Simultaneous determination of chloroquine, proguanil and their metabolites in human biological fluids by high-performance liquid chromatography.
A reversed-phase ion-pair high-performance liquid chromatographic method with ultraviolet detection is described for the simultaneous measurement of chloroquine, proguanil and their major metabolites in human plasma, erythrocytes and urine. After a liquid-solid extraction on a Bond Elut C8 cartridge, the compounds are separated on a C8 Lichrospher 60 RP select B column by isocratic elution; the mobile phase is water-acetonitrile-methanol (78:28:4, v/v/v) with 0.5 M ammonium formate and 0.075 M perchloric acid. The eluent is monitored with an ultraviolet detector at 254 nm. The lower limits of quantification in plasma are near 6.0 ng ml-1 for chloroquine and near 9.0 ng ml-1 for proguanil. No chromatographic interference can be detected from endogenous compounds or from other antimalarial drugs. The method is accurate and precision is good with inter- and intra-assay relative standard deviations lower than 6.8% for plasma samples. N-(2-6 dichlorobenzylidene amino)guanidine is used as an internal standard. The chromatographic procedure takes 35 min and can be used for therapeutic drug monitoring and clinical studies.
Chaulet
J F
JF
Laboratoire de Biochimie du Professeur Lemontey, Hôpital d'Instruction des Armées Desgenettes, Lyon, France.
Grelaud
G
G
Bellemin-Magninot
P
P
Mounier
C
C
Brazier
J L
JL
D000962:Antimalarials / Q000097:blood / Q000652:urine; D002138:Calibration; D002738:Chloroquine / Q000032:analysis* / Q000097:blood / Q000378:metabolism / Q000652:urine; D002851:Chromatography, High Pressure Liquid; D004912:Erythrocytes / Q000737:chemistry*; D006801:Humans; D002727:Proguanil / Q000032:analysis* / Q000097:blood / Q000378:metabolism / Q000652:urine; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D000962:Antimalarials; D002738:Chloroquine; D002727:Proguanil