title: Automated method for the determination of a new matrix metalloproteinase inhibitor in ovine plasma and serum by coupling of restricted access material for on-line sample clean-up to liquid chromatography.
abstract: A fully automated liquid chromatographic method was developed for the determination of Ro 28-2653, a new synthetic inhibitor of matrix metalloproteinases (MMPs), in ovine serum and plasma. The method was based on the coupling of a pre-column packed with restricted access material, namely LiChrospher RP-8 ADS (alkyl diol silica), for sample clean-up to an analytical column containing octyl silica stationary phase. One hundred microl of biological sample, to which 2-propanol was automatically added, were injected onto the ADS pre-column, which was then washed with a washing liquid consisting of a mixture of 25 mM phosphate buffer (pH 7.0) and acetonitrile (90:10; v/v) for 10 min. By rotation of the switching valve, the analyte was then eluted in the back-flush mode with the LC mobile phase composed of a mixture of acetonitrile and 25 mM phosphate buffer (pH 7.0) (57:43; v/v). The UV detection was performed at 395 nm. The main parameters likely to influence the sample preparation technique were investigated. The method was then validated over a concentration range from 17.5 to 1950 ng/ml, the first concentration level corresponding to the lower limit of quantitation. At this concentration level, the mean bias and the R.S.D. value for intermediate precision were -2.4% and 4.2%, respectively.
authors.affiliation: Department of Analytical Pharmaceutical Chemistry, Institute of Pharmacy, University of Liège, CHU, B36, B-4000 Liège 1, Belgium.
authors.forename: P
authors.initials: P
authors.lastname: Chiap
authors.forename: M
authors.initials: M
authors.lastname: Piette
authors.forename: B
authors.initials: B
authors.lastname: Evrard
authors.forename: F
authors.initials: F
authors.lastname: Frankenne
authors.forename: B
authors.initials: B
authors.lastname: Christiaens
authors.forename: G
authors.initials: G
authors.lastname: Piel
authors.forename: D
authors.initials: D
authors.lastname: Cataldo
authors.forename: J-M
authors.initials: JM
authors.lastname: Foidart
authors.forename: L
authors.initials: L
authors.lastname: Delattre
authors.forename: J
authors.initials: J
authors.lastname: Crommen
authors.forename: Ph
authors.initials: P
authors.lastname: Hubert
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D061965:Matrix Metalloproteinase Inhibitors; D011480:Protease Inhibitors / Q000097:blood*; D012680:Sensitivity and Specificity; D012756:Sheep; D013056:Spectrophotometry, Ultraviolet
chemical_list: D061965:Matrix Metalloproteinase Inhibitors; D011480:Protease Inhibitors