Determination of lopinavir and nevirapine by high-performance liquid chromatography after solid-phase extraction: application for the assessment of their transplacental passage at delivery

Details

Serval ID
serval:BIB_FE146774859E
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Determination of lopinavir and nevirapine by high-performance liquid chromatography after solid-phase extraction: application for the assessment of their transplacental passage at delivery
Journal
Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Author(s)
Marzolini  C., Beguin  A., Telenti  A., Schreyer  A., Buclin  T., Biollaz  J., Decosterd  L. A.
ISSN
1570-0232 (Print)
Publication state
Published
Issued date
07/2002
Volume
774
Number
2
Pages
127-40
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Jul 15
Abstract
An adaptation of the HPLC method previously described for the simultaneous assay of amprenavir, ritonavir, indinavir, saquinavir, nelfinavir and efavirenz after solid-phase extraction is proposed here for the separate analysis of the newer PI lopinavir (LPV) and the NNRTI nevirapine (NVP). After viral inactivation by heat (60 degrees C for 60 min), plasma (600 microl), with clozapine added as internal standard, is diluted 1+1 with phosphate buffer pH 7 and subjected to a solid-phase extraction on a C(18) 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. LPV and NVP are eluted with 3 x 500 microl MeOH. The resulting eluate is evaporated under nitrogen at room temperature and is reconstituted in 100 microl MeOH 50%. A 40-microl volume is injected onto a Nucleosil 100, 5 microm C(18) AB column. LPV and NVP are analysed separately using a gradient elution program with solvents constituted of MeCN and phosphate buffer adjusted to pH 5.07 and containing 0.02% sodium heptanesulfonate. LPV and NVP are detected by UV at 201 and 282 nm, respectively. The calibration curves are linear up to 10 microg/ml. The mean absolute recovery of LPV and NVP is 91% and 88%, respectively. The method is precise with mean inter-day C.V.s within 2.1-6.6% and 0.9-1.7% for LPV and NVP, and accurate (range of inter-day deviations -1.1 to +2.4%, and -1.9 to +0.8%, for LPV and NVP, respectively). The method has been validated and is currently applied to the monitoring of LPV and NVP in HIV patients, and has been notably applied in a study aimed at assessing the extent of transplacental passage of nevirapine and PIs, notably lopinavir, at the time of delivery in pregnant HIV-infected women.
Keywords
Calibration Chromatography, High Pressure Liquid/*methods Female HIV Protease Inhibitors/*blood Humans *Maternal-Fetal Exchange Nevirapine/*blood Pregnancy Pyrimidinones/*blood Reverse Transcriptase Inhibitors/*blood Sensitivity and Specificity
Pubmed
Web of science
Create date
25/01/2008 11:41
Last modification date
20/08/2019 17:28
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