Chemistry Department, Kadi Sarva Vishwavidyalaya, Sarva Vidyalaya Campus, Sector 15/23, Gandhinagar 382015, Gujarat, India.
J Pharm Biomed Anal. 2012 Nov;70:462-70. doi: 10.1016/j.jpba.2012.06.027. Epub 2012 Jun 26.
A highly sensitive, selective and rapid bioanalytical method has been developed for the simultaneous determination of bosentan and hydroxybosentan in human plasma by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The analytes and their deuterated analogs were quantitatively extracted from 100 μL human plasma by solid phase extraction. The chromatographic separation of analytes was achieved on a Thermo Hypurity C18 (100 mm × 4.6 mm, 5 μ) analytical column with a resolution factor of 2.4 under isocratic conditions. The method was validated over a dynamic concentration range of 0.4-1600 ng/mL for bosentan and 0.2-250 ng/mL for hydroxybosentan. Ion-suppression effects were investigated by post-column infusion of analytes. The precision (%CV) values for the calculated slopes of calibration curves, which would reflect the relative matrix effect, were less than 1.2% for both the analytes. The intra-batch and inter-batch precision (%CV) across quality control levels was ≤4.0% and the mean relative recovery was >94% for both the analytes. The method was successfully applied to a bioequivalence study of 125 mg tablet formulation (test and reference) in 12 healthy Indian male subjects under fasting condition. The ratios of mean log-transformed values of C(max), AUC(0-t) and AUC(0-inf) and their 90% CIs varied from 91.3 to 104.7%. The percentage change for incurred sample reanalysis (ISR) was within ±13.0%.
已经开发出一种高度敏感、选择性和快速的生物分析方法，用于通过液相色谱-串联质谱（LC-MS/MS）同时测定人血浆中的波生坦和羟基波生坦。通过固相萃取从 100μL 人血浆中定量提取分析物及其氘代类似物。在等度条件下，使用 Thermo Hypurity C18（100mm×4.6mm，5μm）分析柱实现了分析物的色谱分离，分辨率因子为 2.4。该方法在波生坦的动态浓度范围为 0.4-1600ng/mL 和羟基波生坦的 0.2-250ng/mL 进行了验证。通过在柱后注入分析物研究了离子抑制效应。反映相对基质效应的校准曲线计算斜率的精度（%CV）值对于两种分析物均小于 1.2%。批内和批间精密度（%CV）在质控水平内均≤4.0%，两种分析物的平均相对回收率均>94%。该方法成功应用于 12 名健康印度男性空腹条件下服用 125mg 片剂制剂（试验和参比）的生物等效性研究。平均对数转换的 C(max)、AUC(0-t)和 AUC(0-inf)值比值及其 90%置信区间的变化范围为 91.3-104.7%。发生样品重新分析（ISR）的百分比变化在±13.0%范围内。
