title: Development, validation and clinical application of a LC-MS/MS method for the simultaneous quantification of hydroxychloroquine and its active metabolites in human whole blood.
abstract: A rapid, sensitive and specific method using liquid chromatography coupled to tandem mass spectrometry was developed for the simultaneous quantification of hydroxychloroquine (HCQ) and its three major metabolites in human whole blood. The assay, using a sample volume of 100μL, was linear in a dynamic 25-2000ng/mL range (R(2)>0.99) for all four compounds and suitable for the determination of elevated HCQ concentrations up to 20,000ng/mL, after appropriate sample dilution. Inter- and intra-assay precisions were <18.2% and accuracies were between 84% and 113% for any analyte. No matrix effects were observed. The assay was successfully applied to a blood sample obtained from one poisoned patient following a massive HCQ self-ingestion resulting in an estimated concentration of 19,500ng/mL on hospital admission. In this patient, HCQ metabolites were identified and quantified at 1123, 465 and 91ng/mL for monodesethylhydroxychloroquine, desethylchloroquine and bisdesethylchloroquine, respectively. Further investigations are still required to assess the usefulness of the simultaneous measurement of blood concentrations of HCQ and its three active metabolites for monitoring HCQ treatment and managing HCQ poisoning.
authors.affiliation: Laboratoire de Toxicologie Biologique, Hôpital Lariboisière, Assistance Publique-Hôpitaux de Paris, 2 rue Ambroise Paré, 75010 Paris, France.
authors.forename: Marion
authors.initials: M
authors.lastname: Soichot
authors.affiliation: Réanimation Médicale Toxicologique, Hôpital Lariboisière, Assistance Publique-Hôpitaux de Paris, 2 rue Ambroise Paré, 75010 Paris, France; Unité INSERM UMR-S1144, Faculté des Sciences Pharmaceutiques et Biologiques, Université Paris Descartes, 4 avenue de l'Observatoire, 75006 Paris, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Mégarbane
authors.affiliation: Laboratoire de Biochimie, Hôpital Saint Louis, Assistance Publique-Hôpitaux de Paris, 1 avenue Claude Vellefaux, 75010 Paris, France; CNRS UMR 8638-COMETE, Faculté des sciences pharmaceutiques et biologiques, Université Paris Descartes, 4 avenue de l'Observatoire, 75006 Paris, France.
authors.forename: Pascal
authors.initials: P
authors.lastname: Houzé
authors.affiliation: Unité INSERM UMR-S1144, Faculté des Sciences Pharmaceutiques et Biologiques, Université Paris Descartes, 4 avenue de l'Observatoire, 75006 Paris, France.
authors.forename: Lucie
authors.initials: L
authors.lastname: Chevillard
authors.affiliation: Laboratoire de Biochimie, Hôpital Saint Louis, Assistance Publique-Hôpitaux de Paris, 1 avenue Claude Vellefaux, 75010 Paris, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Fonsart
authors.affiliation: Réanimation Médicale Toxicologique, Hôpital Lariboisière, Assistance Publique-Hôpitaux de Paris, 2 rue Ambroise Paré, 75010 Paris, France.
authors.forename: Frédéric J
authors.initials: FJ
authors.lastname: Baud
authors.affiliation: Laboratoire de Toxicologie Biologique, Hôpital Lariboisière, Assistance Publique-Hôpitaux de Paris, 2 rue Ambroise Paré, 75010 Paris, France; CNRS UMR 8638-COMETE, Faculté des sciences pharmaceutiques et biologiques, Université Paris Descartes, 4 avenue de l'Observatoire, 75006 Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Laprévote
authors.affiliation: Laboratoire de Toxicologie Biologique, Hôpital Lariboisière, Assistance Publique-Hôpitaux de Paris, 2 rue Ambroise Paré, 75010 Paris, France; CNRS UMR 8638-COMETE, Faculté des sciences pharmaceutiques et biologiques, Université Paris Descartes, 4 avenue de l'Observatoire, 75006 Paris, France. Electronic address: emmanuel.bourgogne@lrb.aphp.fr.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Bourgogne
mesh_terms: D000328:Adult; D000962:Antimalarials / Q000097:blood* / Q000506:poisoning; D001711:Biotransformation; D002138:Calibration; D002853:Chromatography, Liquid / Q000592:standards; D016903:Drug Monitoring / Q000379:methods; D004355:Drug Stability; D005260:Female; D006801:Humans; D006886:Hydroxychloroquine / Q000031:analogs & derivatives* / Q000097:blood / Q000506:poisoning; D016014:Linear Models; D011041:Poisoning / Q000097:blood / Q000175:diagnosis / Q000628:therapy; D012015:Reference Standards; D015203:Reproducibility of Results; D013406:Suicide, Attempted; D053719:Tandem Mass Spectrometry / Q000592:standards; D013997:Time Factors
chemical_list: D000962:Antimalarials; D006886:Hydroxychloroquine; C049283:cletoquine
keywords: Hydroxychloroquine intoxication; Metabolites; Quantitation; Tandem mass spectrometry; Whole blood