title: A sensitive method for determination of COL-3, a chemically modified tetracycline, in human plasma using high-performance liquid chromatography and ultraviolet detection.
abstract: COL-3, 6-deoxy-6-desmethyl-4-desdimethylamino-tetracycline, is a matrix metalloproteinase inhibitor currently in clinical development. A HPLC-UV method to quantitate COL-3 in human plasma was developed. COL-3 was extracted from plasma using solid-phase extraction cartridges. COL-3 is separated on a Waters Symmetry Shield RP8 (3.9 mm x150 mm, 5 microm) column with EDTA (0.001 M) in sodium acetate (0.01 M, pH 3.5)-acetonitrile mobile phase using a gradient profile at a flow rate of 1 ml/min for 22 min. Carryover was eliminated by using an extended needle wash of methanol:acetonitrile:dichloromethane (1:1:1, v/v/v). Detection of COL-3 and the internal standard, chrysin, was observed at 350 nm. COL-3 and chrysin elute at 8.9 and 9.9 min, respectively. The lower limit of quantitation in human plasma of COL-3 was 75 ng/ml, linearity was observed from 75 to 10,000 ng/ml. A 30,000 ng/ml sample that was diluted 1:50 with plasma was accurately quantitated. This method is rapid, widely applicable, and suitable for quantifying COL-3 in patient samples enabling further clinical pharmacology characterization of COL-3.
authors.affiliation: Division of Medical Oncology, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Bunting-Blaustein Cancer Research Building, 1650 Orleans Street, Room 1M90, Baltimore, MD 21231, USA. mrudek2@jhmi.edu
authors.forename: Michelle A
authors.initials: MA
authors.lastname: Rudek
authors.forename: Carol
authors.initials: C
authors.lastname: Hartke
authors.forename: Yelena
authors.initials: Y
authors.lastname: Zabelina
authors.forename: Ming
authors.initials: M
authors.lastname: Zhao
authors.forename: Pamela
authors.initials: P
authors.lastname: New
authors.forename: Sharyn D
authors.initials: SD
authors.lastname: Baker
mesh_terms: D000704:Analysis of Variance; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D004356:Drug Storage; D006801:Humans; D007202:Indicators and Reagents; D011786:Quality Control; D012015:Reference Standards; D013056:Spectrophotometry, Ultraviolet; D013754:Tetracyclines / Q000032:analysis / Q000097:blood*
chemical_list: D007202:Indicators and Reagents; D013754:Tetracyclines; C117155:tetracycline CMT-3