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Multidimensional Liquid Chromatography using Restricted Access Media Columns: Applications to in vitro metabolism of drugs

Grant number: 09/01304-8
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): July 01, 2009
Effective date (End): June 30, 2012
Field of knowledge:Physical Sciences and Mathematics - Chemistry - Analytical Chemistry
Principal Investigator:Quezia Bezerra Cass
Grantee:Pedro Henrique Viana de Carvalho
Home Institution: Centro de Ciências Exatas e de Tecnologia (CCET). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil


An analytical process involves four main steps: (a) sample preparation; (b) analytical separation; (c) detection system and, (d) data processing. In bioanalytical field, the sample preparation is often considered the time-limiting step. In the past years, several efforts have been conducted in developing faster sample preparation and, consequently, high throughput analysis. Among different strategies, the use of special extraction sorbents, such as restricted access media (RAM), represents an attractive approach, specially allowing the direct and repetitive injection of complex matrices in chromatographic systems. These special sorbents, when integrated in a liquid chromatography system, lead to automation, simplification in sample preparation, reduction of analytes lost; often associated with off line extraction procedures and, consequently speeding up of the sample preparation process.Therefore, this project propose the preparation of different restricted access media bovine serum albumin columns, immobilized on C18, C8, cyano, phenyl, cation exchange or anion exchange derivatized silica supports and their retention evaluation for antituberculosis drugs, as well as, their exclusion efficiency of the microsomes proteins, obtained from liver of rats.Initially, a multidimensional chromatographic method will be developed and validated, using direct injection of biological samples, for the quantification of rifampicin, isoniazid and pirazinamid in microsomal fraction. After that, special attention will be given to the application of the developed method for investigation of the effects of protein-calorie malnutrition on in vitro antituberculosis drugs metabolism in microsomal fractions by multidimensional chromatography.