Chen Fangfang, Gong Zhiyuan, Kelly Barry C
Graduate School of Integrated Sciences and Engineering (NGS), National University of Singapore, Singapore.
Graduate School of Integrated Sciences and Engineering (NGS), National University of Singapore, Singapore; Department of Biological Sciences, National University of Singapore, Singapore.
J Chromatogr A. 2015 Feb 27;1383:104-11. doi: 10.1016/j.chroma.2015.01.033. Epub 2015 Jan 17.
A sensitive analytical method based on liquid-liquid extraction (LLE) and liquid chromatography tandem mass spectrometry (LC-MS/MS) was developed for rapid analysis of 11 pharmaceuticals and personal care products (PPCPs) in fish plasma micro-aliquots (∼20μL). Target PPCPs included, bisphenol A, carbamazepine, diclofenac, fluoxetine, gemfibrozil, ibuprofen, naproxen, risperidone, sertraline, simvastatin and triclosan. A relatively quicker and cheaper LLE procedure exhibited comparable analyte recoveries with solid-phase extraction. Rapid separation and analysis of target compounds in fish plasma extracts was achieved by employing a high efficiency C-18 HPLC column (Agilent Poroshell 120 SB-C18, 2.1mm×50mm, 2.7μm) and fast polarity switching, enabling effective monitoring of positive and negative ions in a single 9min run. With the exception of bisphenol A, which exhibited relatively high background contamination, method detection limits of individual PPCPs ranged between 0.15 and 0.69pg/μL, while method quantification limits were between 0.05 and 2.3pg/μL. Mean matrix effect (ME) values ranged between 65 and 156% for the various target analytes. Isotope dilution quantification using isotopically labelled internal surrogates was utilized to correct for signal suppression or enhancement and analyte losses during sample preparation. The method was evaluated by analysis of 20μL plasma micro-aliquots collected from zebrafish (Danio rerio) from a laboratory bioaccumulation study, which included control group fish (no exposure), as well as fish exposed to environmentally relevant concentrations of PPCPs. Using the developed LC-MS/MS based method, concentrations of the studied PPCPs were consistently detected in the low pg/μL (ppb) range. The method may be useful for investigations requiring fast, reliable concentration measurements of PPCPs in fish plasma. In particular, the method may be applicable for in situ contaminant biomonitoring, as well as bioaccumulation and toxicology studies employing small fishes with low blood compartment volumes.
建立了一种基于液液萃取（LLE）和液相色谱串联质谱（LC-MS/MS）的灵敏分析方法，用于快速分析鱼血浆微量等分试样（约20μL）中的11种药品和个人护理产品（PPCP）。目标PPCP包括双酚A、卡马西平、双氯芬酸、氟西汀、吉非贝齐、布洛芬、萘普生、利培酮、舍曲林、辛伐他汀和三氯生。一种相对更快且更便宜的LLE方法与固相萃取法的分析物回收率相当。通过使用高效C-18 HPLC柱（安捷伦Poroshell 120 SB-C18，2.1mm×50mm，2.7μm）和快速极性切换，实现了鱼血浆提取物中目标化合物的快速分离和分析，能够在单次9分钟运行中有效监测正离子和负离子。除双酚A背景污染相对较高外，各PPCP的方法检测限在0.15至0.69pg/μL之间，而方法定量限在0.05至2.3pg/μL之间。各种目标分析物的平均基质效应（ME）值在65%至156%之间。使用同位素标记的内标进行同位素稀释定量可校正样品制备过程中的信号抑制或增强以及分析物损失。通过分析从实验室生物累积研究中的斑马鱼（Danio rerio）采集的20μL血浆微量等分试样对该方法进行评估，该研究包括对照组鱼（未暴露）以及暴露于环境相关浓度PPCP的鱼。使用所开发 的基于LC-MS/MS的方法，所研究的PPCP浓度始终在低pg/μL（ppb）范围内被检测到。该方法可能有助于需要快速、可靠地测量鱼血浆中PPCP浓度的研究。特别是，该方法可能适用于原位污染物生物监测以及使用血容量低的小鱼进行的生物累积和毒理学研究。
