Molecules 2012, 17(12), 14555-14564; doi:10.3390/molecules171214555
Article

Development and Validation of a Bioanalytical Method for Quantification of 2,6-Bis-(4-hydroxy-3-methoxybenzylidene)-cyclohexanone (BHMC) in Rat Plasma

1,†email, 1,†, 2email, 1email, 1email and 1,* email
Received: 10 October 2012; in revised form: 6 December 2012 / Accepted: 7 December 2012 / Published: 7 December 2012
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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.
Keywords: BHMC; curcuminoid analogue; HPLC; pharmacokinetic; plasma extraction
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Lee, Y.Z.; Ming-Tatt, L.; Lajis, N.H.; Sulaiman, M.R.; Israf, D.A.; Tham, C.L. Development and Validation of a Bioanalytical Method for Quantification of 2,6-Bis-(4-hydroxy-3-methoxybenzylidene)-cyclohexanone (BHMC) in Rat Plasma. Molecules 2012, 17, 14555-14564.

AMA Style

Lee YZ, Ming-Tatt L, Lajis NH, Sulaiman MR, Israf DA, Tham CL. Development and Validation of a Bioanalytical Method for Quantification of 2,6-Bis-(4-hydroxy-3-methoxybenzylidene)-cyclohexanone (BHMC) in Rat Plasma. Molecules. 2012; 17(12):14555-14564.

Chicago/Turabian Style

Lee, Yu Z.; Ming-Tatt, Lee; Lajis, Nordin H.; Sulaiman, Mohd R.; Israf, Daud A.; Tham, Chau L. 2012. "Development and Validation of a Bioanalytical Method for Quantification of 2,6-Bis-(4-hydroxy-3-methoxybenzylidene)-cyclohexanone (BHMC) in Rat Plasma." Molecules 17, no. 12: 14555-14564.


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