Buprenorphine concentration measurements

A commercially available forensic enzyme linked immunosorbent assay kit (Neogen, Lexington, KY) was used to measure buprenorphine concentrations in the dissolution experiments and in the sera of mice in the pharmacokinetic study similar to others^18,26 Standard curves were created using concentrations of 30 ng/ml, 15 ng/ml, 7.5 ng/ml, 3.75 ng/ml, 0 ng/ml of buprenorphine hydrochloride (Buprenex^®). Samples were diluted in supplied diluent buffer before use. Absorbance was read at 450 nm using an iMark Microplate Reader (Biorad, Hercules, CA).