Please use this identifier to cite or link to this item: https://doi.org/10.1016/j.jchromb.2003.10.066
Title: Reversed-phase liquid chromatography method to determine COL-3, a matrix metalloproteinase inhibitor, in biological samples
Authors: Li, J.
Huynh, H.
Chan, E. 
Keywords: COL-3
Metalloproteinase inhibitor
Issue Date: 25-Jan-2004
Citation: Li, J., Huynh, H., Chan, E. (2004-01-25). Reversed-phase liquid chromatography method to determine COL-3, a matrix metalloproteinase inhibitor, in biological samples. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences 799 (2) : 311-321. ScholarBank@NUS Repository. https://doi.org/10.1016/j.jchromb.2003.10.066
Abstract: A reversed-phase high-performance liquid chromatographic (HPLC) method with ultraviolet (UV) detection was developed and validated for the quantification of 6-deoxy-6-demethyl-4-dedimethylamino-tetracycline (COL-3), a matrix metalloproteinase (MMPs) inhibitor, in rat serum. This simple, sensitive, rapid and reproducible assay involved a preliminary serum deproteinization by adding a mixture of acetonitrile-methanol-0.5M oxalic acid (70:20:10 (v/v)), as the combined precipitant and metal blocking agent, into serum samples (2:1 (v/v)). An aliquot (20μl) of the supernatant was injected into the HPLC system linked to a Waters XTerra™ RP18 column (150mm×4.6mm i.d., particle size 5μm). The compound was eluted by a mixture of acetonitrile-methanol-0.01M oxalic acid (40:10:50 (v/v), pH 2.00), as the mobile phase, and detected at the wavelength of 350nm. The total running time was 10min. The low and high concentration calibration curves were linear in the range of 50-1200ng/ml and 1200-12,000ng/ml, respectively. The intra- and inter-day coefficients of variation at three quality control concentrations of 100, 1200, and 12,000ng/ml were all less than 6%, while the percent error ranged from -2.5 to 6.6%. The limit of quantitation (LOQ) for COL-3 in serum was 50ng/ml. This assay was successfully employed to study the serum concentration-time profiles of COL-3 after its intravenous and oral administration in rats. The method with some minor modifications in sample pretreatment was also applicable to the determination of the concentrations of COL-3 in rat bile, urine and feces. © 2003 Elsevier B.V. All rights reserved.
Source Title: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
URI: http://scholarbank.nus.edu.sg/handle/10635/106308
ISSN: 15700232
DOI: 10.1016/j.jchromb.2003.10.066
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