20 minutes vs. 14 hours - Faster preparation method for glucuronides!
27 Jan 2009
I've been doing some reading regarding methods for deglucuronidation of metabolites, specifically 11-nor-9-carboxy-THC glucuronide (THC-COOH glucuronide) in urine. It seems that there are two processes for the removal of the glucuronic acid group from this analyte. The first process involves the enzyme beta-glucuronidase, which is pretty expensive and takes ~14 hours. The second process involves an alkaline hydrolysis reaction using 10N potassium hydroxide and takes about 20 minutes. While I've only tried this for THC-COOH glucuronide in urine, the alkaline hydrolysis method seems to work well. Based on recoveries vs. THC-COOH (not a glucuronide) extracted in parallel with THC-COOH glucuronide, the efficiency of the alkaline hydrolysis reaction is ~100%. Just a tip: when calculating recoveries for glucuronides, be sure to correct for the mass difference of THC-COOH and THC-COOH glucuronide. Now you can relax for 13 hours! Feel free to contact me for more info.