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Method and kit for simultaneously detecting 11 vitamins

A technology of vitamins and kits, applied in the field of vitamin detection, can solve the problems of high cost, low efficiency, and failure to meet the needs of multi-vitamins, and achieve the effect of reducing detection costs and improving detection efficiency

Pending Publication Date: 2022-03-04
BGI CLINICAL LAB (SHENZHEN) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, a more comprehensive understanding of the content of water-soluble vitamins is required, and multiple tests must be carried out, which is time-consuming, inefficient, and expensive
Although liquid chromatography tandem mass spectrometry can theoretically detect multiple water-soluble vitamins at the same time, the number of vitamins that can be mixed and detected in practical applications is limited, and multiple detections are still required to obtain more complete vitamin content information, which is not very good. Meet the needs of multi-vitamin simultaneous detection

Method used

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  • Method and kit for simultaneously detecting 11 vitamins
  • Method and kit for simultaneously detecting 11 vitamins
  • Method and kit for simultaneously detecting 11 vitamins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] In this example, 11 water-soluble ingredients containing thiamine, riboflavin, niacin, niacinamide, pantothenic acid, pyridoxic acid, pyridoxine, biotin, 5-methyltetrahydrofolate, cobalamin and vitamin C were used. A mixed solution of vitamins was tested as a standard. The details of the 11 water-soluble vitamins are shown in Table 3.

[0048] In this example, the method for simultaneously detecting 11 kinds of vitamins includes, when adopting liquid chromatography tandem mass spectrometry (abbreviated LC-MS / MS) to detect the sample to be tested, using an improved aqueous phase and an organic phase; wherein, the aqueous phase consists of a concentration of 0.1% Formic acid with a concentration of 2.0-5.0mM ammonium acetate and water; the organic phase is composed of formic acid with a concentration of 0.1%, ammonium acetate with a concentration of 2.0-5.0mM and an organic solvent; the organic solvent is methanol or acetonitrile.

[0049] The water-soluble vitamin detec...

Embodiment 2

[0065] In this example, on the basis of Example 1, add 30 μL of precipitant to 150 μL of the standard product for protein precipitation and water-soluble vitamin extraction, shake for 5 minutes, then centrifuge at 4000 g for 30 minutes at 4 ° C, take the supernatant, and then 4 ° C, 4000 rpm After centrifugation for 1 min, the supernatant was taken for detection by the same liquid chromatography tandem mass spectrometry as in Example 1, that is, pretreatment of the sample to be tested.

[0066] The precipitant in this example is composed of 10% sulfosalicylic acid and acetonitrile. This example analyzes and compares the influence of different volume ratios of 10% sulfosalicylic acid and acetonitrile on the precipitation and extraction effect of the target test substance, as follows:

[0067] Precipitant 1: 10% sulfosalicylic acid-acetonitrile solution with a volume ratio of 8:2;

[0068] Precipitating agent 2: 10% sulfosalicylic acid-acetonitrile solution with a volume ratio o...

Embodiment 3

[0076] In this example, on the basis of Example 1, the impact of adding different concentration ratios of ammonium acetate in the mobile phase on the target detection substance is analyzed and compared, as follows:

[0077] Mobile phase 1: water phase: ultrapure water + 0.1% formic acid, organic phase: acetonitrile + 0.1% formic acid;

[0078] Mobile phase 2: water phase: ultrapure water + 0.1% formic acid + 2.0mM ammonium acetate, organic phase: acetonitrile + 0.1% formic acid + 2.0mM ammonium acetate;

[0079] Mobile phase 3: aqueous phase: ultrapure water + 0.1% formic acid + 10.0 mM ammonium acetate, organic phase: acetonitrile + 0.1% formic acid + 10.0 mM ammonium acetate.

[0080] In this example, the mobile phases with the above three ratios were used to test the same parameters for the same samples, and some results are shown in Table 6. Table 6 only shows the results of detection of abnormal water-soluble vitamins.

[0081] Table 6 The influence of different mobile ...

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Abstract

The invention discloses a method and a kit for simultaneously detecting 11 vitamins. According to the method for simultaneously detecting 11 vitamins, the 11 vitamins comprise thiamine, riboflavin, nicotinic acid, nicotinamide, pantothenic acid, pyridoxine, pyridoxine, biotin, 5-methyltetrahydrofolic acid, cobalamin and vitamin C; according to the method, an improved water phase and an improved organic phase are used during liquid chromatography-tandem mass spectrometry detection; the water phase consists of formic acid with the concentration of 0.1%, ammonium acetate with the concentration of 2-5mM and water; and the organic phase consists of formic acid with the concentration of 0.1%, ammonium acetate with the concentration of 2-5mM and an organic solvent. According to the method disclosed by the invention, the water phase and the organic phase detected by the liquid chromatography-tandem mass spectrometry are improved, so that the 11 water-soluble vitamins have good separation degree and peak pattern in chromatographic behavior, the 11 water-soluble vitamins are simultaneously detected, the detection efficiency of the water-soluble vitamins is improved, and the detection cost is reduced.

Description

technical field [0001] The application relates to the field of vitamin detection, in particular to a method and a kit for simultaneously detecting 11 kinds of vitamins. Background technique [0002] Vitamins are a class of trace organic substances that participate in the metabolism of the body or regulate energy conversion in the form of coenzymes or prosthetic groups of biological enzymes. According to the difference in solubility, vitamins can be roughly divided into fat-soluble vitamins and water-soluble vitamins. Water-soluble vitamins include vitamin B complex, lipoic acid and vitamin C, mainly thiamine (VB1), riboflavin (VB2), niacin and niacinamide, pyridoxic acid (VB6PA), folic acid, pantothenic acid, biotin, cobalamin and vitamin C etc. [0003] Although the human body has very little demand for vitamins, vitamins cannot be synthesized in the body or the amount of synthesis is insufficient, so they must be supplied by food. Among the common human diseases, most o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/06
CPCG01N30/88G01N30/06
Inventor 邱雪梅林清娜訾金林梁魏媛仪吴志峰石一云
Owner BGI CLINICAL LAB (SHENZHEN) CO LTD
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