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Method for detecting concentrations of vitamin A and vitamin E in human dried blood spots

A detection method and vitamin technology, which can be applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of affecting quantitative accuracy, unstable samples of detection methods, high possibility of pathogenicity and occupational exposure, and improve accuracy. and sensitivity, reducing the risk of pathogenicity and occupational exposure, ensuring stable results

Pending Publication Date: 2022-05-24
WUXI DIAGNOSTICS LAB SHANGHAI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, in the traditional vitamin A and vitamin E concentration detection methods, serum is often used as the source of calibration curves and quality control samples, and the background in serum is large, which is likely to affect the accuracy of quantification. Therefore, a sufficiently accurate sampling volume is required. It is necessary to ensure that the amount of blood collected is large enough; in addition, the samples of traditional detection methods are unstable and require cold chain transportation, and the possibility of pathogenicity and occupational exposure is high

Method used

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  • Method for detecting concentrations of vitamin A and vitamin E in human dried blood spots
  • Method for detecting concentrations of vitamin A and vitamin E in human dried blood spots
  • Method for detecting concentrations of vitamin A and vitamin E in human dried blood spots

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Preparation of calibrators, quality controls and sample extracts

[0042] In a specific embodiment, the specific operations for the preparation of the calibrator, the quality control product and the sample extract are as follows:

[0043] (1) Preparation of standard solutions

[0044]Accurately weigh 25 mg of vitamin VA standard, dissolve it in methanol (0.3% BHT), and prepare a stock solution with a concentration of 2.5 mg / mL, designated as SSC-VA.

[0045] Accurately weigh 100 mg of vitamin VE standard, dissolve it in methanol (0.3% BHT), and prepare a stock solution with a concentration of 10 mg / mL, designated as SSC-VE.

[0046] (2) Preparation of sample extract

[0047] Preparation of VA-D6 stock solution: take 1 mg of vitamin VA-D6 standard, dissolve it with methanol (0.3% BHT), and prepare a stock solution with a concentration of 0.1 mg / mL, numbered as SSC-VA-D6. The preparation batch number is the date of the day, and the prepared stock solution is ...

Embodiment 2

[0058] Example 2 Sample Detection

[0059] 1. Sample pretreatment:

[0060] Pre-treatment of blank samples, the specific operation steps are as follows:

[0061] 1) Add 300 μL of methanol:water (v:v 70:30) solution to the double blank sample, shake and mix at 1500 rpm for 10 min.

[0062] 2) Centrifuge at 12000rpm for 5min.

[0063] 3) Take 150 μL of supernatant to a new 96-well plate.

[0064] 4) Inject 15 μL in the UPLC-MS / MS system.

[0065] The specific steps are as follows:

[0066] 1) Add 250 μL of the sample extract to the sample to be tested, then add the internal standard solution, make up the total volume to 300 μL, and shake and mix at 1500 rpm for 10 min.

[0067] 2) Centrifuge at 12000rpm for 5min.

[0068] 3) Take 150 μL of supernatant to a new 96-well plate.

[0069] 4) Inject 15 μL in the UPLC-MS / MS system.

[0070] 2. Quantitative analysis and detection by ultra-high performance liquid chromatography tandem mass spectrometry UPLC-MS / MS

[0071] Set th...

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Abstract

The invention discloses a method for detecting the concentration of vitamin A and vitamin E in human dried blood spots. The method comprises the following steps: 1) preparing a calibrator; (2) preparing a quality control product, wherein the quality control product comprises a VA group quality control product and a VB group quality control product; (3) respectively preparing the prepared calibrator and the prepared quality control product into dried blood spot sample cards, and setting two blank control sample cards; (4) preparing a sample extracting solution, wherein the components of the sample extracting solution comprise vitamin VA-D6, vitamin VE-D6 and methanol; (5) adding the prepared calibrator, the quality control product, the sample to be detected and the sample extracting solution into a pretreatment container for pretreatment for sample loading analysis for later use; (6) carrying out pretreatment on the blank sample; and 7) carrying out chromatography-mass spectrometry detection on the sample to be detected by using an ultra-high performance liquid chromatography-tandem mass spectrometry UPLC-MS / MS system and adopting an internal and external standard combination method. The detection method provided by the invention has the advantages of short analysis time, accurate detection degree, simple pretreatment, low detection cost and the like.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to a method for detecting the concentrations of vitamin A and vitamin E in human dried blood spots. Background technique [0002] Vitamin A plays a vital role in retinal function (adaptation to low light), is required for epithelial tissue growth and differentiation, and is required for bone growth, reproduction and embryonic development, along with certain carotenoids, Vitamin A enhances immune function and reduces the risk of certain infectious diseases. Degenerative changes in the eyes and skin are often observed in vitamin A deficiency. Vision maladaptation to the dark (night blindness) is an early symptom and can also be accompanied by degenerative changes in the retina. Vitamin A deficiency is the leading cause of blindness in developing countries. Severe or long-term deficiency can lead to dry eyes (dry eye syndrome), which can lead to corneal ulcers, scarring, ...

Claims

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Application Information

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IPC IPC(8): G01N30/88G01N30/06
CPCG01N30/88G01N30/06G01N2030/8813G01N2030/045G01N2030/047
Inventor 方焯刘釜均耿志华徐鸣成
Owner WUXI DIAGNOSTICS LAB SHANGHAI CO LTD
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