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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

UHPLC-MS/MS Method for Determination of Hydroxychloroquine and Its Main Metabolites in Oral Fluid and Whole Blood for Therapeutic Drug Monitoring

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Author(s):
Duarte, Nilo J. C. [1, 2] ; Kupa, Leonard V. K. [3] ; Ferreira-Filho, Julio C. R. [3] ; Fontoura, Nicole [3] ; Chalom, Marc Y. [1] ; Romano, Paschoalina [1, 2] ; Ebner, Persio A. R. [1, 2] ; Silva, Clovis A. A. [3] ; Carvalho, Valdemir M. [4] ; Bonfa, Eloisa [3]
Total Authors: 10
Affiliation:
[1] Univ Sao Paulo, Hosp Clin HCFMUSP, Fac Med, Cent Lab Div, Sao Paulo, SP - Brazil
[2] Univ Sao Paulo, Hosp Clin HCFMUSP, Fac Med, Lab Invest Med LIM 03, Sao Paulo, SP - Brazil
[3] Univ Sao Paulo, Rheumatol Div Hosp Clin HCFMUSP, Fac Med, Sao Paulo, SP - Brazil
[4] Grp Fleury, Sao Paulo, SP - Brazil
Total Affiliations: 4
Document type: Journal article
Source: JOURNAL OF APPLIED LABORATORY MEDICINE; v. 6, n. 4, p. 868-880, JUL 2021.
Web of Science Citations: 0
Abstract

Background: Hydroxychloroquine (HCQ) blood levels are used to monitor efficacy, safety, and patient adherence during treatment. Oral fluid has emerged as an alternative noninvasive, easily accessible, and low-complexity matrix for drug monitoring. However, there is no analytical method to measure HCQ in oral fluid. Therefore, we developed and validated an ultra-high-performance liquid chromatography-tandem mass (UHPLC-MS/MS) method for the measurement of HCQ and its main metabolites in oral fluid and compared to whole blood. Methods: Ten microliters of matrices were used for sample preparation by protein precipitation with acetonitrile followed by online solid phase extraction. The validation process included assessment of lower limit of quantification, linearity, precision, recovery, matrix effect, interferences assessment, carryover, and sample dilution validation. Results: The lower limit of quantification was 50 ng/mL for HCQ and metabolites in both oral fluid and whole blood. The calibration curve was linear from 50 to 2000 ng/mL (r(2) = 0.999). The coefficient of variation for precision assay was 1.2% to 9.7% for intraday and 1.1% to 14.2% for interday for both HCQ and metabolites in oral fluid and whole blood samples at 150, 750, and 1250 ng/mL. The recovery was 85.3% to 118.5% for 150, 750, and 1250 ng/mL of HCQ and metabolites in both oral fluid and whole blood. Dilution factor up to 5-fold was validated for concentrations higher than the upper limit of quantification. Conclusions: The validated method is specific, precise, and accurate to determine the analytical range for therapeutic monitoring of HCQ and its main metabolites in oral fluid and blood. (AU)

FAPESP's process: 15/03756-4 - Assessment of relevance of blood levels of drugs in the monitoring rheumatic autoimmune diseases: safety, effectiveness and adherence to therapy
Grantee:Eloisa Silva Dutra de Oliveira Bonfá
Support Opportunities: Research Projects - Thematic Grants
FAPESP's process: 17/11644-7 - Blood and salivary levels of hydroxychloroquine: relevance in juvenile systemic lupus erythematosus disease (JSLE), in kidney disease in adult SLE activity and in alpha-interferon / TNF production by plasmacytoid dendritic cells
Grantee:Nicole Andressa Gomes Fontoura
Support Opportunities: Scholarships in Brazil - Technical Training Program - Technical Training
FAPESP's process: 17/11516-9 - Assessment of relevance of blood levels of drugs in the monitoring rheumatic autoimmune diseases: safety, effectiveness and adherence to therapy
Grantee:Julio Cesar Rente Ferreira Filho
Support Opportunities: Scholarships in Brazil - Post-Doctoral
FAPESP's process: 19/17272-0 - Relevance of monitoring blood levels compared to salivar levels of drugs used in rheumatic autoimmune diseases: adherence and understanding the possible underlying mechanisms involved in effectiveness and in adverse effects
Grantee:Leonard de Vinci Kanda Kupa
Support Opportunities: Scholarships in Brazil - Post-Doctoral
FAPESP's process: 17/03983-6 - Liquid chromatography coupled to mass spectrometry (2015/03756-4)
Grantee:Eloisa Silva Dutra de Oliveira Bonfá
Support Opportunities: Multi-user Equipment Program