Analysis of an anti-inflammatory steroidal drug, difluprednate, in aqueous humor by combination of semi-micro HPLC and column switching method.
A specific and sensitive method for the determination for difluprednate (DFBA) and its metabolite (deacetylated DFBA, DFB) in aqueous humor was developed. DFBA and DFB were initially absorbed on a Pinkerton-type column, then analyzed by high-performance liquid chromatography using a semi-micro column after column switching. Under the optimized conditions, calibration curves for DFBA and DFB showed good linearity over the range of 1.0-50 and 0.5-50 ng/ml, respectively. We applied the method to the analysis of DFBA and DFB in rabbit aqueous humor, and found that DFBA in rabbit aqueous humor 1 h after instillation of 0.002% DFBA ophthalmic emulsion was not detected, but DFB was present at the concentration of 4.3+/-3.1 ng/ml.
Yasueda
Shin-ichi
S
Senju Pharmaceutical Co., Ltd. 2-5-8, Hiranomachi, Chuo-Ku, Osaka 541-0046, Japan.
Kimura
Masako
M
Ohtori
Akira
A
Kakehi
Kazuaki
K
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000032:analysis* / Q000378:metabolism; D001082:Aqueous Humor / Q000737:chemistry* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D005477:Fluprednisolone / Q000031:analogs & derivatives* / Q000032:analysis* / Q000378:metabolism; D011817:Rabbits; D013678:Technology, Pharmaceutical / Q000295:instrumentation / Q000379:methods*
D000893:Anti-Inflammatory Agents; D005477:Fluprednisolone; C015808:difluprednate