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Method for pretreating fat-soluble vitamin detection sample based on automatic pipetting workstation

A technology of fat-soluble vitamins and workstations, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., and can solve a large number of problems, such as manual operations

Pending Publication Date: 2021-09-14
杭州凯莱谱精准医疗检测技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In order to solve the above problems, the present invention provides a method for pretreatment of samples requiring fat-soluble vitamin detection through an automated liquid pipetting workstation. Through the new application of the automated liquid pipetting workstation, and selection of suitable operating modes and program parameters, Realized the high-throughput, automation and low cost of liquid-liquid extraction of fat-soluble vitamin samples that had been difficult to achieve in the past, solved the problem of a large number of manual operations for liquid-liquid extraction, and can perfectly replace expensive SLE suitable for high-throughput detection Plate extraction method; at the same time, by choosing more suitable extractant, diluent and reconstitution solution, the recovery rate of vitamin K1, which has the lowest content in human serum, is significantly improved, making the subsequent detection results of high performance liquid chromatography tandem mass spectrometry more accurate and more sensitive. High, so that the ability to detect vitamin K1 in blood samples can meet the actual needs of the clinical reference interval, and can be used to more accurately judge whether the normal content of vitamin K1 in the human body is deficient

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  • Method for pretreating fat-soluble vitamin detection sample based on automatic pipetting workstation
  • Method for pretreating fat-soluble vitamin detection sample based on automatic pipetting workstation
  • Method for pretreating fat-soluble vitamin detection sample based on automatic pipetting workstation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1 Sample preparation, pretreatment and detection

[0065] The following preparations are based on the use of solid standard products. If a commercially purchased standard stock solution is used, the preparation method needs to be adjusted according to the actual concentration of the standard stock solution used.

[0066] 1. Sample preparation

[0067] 1. Preparation of stock solution

[0068] 0.1mg / mL BHT methanol solution Preparation: Accurately weigh 5 mg of 2,6-di-tert-butyl-p-cresol BHT and dissolve it in 50 mL of methanol to obtain a BHT methanol solution with a concentration of 0.1 mg / mL.

[0069] primary stock solution The preparation of vitamin A (VA), 25 hydroxyvitamin D3 (VD3), 25 hydroxyvitamin D2 (VD2), vitamin E (VE), vitamin K1 (VK) standard products were accurately weighed, and an appropriate volume of BHT methanol solution was added to obtain Primary stock solution, see Table 1 for details;

[0070] Table 1 Preparation of primary stock sol...

Embodiment 2

[0160] The impact of different extractants of embodiment 2 on detection recovery

[0161] After completing the sample preparation according to the steps in Example 1, take 100 μL of a standard product with a vitamin K1 concentration of 0.1 ng / ml to carry out sample pretreatment according to the sample pretreatment steps in Example 1, and divide it into four parts according to the different extractants taken therein. group, the volume ratio of the extractant to the serum sample was 25:10, after the pretreatment was completed, the liquid chromatography tandem mass spectrometry conditions in Example 1 were used for detection, and the effect of using different extractants on the recovery rate of vitamin K1 in the plasma sample was investigated. The results are shown in Table 24.

[0162] Table 24 The impact of different extractants on the recovery rate of vitamin K1 detection

[0163]

[0164] It can be seen from Table 24 that there are obvious differences in the extraction re...

Embodiment 3

[0167] In the extraction process of embodiment 3, the influence of the mixing volume of the automatic pipetting workstation on the detection recovery rate

[0168] This example adopts the method of sample preparation and pretreatment as provided in Example 1, wherein during the pretreatment process, the parameters related to the "mixing mode" during extraction are set to, and the mixing volume is respectively 0.3mL, 0.6mL and 0.9mL After the pretreatment is completed, the liquid chromatography tandem mass spectrometry conditions in Example 1 are used for detection, and the influence of different mixing volumes on the recovery rate of vitamin K1 in the plasma sample during the extraction process is investigated. The results are shown in Table 25.

[0169] The impact of different mixing volumes during the extraction of table 25 on the detection recovery of vitamin K1

[0170]

[0171] It can be seen from Table 25 that during the extraction process, the automatic pipetting wor...

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Abstract

The invention provides a method for pretreating a fat-soluble vitamin detection sample based on an automatic pipetting workstation. The method is mainly used for detecting fat-soluble vitamins in the sample by high performance liquid chromatography-tandem mass spectrometry, and through a brand new application of the automatic pipetting workstation, proper operation modes and program parameters are selected, a high flux, automation and low cost of liquid-liquid extraction of the fat-soluble vitamin sample are realized, the problem that a large amount of manual operation is needed by liquid-liquid extraction is solved, and an expensive SLE plate extraction method suitable for high flux detection can be perfectly replaced; a mixture of isooctane and isopropanol is used as an extracting agent, isopropanol is used as a dissolving solution, methanol and water are used as a resolvent, and meanwhile, the proportional relation is further optimized so that the recovery rate of the vitamin K1 in a sample is obviously improved, and the subsequent high performance liquid chromatography-tandem mass spectrometry detection result is more accurate and higher in sensitivity; and the detection capability of K1 in the sample can meet the actual demand of a clinical reference interval.

Description

technical field [0001] The invention belongs to the field of vitamin detection, and in particular relates to a sample pretreatment method for fat-soluble vitamin detection, in particular to a pretreatment method for serum samples requiring fat-soluble vitamin detection through an automated liquid transfer workstation. Background technique [0002] Fat-soluble vitamins play an important role in human health, especially the growth and development of children. Vitamin A deficiency can lead to dry eye syndrome and night blindness. Studies have shown that there is a clear correlation between vitamin A levels and cancer risk; the main physiological role of vitamin D It is to maintain the balance of calcium and phosphorus, thus promoting bone health. In addition, studies have shown that vitamin D levels are significantly related to immune system function and the incidence of various diseases, such as the occurrence of tuberculosis, heart disease and endocrine diseases. Vitamin E, ...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88
Inventor 富士山卫晓乐曲亮马金飞刘华芬
Owner 杭州凯莱谱精准医疗检测技术有限公司
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