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 metalloproteinase s (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 µl 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 backflush 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.

Pharmacokinetic study of a new synthetic MMP inhibitor (Ro 28-2653) after IV and oral administration of cyclodextrin solutions

“…The ADS sorbents belong to the group of internal surface reversed-phase supports and have been applied successfully to the clean-up of biological samples prior to LC analysis (Hubert et al, 1999a;Souverain et al, 2004;Yu and Westerlund, 1997). The operating conditions are described in a previous paper (Chiap et al, 2005). The method was fully validated according to a novel approach based on accuracy profiles, taking into account the total measurement error (Hubert et al, 1999b(Hubert et al, , 2003(Hubert et al, , 2004.…”

Matrix Metalloproteinase Inhibitors: A Review on Bioanalytical Methods, Pharmacokinetics and Metabolism

“…In order to avoid traditional time-consuming and laborintensive sample preparation procedures, Chiap et al [18] developed a fully automated method using on-line sample clean-up by coupling of a RP-8 ADS pre-column (alkyl diol silica) with restricted access material (RAM) prior to the analytical column by means of a column-switching system to determine Ro 28-2653 in ovine plasma and serum. Because of the appropriate pore diameter (about 6 nm) of the silica particles, macromolecules, such as proteins, having a molecular mass larger than 15 kDa, are excluded in the void volume of the pre-column and are directly flushed into the waste.…”

“…However, the availability of this radiotracer was significantly reduced due to the first-pass effect partly caused by the carboxylic acid group of SAV03 [53]. In order to reduce the first-pass effect of [ 18 The other case is that the metabolites of a drug cause a therapeutic effect. The therapeutic activity of these active metabolites may be weaker or stronger than that of the corresponding parent drugs.…”