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Method for simultaneously detecting 13 kinds of steroid hormones in serum

A steroid hormone and serum technology, applied in the field of medical analysis and detection, can solve the problems of long derivatization time, time-consuming on-line enrichment and purification, interference of side reaction products, etc., and achieve shortened detection time, low detection limit, increased The effect of large detection throughput

Inactive Publication Date: 2019-08-30
SOUTH CENTRAL UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This pretreatment method is complex in operation, takes a long time for derivatization, has interference from side reaction products, and requires a large investment in equipment. Online enrichment and purification are time-consuming and consumable, and the cost is high.

Method used

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  • Method for simultaneously detecting 13 kinds of steroid hormones in serum
  • Method for simultaneously detecting 13 kinds of steroid hormones in serum
  • Method for simultaneously detecting 13 kinds of steroid hormones in serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] The method for the simultaneous detection of 13 steroid hormones by ultra-high performance liquid chromatography-tandem mass spectrometry provided in this example comprises the following steps:

[0068] 1. Materials

[0069] The experimental samples of the methodological study came from the serum samples of Wuhan Kangshengda Medical Laboratory Co., Ltd. in June 2012.

[0070] 2. Instruments and reagent consumables

[0071] Instruments: Shimadzu 8050 triple quadrupole mass spectrometer, Shimadzu 30A ultra-high performance liquid chromatograph, ultrapure water instrument (Millipore ultrapure water machine), electronic analytical balance (FA1004, Shanghai Sunny Hengping Scientific Instrument Co., Ltd.), High-speed refrigerated centrifuge (GL-20M high-speed refrigerated centrifuge), vortex mixer (US SCILOGEX company), GM-12 water bath nitrogen blowing instrument (Beijing Chengmeng Weiye Technology Co., Ltd.), pipette gun (Dalong Xingchuang Experimental Instrument Co., Ltd...

Embodiment 2

[0102] The ultra-high performance liquid chromatography-tandem mass spectrometry method provided in this example simultaneously detects 13 kinds of steroid hormones, and its steps are the same as those in Example 1, the only difference being:

[0103] The mobile phase gradient elution parameters are:

[0104]

[0105] Compared with Example 1, the MRM mass spectrometry response signal and sensitivity of Example 2 decreased, and the sensitivity (based on the lowest detection limit of quantification) decreased by 15%.

Embodiment 3

[0107] The ultra-high performance liquid chromatography-tandem mass spectrometry method provided in this example simultaneously detects 13 kinds of steroid hormones, and its steps are the same as those in Example 1, the only difference being that the Shimadzu 8050 triple quadrupole mass spectrometer is used When performing mass spectrometry detection, the installation position of the electrospray ion source nozzle is the factory setting (position 0), instead of the +3 position after adjustment and installation in Example 1.

[0108] Compared with Example 1, the MRM mass spectrometry response signal intensity in Example 3 is reduced by 35-45%, thereby reducing its detection sensitivity and further affecting the lowest detection limit of steroid hormones.

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Abstract

The invention provides a method for simultaneously detecting 13 kinds of steroid hormones in serum. The method includes the following steps: a to-be-tested serum sample is mixed with an internal standard solution, a detection liquid is obtained by liquid-liquid extraction, and detection is performed by ultra-high-performance liquid chromatography-tandem mass spectrometry; liquid chromatography conditions are that: a mobile phase A is a formic acid aqueous solution of 0.1%, and a mobile phase B is a formic acid methanol solution of 0.1%; mass spectrometry conditions are that: a positive ion mode employs a multi-reaction monitoring mass spectrometry scanning mode; and the 13 kinds of steroid hormones include pregnenolone, progesterone, 17-hydroxypregnenolone, 17[alpha]-hydroxyprogesterone, 11[alpha]-hydroxyprogesterone, 21-deoxycortisol, 11-deoxycorticosterone, corticosterone, cortisol, 11-deoxycortisol, cortisone, aldosterone, and estrone. The method simultaneously detects the 13 kindsof steroid hormones with the detection time of only 6 minutes, and is easy to operate and takes less time; the sensitivity is high, the accuracy is good, the specificity is strong, a detection range is wide, and an application prospect in medical and biochemical detection fields is broad.

Description

technical field [0001] The invention relates to the technical field of medical analysis and detection, in particular to a method for simultaneously detecting 13 steroid hormones in serum. Background technique [0002] The endocrine system directly controls important physiological processes in humans, such as growth, development, reproduction, and metabolism. Steroid hormone drugs are a large family in the endocrine system. Specifically, they refer to hormone drugs with steroidal structure in their molecular structure. They are also a clinically important class of drugs, mainly including adrenocortical hormones and sex hormones. Steroid hormones are synthesized from the backbone of cholesterol and converted into different types of steroids by various enzymes. They are all a class of tetracyclic aliphatic hydrocarbon compounds with cyclopentane polyhydrophenanthrene nuclei, usually in the number and position of carbonyl and hydroxyl functional groups Subtle changes in their s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/009G01N2030/045G01N2030/8813
Inventor 王献刘琪刘杨茜
Owner SOUTH CENTRAL UNIVERSITY FOR NATIONALITIES
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