Validation of a liquid chromatography tandem mass spectrometry method for the simultaneous determination of hydroxychloroquine and metabolites in human whole blood.
Identifieur interne : 000C43 ( Main/Corpus ); précédent : 000C42; suivant : 000C44Validation of a liquid chromatography tandem mass spectrometry method for the simultaneous determination of hydroxychloroquine and metabolites in human whole blood.
Auteurs : Donna Austin ; Catharine John ; Beverley J. Hunt ; Rachel S. CarlingSource :
- Clinical chemistry and laboratory medicine [ 1437-4331 ] ; 2020.
English descriptors
- KwdEn :
- MESH :
- chemical , blood : Hydroxychloroquine.
- chemical , metabolism : Hydroxychloroquine.
- methods : Blood Chemical Analysis.
- Calibration, Chromatography, High Pressure Liquid, Humans, Limit of Detection, Tandem Mass Spectrometry, Time Factors.
Abstract
Objectives Hydroxychloroquine (HCQ) is an anti-malarial and immunomodulatory drug reported to inhibit the Corona virus, SARS-CoV-2, in vitro. At present there is insufficient evidence from clinical trials to determine the safety and efficacy of HCQ as a treatment for COVID-19. However, since the World Health Organisation declared COVID-19 a pandemic in March 2020, the US Food and Drug Administration issued an Emergency Use Authorisation to allow HCQ and Chloroquine (CQ) to be distributed and used for certain hospitalised patients with COVID-19 and numerous clinical trials are underway around the world, including the UK based RECOVERY trial, with over 1000 volunteers. The validation of a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of HCQ and two of its major metabolites, desethylchloroquine (DCQ) and di-desethylchloroquine (DDCQ), in whole blood is described. Methods Blood samples were deproteinised using acetonitrile. HCQ, DCQ and DDCQ were chromatographically separated on a biphenyl column with gradient elution, at a flow rate of 500 μL/min. The analysis time was 8 min. Results For each analyte linear calibration curves were obtained over the concentration range 50-2000 μg/L, the lower limit of quantification (LLOQ) was 13 μg/L, the inter-assay relative standard deviation (RSD) was <10% at 25, 800 and 1750 μg/L and mean recoveries were 80, 81, 78 and 62% for HCQ, d4-HCQ, DCQ and DDCQ, respectively. Conclusion This method has acceptable analytical performance and is applicable to the therapeutic monitoring of HCQ, evaluating the pharmacokinetics of HCQ in COVID-19 patients and supporting clinical trials.
DOI: 10.1515/cclm-2020-0610
PubMed: 32915766
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Validation of a liquid chromatography tandem mass spectrometry method for the simultaneous determination of hydroxychloroquine and metabolites in human whole blood.</title><author><name sortKey="Austin, Donna" sort="Austin, Donna" uniqKey="Austin D" first="Donna" last="Austin">Donna Austin</name><affiliation><nlm:affiliation>Biochemical Sciences, Viapath, Guys & St Thomas' NHSFT, London, UK.</nlm:affiliation></affiliation></author><author><name sortKey="John, Catharine" sort="John, Catharine" uniqKey="John C" first="Catharine" last="John">Catharine John</name><affiliation><nlm:affiliation>Biochemical Sciences, Viapath, Guys & St Thomas' NHSFT, London, UK.</nlm:affiliation></affiliation></author><author><name sortKey="Hunt, Beverley J" sort="Hunt, Beverley J" uniqKey="Hunt B" first="Beverley J" last="Hunt">Beverley J. Hunt</name><affiliation><nlm:affiliation>Thrombosis & Haemophilia Centre, Guy's & St Thomas' NHSFT, London, UK.</nlm:affiliation></affiliation></author><author><name sortKey="Carling, Rachel S" sort="Carling, Rachel S" uniqKey="Carling R" first="Rachel S" last="Carling">Rachel S. Carling</name><affiliation><nlm:affiliation>Biochemical Sciences, Viapath, Guys & St Thomas' NHSFT, London, UK.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>GKT Medical School, Kings College London, London, UK.</nlm:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2020">2020</date><idno type="RBID">pubmed:32915766</idno><idno type="pmid">32915766</idno><idno type="doi">10.1515/cclm-2020-0610</idno><idno type="wicri:Area/Main/Corpus">000C43</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000C43</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Validation of a liquid chromatography tandem mass spectrometry method for the simultaneous determination of hydroxychloroquine and metabolites in human whole blood.</title><author><name sortKey="Austin, Donna" sort="Austin, Donna" uniqKey="Austin D" first="Donna" last="Austin">Donna Austin</name><affiliation><nlm:affiliation>Biochemical Sciences, Viapath, Guys & St Thomas' NHSFT, London, UK.