Arafat Tawfiq, Arafat Basil, awad Riad, awwad Ahmad Abu
University of Petra, Faculty of Pharmacy, Amman, Jordan.
Al-Ahliyya Amman University, Faculty of Pharmacy and Medical Sciences, Amman, Jordan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Dec 1;972:81-8. doi: 10.1016/j.jchromb.2014.09.037. Epub 2014 Oct 2.
A simple and sensitive liquid chromatography-tandem mass spectrometric method for quantification of loperamide in human plasma and saliva was developed and validated, and then successfully applied in pharmacokinetic clinical study to investigate and correlate bioavailability of Imodium(®) 2mg quartet tablet dose in both human plasma and saliva. Loperamide with labeled internal standard was extracted from its biological matrix by methanol as protein direct precipitant in single extraction step. Adequate chromatographic separation for analytes from plasma and saliva matrices was achieved using ACE C18 (50mm×2.1mm, 5μm) column, eluted by water/methanol/formic acid (30:70:0.1%, v/v), delivered isocratically at constant flow rate of 0.75ml/min. The method validation intends to investigate specificity, sensitivity, linearity, precision, accuracy, recovery, matrix effect and stability according to European guideline, and partial validation was applied on saliva, specificity, matrix effect, recovery, sensitivity, within and between day precision and accuracy. The calibration curve was linear through the range of 20-3000pg/ml in both plasma and saliva using a 50μl sample volume. The partial validation sections outcome in saliva was so close to those in plasma. The within- and between-day precisions were all below 8.7% for plasma and below 11.4% for saliva. Accuracies ranged from 94 to 105% for both matrices. In this study, 26 healthy volunteers participated in the clinical study, and 6 of gave their saliva samples in addition to plasma at the same time schedule. The pharmacokinetic parameters of Cmax, AUC0-t and AUC0-∞, Tmax and T1/2 in both plasma and saliva were calculated and correlated.
建立并验证了一种简单灵敏的液相色谱 - 串联质谱法，用于定量测定人血浆和唾液中的洛哌丁胺，然后成功应用于药代动力学临床研究，以研究和关联易蒙停（®）2mg四联片剂量在人血浆和唾液中的生物利用度。将带有标记内标的洛哌丁胺通过甲醇作为蛋白质直接沉淀剂，在单一萃取步骤中从其生物基质中萃取出来。使用ACE C18（50mm×2.1mm，5μm）色谱柱实现了血浆和唾液基质中分析物的充分色谱分离，以水/甲醇/甲酸（30:70:0.1%，v/v）洗脱，以0.75ml/min的恒定流速等度输送。该方法的验证旨在根据欧洲指南研究特异性、灵敏度、线性、精密度、准确度、回收率、基质效应和稳定性，并对唾液进行部分验证，包括特异性、基质效应、回收率、灵敏度、日内和日间精密度及准确度。使用50μl样品体积时，血浆和唾液中校准曲线在20 - 3000pg/ml范围内呈线性。唾液中的部分验证结果与血浆中的非常接近。血浆的日内和日间精密度均低于8.7%，唾液的低于11.4%。两种基质的准确度范围均为94%至105%。在本研究中，26名健康志愿者参与了临床研究，其中6人除了在相同时间采集血浆外还提供了唾液样本。计算并关联了血浆和唾液中Cmax、AUC0 - t、AUC0 - ∞、Tmax和T1/2的药代动力学参数。
