摘要
目的建立HPLC-MS/MS法同时测定肝微粒体孵育体系中地塞米松及其代谢产物的浓度。方法选用Kromasil C18色谱柱（2.1 mm×150 mm,5μm）,流动相为乙腈-0.01%甲酸水溶液（45∶55）,流速为0.35 m L·min-1,柱温为40℃,用正离子扫描,多反应监测方式测定,考察专属性、标准曲线与定量下限、精密度与回收率、基质效应和稳定性。结果地塞米松和6β-羟基地塞米松在20~1000,1~50 ng·m L-1内线性关系良好（r=0.998 0,r=0.998 5）,定量下限分别为20,1 ng·m L-1,其标准曲线回归方程分别为y=3.93×10-3x＋9.88×10-3,y=1.88×10-2x＋2.21×10-4。地塞米松和6β-羟基地塞米松的绝对回收率分别73.87%~92.18%,86.33%~87.22%,日内、日间相对标准偏差均在15%以内。结论本研究方法简便、快速、准确,适用于测定肝微粒体孵育样品中地塞米松及其代谢产物的浓度。
Objective To establish a HPLC-MS / MS method for simultaneous determinations of dexamethasone and its metabolite in human liver microsomal incubation system.Methods Kromasil C18column（2.1 mm × 150 mm,5 μm） was used at 40 ℃,the mobile phase was a mixed system consisting of acetonitrile and 0.01% formic acid（45 ∶ 55）and the flow rate was 0.35 m L·min-1.The detection of the samples was made using positive ion scan and multiple reaction monitoring mode.The specificity,standard curve and lower limit of quantification,precision and recovery rate and stability as well as the matrix effect were investigated.Results The liner calibration curve of dexamethasone and 6-beta-hydroxydexamethasone obtained a concentration range of 20-1000ng·m L-1and 1-50 ng · m L-1（r = 0.998 0,r = 0.998 5） and the lower limit of quantification was 20 and 1 ng · m L-1,the regression equation was y = 3.93 × 10-3x ＋ 9.88 × 10-3,y = 1.88 × 10-2x ＋2.21 × 10-4.The average recovery rates of dexamethasone and6-beta-hydroxydexamethasone were 73.87%-92.18% and86.33%-87.22%.Intra-day and inter-day relative standard deviations（RSD） of dexamethasone and 6-beta-hydroxydexamethasone were less than 15%.Conclusion The method is simple,rapid andaccurate,suitable for measuring the concentration of dexamethasone and its metabolite in human liver microsomal.
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2015年第13期1292-1295,共4页
The Chinese Journal of Clinical Pharmacology
