Published:Journal of Chromatographic Science,
ISSN 0021-9665Volume
41, Number 2, February 2003, pp. 80-86
Performance Analysis of a Rapid HPLC Determination with the
Solvent Demixing Extraction of HIV Antiproteases and Efavirenz in Plasma
M. Cociglio, D. Hillaire-Buys*, H. Peyrière, and R.
Alric
CHU Montpellier, Service de Pharmacologie Médicale, UF de Pharmacomonitorage,
Hôpital Lapeyronie, F-34295 Montpellier cedex 5, France
A rapid and simple high-performance liquid chromatographic method
without internal standardization is evaluated for the drug-level monitoring
of most marketed antiproteases and efavirenz. Following plasma deproteinization
with acetonitrile, the analytes are extracted into the solvent while it is
demixed by the addition of a saturating amount of neutral salt. The organic
supernatant is diluted by half with water up to the polarity of the mobile
phase before being injected. The isocratic mobile phase is unbuffered water–acetonitrile
(52:48), and the stationary phase is LiChrospher 100 RP-8 (5 µm). Analytes
are eluted between 4 min (amprenavir and indinavir) and 20 min (nelfinavir).
A spreadsheet program including analysis of variance (ANOVA) and regression
is used for both the overall validation of milligrams-per-liter determinations
and the performance evaluation of analytical steps from chromatographic raw
data. Extraction shows acceptable 5% repeatability and nearly 100% recovery,
although it is somewhat concentration-dependent. The calibration function is
better fitted by bilogarithmic than arithmetical regression, and the ANOVA
of raw data is found quite predictive of the quality of the final determinations.
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