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Method for detection of 25-hydroxyvitamin D in serum

A hydroxyvitamin and detection method technology, which is applied in the field of detection of 25-hydroxyvitamin D in serum, can solve the problems of high operator requirements, long processing time, and high reagent cost, and achieve improved detection throughput, short processing time, and shortened The effect of detection time

Active Publication Date: 2017-03-22
GUANGZHOU BIOHOP TECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these two methods have the following problems: method 1: complex pretreatment, long processing time (about 2 hours for one sample), low throughput, high reagent cost, and high requirements for operators; method 2: large investment in instruments, and The cost of consumables for online extraction is high

Method used

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  • Method for detection of 25-hydroxyvitamin D in serum
  • Method for detection of 25-hydroxyvitamin D in serum
  • Method for detection of 25-hydroxyvitamin D in serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The detection method of 25-hydroxyvitamin D of the present embodiment comprises the following steps:

[0037] 1. Sample pretreatment

[0038] Pour 200 μL of human serum sample into a clean centrifuge tube, add 400 μL of acetonitrile solution containing internal standards (112 μg / L 6d-25 hydroxyvitamin D3 and 50 μg / L 3d-25 hydroxyvitamin D2), vortex and mix for 60 seconds , in a centrifuge at 12000r / min, centrifuged at 4°C for 5min, take 200μL of the supernatant into a brown sampling bottle, then add 400μL of acetonitrile, vortex and mix for 60s, the sample to be tested is obtained, and loaded into the liquid chromatography in the autosampler.

[0039] 2. Enrichment, separation and detection

[0040] The autosampler automatically loads 50 μL of the sample to be tested into the two-dimensional liquid chromatography system, and the sample to be tested is enriched, separated and detected by two-dimensional liquid chromatography in series with quadrupole mass spectrometer ...

Embodiment 2

[0065] Embodiment 2 Methodological verification experiment

[0066] In this example, a methodological verification experiment is carried out on the detection method of 25-hydroxyvitamin D in Example 1.

[0067] 1. Precision experiment

[0068] 1.1 Intra-assay precision (Intra-assay)

[0069] a. The actual serum samples were spiked to low, medium and high levels respectively, and the obtained samples were subjected to intra-batch precision experiments;

[0070] b. 20 samples of each concentration level were processed in parallel, and each sample was injected once;

[0071] c. Calculate the average value and relative standard deviation (RSD) of the test results, and the RSD is required to be less than 20%;

[0072] d. Tables 5 and 6 are the intra-assay precision data, and the intra-assay precision is less than 5%, meeting the requirements.

[0073] Table 5 Intra-assay precision test results of 25-hydroxyvitamin D2

[0074]

[0075]

[0076] Table 6 25-hydroxyvitamin D...

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Abstract

The invention relates to a method for detection of 25-hydroxyvitamin D in serum. The method comprises sample pretreatment: adding an acetonitrile solution containing a 25-hydroxyvitamin D internal standard substance into a serum sample, carrying out protein precipitation, carrying out centrifugation, taking the supernatant and diluting the supernatant through acetonitrile to obtain a sample to be detected, wherein a volume ratio of the serum sample to the acetonitrile solution is 1: 1-4 and a volume ratio of the supernatant to acetonitrile is 1: 1-4, and enrichment, separation and detection: carrying out enrichment, separation and detection on the sample to be detected through a two-dimensional liquid chromatography-tandem quadrupole mass spectrometer. The method is simple, greatly shortens the detection time, improves the detection flux of the sample and has a high detection precision, a low cost, specificity and strong matrix interference resistance.

Description

technical field [0001] The invention relates to the technical field of analysis and detection, in particular to a method for detecting 25-hydroxyvitamin D in serum. Background technique [0002] Vitamin D is a fat-soluble vitamin, a sterol derivative, and a compound necessary to maintain human health. Vitamin D includes 5 compounds, and the ones closely related to health are vitamin D2 and vitamin D3. Vitamin D2 and vitamin D3 are oxidized by monooxygenase (25-hydroxyl oxidase) in the liver to form 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3. The concentration of 25-hydroxyvitamin D in the blood is high and the circulation period is long, so it becomes The concentration indicator of vitamin D in the human body is an evaluation index of the nutritional status of vitamin D in the human body. [0003] Vitamin D3 can be formed by the activation of animal provitamin (7-dehydrocholesterol) by ultraviolet 270-300nm, and vitamin D2 is formed by the activation of plant vitamin (e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/88
CPCG01N30/02G01N30/88G01N2030/8822
Inventor 何翠红李玉冬段倩倩梁田熊晓明
Owner GUANGZHOU BIOHOP TECH INC
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