Simple and rapid high-performance liquid chromatographic method for nelfinavir, M8 nelfinavir metabolite, ritonavir and saquinavir assay in plasma.
A simple reversed-phase liquid chromatographic method has been developed to determine protease inhibitors concentrations in plasma. Plasma samples (250 micro l) containing protease inhibitors were prepared by a simple deproteinization (recovery: 92, 91, 91 and 90.5% for ritonavir, saquinavir, nelfinavir and M8 nelfinavir metabolite, respectively). Chromatography was accomplished using a Hypersil octadecylsilyl column (100 x 4.6 mm I.D.) and a mobile phase composed of acetonitrile, tetrahydrofuran and dihydrogenophosphate buffer (pH 4) (32:10:58, v/v). Ultraviolet detection at 210 nm was used. The limit of detection was 200 ng/ml for ritonavir, saquinavir, nelfinavir and M8 nelfinavir metabolite. Calibration curves were linear up to 20000 ng/ml, with correlation coefficients better than 0.997 for all compounds. Intra- and inter-day coefficients of variation of the assay were <or=6% for all compounds. This method was used to analyse protease inhibitors plasma concentrations after oral administration within the framework of therapeutic drug monitoring and pharmacokinetic studies in AIDS patients.
Janoly
A
A
Pharmacy Department, Debrousse Hospital, 29 Rue Soeur Bouvier, 69005, Lyon, France.
Bleyzac
N
N
Favetta
P
P
Gagneu
M C
MC
Bourhis
Y
Y
Coudray
S
S
Oger
I
I
Aulagner
G
G
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D017320:HIV Protease Inhibitors / Q000097:blood* / Q000493:pharmacokinetics; D006801:Humans; D019888:Nelfinavir / Q000097:blood* / Q000493:pharmacokinetics; D012015:Reference Standards; D019438:Ritonavir / Q000097:blood* / Q000493:pharmacokinetics; D019258:Saquinavir / Q000097:blood* / Q000493:pharmacokinetics; D012680:Sensitivity and Specificity
D017320:HIV Protease Inhibitors; D019888:Nelfinavir; D019258:Saquinavir; D019438:Ritonavir