摘要
目的：建立测定人血浆中25羟基维生素D[25（OH）D]的方法,并将其应用于临床。方法：血浆样品经液-液萃取后,采用超高效液相色谱-串联质谱法测定。以氘代25（OH）D3为内标,色谱柱为Zorbax Eclipse Plus C18,流动相为水（含0.1%甲酸）-甲醇（含0.1%甲酸）,梯度洗脱,流速为0.35 ml/min,柱温30℃采用电喷雾离子源,以多反应离子监测方式进行正离子扫描;用于定量分析的离子对分别为m/z 413.2→395.2[25（OH）D2]、m/z 401.2→383.2[25（OH）D3]、m/z 404.3→368.2（内标）。结果：25（OH）D2与25（OH）D3血药浓度在2-80 ng/ml范围内线性关系均良好（r分别为0.998 0、0.999 9,n=3）,其定量下限均为2 ng/ml,日间、日内RSD为0.78%-9.42%,相对偏差为-2.90%-1.95%。采用该法检测78例健康受试者及98例结核患者体内25（OH）D平均血药浓度分别为（21.53±6.33）、（8.84±4.05）ng/ml,差异有统计学意义（P〈0.01）。结论：该方法准确、可靠、灵敏度高,适用于血浆中25（OH）D的定量分析及人体药动学研究。
OBJECTIVE：To establish the method for the determination of 25-hydroxyvitamin D [25（OH）D] in human plasma,and apply it in the clinic. METHODS：After liquid-liquid extraction,UPLC-MS/MS method was used to determine plasma sample.Using deuteron-25（OH）D3as internal standard,the determination was performed on Zorbax Eclipse Plus C18 column with mobile phase consisted of water（containing 0.1% formic acid）-methanol（containing 0.1% formic acid）in gradient mode at flow rate of0.35 ml/min,and the column temperature was 30 ℃. By ESI,positive ion scanning was conducted in MRM mode;the monitoring transition ion-pair was m/z 413.2→395.2 for 25（OH）D2,m/z 401.2→383.2 for 25（OH）D3and m/z 404.3→368.2 for internal standard. RESULTS：The linear range of 25（OH）D2and 25（OH）D3were both 2-80 ng/ml（r=0.998 0 and 0.999 9,n=3）;the limit of quantitation was 2 ng/ml. RSDs of inter-day and intra-day were 0.78%-9.42%,and relative deviation were-2.90%-1.95%,respectively. Average plasma concentration of 25（OH）D in 78 health volunteers and 98 tuberculosis patients were（21.53±6.33）and（8.84±4.05）ng/ml,with statistical significance（P〈0.01）. CONCLUSIONS：The method is accurate,reliable and sensitive. It is suitable for quantitative analysis and pharmacokinetic study of 25（OH）D in human plasma.
出处
《中国药房》
CAS
北大核心
2016年第14期1907-1910,共4页
China Pharmacy
基金
北京市医院管理局重点医学专业发展计划项目(No.ZYLX201304)
