摘要
目的建立高效液相色谱-质谱联用法同时测定人血浆中替格瑞洛及其活性代谢产物AR-C124910XX的浓度。方法血浆样品用乙腈沉淀蛋白后,以伊曲康唑为内标,用Eclipse Plus-C_(18)色谱柱(2.1 mm×10 mm,3.5μm),以乙腈-10 mmol·L^(-1)醋酸铵溶液(均含0.15%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10μL,总分析时间为4 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。考察其专属性、标准曲线与定量下限、精密度与准确度、提取回收率、基质效应和稳定性。结果血浆中替格瑞洛的标准曲线方程为:y=3.79×10^(-2)x+5.27×10^(-3)(r=0.997 6),线性范围为2~2000μg·L^(-1),线性相关良好,定量下限为2μg·L^(-1);AR-C124910XX标准曲线方程为:y=4.58×10^(-2)x+6.15×10^(-3)(r=0.996 5),线性范围为1~1000μg·L^(-1),线性相关良好,定量下限为1μg·L^(-1)。替格瑞洛和AR-C124910XX低、中、高3个质量浓度的准确度在93.6%~108.7%;日内、日间的相对标准偏差(RSD)均小于15%;提取回收率在84.7%~91.3%。结论液质联用法快速、灵敏、准确,专属性强,重复性好,适用于人血浆中替格瑞洛及其活性代谢产物AR-C124910XX的浓度测定。
Objective To establish an HPLC- MS / MS method for simultaneous determination of ticagrelor and its active metabolite（ AR-C124910XX） in human plasma. Methods The plasma samples were precipitated protein by acetonitrile,with itraconazoleas an internal standard. The separation was achieved on Eclipse Plus- C（18）column（ 2. 1 mm× 10 mm,3. 5 μm） and eluted with linear gradient by mobile phase consisted of acetonitrile and 10 mmol·L^-1ammonium acetate,both of which were acidified with 0. 15% formicacid,at the flow rate of 0. 3mL ·min^-1. The injection volume was 10 μL and the column temperature was 30 ℃. The total time of the analysis was 4. 0 minutes. Detection of the analytes were achieved using positive ion electrospray ionization（ ESI） in the multiple reaction monitoring（ MRM） mode. The specificity,standard curve and lower limit of quantitation,precision and accuracy,extraction recoveries,the matrix effect and stability were investigated.Results The linear range of ticagrelor was 2- 2000 μg · L^-1. The linear regression equation was y = 3. 79 × 10^-2x ＋ 5. 27 × 10^-3（ r =0. 997 6）with the lower limit of quantitation（ LLOQ） of 2 μg·L^-1. While liner range of AR- C124910 XX was 1- 1000 μg·L^-1（ r =0. 996 5）,and the standard curve was y =4. 58 ×10^-2x ＋ 6. 15 × 10^-3（ r =0. 996 5） with LLOQ of 1 μg·L^-1. The accuracy of low,medium and high concentrations of ticagrelor and AR- C124910 XX was 93. 6%- 108. 7%. The intra- day and inter- day relative standard deviations（ RSD） were both less than 15%. The extraction recoveries were84. 7%- 91. 3%. Conclusion The established method is rapid,sensitive,accurate,specific and reliable,and suitable for simultaneous determination of ticagrelor and its active metabolite（ AR- C124910XX） in human plasma.
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2016年第13期1223-1226,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)
浙江省自然科学基金资助项目(LQ15H310003)
