Determination of lopinavir and nevirapine by high-performance liquid chromatography after solid-phase extraction: application for the assessment of their transplacental passage at delivery
Détails
ID Serval
serval:BIB_FE146774859E
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Determination of lopinavir and nevirapine by high-performance liquid chromatography after solid-phase extraction: application for the assessment of their transplacental passage at delivery
Périodique
Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
ISSN
1570-0232 (Print)
Statut éditorial
Publié
Date de publication
07/2002
Volume
774
Numéro
2
Pages
127-40
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Jul 15
Research Support, Non-U.S. Gov't --- Old month value: Jul 15
Résumé
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.
Mots-clé
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
Création de la notice
25/01/2008 11:41
Dernière modification de la notice
25/08/2023 22:20