Optimization and performance evaluation of an LC-MS/MS method for the determination of phytosterols in human plasma
10.3760/cma.j.cn114452-20240306-00113
- VernacularTitle:液相色谱串联质谱系统检测血浆植物固醇方法的优化与性能评价
- Author:
Yuxuan ZHANG
1
;
Luya WANG
;
Weimin WAN
;
Hu DING
;
Junfang WU
Author Information
1. 华中科技大学同济医学院附属同济医院心血管内科，武汉 430000
- Publication Type:Journal Article
- Keywords:
Phytosterol;
Liquid chromatography/mass spectrometry;
Plasma;
Sitosterolemia
- From:
Chinese Journal of Laboratory Medicine
2025;48(1):125-132
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish and evaluate the performance of a stable and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of six phytosterols in human plasma.Methods:The methodology was established, evaluated, and applied. A number of 136 healthy subjects were recruited from the Physical Examination Center of Suzhou Municipal Hospital from October 2023 to December 2023, meanwhile plasma phytosterols′ levels were validated in 5 patients with sitosterolemia. Taking deuterium isotope as internal standards, the clear separation was achieved using a Phenomenex C18 (2.1×100 mm, 2.6 μm) column. The mobile phase consisted of water and methanol at a flow rate of 0.4 ml/min with gradient elution, and the column temperature was 25 ℃. The experiments were performed on AB Sciex 6500+triple quadrupole mass spectrometry, and the linearity, limit of quantification, rate of recovery, precision, and stability of the method were evaluated. The testing method was further validated in the sitosterolemia patients.Results:The method had in-run and between-run coefficient of variation ranging 1.8%-12.5% and 2.6%-13.6%, respectively, which accorded with the precision requirements. The rates of recovery were 85.0%-105.1%, which accorded with the accuracy requirements. The samples showed good stability before and after handling stored at 4 ℃ for 72 hours. The plasma levels of sitosterol, campesterol, stigmasterol and cholestanol in sitosterolemia patients were apparently higher compared with those in the healthy group ( P<0.001). Conclusion:LC-MS/MS method for the quantitative measurement of plant sterols in human plasma has been established, which has convenient preprocessing and passed the performance evaluation including linearity, lower limit of quantification, recovery rate, precision and stability, and shown to be suitable for clinical application.
