HPLC-MS/MS studies of brimonidine in rabbit aqueous humor by microdialysis.

Aim: The pharmacokinetic study of the brimonidine tartrate in situ gel in the anterior chamber of the rabbit eye was studied by microdialysis technique, and samples were analyzed by HPLC-MS/MS. Materials & methods: It was monitored in ESI mode at transition 291.9→212.0 and 296.0→216.0 for brimonidine and internal standard, respectively. Acetonitrile and 0.1% aqueous formic acid (50:50, v/v) were used as the mobile phase at 0.4 ml/min. Results & conclusion: It showed a good linear correlation between 5 and 5000 ng/ml in microdialysis solution, and the inter- and intra-day precision (relative standard deviation) was less than 4.0%. The pharmacokinetic study showed that the AUC(0-t) of in situ gel was 3.5-times than that of eyedrops, which significantly improve the bioavailability of brimonidine.