Determination of ofloxacin and moxifloxacin and their penetration in human aqueous and vitreous humor by using high-performance liquid chromatography fluorescence detection.
Information on comparing the penetration of ofloxacin and moxifloxacin in the human eye is unavailable, although these two antibiotics are commonly used in ophthalmic surgery. There is a need for a rapid, reliable, and sensitive methodology for their determination in ocular fluids. We developed a robust HPLC procedure with fluorescence detection for simultaneous analysis of ofloxacin and moxifloxacin in human and rabbit aqueous and vitreous samples. The linearity of the method ranged from 10 ng/ml to 100 microg/ml with r(2) > 0.996. Most inter- and intrabatch imprecision was about 5% (range 1.6-7.6%), recoveries between 95 and 104%, and accuracies between 93 and 104% at 0.1 and 1 microg/ml. The detection limits of both compounds were 10 ng/ml (0.028 nmol/ml for ofloxacin and 0.023 nmol/ml for moxifloxacin). No sample treatment was necessary for aqueous humor and only acetonitrile precipitation was required for vitreous humor. The chromatographic time was short, 22 min. We applied this method to study penetrations of ofloxacin and moxifloxacin in aqueous and vitreous humors of human and rabbits. There was no significant difference of penetration between the two antibiotics into aqueous and vitreous but ofloxacin was found at significantly higher concentrations in aqueous than in vitreous. We also detected contralateral transfer of the antibiotics in rabbit eyes.
Chan
Kwok Ping
KP
Department of Ophthalmology & Visual Sciences, The Chinese University of Hong Kong, China.
Chu
Kai On
KO
Lai
Wico Wai-Kwan
WW
Choy
Kwong Wai
KW
Wang
Chi Chiu
CC
Lam
Dennis Shun-Chiu
DS
Pang
Chi Pui
CP
D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis*; D001082:Aqueous Humor / Q000737:chemistry*; D001372:Aza Compounds / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005453:Fluorescence; D024841:Fluoroquinolones; D006801:Humans; D000077266:Moxifloxacin; D015242:Ofloxacin / Q000032:analysis*; D011804:Quinolines / Q000032:analysis*; D011817:Rabbits; D014822:Vitreous Body / Q000737:chemistry*
D000900:Anti-Bacterial Agents; D001372:Aza Compounds; D024841:Fluoroquinolones; D011804:Quinolines; D015242:Ofloxacin; D000077266:Moxifloxacin