Ramanujam N, Sivaselvakumar M, Ramalingam S
Department of Pharmacology, PSG Institute of Medical Sciences and Research, Coimbatore, Tamilnadu, India.
Department of Molecular Medicine and Therapeutics, PSG Institute of Medical Sciences, Coimbatore, Tamilnadu, India.
Biomed Chromatogr. 2017 Nov;31(11). doi: 10.1002/bmc.4000. Epub 2017 Jun 5.
A simple, sensitive and reproducible ultra-performance liquid chromatography (UPLC) method has been developed and validated for simultaneous estimation of polychlorinated biphenyl (PCB) 77 and PCB 180 in mouse plasma. The sample preparation was performed by simple liquid-liquid extraction technique. The analytes were chromatographed on a Waters Acquity H class UPLC system using isocratic mobile phase conditions at a flow rate of 0.3 mL/min and Acquity UPLC BEH shield RP column maintained at 35°C. Quantification was performed on a photodiode array detector set at 215 nm and PCB 101 was used as internal standard (IS). PCB 77, PCB 180, and IS retention times were 2.6, 4.7 and 2.8 min, respectively, and the total run time was 6 min. The method was validated for specificity, selectivity, recovery, linearity, accuracy, precision and sample stability. The calibration curve was linear over the concentration range 10-3000 ng/mL for PCB 77 and PCB 180. Intra- and inter-day precisions for PCBs 77 and 180 were found to be good with CV <4.64%, and the accuracy ranged from 98.90 to 102.33% in mouse plasma. The validated UPLC method was successfully applied to the pharmacokinetic study of PCBs 77 and 180 in mouse plasma.
已开发并验证了一种简单、灵敏且可重复的超高效液相色谱(UPLC)方法,用于同时测定小鼠血浆中的多氯联苯(PCB)77和PCB 180。采用简单的液-液萃取技术进行样品制备。使用等度流动相条件,流速为0.3 mL/min,在Waters Acquity H class UPLC系统上对分析物进行色谱分离,并使用保持在35°C的Acquity UPLC BEH shield RP柱。在设置为215 nm的光电二极管阵列检测器上进行定量分析,并使用PCB 101作为内标(IS)。PCB 77、PCB 180和IS的保留时间分别为2.6、4.7和2.8分钟,总运行时间为6分钟。该方法在特异性、选择性、回收率、线性、准确度、精密度和样品稳定性方面得到了验证。PCB 77和PCB 180在10-3000 ng/mL浓度范围内校准曲线呈线性。发现PCB 77和180的日内和日间精密度良好,CV<4.64%,在小鼠血浆中的准确度范围为98.90%至102.33%。经过验证的UPLC方法成功应用于小鼠血浆中PCB 77和180的药代动力学研究。