title: Development and validation of a bioanalytical method for quantification of 2,6-bis-(4-hydroxy-3-methoxybenzylidene)-cyclohexanone (BHMC) in rat plasma.
abstract: A sensitive and accurate high performance liquid chromatography with ultraviolet/visible light detection (HPLC-UV/VIS) method for the quantification of 2,6-bis-(4-hydroxy-3-methoxybenzylidene)-cyclohexanone (BHMC) in rat plasma was developed and validated. BHMC and the internal standard, harmaline, were extracted from plasma samples by a simple liquid-liquid extraction using 95% ethyl acetate and 5% methanol. Plasma concentration of BHMC and internal standard were analyzed by reversed phase chromatography using a C₁₈ column (150 × 4.6 mm I.D., particle size 5 µm) and elution with a gradient mobile phase of water and methanol at a flow rate of 1.0 mL/min. Detection of BHMC and internal standard was done at a wavelength of 380 nm. The limit of quantification was 0.02 µg/mL. The calibration curves was linear (R² > 0.999) over the concentration range of 0.02-2.5 µg/mL. Intra- and inter-day precision were less than 2% coefficient of variation. The validated method was then applied to a pharmacokinetic study in rats by intravenous administration of BHMC at a single dose of 10 mg/kg. Pharmacokinetic parameters such as half-life, maximum plasma concentration, volume of distribution, clearance and elimination rate constant for BHMC were calculated.
authors.affiliation: Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
authors.forename: Yu Zhao
authors.initials: YZ
authors.lastname: Lee
authors.forename: Lee
authors.initials: L
authors.lastname: Ming-Tatt
authors.forename: Nordin Hj
authors.initials: NH
authors.lastname: Lajis
authors.forename: Mohd Roslan
authors.initials: MR
authors.lastname: Sulaiman
authors.forename: Daud Ahmad
authors.initials: DA
authors.lastname: Israf
authors.forename: Chau Ling
authors.initials: CL
authors.lastname: Tham
mesh_terms: D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D003474:Curcumin / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000493:pharmacokinetics; D003512:Cyclohexanones / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification* / Q000493:pharmacokinetics; D051381:Rats; D012015:Reference Standards; D013056:Spectrophotometry, Ultraviolet / Q000379:methods
chemical_list: C548340:2,6-bis-4-(hydroxy-3-methoxybenzylidene)cyclohexanone; D003512:Cyclohexanones; C036468:cyclohexanone; D003474:Curcumin