[Development and validation of laboratory methods for antiretroviral quantitation using HPLC].
OBJECTIVE
Development, validation and error characterization of three analytical methods, by high performance liquid chromatography (HPLC), for the quantitative analysis of ritonavir, saquinavir and abacavir in human plasma.


METHODS
Reagents and instrumentation used, preparation of different standards, sample extraction procedure from biologic matrix, and analytical conditions assayed were detailed to set up three analytical methods. In addition, the validation and the determination of analytical error were also described.


RESULTS
The analytical methods developed for ritonavir, saquinavir and abacavir in human plasma were selective, linear (r2>0.99), precise (coefficients of variation<15%) and accurate (relative errors<15%) over the concentration range selected. The recovery was more than 95% in all methods. Antiretroviral drugs were stable in the storage conditions assayed according to the routine laboratory. The error function discriminated for each analytical method validated was linear in saquinavir (SD=4.84+7.14.10(-2)C) and abacavir (SD=-1.072+3.70.10(-2)C), and non-linear in ritonavir (SD=39.98+2.40.10(-5)C2).


CONCLUSIONS
Three analytical methods were developed and subsequently validated, with validation parameters being within the specifications and attributes of quality established. The error function characterized for each validated method can be used as a heteroscedastic weighting method in the parameter estimation by non-linear regression analysis in clinical pharmacokinetic studies of antiretroviral drugs assayed.
Mariño
E L
EL
Unidad de Farmacia Clínica y Farmacoterapia, Departamento de Farmacia y Tecnología Farmacéutica, Facultad de Farmacia, Universidad de Barcelona, Spain. emarino@ub.edu
Albert
V
V
Ferrer
M S M
MS
Modamio
P
P
Lastra
C F
CF
D019380:Anti-HIV Agents / Q000097:blood*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D015224:Dideoxynucleosides / Q000097:blood*; D016903:Drug Monitoring / Q000379:methods*; D015658:HIV Infections / Q000097:blood / Q000188:drug therapy; D017320:HIV Protease Inhibitors / Q000097:blood; D006801:Humans; D011786:Quality Control; D018894:Reverse Transcriptase Inhibitors / Q000097:blood; D019438:Ritonavir / Q000097:blood*; D019258:Saquinavir / Q000097:blood*; D012680:Sensitivity and Specificity; D000099227:Abacavir
D019380:Anti-HIV Agents; D015224:Dideoxynucleosides; D017320:HIV Protease Inhibitors; D018894:Reverse Transcriptase Inhibitors; D019438:Ritonavir; D019258:Saquinavir; D000099227:Abacavir