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Method for rapidly detecting steroid hormones in urea through UPLC-MS/MS technology

A steroidal, rapid technology, applied in the field of analytical chemistry and medicine, to achieve the effect of high precision, wide coverage and high sensitivity

Inactive Publication Date: 2016-12-07
WUXI NO 4 PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In summary, there is no report on the application of UPLC-MS / MS technology at home and abroad to rapidly (enzymolysis time less than 7h) detect 5 kinds of urosteroid hormones at the same time

Method used

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  • Method for rapidly detecting steroid hormones in urea through UPLC-MS/MS technology
  • Method for rapidly detecting steroid hormones in urea through UPLC-MS/MS technology
  • Method for rapidly detecting steroid hormones in urea through UPLC-MS/MS technology

Examples

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Embodiment 1

[0042]Take a mixed methanol solution of five steroid hormones (estriol, hydrocortisone, 17β-estradiol, testosterone and progesterone) frozen and thawed at -20°C, with a concentration of 100 ng / mL. Take 500 μL of urine, add 10 μL of testosterone-d3 as an internal standard, 1.0 mL of acetic acid-sodium acetate buffer, 1800-2200 U β-glucuronidase aqueous solution, and pass through the SPE cartridge in a water bath at 50-60 °C for 3 hours. The specific process of the SPE cartridge is as follows: 1) The SPE cartridge is first equilibrated with 10 mL of methanol; 2) Then washed with 10 mL of deionized water; 3) Sample loading: the urine sample after enzymatic hydrolysis is loaded onto the SPE cartridge; 4) Rinse: wash once with 2 mL of 20% methanol aqueous solution under positive pressure; 5) Elution: use 5 mL of 100% methanol solution under positive pressure to elute twice. Nitrogen blow-drying temperature is 60 ℃, time is about 15min. The residue was vortex-dissolved with 200 μL ...

Embodiment 2

[0044] Take 500 μL of female urine that was frozen and thawed at -20°C, add 10 μL of testosterone-d3 as an internal standard, 1.0 mL of acetic acid-sodium acetate buffer, and 1800-2200 U β-glucuronidase aqueous solution. SPE cartridges. The specific process of the SPE cartridge is as follows: 1) The SPE cartridge is first equilibrated with 10 mL of methanol; 2) Then washed with 10 mL of deionized water; 3) Sample loading: the urine sample after enzymatic hydrolysis is loaded onto the SPE cartridge; 4) Rinse: wash once with 2 mL of 20% methanol aqueous solution under positive pressure; 5) Elution: use 5 mL of 100% methanol solution under positive pressure to elute twice. Nitrogen blow-drying temperature is 60 ℃, time is about 15min. The residue was vortex-dissolved with 200 μL of 30% acetonitrile-water solution, filtered through a 0.22 μm filter membrane, and waited for UPLC-MS / MS injection analysis. The results are attached figure 2 as shown in b. Five steroid hormones ca...

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Abstract

The invention relates to a method for rapidly detecting steroid hormones in urea through an UPLC-MS / MS technology, and belongs to the field of analysis chemistry and medical science. The concentrations of estriol, hydrocortisone, 17beta-estradiol, testosterone and progesterone in human urea are simultaneously detected through the UPLC-MS / MS technology by adopting a beta-glucuronidase enzymatic hydrolysis technology, the detection limit is 0.1-1.0 ng / mL, and the quantitation limit is 0.3-3.0 ng / mL. The method is fast, sensitive and accurate, can meet requirement of simultaneous detection of above five steroid hormones in urea, and has wide application prospect.

Description

technical field [0001] The invention relates to a method for rapidly detecting urinary steroid hormones by UPLC-MS / MS (ultra-high performance liquid chromatography-tandem mass spectrometry), belonging to the fields of analytical chemistry and medicine. Background technique [0002] Blood steroid hormones such as estriol, hydrocortisone, 17β-estradiol, testosterone and progesterone are common clinical detection indicators for the diagnosis and treatment of many human endocrine diseases. Currently, the detection methods include immunoassay and chromatography-mass spectrometry Wait. Immunoassay methods such as immunofluorescence analysis are currently widely used in clinical practice, and can simultaneously measure multiple steroid hormones in human blood, but they are extremely susceptible to background interference and cross-reactions, and often only detect free hormones, etc., so sometimes they cannot truly reflect the in vivo Actual level; chromatographic mass spectrometry...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/062
Inventor 徐庄剑马亚萍夏冰洋
Owner WUXI NO 4 PEOPLES HOSPITAL
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