Marchioni Camila, de Souza Israel Donizeti, Grecco Caroline Fernandes, Crippa José Alexandre, Tumas Vitor, Queiroz Maria Eugênia Costa
Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo (USP), Avenida Bandeirantes, 3900, São Paulo, Ribeirão Preto, CEP 14040-901, Brazil.
Departamento de Química, Universidade de São Paulo (USP), Avenida Bandeirantes, 3900, São Paulo, Ribeirão Preto, CEP 14040-901, Brazil.
Anal Bioanal Chem. 2017 May;409(14):3587-3596. doi: 10.1007/s00216-017-0300-3. Epub 2017 Mar 23.
This study reports a fast, sensitive, and selective column switching ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method to determine the endocannabinoids (eCBs), anandamide (AEA), and 2-arachidonoylglycerol (2-AG) in plasma samples. This bidimensional system used a restricted access media column (RP-8 ADS, 25 mm × 4 mm × 25 μM) in the first dimension and a core-shell Kinetex C18 (100 mm × 2, 1.7 mm × 1 μM) column in the second dimension, followed by detection in a mass spectrometer triple quadrupole (multiple reactions monitoring mode) operating in the positive mode. RP-8 ADS was used for trace enrichment of eCBs (reverse phase partitioning) and macromolecular matrix size exclusion; the core-shell column was used for the chromatographic separation. The column switching UHPLC-MS/MS method presented a linear range spanning from 0.1 ng mL (LOQ) to 6 ng mL for AEA and from 0.04 ng mL (LOQ) to 10 ng mL for 2-AG. Excluding the LLOQ values, the precision assays provided coefficients of variation lower than 8% and accuracy with relative standard error values lower than 14%. Neither carryover nor matrix effects were detected. This high-throughput column switching method compared to conventional methods is time saving as it involves fewer steps, consumes less solvent, and presents lower LLOQ. The column switching UHPLC-MS/MS method was successfully applied to determine AEA and 2-AG in plasma samples obtained from Alzheimer's disease patients. Graphical abstract A column switching ultra high-performance liquid chromatography-tandem mass spectrometry method using RP-8 ADS column and core shell column to determine endocannabinoids in plasma samples.
本研究报告了一种快速、灵敏且具有选择性的柱切换超高效液相色谱-串联质谱（UHPLC-MS/MS）方法，用于测定血浆样品中的内源性大麻素（eCBs）、花生四烯乙醇胺（AEA）和2-花生四烯酸甘油酯（2-AG）。该二维系统在第一维使用了限制进入介质柱（RP-8 ADS，25 mm×4 mm×25 μM），在第二维使用了核壳型Kinetex C18柱（100 mm×2，1.7 mm×1 μM），随后在正模式下运行的质谱仪三重四极杆（多反应监测模式）中进行检测。RP-8 ADS用于eCBs的痕量富集（反相分配）和大分子基质尺寸排阻；核壳柱用于色谱分离。柱切换UHPLC-MS/MS方法的线性范围为：AEA从0.1 ng/mL（定量下限）到6 ng/mL，2-AG从0.04 ng/mL（定量下限）到10 ng/mL。除定量下限值外，精密度测定的变异系数低于8%，准确度的相对标准误差值低于14%。未检测到残留和基质效应。与传统方法相比，这种高通量柱切换方法节省时间，因为它涉及的步骤更少，消耗的溶剂更少，且定量下限更低。柱切换UHPLC-MS/MS方法已成功应用于测定从阿尔茨海默病患者获得的血浆样品中的AEA和2-AG。图形摘要：一种使用RP-8 ADS柱和核壳柱的柱切换超高效液相色谱-串联质谱方法，用于测定血浆样品中的内源性大麻素。
