液相色谱串联质谱法分析血清中多囊卵巢综合征相关甾体激素的性能评价  

Performance evaluation of LC-MS/MS in the analysis of serum steroids related to polycystic ovary syndrome

作　　者：郑东 黄惠芳[1] 尹晶平[1] 丰斌[1] 张政 邱骏[1] ZHENG Dong;HUANG Huifang;YIN Jingping;FENG Bin;ZHANG Zheng;QIU Jun(Centre of Clinical Laboratory,the First Affiliated Hospital of Soochow University,Suzhou 215006,Jiangsu,China)

机构地区：[1]苏州大学附属第一医院临床检测中心,江苏苏州215006

出　　处：《临床检验杂志》2022年第9期646-653,共8页Chinese Journal of Clinical Laboratory Science

基　　金：血液和血管疾病诊疗药物技术教育部工程研究中心课题(GCZ202001)。

摘　　要：目的建立液相色谱串联质谱法定量检测血清中17-α-羟孕酮、雄烯二酮、双氢睾酮、脱氢表雄酮、脱氢表雄酮硫酸酯的方法,并对其进行性能分析评价。方法采用液相色谱串联质谱法定量检测血清中17-α-羟孕酮、雄烯二酮、双氢睾酮、脱氢表雄酮、脱氢表雄酮硫酸酯的含量。根据《中国药典》(2015年版)通则中9012“生物样品定量分析方法验证指导原则”和《液相色谱串联质谱临床检测方法的开发与验证》对所建方法的线性范围、检出限、精密度、样本稳定性、回收率和携带污染率等基本性能进行确认。结果液相色谱串联质谱法定量检测血清中17-α-羟孕酮、雄烯二酮、双氢睾酮、脱氢表雄酮、脱氢表雄酮硫酸酯的线性范围分别为0.16~40 ng/mL、0.16~40 ng/mL、32~8000 pg/mL、0.16~40 ng/mL、2~500μg/dL,检出限分别为0.008 ng/mL、0.008 ng/mL、0.0032 ng/mL、0.008 ng/mL、1 ng/mL,定量下限分别为0.032 ng/mL、0.016 ng/mL、0.0064 ng/mL、0.016 ng/mL、4 ng/mL,批内变异系数(CV)和批间CV均小于15%,回收率分别为89.79%~101.23%、87.23%~98.33%、91.39%~101.10%、86.99%~94.30%、86.35%~108.35%。同时,血清中各激素在-20℃保存条件下7 d内是稳定的。结论建立的液相色谱串联质谱法基本性能符合评价标准,能够灵敏且准确地检测血清中17-α-羟孕酮、雄烯二酮、双氢睾酮、脱氢表雄酮和脱氢表雄酮硫酸酯的含量。Objective To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for the quantitative determination of serum 17-α-hydroxyprogesterone,androstenedione,dihydrotestosterone,dehydroepiandrosterone and dehydroepiandrosterone sulfate,and evaluate their performance.Methods The contents of serum 17-α-hydroxyprogesterone,androstenedione,dihydrotestosterone,dehydroepiandrosterone and dehydroepiandrosterone sulfate were detected by the established LC-MS/MS.The linear range,limit of detection,precision,sample stability,recovery rate and contamination-carrying rate of the established method were evaluated according to 9012"Guiding Principles for Verification of Quantitative Analysis Methods of Biological Samples"in The General Rules of Chinese Pharmacopoeia(2015 edition)and“Consensus of method development and validation of liquid chromatography-tandem mass spectrometry in clinical laboratories”.Results The linear ranges of serum 17-α-hydroxyprogesterone,androstenedione,dihydrotestosterone,dehydroepiandrosterone and dehydroepiandrosterone sulfate were 0.16-40 ng/mL,0.16-40 ng/mL,32-8000 pg/mL,0.16-40 ng/mL and 2-500μg/dL,respectively,and their detection limits were 0.008 ng/mL,0.008 ng/mL,0.0032 ng/mL,0.008 ng/mL and 1 ng/mL,respectively.The lower limits of quantitation for serum 17-α-hydroxyprogesterone,androstenedione,dihydrotestosterone,dehydroepiandrosterone and dehydroepiandrosterone sulfate were 0.032 ng/mL,0.016 ng/mL,0.0064 ng/mL,0.016 ng/mL and 4 ng/mL,respectively,and their recovery rates were 89.79%-101.23%,87.23%-98.33%,91.39%-101.10%,86.99%-94.30%and 86.35%-108.35%,respectively.The intra-batch coefficients of variation(CV)and inter-batch CV were all less than 15%.In addition,the five steroids in serum were stable within 7 days under the condition of-20℃.Conclusion The basic performance of the established LC-MS/MS meets the evaluation criteria,and it can detect the contents of serum 17-α-hydroxyprogesterone,androstenedione,dihydrotestosterone,dehydroepiandrosterone and dehydroepia

关 键 词：液相色谱串联质谱法 性能评价 17-α-羟孕酮 雄烯二酮 双氢睾酮 脱氢表雄酮 脱氢表雄酮硫酸酯 

分 类 号：R446[医药卫生—诊断学]