Evaluation of the performance of equilibrium dialysis combined with liquid chromatography-tandem mass spectrometry for the detection of free testosterone
10.3760/cma.j.cn114452-20241015-00562
- VernacularTitle:平衡透析联合液相色谱-串联质谱检测游离睾酮的性能评价
- Author:
Peng LIU
1
;
Zhaozhao WU
;
Ping HE
;
Qi YU
;
Jingran ZHEN
;
Wei QIAN
;
Jiaqi WANG
;
Qiang GAO
;
Sili LIN
;
Chaochao WU
Author Information
1. 杭州佰辰医疗器械有限公司，杭州 311103
- Publication Type:Journal Article
- Keywords:
Testosterone;
Magnetic beads extraction-in situ derivatization;
Liquid chromatography-tandem mass spectrometry
- From:
Chinese Journal of Laboratory Medicine
2025;48(8):1048-1054
- CountryChina
- Language:Chinese
-
Abstract:
Objective:This study aimed to evaluate to evaluate the performance of equilibrium dialysis combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the accurate measurement of free testosterone in clinical samples. Mthods We conducted a prospective observational study using 161 serum samples from healthy women of reproductive 26(24, 32)years at the Gynecology Outpatient department of Peking Union Medical College Hospital from June to September 2024, and their concentrations were determined. In this study, after equilibrium dialysis of serum samples, free testosterone was extracted from the dialysate using a magnetic bead-based method. It was then directly derivatized using hydroxylamine hydrochloride in situ after elution from the magnetic bead and further quantified by LC-MS/MS.Method:validation assessed linearity, limit of quantification (LOQ), precision, accuracy, matrix effects, and carryover according to established guidelines. Data were analyzed using Origin 2019 and WPS Office 2019.Results:The method demonstrated excellent linearity ( R2>0.99) across 1-250 pg/ml with an LOQ of 1 pg/ml. The coefficients of variation for both intra-day and inter-day imprecision were less than 10% while recovery rates ranged from 92.60% to 99.10%. Matrix effect deviations were all within the range of 6% and carryover was negligible. Conclusions:In this study, the established method of magnetic bead-based extraction followed by in situ derivatization combined with liquid chromatography-tandem mass spectrometry performed well, and could be further applied to the detection of free testosterone concentration in childbearing age women.