</nlm:affiliation></affiliation></author><author><name sortKey="John, Catharine" sort="John, Catharine" uniqKey="John C" first="Catharine" last="John">Catharine John</name><affiliation><nlm:affiliation>Biochemical Sciences, Viapath, Guys & St Thomas' NHSFT, London, UK.</nlm:affiliation></affiliation></author><author><name sortKey="Hunt, Beverley J" sort="Hunt, Beverley J" uniqKey="Hunt B" first="Beverley J" last="Hunt">Beverley J. Hunt</name><affiliation><nlm:affiliation>Thrombosis & Haemophilia Centre, Guy's & St Thomas' NHSFT, London, UK.</nlm:affiliation></affiliation></author><author><name sortKey="Carling, Rachel S" sort="Carling, Rachel S" uniqKey="Carling R" first="Rachel S" last="Carling">Rachel S. Carling</name><affiliation><nlm:affiliation>Biochemical Sciences, Viapath, Guys & St Thomas' NHSFT, London, UK.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>GKT Medical School, Kings College London, London, UK.</nlm:affiliation></affiliation></author></analytic><series><title level="j">Clinical chemistry and laboratory medicine</title><idno type="eISSN">1437-4331</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Blood Chemical Analysis (methods)</term><term>Calibration (MeSH)</term><term>Chromatography, High Pressure Liquid (MeSH)</term><term>Humans (MeSH)</term><term>Hydroxychloroquine (blood)</term><term>Hydroxychloroquine (metabolism)</term><term>Limit of Detection (MeSH)</term><term>Tandem Mass Spectrometry (MeSH)</term><term>Time Factors (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>Hydroxychloroquine</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Hydroxychloroquine</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Blood Chemical Analysis</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Calibration</term><term>Chromatography, High Pressure Liquid</term><term>Humans</term><term>Limit of Detection</term><term>Tandem Mass Spectrometry</term><term>Time Factors</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Objectives Hydroxychloroquine (HCQ) is an anti-malarial and immunomodulatory drug reported to inhibit the Corona virus, SARS-CoV-2, in vitro. At present there is insufficient evidence from clinical trials to determine the safety and efficacy of HCQ as a treatment for COVID-19. However, since the World Health Organisation declared COVID-19 a pandemic in March 2020, the US Food and Drug Administration issued an Emergency Use Authorisation to allow HCQ and Chloroquine (CQ) to be distributed and used for certain hospitalised patients with COVID-19 and numerous clinical trials are underway around the world, including the UK based RECOVERY trial, with over 1000 volunteers. The validation of a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of HCQ and two of its major metabolites, desethylchloroquine (DCQ) and di-desethylchloroquine (DDCQ), in whole blood is described. Methods Blood samples were deproteinised using acetonitrile. HCQ, DCQ and DDCQ were chromatographically separated on a biphenyl column with gradient elution, at a flow rate of 500 μL/min. The analysis time was 8 min. Results For each analyte linear calibration curves were obtained over the concentration range 50-2000 μg/L, the lower limit of quantification (LLOQ) was 13 μg/L, the inter-assay relative standard deviation (RSD) was <10% at 25, 800 and 1750 μg/L and mean recoveries were 80, 81, 78 and 62% for HCQ, d4-HCQ, DCQ and DDCQ, respectively. Conclusion This method has acceptable analytical performance and is applicable to the therapeutic monitoring of HCQ, evaluating the pharmacokinetics of HCQ in COVID-19 patients and supporting clinical trials.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">32915766</PMID><DateCompleted><Year>2020</Year><Month>11</Month><Day>30</Day></DateCompleted><DateRevised><Year>2021</Year><Month>02</Month><Day>12</Day></DateRevised><Article PubModel="Electronic"><Journal><ISSN IssnType="Electronic">1437-4331</ISSN><JournalIssue CitedMedium="Internet"><Volume>58</Volume><Issue>12</Issue><PubDate><Year>2020</Year><Month>09</Month><Day>13</Day></PubDate></JournalIssue><Title>Clinical chemistry and laboratory medicine</Title><ISOAbbreviation>Clin Chem Lab Med</ISOAbbreviation></Journal><ArticleTitle>Validation of a liquid chromatography tandem mass spectrometry method for the simultaneous determination of hydroxychloroquine and metabolites in human whole blood.