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Method for simultaneously detecting multiple fat-soluble vitamins

A fat-soluble vitamin and vitamin technology, which can be used in measuring devices, instruments, scientific instruments, etc., can solve problems such as difficulties, large limitations, and increase testing costs, and achieve the goal of reducing testing costs, improving compliance, and reducing sample blood volume. Effect

Pending Publication Date: 2020-11-27
大连和合医学检验实验室有限公司
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Problems solved by technology

At present, with the development of liquid chromatography and mass spectrometry, methods for detecting vitamins based on liquid chromatography and liquid chromatography-mass spectrometry have emerged. The Chinese patent, publication number: CN110763788A, discloses a method for quantitatively detecting five lipids in plasma. Kits and methods for soluble vitamins, however, the required blood collection volume is as high as 0.25mL, which is still difficult in the clinical blood collection process, especially for people with poor compliance (infants and young children), and SLE extraction is used in the pretreatment process board, significantly increasing the cost of detection; Chinese patent, publication number: CN110927310A, discloses a method for simultaneously detecting the content of 25 hydroxy-vitamin D3 and 25 hydroxy-vitamin D2 in trace blood, this method can only detect vitamin D, the limitation Big

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  • Method for simultaneously detecting multiple fat-soluble vitamins
  • Method for simultaneously detecting multiple fat-soluble vitamins
  • Method for simultaneously detecting multiple fat-soluble vitamins

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Experimental program
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Effect test

Embodiment 1

[0029] Step 1: Preparation of standard solution: preparation containing retinol (VA), 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, α-tocopherol (VE) and VK1, VK2 concentrations were 1000, 40, 160 . Take 200 μL of the solution to 2# tube, mix well, then pipette 200 μL to 3#, mix well until the 9# tube is completely mixed, and get 9 standard mixed solutions with different concentrations.

[0030] Step 2: Preparation of internal standard solution: preparation containing d3-retinol, d6-α tocopherol, d3-25-hydroxyvitamin D2, d6-25-hydroxyvitamin D3, d7-VK1, d7-K1, d7-K2 Mixed internal standard solutions with concentrations of 3750, 5000, 25, 25, 833, and 208 ng / mL respectively, and the solvent is chromatographic ethanol.

[0031] Step 3: Pretreatment: Use a pipette gun to pipette 20 μL of 1#-9# standard solution into a 2.0mL plastic centrifuge tube, add 20 μL of internal standard solution, 180 μL of protein precipitation solution (methanol: acetonitrile = 1:1, v / v), vortexed at 20...

Embodiment 2

[0054] Step 1: Collection of blood samples: Collect venous blood from 64 healthy adults using yellow cap blood collection tubes with gel-promoting gel. The blood is centrifuged at 4000rpm for 10 minutes to obtain serum. Mix the sera of 64 persons evenly to obtain mixed serum and store in separate packages. at -80°C. Take another 50mL of mixed serum, add 32.5, 32.5, 10.35, 20.5, 0.5, 0.5μL of vitamin A, E, D2, D3, K1, K2 solutions with concentrations of 1000, 20000, 100, 100, 10, 10ug / mL ( That is, 0.65, 13, 0.0207, 0.04106, 0.0001, 0.0001ug of vitamins A, E, D2, D3, K1, K2) were added to each 1mL of mixed serum, and mixed thoroughly to obtain standard supplemented serum, which was stored in -80 ℃.

[0055] Step 2: Preparation of internal standard solution: preparation containing d3-retinol, d6-α tocopherol, d3-25-hydroxyvitamin D2, d6-25-hydroxyvitamin D3, d7-VK1, d7-K1, d7-K2 Mixed internal standard solutions with concentrations of 3750, 5000, 25, 25, 833, and 208 ng / mL res...

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Abstract

The invention discloses the technical field of fat-soluble vitamin detection and particularly relates to a method for simultaneously detecting multiple fat-soluble vitamins. The detection method comprises the following steps: step 1, fully and uniformly mixing 10-200 microlitres of sample plasma with a protein precipitation solution and a vitamin deuterated internal standard substance at 100-3000rpm for 1-5 min to obtain a solution A, and centrifuging the solution A at 0-8 DEG C at 200-12000 rpm to obtain a supernatant; and step 2, analyzing the fat-soluble vitamin of the supernatant by usinga liquid chromatograph-mass spectrometer, and calculating the content of the fat-soluble vitamin in the sample plasma by using an internal standard method. Compared with the prior art, the method hasthe advantages that more types of vitamins are detected, the sample can be enriched without the assistance of SLE, SPE and other materials, the detection cost is effectively reduced, the required blood volume is less than that of the existing method, the compliance of patients, especially infants, is better due to the reduction of the blood volume of the sample, and the method has a wide application prospect.

Description

technical field [0001] The invention relates to the technical field of detection of fat-soluble vitamins, and specifically relates to a method for simultaneously detecting six kinds of fat-soluble vitamins in blood plasma (clear). Background technique [0002] Vitamins, divided into fat-soluble vitamins and water-soluble vitamins, are a type of organic substances necessary to maintain human life activities, and are also important active substances to maintain human health. Vitamin deficiency or excess will lead to the occurrence of nutritional diseases; [0003] Vitamin A has various physiological functions such as promoting growth and reproduction, maintaining bone, epithelial tissue, vision and normal secretion of mucosal epithelium; vitamin D, also known as anti-rickets vitamin, is a sterol derivative, and the most important member of its family members is 25 hydroxy-vitamin D3 (cholecalciferol) and 25 hydroxy-vitamin D2 (ergocalciferol), both are fat-soluble vitamins, w...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/34G01N30/32G01N30/16
CPCG01N30/02G01N30/06G01N30/34G01N30/32G01N30/16G01N2030/045G01N2030/062G01N2030/324
Inventor 晏嘉泽李世阳赵旭生徐恒宇隋文雯刘丽华刘韵姚玉涵万媛林琳
Owner 大连和合医学检验实验室有限公司
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