Analytical performance of a candidate reference measurement procedure for serum 17α-hydroxyprogesterone based on liquid chromatography tandem mass spectrometry
10.3760/cma.j.cn114452-20220321-00162
- VernacularTitle:同位素稀释液相色谱串联质谱法测定血清17α-羟孕酮候选参考测量程序的研究
- Author:
Keying YU
1
;
Hewei SUN
;
Zhonggan JIN
;
Sujie ZHANG
;
Qing LI
;
Yi JU
Author Information
1. 上海市临床检验中心参考测量实验室，上海 200126
- Keywords:
17-alpha-Hydroxyprogesterone;
Mass spectrometry;
Radioisotope dilution technique;
Reference measurement procedure
- From:
Chinese Journal of Laboratory Medicine
2022;45(5):449-455
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To determine the analytical performance of a candidate reference measurement procedure for 17α-hydroxyprogesterone in human serum by liquid chromatography-tandem mass spectrometry (LC-MS/MS).Methods:The serum spiked with a deuterium-labeled internal standard was extracted from serum from individual undergoing physical examination by liquid-liquid extraction with n-hexane/ethyl acetate (3∶2, v/v), separated by C18 reversed-phase chromatography and detected by positive electrospray ionization mass spectrometry. According to the Clinical and Laboratory Standards Institute (CLSI) C62-A documents, the analytical performance including linearity, limit of detection,limit of quantitation,relative matrix effect,precision and trueness,carry-over and specificity was evaluated.Results:The linear range of 17α-hydroxyprogesterone by LC-MS/MS was 0.21-119.67 μg/L. The limit of detection and limit of quantitation were 5.218 ng/L and 0.116 μg/L. The relative matrix effects were -0.02%, -0.40% and -0.90% for sera and solution mixtures in 3 different ratios (50∶50, 80:20 and 20∶80). The coefficients of variation ( CVs) of intra-assay were 1.73%-2.11%, 0.98%-1.71%, 0.47%-0.87% at 0.164 μg/L, 14.81 μg/L, 81.63 μg/L and the CVs of inter-assay were 1.82%, 1.03%, 0.80% at above three concentrations. The average recovery rates of 3 levels (0.5, 20 and 100 μg/L) were 100.4%, 101.7%, 102.8%, respectively. The measured values of GBW09829 of National Institute of Metrology were within the specified uncertainty range. Conclusion:The candidate reference measurement procedure for 17α-hydroxyprogesterone in human by LC-MS/MS is established with good accuracy and precision, which can be clinically used for measurement traceability.
