Development and validation of a fast U-HPLC-fluorescent method for the quantification of hydroxychloroquine and its metabolites in patients with lupus.
Identifieur interne : 000021 ( France/Analysis ); précédent : 000020; suivant : 000022Development and validation of a fast U-HPLC-fluorescent method for the quantification of hydroxychloroquine and its metabolites in patients with lupus.
Auteurs : Gaëlle Noé [France] ; Zahir Amoura [France] ; David Combarel [France] ; Lesli Lori [France] ; Nadine Tissot [France] ; Aguila Seycha [France] ; Christian Funck-Brentano [France] ; Noel Zahr [France]Source :
- Therapeutic Drug Monitoring [ 0163-4356 ] ; 2019.
Abstract
BACKGROUND:Hydroxychloroquine (HCQ) is approved for the treatment of systemic lupus erythematosus (SLE). Therapeutic drug monitoring (TDM) of HCQ is necessary to detect non-adherence and to improve treatment efficacy in SLE patients. Liquid chromatographic-tandem mass spectroscopy and HPLC-fluorescent methods are currently used to measure whole blood concentrations of hydroxychloroquine and its two main metabolites desethylhydroxychloroquine (DHCQ) and desethylchloroquine (DCQ) in SLE patients. This study reports the development and validation of an ultra-high performance liquid chromatography (UHPLC) method with fluorescence detection for the simultaneous quantification of HCQ and its metabolites in whole blood.METHODS:After adding chloroquine (internal standard) to the samples, a single-step protein precipitation and a subsequent filtration were employed for blood sample preparation. Analytes were separated under isocratic elution on a U-HPLC RP18 column with a total run time of seven minutes. The mobile phase consisted of piperazine buffer (46.4 mM, pH=9.8) and acetonitrile (68:32, v/v), was delivered at a flow rate of 0.4 mL/min. Fluorescence excitation and emission wavelengths were 335 and 390 nm, respectively. Assay performance parameters were evaluated per FDA bioanalytical guidelines.RESULTS:The calibration curve was linear from 125 to 4000 ng/mL for HCQ. The lower limit of quantification was 10 ng/mL for all analytes. For HCQ, DCQ and DHCQ, accuracies and imprecisions ranged from -7.90 to 7.85% and 1.14 to 8.78%, respectively.CONCLUSION:A sensitive, accurate and fast U-HPLC-fluorescent method was validated and successfully applied to quantify whole blood concentrations to perform TDM of HCQ in pediatric and adult lupus patients.
Url:
DOI: 10.1097/FTD.0000000000000614
Affiliations:
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Hal:hal-02153408Le document en format XML
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Hôpitaux de Paris (AP-HP)</orgName> <orgName type="acronym">APHP</orgName> <desc> <address> <country key="FR"></country> </address> </desc> </org></tutelle><tutelle active="#struct-353778" type="direct"><org type="institution" xml:id="struct-353778" status="VALID"> <orgName>CHU Pitié-Salpêtrière [APHP]</orgName> <desc> <address> <addrLine>47-83 Boulevard de l'Hôpital, 75013 Paris</addrLine> <country key="FR"></country> </address> <ref type="url">http://www.aphp.fr/contenu/hopital-universitaire-pitie-salpetriere-0</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>France</country></affiliation></author><author><name sortKey="Tissot, Nadine" sort="Tissot, Nadine" uniqKey="Tissot N" first="Nadine" last="Tissot">Nadine Tissot</name><affiliation wicri:level="1"><hal:affiliation type="department" xml:id="struct-41012" status="VALID"> <orgName>Service de pharmacologie biologique [CHU Pitié-Salpâtrière]</orgName> <desc> <address> <addrLine>47-83, boulevard de l'Hôpital 75651 PARIS Cedex 13</addrLine> <country key="FR"></country> </address> </desc> <listRelation> <relation active="#struct-300068" type="direct"></relation> <relation active="#struct-353778" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-300068" type="direct"><org type="regroupinstitution" xml:id="struct-300068" status="VALID"> <orgName>Assistance publique - Hôpitaux de Paris (AP-HP)</orgName> <orgName type="acronym">APHP</orgName> <desc> <address> <country key="FR"></country> </address> </desc> </org></tutelle><tutelle