Determination of the new HIV-protease inhibitor atazanavir by liquid chromatography after solid-phase extraction

Details

Serval ID
serval:BIB_21D53DE5F776
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Determination of the new HIV-protease inhibitor atazanavir by liquid chromatography after solid-phase extraction
Journal
Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Author(s)
Colombo  S., Guignard  N., Marzolini  C., Telenti  A., Biollaz  J., Decosterd  L. A.
ISSN
1570-0232 (Print)
Publication state
Published
Issued date
10/2004
Volume
810
Number
1
Pages
25-34
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Oct 15
Abstract
An HPLC method previously described for the simultaneous assay of amprenavir, ritonavir, indinavir, saquinavir, nelfinavir and efavirenz is proposed here for the simultaneous analysis of the new HIV protease inhibitor atazanavir (ATV) in human plasma, by off-line solid-phase extraction (SPE) followed by HPLC coupled with UV-diode array detection. After viral inactivation by heat (60 degrees C for 60 min), plasma (600 microl) with clozapine (internal standard) is diluted 1 + 1 with phosphate buffer pH 7 and subjected to a SPE on a C18 cartridge. Matrix components are eliminated with 2 x 500 microl of a solution of 0.1% H(3)PO(4) neutralised with NaOH to pH 7. ATV is eluted with 3 x 500 microl MeOH. The resulting eluate is evaporated under nitrogen at room temperature and is reconstituted in 100 microl MeOH/H(2)O 50/50. A 40 microl volume is injected onto a Nucleosil 100-5 microm C18 AB column. ATV is analysed by UV detection at 201 nm using a gradient elution program with solvents constituted of MeCN and phosphate buffer adjusted to pH 5.14. The mobile phase also contains 0.02% sodium heptanesulfonate, enabling an excellent separation of ATV from the other HIV protease inhibitors (PIs) amprenavir, indinavir, saquinavir, ritonavir, lopinavir, nelfinavir and the non-nucleoside reverse transcriptase inhibitors (NNRTIs) efavirenz and nevirapine. The calibration curves are linear up to 10 microg/ml, with a lower limit of quantification of 0.2 microg/ml. The mean absolute recovery of ATV is 96.4 +/- 3.2%. The method is precise with mean inter-day CVs within 1.1-6.1%, and accurate (range of inter-day deviations +0.3 to +2.3%). The method has been validated and is currently applied to the monitoring of ATV in HIV patients.
Keywords
Buffers Calibration Chromatography, High Pressure Liquid Freezing HIV Protease Inhibitors/*analysis/blood Humans Indicators and Reagents Oligopeptides/*analysis/blood Pyridines/*analysis/blood Reference Standards Reproducibility of Results Solutions Specimen Handling Spectrophotometry, Ultraviolet
Pubmed
Web of science
Create date
25/01/2008 11:40
Last modification date
20/08/2019 13:58
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