Da Silva Bianca Ferreira, Ahmadireskety Atiye, Aristizabal-Henao Juan J, Bowden John A
College of Veterinary Medicine, Department of Physiological Sciences, University of Florida, Gainesville, FL, USA.
Department of Chemistry, University of Florida, Gainesville, FL, USA.
MethodsX. 2020 Oct 17;7:101111. doi: 10.1016/j.mex.2020.101111. eCollection 2020.
Per- and polyfluoroalkyl substances (PFAS) are synthetic organic compounds that over the past several years, have witnessed a dramatic increase in scientific attention. As PFAS are predominantly accumulated in plasma, monitoring individual burden levels in plasma are typically achieved via some combination of protein precipitation and/or solid phase extraction (SPE), either in online or offline modes. This work describes an updated PFAS extraction workflow, using 96-well plate technology and protein precipitation that is rapid, simple, inexpensive, and amenable for large cohort studies. In brief, plasma proteins were precipitated using methanol and the resulting centrifuged supernatant was directly analyzed using UHPLC-MS/MS. We monitored 51 PFAS, which were quantified via isotope dilution and the effectiveness of the method was demonstrated by using NIST blood-based Standard Reference Materials (SRMs). This method resulted in recoveries ranging between 70 and 89% for all analytes. The 96-well design exhibited low limits of detection and only required sample volumes of 100 µL, thus resulting in an amenable method for high-throughput plasma/serum PFAS screening. • PFAS were directly quantified in plasma and serum samples; • No SPE needed after protein precipitation; • SRMs can be used to validate PFAS measurement in plasma/serum.
全氟和多氟烷基物质(PFAS)是合成有机化合物,在过去几年中,其受到的科学关注急剧增加。由于PFAS主要在血浆中蓄积,监测血浆中的个体负担水平通常通过蛋白质沉淀和/或固相萃取(SPE)的某种组合来实现,采用在线或离线模式均可。本研究描述了一种更新的PFAS提取工作流程,使用96孔板技术和蛋白质沉淀法,该方法快速、简单、廉价,适用于大规模队列研究。简而言之,使用甲醇沉淀血浆蛋白,所得离心上清液直接用超高效液相色谱-串联质谱法(UHPLC-MS/MS)分析。我们监测了51种PFAS,通过同位素稀释进行定量,并使用基于NIST血液的标准参考物质(SRM)证明了该方法的有效性。该方法对所有分析物的回收率在70%至89%之间。96孔设计的检测限较低,仅需100µL样品体积,因此成为一种适用于高通量血浆/血清PFAS筛查的方法。• PFAS可在血浆和血清样品中直接定量;• 蛋白质沉淀后无需SPE;• SRM可用于验证血浆/血清中PFAS的测量。