Simultaneous determination of monodesethylchloroquine, chloroquine, cycloguanil and proguanil on dried blood spots by reverse-phase liquid chromatography

被引:57
作者
Lejeune, Delphine
Souletie, Isabelle
Houze, Sandrine
Le Bricon, Thierry
Le Bras, Jacques
Gourmel, Bernard
Houze, Pascal
机构
[1] Hop St Louis, Lab Biochim A, F-75010 Paris, France
[2] Hop Bichat Claude Bernard, AP HP, Lab Parasitol, F-75010 Paris, France
[3] Hop Bichat Claude Bernard, AP HP, Ctr Natl Reference Paludisme, F-75010 Paris, France
[4] Univ Paris 05, CNRS, UMR 7157, INSERM,U705,Hop F Widal, F-75475 Paris 10, France
[5] Univ Paris 07, Hop F Widal, F-75475 Paris 10, France
关键词
chloroquine; proguanil; dried blood spot; reverse-phase chromatography;
D O I
10.1016/j.jpba.2006.09.036
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A method for simultaneous analysis of chloroquine, proguanil and their metabolites from a whole blood sample (80 mu L) dried on a filterpaper was developed. Sample preparation included a liquid extraction from the filter paper, followed by a solid-phase extraction (C18 Bond Elut (R) cartridge). Separation was obtained by reverse-phase liquid chromatography (HPLC) using a gradient elution on an X-Terra (R) column; UV detection was made at 254 nm. This assay was linear between 150 and 2500 ng mL(-1) for chloroquine (and metabolite) and 300 and 2500 ng mL(-1) for proguanil and cycloguanil. The lower limit of quantification was close to 50 ng mL-1 for chloroquine (and its metabolite) and 100 ng mL-1 for proguanil (and its metabolite). No chromatographic interference from endogenous compounds or other tested anti-malarial drugs was evidenced. Chromatographic separation takes about 40 min with a coefficient of variation below 10.3% for within- and between-batch precision. The paper sampling method was validated in 10 healthy subjects treated by Savarine (R). The stability of compounds and metabolites on the filter paper was evaluated at four temperatures (-20, +4, 20 and 50 degrees C) and for 1, 5 and 20 days. Cycloguanil concentrations were not influenced by storage conditions, whereas, high temperatures and prolonged storage decreased chloroquine and proguanil levels. The proposed HPLC assay is accurate, precise and cost-effective; it can be used for pharmacokinetic and epidemiological studies on anti-malarial treatments. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:1106 / 1115
页数:10
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