized and applied, and is complicated to operate, so as to achieve a complete three-dimensional spatial structure, The effect of low cost and simple method of testing

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

AI Technical Summary

Benefits of technology

In this patented technology, we use an improved way called enzymology that uses certain compounds found naturally within foods like chickpeas (chlorophyll), lithocholic acid, ellagitannins). These natural ingredients have unique properties making them ideal materials for testing purposes. They work very effectively when mixed together properly without causing any unwanted side products during processing. Overall, these techniques provide precise measurements while minimizing environmental concerns associated with traditional methods.

Problems solved by technology

Technological Problem addressed in this patents relates to accurately identifying low levels of calcium from unknown sources within biologically relevant tissue samples without causing errors associated with current analyte measurement techniques like radioactive scintigraphy and X ray absorption measurements. Current solutions require expensive equipment while maintain accuracy over longer periods. Chemistry tests provide better results compared to liquid chromatography and gas phase diffusion coefficients. However these chemistries involve complex operations and often result in interferences between various components making their interpretation difficult.

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment one (single agent)

[0051] Prepare the potassium ion diagnostic kit according to the following ingredients and dosage:

[0052] Glycine buffer 80mmol / l,

[0053] Adenosine diphosphate 2mmol / l,

[0054] Phosphoenolpyruvate 1mmol / l,

[0055] thio-NADH 0.2mmol / l,

[0056] Pyruvate kinase 6000U / l,

[0057] Lactate dehydrogenase 10000U / l,

[0058] Ethylene glycol 50% (accounting for the total volume of the reagent).

[0059] 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 negative reaction (absorbance decreases, the same below).

[0060] Add the sample and the prepared single agent, and the two are automatically mixed inside the analyzer, and the decrease of the absorbance at 340nm is detected and recorded. According to the method o...

Embodiment 2

[0061] Embodiment two (two doses)

[0062] Prepare the potassium ion diagnostic kit according to the following ingredients and dosage:

[0063] Reagent I——

[0064] Imidazole ~ hydrochloric acid buffer 100mmol / l,

[0065] NADPH 0.25mmol / l,

[0066] Lactate dehydrogenase 20000U / l,

[0067] Glycerol 50% (accounting for the total volume of reagent I);

[0068] Reagent II——

[0069] Imidazole ~ hydrochloric acid buffer 100mmol / l,

[0070] Adenosine diphosphate 5mmol / l,

[0071] Phosphoenolpyruvate 3mmol / l,

[0072] Pyruvate kinase 10000U / l,

[0073] Ethylene glycol 50% (accounting for the total volume of reagent II).

[0074] 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 1:25, the ratio of the amount of reagent I to reagent II is 4:1, a...

Embodiment 3

[0076] Embodiment three (three doses)

[0077] Prepare the potassium ion diagnostic kit according to the following ingredients and dosage:

[0078] Reagent I——

[0079] Triethanolamine buffer 120mmol / l,

[0080] thio-NADPH 0.3mmol / l,

[0081] Lactate dehydrogenase 30000U / l,

[0082] Propylene glycol 50% (accounting for reagent I total volume);

[0083] Reagent II——

[0084] Triethanolamine buffer 120mmol / l,

[0085] Adenosine diphosphate 8mmol / l,

[0086] Phosphoenolpyruvate 5mmol / l,

[0087] Propylene glycol 20mmol / l;

[0088] Reagent III——

[0089] Triethanolamine buffer 120mmol / l,

[0090] Pyruvate kinase 12000U / l,

[0091] Ethylene glycol 50% (accounting for the total volume of reagent III).

[0092]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 ion sample and reagent I, reagent II and reagent III The ratio of the tot...

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Abstract

The invention relates to a method to measure potassium ion content by enzyme method and potassium ion diagnosing kit. The adenosine diphosphate and phosphoric acid enol pyruvic acid take reaction to form into pyruvic acid by utilizing the characteristic of potassium ion activate pyruvate kinase, and pyruvic acid would take reaction with lactate dehydrogenase to form into lactic acid, and at the same time the reduced coenzyme would oxygenated into oxygenated type coenzyme, by measuring the rate of decent of dominant wavelength at 340nm absorbance to quantitative response the content of potassium ion in sample. The invention has high specificity, and has no impact from the sodion in the sample.

Description

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Claims

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Application Information

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Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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