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Liquid chromatography-mass spectrometry tandem detection method for detecting 25 (OH) D2 and 25 (OH) D3 in serum

A technology of liquid chromatography mass spectrometry and detection methods, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of affecting the service life of chromatographic columns and mass spectrometer instruments, using a large amount of organic reagents, and harming the environment by personnel, so as to reduce personnel Effect of injury and environmental pollution, operator health and environmental friendliness, high sensitivity

Pending Publication Date: 2022-06-28
NAT INST OF METROLOGY CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Usually, derivatization takes a long time, and nitrogen drying requires a large amount of organic reagents, causing personal injury and environmental pollution.
Fatty impurities are not removed, affecting the service life of chromatographic columns and mass spectrometers

Method used

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  • Liquid chromatography-mass spectrometry tandem detection method for detecting 25 (OH) D2 and 25 (OH) D3 in serum
  • Liquid chromatography-mass spectrometry tandem detection method for detecting 25 (OH) D2 and 25 (OH) D3 in serum
  • Liquid chromatography-mass spectrometry tandem detection method for detecting 25 (OH) D2 and 25 (OH) D3 in serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] This example provides detection of 25(OH)D in serum 2 and 25(OH)D 3 The liquid chromatography-mass spectrometry tandem detection method comprises the following steps:

[0052] 1. Materials and Reagents

[0053]

[0054] 2. Instruments and equipment

[0055]

[0056] 3. Method:

[0057] (1) Chromatographic conditions:

[0058] The chromatographic column is HSS T3 (1.8μm, 2.1×100mm), the mobile phase A is 0.1% formic acid-water solution (ultrapure water), and B is 0.1% formic acid methanol solution; the mobile phase is 0.4mL / min, and the column temperature is 40℃ , gradient elution;

[0059] The mobile phase gradient elution parameters are:

[0060]

[0061] (2) Mass spectrometry conditions:

[0062] The ionization mode is positive ion mode, and the detection adopts the mass spectrometry scanning mode of multiple reaction monitoring. The ionization source is an electrospray ionization source (ESI), wherein the curtain gas (CUR) flow is 20 psi, the collisi...

Embodiment 2

[0082] The detection of 25(OH)D in serum provided in this example 2 and 25(OH)D 3 The liquid chromatography mass spectrometry tandem detection method, the steps are the same as those of Example 1, the difference is only:

[0083] The chromatographic column is BEH C18 (1.8 μm, 2.1×100 mm), the mobile phase A is 0.1% formic acid-aqueous solution (ultra pure water), and B is 0.1% formic acid methanol solution.

[0084] The detection of the two hydroxyvitamins was completed, and the measured sensitivity was reduced by 30%, and the resolution was similar to that of Example 1.

Embodiment 3

[0095] The detection of 25(OH)D in serum provided in this example 2 and 25(OH)D 3 The liquid chromatography mass spectrometry tandem detection method, the steps are the same as those of Example 1, the difference is only:

[0096] Prepare pre-activated lipid dispersion solid phase extraction packing (Bond Elut EMR-Lipid) EP tubes: Pre-weigh different masses of Bond Elut EMR-Lipid packings in EP tubes, respectively without adding packing, weighing 50mg, 100mg and Add 200 mg of filler, add 100 μL of pure water, and vortex for 5 seconds.

[0097] The degree of purification of the samples by different fillers was evaluated by the matrix effect.

[0098] The formula for calculating the matrix effect is:

[0099]

[0100] The specific results are figure 2 shown in (a) and (b). The none in the figure represents no filler, and 50mg, 100mg, and 200mg represent weighing 50mg, 100mg, and 200mg, respectively. 25(OH)D 2 Matrix effect reduced from 61.3% to 17.2%, 25(OH)D 3 The matr...

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Abstract

The invention relates to a liquid chromatography-mass spectrometry tandem detection method for detecting 25 (OH) D2 and 25 (OH) D3 in serum, which comprises the following steps: mixing a serum sample to be detected with an internal standard solution, carrying out lipid removal dispersive solid-phase extraction pretreatment to obtain a detection solution, and detecting the detection solution by adopting an ultra-high performance liquid chromatography-mass spectrometry tandem technology, the method disclosed by the invention is simple to operate, low in time consumption, high in sensitivity, high in precision, low in use amount of organic reagents, friendly to health of operators and environment and wide in practical application prospect.

Description

technical field [0001] The present invention relates to the technical field of medical analysis and detection, and more particularly, to a method for detecting 25(OH)D in serum 2 and 25(OH)D 3 A liquid chromatography-mass spectrometry tandem detection method. Background technique [0002] Vitamin D (VD) is an essential nutrient element for the human body. With the deepening of people's awareness of the importance of vitamin D and health, the demand for accurate, rapid and efficient determination of vitamin D levels has increased rapidly. 25-hydroxyvitamin D (25(OH)D) is the main form of vitamin D in the human body. Because of its long half-life and stability, it is usually used as an index to evaluate the level of vitamin D. According to the detection principle, the commonly used methods for the determination of serum 25(OH)D are divided into immunoassay and chromatography. Immunoassay is based on the principle of antigen-antibody binding reaction. Different types of 25(O...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/72G01N30/06G01N30/14G01N30/34G01N30/60G01N30/86
CPCG01N30/02G01N30/7266G01N30/06G01N30/14G01N30/6052G01N30/34G01N30/86G01N2030/027G01N2030/045G01N2030/047G01N2030/065G01N2030/143
Inventor 乔晓婷谢洁易可可江游黄泽建刘梅英戴新华方向
Owner NAT INST OF METROLOGY CHINA
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