A Purification and Isolation of an Endocannabinoid
Ashby, Taylor J.
2-Arachidonoylglycerol, or 2-AG, is an endocannabinoid that primarily functions in the central nervous system. Cayman Chemical, located in Ann Arbor, Michigan, supplies 2-AG for research and medical purposes. The product is sold by Cayman Chemical at 95% purity, or in a 9:1 ratio of 2-AG to 1-AG. Due to its chemical instability, 2-AG quickly isomerizes into 1-AG, 1-Arachidonoylglycerol, which has the same molecular formula and R groups, save for the position of two alcohol groups. The isomerization of 2-AG into 1-AG happens both inside and outside of the body, typically inhibiting the large role it plays in activating cannabinoid receptors in the brain and central nervous system. A purification method was developed using normal phase preparative High Performance Liquid Chromatography to separate and isolate the 2-AG peak to at least 95% purity so that it can be sold for further use. The sample was initially 77% pure and was dissolved in 75% Heptane and 25% ethanol, which was the same ratio used for the mobile phase. There were two columns used to complete the purification, one being a nonpolar normal phase preparative column that was free of acid and one of the same type that had been exposed to acid. The sample was injected at 200 microliters and adjusted to create the best resolution, peak shape, and peak height as the project continued. The final purity was an average of 97% pure 2-Arachidonoylglycerol.
v, 20 p.
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