Khatal Laxman, Gaur Ashwani, Naphade Ashish, Kandikere Vishwottam, Mookhtiar Kasim
Advinus Therapeutics Ltd, Pune, India.
Biomed Chromatogr. 2016 Oct;30(10):1676-85. doi: 10.1002/bmc.3740. Epub 2016 May 19.
Measurement of test article concentration in tissue samples has been an important part of pharmacokinetic study and has helped to co-relate pharmacokinetic/pharmacodynamic relationships since the 1950s. Bioanalysis of tissue samples using LC-MS/MS comes with unique challenges in terms of sample handling and inconsistent analyte response owing to nonvolatile matrix components. Matrix effect is a phenomenon where the target analyte response is either suppressed or enhanced in the presence of matrix components. Based on previous reports electrospray ionization (ESI) mode of ionization is believed to be more affected by matrix components than atmospheric pressure chemical ionization (APCI) or atmospheric pressure photoionization. To explore the impact of ionization source with respect to bioanalysis of tissue samples, five structurally diverse compounds - atenolol, verapamil, diclofenac, propranolol and flufenamic acid - were selected. Quality control standards were spiked into 10 different biological matrices like whole blood, liver, heart, brain, spleen, kidney, skeletal muscle, eye and skin tissue and were quantified against calibration standards prepared in rat plasma. Quantitative bioanalysis was performed utilizing both APCI and ESI mode and results were compared. Quality control standards when analyzed with APCI mode were found to be more consistent in terms of accuracy and precision as compared with ESI mode. Additionally, for some instances, up to 20-fold broader dynamic linearity range was observed with APCI mode as compared with ESI mode. As phospholid interferences have poor response in APCI mode, protein precipitation extraction technique can be used for multimatrix quantitation, which is more amenable to automation. The approach of multiple biological matrix quantitation against a single calibration curve helps bioanalysts to reduce turnaround time. Copyright © 2016 John Wiley & Sons, Ltd.
自20世纪50年代以来，测量组织样本中的受试物浓度一直是药代动力学研究的重要组成部分，并有助于建立药代动力学/药效学之间的相互关系。使用液相色谱-串联质谱法（LC-MS/MS）对组织样本进行生物分析，在样本处理方面存在独特的挑战，并且由于非挥发性基质成分，分析物响应不一致。基质效应是指在基质成分存在的情况下，目标分析物响应被抑制或增强的现象。根据以往的报道，电喷雾电离（ESI）模式的电离被认为比大气压化学电离（APCI）或大气压光电离更容易受到基质成分的影响。为了探讨电离源对组织样本生物分析的影响，选择了五种结构不同的化合物——阿替洛尔、维拉帕米、双氯芬酸、普萘洛尔和氟芬那酸。将质量控制标准品添加到10种不同的生物基质中，如全血、肝脏、心脏、大脑、脾脏、肾脏、骨骼肌、眼睛和皮肤组织，并根据在大鼠血浆中制备的校准标准品进行定量。利用APCI和ESI模式进行定量生物分析，并比较结果。与ESI模式相比，用APCI模式分析质量控制标准品时，在准确性和精密度方面更一致。此外，在某些情况下，与ESI模式相比，APCI模式观察到的动态线性范围宽达20倍。由于磷脂干扰在APCI模式下响应较差，蛋白质沉淀提取技术可用于多基质定量，这更适合自动化。针对单一校准曲线进行多种生物基质定量的方法有助于生物分析人员减少周转时间。版权所有© 2016约翰威立父子有限公司。
