Quantification of endocannabinoids in rat biological samples by GC/MS: technical and theoretical considerations.
In the last several years, interest has increased significantly about the endocannabinoids anandamide and 2-arachidonylglycerol, two lipid messengers that activate cannabinoid receptors. Quantification of these compounds in biological samples presents numerous technical challenges. Because of their low abundance, endocannabinoids are usually quantified by isotope dilution assays using mass spectrometry coupled to either gas chromatography or high-performance liquid chromatography. Although endocannabinoid levels in biological fluids, such as plasma and cerebrospinal fluid, can be directly determined by these techniques, the complex lipid profile of brain tissue samples mandates purification of lipid extracts before GC/MS analysis; this step is not necessary when using HPLC/MS. We have found that when silica gel chromatography is used for endocannabinoid purification, poor recovery and loss of deuterium from the internal standards lead to inaccurate estimation of endocannabinoid levels. By contrast, purification strategies using C(18) solid-phase extraction permits precise and reproducible GC/MS quantification of endocannabinoids in tissue samples.
Hardison
Sarah
S
Department of Pharmacology, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229, USA.
Weintraub
Susan T
ST
Giuffrida
Andrea
A
D000818:Animals; D001921:Brain / Q000378:metabolism; D063385:Cannabinoid Receptor Modulators / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000592:standards; D003903:Deuterium / Q000737:chemistry; D063388:Endocannabinoids; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods / Q000592:standards; D008055:Lipids / Q000737:chemistry; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
D063385:Cannabinoid Receptor Modulators; D063388:Endocannabinoids; D008055:Lipids; D003903:Deuterium