Kit for detecting folic acids in blood spots through high performance liquid chromatography-tandem mass spectrometry

A high-performance liquid chromatography and technical detection technology, applied in the field of quantitative detection of folic acid, can solve the problems of complex matrix of blood spot samples

Inactive Publication Date: 2019-08-20
上海可力梅塔生物医药科技有限公司 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the complex matrix of blood spot samples, it needs to be extracted for detection

Method used

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  • Kit for detecting folic acids in blood spots through high performance liquid chromatography-tandem mass spectrometry
  • Kit for detecting folic acids in blood spots through high performance liquid chromatography-tandem mass spectrometry
  • Kit for detecting folic acids in blood spots through high performance liquid chromatography-tandem mass spectrometry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] The components of the analysis kit are shown in Table 5.

[0085] Table 5 Preparation of Analysis Kit Components

[0086]

[0087]

[0088] The preparation method of the above-mentioned kit is as follows:

[0089] (1) Standard product:

[0090] a. Wash the rabbit erythrocytes 3 times with normal saline, and set aside;

[0091] b. Prepare 50 mg / mL human serum albumin solution containing 100 μg / mL combined antioxidants:

[0092] Weigh 2 g of human serum albumin powder, add 40 mL of normal saline with 100 μg / mL combined with antioxidants, mix well and prepare a human serum albumin solution with a concentration of 50 mg / mL;

[0093] c. Fully mix rabbit red blood cells and human serum albumin solution according to the volume ratio of 55:45 to make a matrix;

[0094] d. Prepare 100 μg / mL combined antioxidant aqueous solution;

[0095] Stock solution configuration: Weigh 50 mg of folic acid and 50 mg of 5-methyltetrahydrofolate, respectively add 1 L of 100 μg / mL joi...

Embodiment 2

[0110] 1. Sample collection

[0111] (1) Because food affects the measurement results, the subjects need to take samples on an empty stomach, and the blood spot collectors wash their hands and wear sterile gloves;

[0112] (2) The blood was collected and dropped on the filter paper made of folic acid, so that the blood naturally penetrated to the back of the filter paper, avoiding repeated bleeding, and at least 3 blood spots were collected;

[0113] (3) Place the blood spots in the air and let them dry naturally until dark brown, avoiding sunlight and ultraviolet radiation, baking, volatile chemical substances and other pollution;

[0114] (4) All blood spots should be treated as specimens of blood-borne infectious diseases, and specimens of special infectious diseases, such as AIDS, should be indicated and packaged separately;

[0115] (5) Qualified dried blood spots should be: at least 3 blood spots, and the diameter of each blood spot is greater than 8 mm; the blood dropl...

Embodiment 3

[0134] 1. Method verification

[0135] (1) Ion flow chromatograms of the standard substance and isotopic internal standard of folic acid and 5-methyltetrahydrofolate

[0136] figure 1 The chromatograms of 5-methyltetrahydrofolate standard, 5-methyltetrahydrofolate-d3 isotope chromatogram, the chromatogram of folic acid standard, folic acid- 13 C5 isotope internal standard chromatogram, these four substances can meet the detection and quantification requirements of LC-MS / MS.

[0137] (2) Standard curve

[0138] Using the isotope internal standard quantification method, draw the standard curve with the 3 concentrations of the 2 standard substances as the abscissa (x), and the ratio of the peak area of ​​each concentration sample of the 2 standard substances to the internal standard as the ordinate (y). , and calculate the content of the above-mentioned folic acid and 5-methyltetrahydrofolate. The linear fitting equations of folic acid and 5-methyltetrahydrofolate in their r...

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Abstract

The invention discloses a kit for detecting folic acids in blood spots through a high performance liquid chromatography-tandem mass spectrometry. The kit comprises a standard product, a quality control product, a mixed internal standard solution, an eluent A, an eluent B, an extract, an extract additive, a combined antioxidant and a complex solution. The folic acids comprise a folic acid and 5-methyltetrahydrofolate. According to the invention, the kit is used for detection, which has the advantages of high sensitivity, high specificity, accuracy, simple pretreatment method, short time and greatly saved time cost; and the precision and recovery rate basically meet requirements.

Description

technical field [0001] The invention relates to quantitative detection of folic acid, a method for detecting folic acid in human blood spots by liquid chromatography-tandem mass spectrometry, and a finished kit for detecting folic acid in dried blood spots. Background technique [0002] Folic acid, also known as vitamin B9, is a water-soluble vitamin. In fact, it is one of the complexes of vitamin B, which is equivalent to pteroyl glutamic acid. Epidemiological and experimental studies have shown that folic acid deficiency and metabolic disorders seriously affect human health and are the cause of coexistence of various diseases, such as megaloblastic anemia, leukopenia, cardiovascular disease, congenital heart disease and neural tube defects, etc. . In addition, folic acid is especially important for pregnant women, and it is a consensus that pregnant women need to supplement folic acid. If folic acid is lacking in the first 3 months of pregnancy, it can lead to fetal neur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
CPCG01N30/02G01N2030/027
Inventor 朱监宝黄泽博官培龙于嘉屏
Owner 上海可力梅塔生物医药科技有限公司
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