Chemical stability of 2-arachidonylglycerol under biological conditions.

Rouzer CA, Ghebreselasie K, Marnett LJ
Chem Phys Lipids. 2002 119 (1-2): 69-82

PMID: 12270674 · DOI:10.1016/s0009-3084(02)00068-3

Recent evidence indicates that 2-arachidonylglycerol (2-AG) is a potent and specific ligand for the central and peripheral cannabinoid receptors. Therefore, the chemical stability of this molecule under biological conditions is of interest. A method for the isolation and detection of 2-AG using HPLC with evaporative light scattering detection is described. The method provides an extraction recovery from aqueous media of 78%, and a limit of detection of 60 ng on column. Incubation of 2-AG in culture medium or biological buffers indicated that it is stable to oxidation and ester hydrolysis for up to 6 h at 37 degrees C. However, gradual disappearance of the compound was noted due to adherence to glass and plastic surfaces. During incubation in RPMI culture medium, 2-AG rearranged to 1(3)-arachidonylglycerol (1(3)-AG) in a first order process with a half-life of 10 min in the absence of serum and 2.3 min in the presence of 10% fetal calf serum. Further studies indicated that the acyl migration reaction is base catalyzed (k(cat)=78,000/min M), and that the reaction is affected slightly by changes in buffer (Tris) concentration and not at all by changes in ionic strength. The results indicate that 2-AG is readily converted to 1(3)-AG under conditions commonly used to study receptor-ligand interactions, findings that have significant implications for the interpretation of relative ligand potency between the two isomers.

MeSH Terms (11)

Arachidonic Acids Buffers Chromatography, High Pressure Liquid Culture Media Drug Stability Endocannabinoids Glycerides Ligands Oxidation-Reduction Receptors, Cannabinoid Receptors, Drug

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