title: Quantification of aesculin in rabbit plasma and ocular tissues by high performance liquid chromatography using fluorescent detection: application to a pharmacokinetic study.
abstract: A simple and sensitive high performance liquid chromatography method with fluorescence detection (HPLC-FD) was described for the determination of aesculin (AL) at low concentrations in rabbit plasma and ocular tissues. After deproteinization by methanol using pazufloxacin mesilate (PM) as an internal standard (I.S.), supernatants were evaporated to dryness at 40°C under a gentle stream of nitrogen. The residue was reconstituted in mobile phase and a volume of 20μL was injected into the HPLC for analysis. Analytes were separated on an Ultimate XB-C18 column (250mm × 4.6mm i.d., 5μm particle size) and protected by a ODS guard column (10mm × 4.0mm i.d., 5μm particle size), using acetonitrile-0.1% triethylamine in water (adjusted to pH 3.0 using phosphoric acid) (12:88, v/v) as mobile phase with a flow rate of 1.0mL/min. The wavelengths of fluorescence detector (FD) were set at 344nm for excitation and 466nm for emission. The lower limit of quantitation (LOQ) for AL was 0.80ng/mL for plasma and vitreous body, 1.59ng/mL for aqueous humor, and 6.55ng/g for iris and 1.66ng/g for retina. The method was used in the study of AL concentrations in plasma and ocular tissues after topical administration of AL eye drops.
authors.affiliation: College of Materials and Chemistry, Chengdu University of Technology, Sichuan 610059, People's Republic of China.
authors.forename: QiuHong
authors.initials: Q
authors.lastname: Chen
authors.forename: Ying
authors.initials: Y
authors.lastname: Zeng
authors.forename: JianChao
authors.initials: J
authors.lastname: Kuang
authors.forename: Ye
authors.initials: Y
authors.lastname: Li
authors.forename: XiaoHui
authors.initials: X
authors.lastname: Li
authors.forename: Yu
authors.initials: Y
authors.lastname: Zheng
authors.forename: Hua
authors.initials: H
authors.lastname: Hou
authors.forename: ShiXiang
authors.initials: S
authors.lastname: Hou
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics; D001082:Aqueous Humor / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D004929:Esculin / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics*; D005123:Eye / Q000378:metabolism*; D005240:Feasibility Studies; D006207:Half-Life; D007498:Iris / Q000378:metabolism; D057230:Limit of Detection; D008657:Metabolic Clearance Rate; D009883:Ophthalmic Solutions / Q000493:pharmacokinetics*; D011817:Rabbits; D015203:Reproducibility of Results; D012160:Retina / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013678:Technology, Pharmaceutical; D014018:Tissue Distribution; D014822:Vitreous Body / Q000378:metabolism
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D009883:Ophthalmic Solutions; D004929:Esculin