摘要
目的：建立LC－MS／MS法同时测定人血浆中阿昔洛韦、更昔洛韦、拉米夫定、阿德福韦和恩替卡韦的浓度。方法血浆样品用乙腈沉淀蛋白后，以喷昔洛韦为内标，用Poreshell C18色谱柱（3.0 mm ×50 mm，2.7μm），以甲醇－10 mmol· L－1醋酸铵溶液（均含0.2％甲酸）为流动相进行梯度洗脱，流速为0.3 mL· min－1，柱温为30℃，进样量为10μL，总分析时间为5 min，用电喷雾离子化源，正离子方式，多反应监测（ MRM）扫描方式进行监测，检测专属性、标准曲线与定量下限、精密度与回收率、基质效应和稳定性。结果血浆中阿昔洛韦、更昔洛韦、拉米夫定的线性范围为10.0～5000.0μg · L－1（ r＞0.99），定量下限为10.0μg· L－1；阿德福韦线性范围为0.50～100.00μg· L－1（r＞0.9968），定量下限为0.50μg· L－1；恩替卡韦的线性范围为0.02～20.00μg· L－1，定量下限为0.02μg· L－1（r＞0.9980）。低、中、高3个质量浓度样本的平均回收率在93.3％～109.8％，日内、日间RSD均小于15％，提取回收率在85.6％～92.9％。结论该方法快速、灵敏、准确，且专属性强、重复性好，适用于同时测定人血浆中阿昔洛韦、更昔洛韦、拉米夫定、阿德福韦和恩替卡韦的浓度。
Objective To develop an LC -MS/MS method for simul-taneous determination of aciclovir, ganciclovir, lamivudine, adefovir and entecavir concentration in human plasma.Methods The plasma samples were precipitated by acetonitrile, with penciclovir as an internal standard.The separation was achieved on Poreshell C18 （ 3.0 mm ×50 mm, 2.7 μm ） and eluted with linear gradient using methanol and 10 mmol· L-1 ammonium acetate, both of which were acidified with 0.2%formic acid at the flow rate of 0.3 mL· min-1.The injection volume was 10 μL and the column temperature was 30 ℃.The total time of the analysis was 5 min.Detection of the analytes were achieved by positive ion electrospray ionization in the multiple reaction monitoring （ MRM ） mode.The specificity, lower limit of quantitation and standard curve, precision and recovery rate and stability as well as the matrix effect were investigated.Results The linear calibration curves of aclciclovir, ganciclovir and lamivudine were obtained, and the concentration ranged from 10.0 to 5000.0 μg · L-1 （ r 〉0.99 ） , with the lower limit of＆amp;nbsp;quantitation of 10.0μg· L-1 .While the standard curves of adefovir and entecavir were linear over the range of 0.50 to 100.00μg· L-1（r=0.996 8） and 0.02 to 20.00μg· L-1（r=0.998 0）, with the lower limit of quantitation of 0.50μg· L-1 and 0.02 μg· L-1 , respectively.The average recovery was between 93.3% and 109.8%.Intra-day and inter-day relative standard deviations （ RSD） were both below 15%.The recoveries of low, middle and high concen-trations were from 85.6%to 92.9%.Conclusion The established method is rapid, sensitive, accurate, specific and reliable, suitable for simultaneous determination of aciclovir, ganciclovir, lamivudine, adefovir and entecavir in human plasma.
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2015年第9期747-750,共4页
The Chinese Journal of Clinical Pharmacology
基金
国家科技重大专项基金资助项目(2013ZX09303005)