active="#struct-353778" type="direct"><org type="institution" xml:id="struct-353778" status="VALID"> <orgName>CHU Pitié-Salpêtrière [APHP]</orgName> <desc> <address> <addrLine>47-83 Boulevard de l'Hôpital, 75013 Paris</addrLine> <country key="FR"></country> </address> <ref type="url">http://www.aphp.fr/contenu/hopital-universitaire-pitie-salpetriere-0</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>France</country></affiliation></author><author><name sortKey="Seycha, Aguila" sort="Seycha, Aguila" uniqKey="Seycha A" first="Aguila" last="Seycha">Aguila Seycha</name><affiliation wicri:level="1"><hal:affiliation type="department" xml:id="struct-41012" status="VALID"> <orgName>Service de pharmacologie biologique [CHU Pitié-Salpâtrière]</orgName> <desc> <address> <addrLine>47-83, boulevard de l'Hôpital 75651 PARIS Cedex 13</addrLine> <country key="FR"></country> </address> </desc> <listRelation> <relation active="#struct-300068" type="direct"></relation> <relation active="#struct-353778" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-300068" type="direct"><org type="regroupinstitution" xml:id="struct-300068" status="VALID"> <orgName>Assistance publique - Hôpitaux de Paris (AP-HP)</orgName> <orgName type="acronym">APHP</orgName> <desc> <address> <country key="FR"></country> </address> </desc> </org></tutelle><tutelle active="#struct-353778" type="direct"><org type="institution" xml:id="struct-353778" status="VALID"> <orgName>CHU Pitié-Salpêtrière [APHP]</orgName> <desc> <address> <addrLine>47-83 Boulevard de l'Hôpital, 75013 Paris</addrLine> <country key="FR"></country> </address> <ref type="url">http://www.aphp.fr/contenu/hopital-universitaire-pitie-salpetriere-0</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>France</country></affiliation></author><author><name sortKey="Funck Brentano, Christian" sort="Funck Brentano, Christian" uniqKey="Funck Brentano C" first="Christian" last="Funck-Brentano">Christian Funck-Brentano</name><affiliation wicri:level="1"><hal:affiliation type="department" xml:id="struct-457063" status="VALID"> <orgName>CIC Paris Est</orgName> <desc> <address> <addrLine>Bâtiment Antonin Gosset47-83, boulevard de l'Hôpital75651 PARIS Cedex 13</addrLine> <country key="FR"></country> </address> </desc> <listRelation> <relation active="#struct-300068" type="direct"></relation> <relation name="CIC1421" active="#struct-303623" type="direct"></relation> <relation active="#struct-353778" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-300068" type="direct"><org type="regroupinstitution" xml:id="struct-300068" status="VALID"> <orgName>Assistance publique - Hôpitaux de Paris (AP-HP)</orgName> <orgName type="acronym">APHP</orgName> <desc> <address> <country key="FR"></country> </address> </desc> </org></tutelle><tutelle name="CIC1421" active="#struct-303623" type="direct"><org type="institution" xml:id="struct-303623" status="VALID"> <idno type="IdRef">026388278</idno> <orgName>Institut National de la Santé et de la Recherche Médicale</orgName> <orgName type="acronym">INSERM</orgName> <desc> <address> <addrLine>101, rue de Tolbiac, 75013 Paris </addrLine> <country key="FR"></country> </address> <ref type="url">http://www.inserm.fr</ref> </desc> </org></tutelle><tutelle active="#struct-353778" type="direct"><org type="institution" xml:id="struct-353778" status="VALID"> <orgName>CHU Pitié-Salpêtrière [APHP]</orgName> <desc> <address> <addrLine>47-83 Boulevard de l'Hôpital, 75013 Paris</addrLine> <country key="FR"></country> </address> <ref type="url">http://www.aphp.fr/contenu/hopital-universitaire-pitie-salpetriere-0</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>France</country></affiliation></author><author><name sortKey="Zahr, Noel" sort="Zahr, Noel" uniqKey="Zahr N" first="Noel" last="Zahr">Noel Zahr</name><affiliation wicri:level="1"><hal:affiliation type="department" xml:id="struct-41012" status="VALID"> <orgName>Service de pharmacologie biologique [CHU Pitié-Salpâtrière]</orgName> <desc> <address> <addrLine>47-83, boulevard de l'Hôpital 75651 PARIS Cedex 13</addrLine> <country key="FR"></country> </address> </desc> <listRelation> <relation