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Detection kit and detection method of vitamin B12 and sample pretreatment method of vitamin B12 detection

A detection method, B12 technology, applied in biological testing, preparation of test samples, material inspection products, etc., can solve the problems of different stability, deviation of test results, etc.

Active Publication Date: 2017-09-29
SHENZHEN MINDRAY BIO MEDICAL ELECTRONICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the vitamin B12 released from the binding protein has various forms, the different R groups will lead to different stability. When the above analysis method is used for detection, the detection results may be different due to the different stability of vitamin B12. deviation

Method used

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  • Detection kit and detection method of vitamin B12 and sample pretreatment method of vitamin B12 detection
  • Detection kit and detection method of vitamin B12 and sample pretreatment method of vitamin B12 detection
  • Detection kit and detection method of vitamin B12 and sample pretreatment method of vitamin B12 detection

Examples

Experimental program
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Embodiment 1

[0059] A sample pretreatment method for vitamin B12 detection, comprising the following steps:

[0060] Take 50 μl of the serum sample to be tested, add 50 μl of vitamin B12 pretreatment agent, the pretreatment agent is sodium hydroxide concentration of 0.5mol / L and K 3 Co(CN) 6 Concentration is the aqueous solution of 1.5g / L (namely every milliliter of sample solution to be tested adds 1.5mg complex), and DTT concentration is the sodium citrate buffer solution of 1.028g / L, react at 37 ℃ for 12.5min, make it The vitamin B12 is released from the binding protein, and with K 3 Co(CN) 6 The traces of cyanide ions dissociated in the solution combine to form stable cyanocobalamin for determination, such as figure 2 Shown; K 3 Co(CN) 6 The overall stability constant β of the cyanide ion in aqueous solution is 10 42 .

[0061] A vitamin B12 detection method. After the serum sample to be tested is processed by the pretreatment method of this embodiment, the content of cyanocoba...

Embodiment 2

[0094] A sample pretreatment method for vitamin B12 detection is basically the same as the method in Example 1, except that:

[0095] In this embodiment, hexacyanoferrate (potassium hexacyanoferrate) and hexacyanoferrate (potassium hexacyanoferrate) are respectively used as complexes to pretreat the sample, wherein, hexacyanoferrate The addition of potassium hexacyanoferrate is 50 μl of potassium hexacyanoferrate aqueous solution of 5 g / L (that is, every milliliter of the sample solution to be tested adds 5 mg of the complex), and the addition of potassium hexacyanoferrite is 3 g / L. 50 μl of an aqueous hexacyanoferrous salt solution at a concentration of L (that is, 3 mg of the complex is added to each milliliter of the sample solution to be tested). The overall stability constant β of potassium hexacyanoferrate in aqueous solution is 10 42 , the total stability constant β of potassium hexacyanoferrite in aqueous solution is 10 35 .

[0096] A method for detecting vitamin B...

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Abstract

The invention relates to a detection method of vitamin B12 and a sample pretreatment method of vitamin B12, and belongs to the technical field of detection of biological samples. The pretreatment method comprises the following steps of obtaining a to-be-detected sample solution, and adding a protein releasing agent and a complexing agent, wherein the protein releasing agent is used for releasing the vitamin B12 of a to-be-detected sample from binding protein; the complexing agent is a complex with cyanide ions, and the total stability constant beta of the complex in a water solution is 1010 to 1045; dissociating the cyanide ions from the complex in the solution, binding the cyanide ions and the released vitamin B12, and forming stable cyanocobalamine for measuring. The detection method and sample pretreatment method have the advantages that the hypertoxic cyanide is replaced with the low-toxic complex with the cyanide ions, the dissociated trace cyanide ions can react with the vitamin B12 in the to-be-detected sample, and the stable cyanocobalamine is produced; the requirement of sample treatment is met, the sample can be fully treated, and the larger toxicity is not produced.

Description

technical field [0001] The invention relates to the technical field of biological sample detection, in particular to a vitamin B12 detection kit and method and a sample pretreatment method thereof. Background technique [0002] Vitamin B12, also known as cobalamin, is the only vitamin containing metal elements. Its main physiological function is to participate in the manufacture of bone marrow red blood cells, prevent pernicious anemia, and prevent brain nerves from being damaged. Vitamin B12 is the latest to be discovered among the B vitamins so far. [0003] The structure of vitamin B12 is shown below: [0004] [0005] Its characteristics are: 1. The molecular center is a trivalent cobalt ion; 2. It has a corrin ring (a structure similar to a porphyrin ring), and the 4 N atoms of the corrin ring form 4 ligands with the trivalent cobalt ion. 3. An N atom of the dimethyl indazole structure forms a coordination bond with a trivalent cobalt ion; 4. The indazole and the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/82G01N1/28
CPCG01N1/28G01N33/82
Inventor 何飞唐涛张裕平黄佳俊刘挺李可
Owner SHENZHEN MINDRAY BIO MEDICAL ELECTRONICS CO LTD
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