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Chemical Derivatization-Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry for the Simultaneous Detection of 18 Steroid Hormones in Serum

一种超高效液相、类固醇激素的技术，应用在化学衍生-超高效液相色谱-串联质谱法领域，能够解决不能有效提高ESI-MS检测灵敏度、激素定量覆盖率低、反应活性低等问题，达到良好离子化效率、缩短检测时间、增大检测通量的效果

Active Publication Date: 2020-10-30
SPECTRUM (WUHAN) MEDICAL BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, sulfonyl chloride derivatization reagents can only derivatize steroid hormones containing phenolic hydroxyl groups, and their quantitative coverage of hormones is low; - After MS / MS detection, the lowest average detection limit can reach the level of 10pg / mL
In the above-mentioned derivatization methods, the derivatization reaction of hydroxylamine solution is a derivatization reagent that can simultaneously derivatize estrogen, androgen, corticosteroids and progesterone. However, although conventional hydroxylamine reagents can selectively react with the carbonyl group of steroid hormones , but its reactivity is low, it is difficult to completely label with the 20-position carbonyl steroid hormone, and hydroxylamine does not contain a proton affinity part (proton affinity 193.8kcal / mol), which cannot effectively improve the detection sensitivity of ESI-MS; and Benzoic acid reagents need activators, such as 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) and imidazole, etc., and can only react with the labeling reagent after activating the target, and the side reaction Many, poor specificity

Method used

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  • Chemical Derivatization-Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry for the Simultaneous Detection of 18 Steroid Hormones in Serum
  • Chemical Derivatization-Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry for the Simultaneous Detection of 18 Steroid Hormones in Serum
  • Chemical Derivatization-Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry for the Simultaneous Detection of 18 Steroid Hormones in Serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] The chemical derivation-ultra high performance liquid chromatography-tandem mass spectrometry method provided in this example simultaneously detects 18 steroid hormones, including 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 C...

Embodiment 2

[0100] The chemical derivation-ultra high performance liquid chromatography-tandem mass spectrometry method provided in this example simultaneously detects 18 steroid hormones, the steps of which are the same as those in Example 1, the only difference being:

[0101] The mobile phase gradient elution parameters are:

[0102]

[0103] Compared with Example 1 in Example 2, the MRM mass spectrometry response signals and sensitivities of the 18 steroid hormones decreased, specifically, the sensitivity (based on the lowest detection limit of quantification) decreased by 8.5-15.0%.

experiment example

[0108] The sensitivity comparative analysis of the detection methods of 18 steroid hormones in Example 1 and Comparative Example 1 was carried out.

[0109] The results of the sensitivity comparison experiment are shown in Table 3.

[0110] Table 3 Sensitivity comparison experiment results

[0111]

[0112] Comparative analysis of the sensitivity comparison experiment results of Example 1 and Comparative Example 1 in Table 1 shows that the sensitivity of chemical derivation-ultra high performance liquid chromatography-tandem mass spectrometry in Example 1 of the application to detect 18 steroid hormones is significantly improved. (Based on the lowest detection limit of quantitation) increased by 2-454 times.

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Abstract

The invention provides a chemical derivatization-ultra-high performance liquid chromatography-tandem mass spectrometry method for simultaneously detecting 18 steroid hormones in serum, which is characterized in that a novel derivatization reagent 3-pyridylmethoxylamine is adopted for derivatization and then ultra-high performance liquid chromatography-tandem mass spectrometry is used for detection. According to the method, only 10 minutes are needed for detecting 18 steroid hormones at the same time, the hydrophobicity index logP is greatly improved after derivatization, the peak shape and theseparation degree in a high performance liquid phase reversed phase chromatographic column are improved, and the ionization efficiency is better, so that the mass spectrum detection sensitivity is improved. The method has the characteristics of simplicity and convenience in operation, good accuracy, strong specificity, wide detection range and the like.

Description

technical field [0001] The invention relates to the technical field of medical analysis and detection, more specifically, to a chemical derivation-ultra-high performance liquid chromatography-tandem mass spectrometry method for simultaneously detecting 18 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 hormones play an important role in enhancing immune function, sodium, salt, water balance, inflammation and sexual characteristics. The level of human endocrine hormones plays an important role in disease diagnosis and disease assessment, such as adrenal hyperplasia, polycystic ovary syndrome, and adrenal insufficiency. [0003] However, steroid hormones exert strong physiological effects by binding to receptors in the human body usually at very low concentrations at nanomolar or picomolar levels. Due to the extremely...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/009G01N2030/045G01N2030/8813
Inventor 王献肖华明刘杨茜
Owner SPECTRUM (WUHAN) MEDICAL BIOTECHNOLOGY CO LTD