Sharma Kuldeep, Singh Radhe Raman, Kandaswamy Murugesh, Mithra Chandan, Giri Sanjeev, Rajagopal Sriram, Mullangi Ramesh
Jubilant Biosys, Industrial Suburb, 2nd Stage, Yeshwanthpur, Bangalore-560 022, India.
Bioanalysis. 2011 Jan;3(2):181-96. doi: 10.4155/bio.10.192.
An LC-MS/MS-ESI method has been validated for simultaneous estimation of the three endocannabinoids; N-arachidonoylethanolamine (AEA), N-oleoylethanolamine (OEA) and palmitoylethanolamide (PEA), in surrogate matrix using AEA-d (4) as an internal standard with highest sensitivity over the existing methods. Simple precipitation was used to extract analytes and these were subsequently analyzed on a monolithic column. Linear response function was established over the concentration range 12.3 to 1225 pg/ml for AEA (r > 0.994); 0.70 to 641 ng/ml for OEA (r > 0.999) and 0.54 to 321 ng/ml (r > 0.998) for PEA. The intra- and inter-day precision values met the acceptance to criteria as per US FDA guidelines. Analytes were found to be stable in the battery of stability studies. The method was applied to quantify endogenous levels of analytes in rat plasma.
已验证一种液相色谱-串联质谱-电喷雾电离（LC-MS/MS-ESI）方法，可使用氘代花生四烯酸乙醇胺（AEA-d(4)）作为内标，在替代基质中同时测定三种内源性大麻素；N-花生四烯酰乙醇胺（AEA）、N-油酰乙醇胺（OEA）和棕榈酰乙醇胺（PEA），该方法比现有方法具有更高的灵敏度。采用简单沉淀法提取分析物，随后在整体柱上进行分析。建立了AEA在12.3至1225 pg/ml浓度范围内的线性响应函数（r > 0.994）；OEA在0.70至641 ng/ml浓度范围内的线性响应函数（r > 0.999）以及PEA在0.54至321 ng/ml浓度范围内的线性响应函数（r > 0.998）。日内和日间精密度值符合美国食品药品监督管理局（US FDA）指南的验收标准。在一系列稳定性研究中发现分析物是稳定的。该方法用于定量大鼠血浆中分析物的内源性水平。
