Sengupta Pinaki, Sarkar Amlan Kanti, Bhaumik Uttam, Chatterjee Bappaditya, Roy Bikash, Chakraborty Uday Shankar, Pal Tapan Kumar
Bioequivalence Study Centre, Department of Pharmaceutical Technology, Jadavpur University, Kolkata 700 032, India.
Biomed Chromatogr. 2010 Dec;24(12):1342-9. doi: 10.1002/bmc.1447.
A simple, high-throughput and specific high-performance liquid chromatography tandem mass spectrometry method has been developed and validated according to the FDA guidelines for simultaneous quantification of olmesartan and pioglitazone in rat plasma. The bioanalytical method consists of liquid-liquid extraction and quantitation by triple quadrupole mass spectrometry using electrospray ionization technique, operating in multiple reaction monitoring and positive ion modes. The compounds were eluted isocratically on a C(18) column with a mobile phase consisting of a mixture of methanol and water (containing 0.5% formic acid) in a ratio of 9:1. The response to olmesartan and pioglitazone was linear over the range 0.01-10 µg/mL. The validation results demonstrated that the method had satisfactory precision and accuracy across the calibration range. Intra- and inter-day precisions ranged from 0.66 to 3.32 and from 0.94 to 2.93% (%CV), respectively. The accuracy determined at three quality control levels was within 91.27-107.28%. There was no evidence of instability of the analytes in rat plasma following the stability studies. The method proved highly reproducible and sensitive and was successfully applied in a pharmacokinetic study after single dose oral administration of olmesartan and pioglitazone to the rat.
已根据美国食品药品监督管理局（FDA）的指南开发并验证了一种简单、高通量且特异的高效液相色谱串联质谱法，用于同时定量大鼠血浆中的奥美沙坦和吡格列酮。该生物分析方法包括液液萃取，并采用电喷雾电离技术通过三重四极杆质谱进行定量，以多反应监测和正离子模式运行。化合物在C(18)柱上以等度洗脱，流动相由甲醇和水（含0.5%甲酸）按9:1的比例混合而成。奥美沙坦和吡格列酮在0.01 - 10μg/mL范围内呈线性响应。验证结果表明，该方法在校准范围内具有令人满意的精密度和准确度。日内和日间精密度分别在0.66%至3.32%和0.94%至2.93%（%CV）之间。在三个质量控制水平下测定的准确度在91.27%至107.28%之间。稳定性研究后未发现大鼠血浆中分析物有不稳定的迹象。该方法被证明具有高度的重现性和灵敏性，并成功应用于大鼠单次口服奥美沙坦和吡格列酮后的药代动力学研究。