</ArticleTitle><Pagination><MedlinePgn>2047-2061</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1515/cclm-2020-0610</ELocationID><Abstract><AbstractText>Objectives Hydroxychloroquine (HCQ) is an anti-malarial and immunomodulatory drug reported to inhibit the Corona virus, SARS-CoV-2, in vitro. At present there is insufficient evidence from clinical trials to determine the safety and efficacy of HCQ as a treatment for COVID-19. However, since the World Health Organisation declared COVID-19 a pandemic in March 2020, the US Food and Drug Administration issued an Emergency Use Authorisation to allow HCQ and Chloroquine (CQ) to be distributed and used for certain hospitalised patients with COVID-19 and numerous clinical trials are underway around the world, including the UK based RECOVERY trial, with over 1000 volunteers. The validation of a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of HCQ and two of its major metabolites, desethylchloroquine (DCQ) and di-desethylchloroquine (DDCQ), in whole blood is described. Methods Blood samples were deproteinised using acetonitrile. HCQ, DCQ and DDCQ were chromatographically separated on a biphenyl column with gradient elution, at a flow rate of 500 μL/min. The analysis time was 8 min. Results For each analyte linear calibration curves were obtained over the concentration range 50-2000 μg/L, the lower limit of quantification (LLOQ) was 13 μg/L, the inter-assay relative standard deviation (RSD) was <10% at 25, 800 and 1750 μg/L and mean recoveries were 80, 81, 78 and 62% for HCQ, d4-HCQ, DCQ and DDCQ, respectively. Conclusion This method has acceptable analytical performance and is applicable to the therapeutic monitoring of HCQ, evaluating the pharmacokinetics of HCQ in COVID-19 patients and supporting clinical trials.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Austin</LastName><ForeName>Donna</ForeName><Initials>D</Initials><AffiliationInfo><Affiliation>Biochemical Sciences, Viapath, Guys & St Thomas' NHSFT, London, UK.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>John</LastName><ForeName>Catharine</ForeName><Initials>C</Initials><AffiliationInfo><Affiliation>Biochemical Sciences, Viapath, Guys & St Thomas' NHSFT, London, UK.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hunt</LastName><ForeName>Beverley J</ForeName><Initials>BJ</Initials><AffiliationInfo><Affiliation>Thrombosis & Haemophilia Centre, Guy's & St Thomas' NHSFT, London, UK.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Carling</LastName><ForeName>Rachel S</ForeName><Initials>RS</Initials><Identifier Source="ORCID">https://orcid.org/0000-0003-1988-3088</Identifier><AffiliationInfo><Affiliation>Biochemical Sciences, Viapath, Guys & St Thomas' NHSFT, London, UK.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>GKT Medical School, Kings College London, London, UK.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D023361">Validation Study</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2020</Year><Month>09</Month><Day>13</Day></ArticleDate></Article><MedlineJournalInfo><Country>Germany</Country><MedlineTA>Clin Chem Lab Med</MedlineTA><NlmUniqueID>9806306</NlmUniqueID><ISSNLinking>1434-6621</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>4QWG6N8QKH</RegistryNumber><NameOfSubstance UI="D006886">Hydroxychloroquine</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D001774" MajorTopicYN="N">Blood Chemical Analysis</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D002138" MajorTopicYN="N">Calibration</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002851" MajorTopicYN="Y">Chromatography, High Pressure Liquid</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006886" MajorTopicYN="N">Hydroxychloroquine</DescriptorName><QualifierName UI="Q000097" MajorTopicYN="Y">blood</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D057230" MajorTopicYN="N">Limit of Detection</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D053719" MajorTopicYN="Y">Tandem Mass Spectrometry</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D013997" MajorTopicYN="N">Time Factors</DescriptorName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="Y">human whole blood</Keyword><Keyword MajorTopicYN="Y">hydroxychloroquine</Keyword><Keyword MajorTopicYN="Y">liquid chromatography tandem mass spectrometry</Keyword><Keyword MajorTopicYN="Y">method validation</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2020</Year><Month>04</Month><Day>28</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2020</Year><Month>08</Month><Day>29</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2020</Year><Month>9</Month><Day>12</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>12</Month><Day>1</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2020</Year><Month>9</Month><Day>11</Day><Hour>17</Hour><Minute>12</Minute></PubMedPubDate></History><PublicationStatus>epublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">32915766</ArticleId><ArticleId IdType="doi">10.1515/cclm-2020-0610</ArticleId><ArticleId IdType="pii">cclm-2020-0610</ArticleId></ArticleIdList><ReferenceList><Title>References</Title><Reference><Citation>Kalia, S, Dutz, J. 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