南方医科大学学报

南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (2): 301-315.doi: 10.12122/j.issn.1673-4254.2026.02.08

• • 上一篇    

基于液相色谱-质谱串联法技术的32种类固醇激素的检测方法建立及其在妊娠全周期变化应用

周祺淇(), 陈君贺, 聂城涛, 王前, 夏方博(), 周宏伟()   

  1. 南方医科大学珠江医院检验医学部，广东 广州 510280
  • 收稿日期:2025-10-21 出版日期:2026-02-20 发布日期:2026-03-10
  • 通讯作者: 夏方博,周宏伟 E-mail:zqq19950928@126.com;xiafb08@163.com;hzhou@smu.edu.cn
  • 作者简介:周祺淇，主管技师，在读硕士研究生，E-mail: zqq19950928@126.com
  • 基金资助:
    国家自然科学基金(82102501)

Development of an LC-MS/MS method for 32 steroid hormones and exploration of their gestational variations

Qiqi ZHOU(), Junhe CHEN, Chengtao NIE, Qian WANG, Fangbo XIA(), Hongwei ZHOU()   

  1. Department of Laboratory Medicine, Zhujiang Hospital of Southern Medical University, Guangzhou 510280, China
  • Received:2025-10-21 Online:2026-02-20 Published:2026-03-10
  • Contact: Fangbo XIA, Hongwei ZHOU E-mail:zqq19950928@126.com;xiafb08@163.com;hzhou@smu.edu.cn
  • Supported by:
    National Science Foundation of China(82102501)

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摘要：

目的 建立一种人血清中类固醇激素的高灵敏度、高特异性液相色谱-质谱串联法（LC-MS/MS）分析方法。 方法 采用固相萃取（SPE）法进行样本前处理，优化LC-MS/MS确保痕量激素的准确检测。经验证方法的线性范围、正确度、精密度等指标后，将该方法应用于正常妊娠人群血清样本的检测，分析孕早、中、晚期血清中32种类固醇激素的动态变化规律。 结果 线性范围为0.001~7500 ng/mL，检出限（LOD）为0.0007 ng/mL，日内和日间变异系数（CV）均小于10%，加标回收率在为86.21%~112.67%，方法的萃取效率为85.05%~106.3%，基质效应为85.60%~113.22%。将该方法应用于妊娠期类固醇激素动态分析发现，多数激素（如11-脱氧皮质醇、11β-羟孕酮、硫酸脱氢表雄酮、雌三醇等）水平随妊娠进展显著上升（P<0.05），提示其与胎儿-胎盘功能发育密切相关；部分激素在孕晚期升高（P<0.05），可能与母体适应性调节及胎儿肾上腺功能成熟有关。 结论 本研究基于LC-MS/MS技术建立了32种类固醇激素高灵敏度检测方法，揭示了妊娠期间类固醇激素的动态变化规律，为相关生理与病理机制研究提供了基础。

关键词: 类固醇激素, 液相色谱-串联质谱技术, 固相萃取

Abstract:

Objective To establish a highly sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based analytical method for detecting steroid hormones in human serum. Methods Solid-phase extraction (SPE) was used for sample pretreatment. Chromatographic conditions were optimized to achieve baseline separation, and mass spectrometry parameters were adjusted to ensure accurate detection of trace amounts of hormones. After validation of the linear range, accuracy, and precision, the method was applied in detection of serum samples from normal pregnant women for analyzing dynamic variation characteristics of 32 steroid hormones during the first, second, and third trimesters of pregnancy. Results The linear range of the method was 0.001-7500 ng/mL, with a limit of detection (LOD) of 0.0007 ng/mL. Both intra-day and inter-day coefficients of variation (CV) of the method were less than 10%, the spiked recovery ranged from 86.21% to 112.67%, the extraction efficiency was 85.05%-106.3%, and the matrix effects were 85.60%-113.22%. The results of detection using this method for serum steroid hormones during pregnancy revealed significant elevation of most of the hormones (such as 11-deoxycortisol, 11β-hydroxyprogesterone, dehydroepiandrosterone sulfate, and estriol) with the progression of pregnancy, indicating a close correlation with the development of fetal-placental function. Some hormones increased in the third trimester, possibly due to maternal adaptive regulation and maturation of fetal adrenal function. Conclusion The detection results using the method established in this study reveal dynamic variations of steroid hormones during pregnancy, which provides clues for further physiological and pathological studies of these hormones.

Key words: steroid hormones, liquid chromatography-tandem mass spectrometry, solid-phase extraction