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Citation: TONG Hongbin, TONG Haijiang, LU Dezhao. Monitoring of dynamic changes of 15 steroids in serum by liquid chromatography-tandem mass spectrometry[J]. Chinese Journal of Chromatography, ;2017, 35(4): 421-426. doi: 10.3724/SP.J.1123.2016.09015 shu

Monitoring of dynamic changes of 15 steroids in serum by liquid chromatography-tandem mass spectrometry

1.
Zhejiang Chinese Medical University, Hangzhou 310053, China;
2.
Department of Clinical Laboratory, the Second Hospital of Shaoxing, Shaoxing 312000, China

Received Date: 5 September 2016

Fund Project: Project of Medical and Health Technology Program of Zhejiang Province (No. 2014KYB285)

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the determination of 15 steroids. Acetonitrile containing internal standard was added to the serum samples to precipitate the proteins. Then the derivatization of the samples was conducted with 100 mmol/L hydroxylamine hydrochloride. The separation of the samples was performed on an Agilent-C18 column (50 mm×3.0 mm, 2.7 μm), and quantified with internal standard method. The steroids were determined by multiple-reaction monitoring (MRM) with electrospray ionization (positive mode) MS. The 13 steroids showed linear relationships from 0.05 to 20 ng/mL (R²≥0.9956) with limits of detection no more than 1 ng/mL; cortisol and dehydroepiandrosterone sulfate showed linear relationships from 50 to 2000 ng/mL (R²≥0.9979) with limits of detection less than 0.5 ng/mL. The method was used for the monitoring of the dynamic changes of steroids in a menstrual cycle of women. The 15 steroids fluctuated with different properties, in which androgens reached the peak in late follicular phase, progesterone (estrogens) reached the peak in luteal phase, while the changes of cortical steroids were not so evident. The method is sensitive and repeatable. The data in this project can provide some references for clinical practice.
- liquid chromatography-tandem mass spectrometry (LC-MS/MS),
- steroids,
- menstrual cycle
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