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A robust capillary liquid chromatography/tandem mass spectrometry method for quantitation of neuromodulatory endocannabinoids
Author(s) -
Qi Ming,
Morena Maria,
Vecchiarelli Haley A.,
Hill Matthew N.,
Schriemer David C.
Publication year - 2015
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.7277
Subject(s) - chemistry , chromatography , tandem mass spectrometry , mass spectrometry , endocannabinoid system , detection limit , liquid chromatography–mass spectrometry , capillary action , microscale chemistry , tandem , elution , analytical chemistry (journal) , biochemistry , materials science , receptor , mathematics education , mathematics , composite material
Rationale Methods for quantifying anandamide (AEA) and 2‐arachidonoyl glycerol (2‐AG) are needed to support programs investigating molecular mechanisms of the central nervous system. Existing methods, while useful, are not well adapted to efficiently process large numbers of very small tissue samples. A unique challenge involves the disparity in endogenous levels of AEA (pmol/g tissue) and 2‐AG (nmol/g tissue). Methods A simplified one‐step solvent extraction procedure was developed for recovering endocannabinoids from rat brain tissues, and combined with capillary liquid chromatography/tandem mass spectrometry (LC/MS/MS). Various multiple reaction monitoring (MRM)‐based methods were evaluated for limit of detection (LOD) and robustness. Results The optimized simultaneous quantitation method achieves an LOQ of 50 amol for AEA and 25 fmol for 2‐AG, both with a linearity over 3 orders of magnitude, and elution times under 3 min. Accuracy, expressed as relative error (RE), is less than 12% for AEA and less than 6% for 2‐AG. Precision, expressed as relative standard deviation (RSD), is less than 6% for AEA and less than 3% for 2‐AG. Sample handling routines are sufficiently robust to support the automated analysis of thousands of samples from a range of tissue types. Conclusions The microscale method is a sensitive, economical and robust alternative to the larger scale LC/MS methods currently implemented for quantitation of AEA and 2‐AG. Copyright © 2015 John Wiley & Sons, Ltd.