active="#struct-300068" type="direct"></relation> <relation active="#struct-353778" type="direct"></relation> </listRelation> <tutelles><tutelle active="#struct-300068" type="direct"><org type="regroupinstitution" xml:id="struct-300068" status="VALID"> <orgName>Assistance publique - Hôpitaux de Paris (AP-HP)</orgName> <orgName type="acronym">APHP</orgName> <desc> <address> <country key="FR"></country> </address> </desc> </org></tutelle><tutelle active="#struct-353778" type="direct"><org type="institution" xml:id="struct-353778" status="VALID"> <orgName>CHU Pitié-Salpêtrière [APHP]</orgName> <desc> <address> <addrLine>47-83 Boulevard de l'Hôpital, 75013 Paris</addrLine> <country key="FR"></country> </address> <ref type="url">http://www.aphp.fr/contenu/hopital-universitaire-pitie-salpetriere-0</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>France</country></affiliation></author></analytic><idno type="DOI">10.1097/FTD.0000000000000614</idno><series><title level="j">Therapeutic Drug Monitoring</title><idno type="ISSN">0163-4356</idno><imprint><date type="datePub">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"> <p>BACKGROUND:Hydroxychloroquine (HCQ) is approved for the treatment of systemic lupus erythematosus (SLE). Therapeutic drug monitoring (TDM) of HCQ is necessary to detect non-adherence and to improve treatment efficacy in SLE patients. Liquid chromatographic-tandem mass spectroscopy and HPLC-fluorescent methods are currently used to measure whole blood concentrations of hydroxychloroquine and its two main metabolites desethylhydroxychloroquine (DHCQ) and desethylchloroquine (DCQ) in SLE patients. This study reports the development and validation of an ultra-high performance liquid chromatography (UHPLC) method with fluorescence detection for the simultaneous quantification of HCQ and its metabolites in whole blood.METHODS:After adding chloroquine (internal standard) to the samples, a single-step protein precipitation and a subsequent filtration were employed for blood sample preparation. Analytes were separated under isocratic elution on a U-HPLC RP18 column with a total run time of seven minutes. The mobile phase consisted of piperazine buffer (46.4 mM, pH=9.8) and acetonitrile (68:32, v/v), was delivered at a flow rate of 0.4 mL/min. Fluorescence excitation and emission wavelengths were 335 and 390 nm, respectively. Assay performance parameters were evaluated per FDA bioanalytical guidelines.RESULTS:The calibration curve was linear from 125 to 4000 ng/mL for HCQ. The lower limit of quantification was 10 ng/mL for all analytes. For HCQ, DCQ and DHCQ, accuracies and imprecisions ranged from -7.90 to 7.85% and 1.14 to 8.78%, respectively.CONCLUSION:A sensitive, accurate and fast U-HPLC-fluorescent method was validated and successfully applied to quantify whole blood concentrations to perform TDM of HCQ in pediatric and adult lupus patients.</p> </div></front></TEI><affiliations><list><country><li>France</li></country></list><tree><country name="France"><noRegion><name sortKey="Noe, Gaelle" sort="Noe, Gaelle" uniqKey="Noe G" first="Gaëlle" last="Noé">Gaëlle Noé</name></noRegion><name sortKey="Amoura, Zahir" sort="Amoura, Zahir" uniqKey="Amoura Z" first="Zahir" last="Amoura">Zahir Amoura</name><name sortKey="Combarel, David" sort="Combarel, David" uniqKey="Combarel D" first="David" last="Combarel">David Combarel</name><name sortKey="Funck Brentano, Christian" sort="Funck Brentano, Christian" uniqKey="Funck Brentano C" first="Christian" last="Funck-Brentano">Christian Funck-Brentano</name><name sortKey="Lori, Lesli" sort="Lori, Lesli" uniqKey="Lori L" first="Lesli" last="Lori">Lesli Lori</name><name sortKey="Seycha, Aguila" sort="Seycha, Aguila" uniqKey="Seycha A" first="Aguila" last="Seycha">Aguila Seycha</name><name sortKey="Tissot, Nadine" sort="Tissot, Nadine" uniqKey="Tissot N" first="Nadine" last="Tissot">Nadine Tissot</name><name sortKey="Zahr, Noel" sort="Zahr, Noel" uniqKey="Zahr N" first="Noel" last="Zahr">Noel Zahr</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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