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Présenté au congrès hiv therapy
Evaluation of assays for anti-HIV drugs in serum in France :
preliminary results of a quality control survey.
C. Palette1,2, A.M. Taburet3, P. Marquet4, B. Diquet5, G. Peytavin6, A. Vassault1,7.
1 : ASQUALAB Hôpital Corentin Celton - 92130 Issy les Moulineaux, 2 : Hôpital A. Mignot (Biochimie-Pharmacologie) - 78157 Le Chesnay, 3 :
Hôpital Bicêtre (Pharmacie) - 94270 Le Kremlin Bicêtre, Hôpital Dupuytren (Pharmacologie) - 87042 Limoges, 5 : Hôpital Pitié-Salpétrière
(Pharmacologie)- 75013 Paris, 6 : Hôpital Bichat (Pharmacie) - 75018 Paris, Hôpital Necker (Biochimie) - 75015 Paris.
The recent development of new HIV treatment combining various protease inhibitors and non-nucleoside reverse transcriptase inhibitors (NNRTI)
has emphasised the necessity for drug determination in serum for therapeutic drug monitoring (TDM). The need for inter-laboratory quality control
survey has been first raised by the members of the pharmacology group of the French Research Agency against AIDS (ANRS). ASQUALAB,
organiser of QC survey in the field of TDM, has been in charge of the development, design and organisation of this survey.
The aim of this study is to list and evaluate the assays current used for the different HIV drugs through the results of an external quality control. This
exercise is intended to lead to improvement of the analytical methods used and possibly to the development of standard techniques.
MATERIALS, DESIGN AND METHODS
Human serum pool was spiked with the five VIH-protease inhibitors (Amprenavir, Indinavir, Nelfinavir,
Table 2 : Number of participants for each drug
Ritonavir and Saquinavir) and two non-nucleoside
Number of labs not performing Number of unreported results
Efavirenz), then lyophilised (Randox). Twelve samples
were shipped to 27 laboratories during February 2 000.
The theoretical concentration ranges for the first survey
Some of laboratories sent no result.
Table 3 : Characteristics of the methods used
Table 1 : Drugs targed concentrations
H.P.L.C. has been used by all the participants.
Table 4 : Mean, SD and CV for each drug
This first two samples had to be assayed before end of
Consensus mean determination
: data were computed
according the usual procedure of QCs. Outliers,
identified by a statistical treatment, were omitted from
Protease inhibitors are the most often determined drugs
because of their short half-life, drug interactions and
high inter-individual variability in metabolism in human. May be NNRTIs are seldom assayed because of lessinter-individual variability and difficulty in obtaining
For Ritonavir V02, Saquinavir V01, Nevirapine V01,
Efavirenz V01, a limited number of laboratories were
The inter-laboratory variability illustrates the difficulties encountered by the
able to participate : the concentrations were found minus
laboratories in the implementation of new HPLC methods when no internal nor
than the limit of quantification for most of the other
external QC are available. This first exercise demonstrates the need for development
laboratories. Limits differs widely from one laboratory
of standard methods and will help improve assay quality. The spiked concentrations
were chosen to include both the therapeutic range of the anti-HIV drugs and the
In this first round-robin, the criterium for quality
lowest concentrations found in pharmaceutical studies. This is a useful tool for
assessment was to be within ± 20% of the
increasing inter-laboratory consistency of the results and consequently to improve
interlaboratory mean for the two levels.
ASQUALAB : Assurance de Qualité des Laboratoires
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