Measurement of hydroxychloroquine in blood from SLE patients using LC-HRMS—evaluation of whole blood, plasma, and serum as sample matrices
Open accessAbstract
Background: Hydroxychloroquine (HCQ) is the standard of care in the treatment of systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and other inflammatory rheumatic diseases and potentially for the treatment in COVID-19 patients. Determination of HCQ for therapeutic drug monitoring (TDM) can be performed in whole blood (WB), serum, and plasma. Direct comparisons of WB, serum, and plasma levels of HCQ in patients with SLE have not previously been reported. We describe a method for the determination of HCQ in human blood using liquid chromatography-high-resolution mass spectrometry (LC-HRMS) and compare the suitability of the three sample matrices. Methods: A method for the determination of HCQ in human blood using LC-HRMS was developed, validated, and applied for the determination of HCQ levels in WB, serum, and plasma from 26 SLE patients. The reproducibility of the method, in the three matrices, was evaluated using quality control samples and repeated preparations and measurements of patient samples. The performance of the developed method for HCQ measurement in serum was further evaluated by comparison with two previously reported extraction methods. Results: The performance of the presented method demonstrated high accuracy and precision. A large range of HCQ concentrations was observed for the SLE patients in all three matrices (WB, serum, and plasma). The mean levels in WB were approximately two-fold the levels in serum and plasma (813 ng/mL compared to 436 ng/mL and 362 ng/mL, respectively). Spiked quality controls showed high reproducibility for all matrices (coefficient of variation, CV, approx. 5%), whereas in patient samples, equally high-precision was only found using WB as the matrix (CV 3%). The CV for serum and plasma was 14% and 39%, respectively. Two alternative methods applied to serum samples did not demonstrate improved precision. Conclusions: A LC-HRMS method for the measurement of HCQ in human blood was developed and validated. Whole blood was found to be the superior sample matrix in terms of sample reproducibility. Thus, whole blood samples should be used for HCQ analysis when patients are monitored for HCQ treatment effects. The assay is in clinical use to monitor levels of HCQ in patients.
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Whole blood is preferred over plasma for measuring hydroxychloroquine because the drug binds to red blood cells and is sequestered in leukocytes.
“Hydroxychloroquine blood levels are preferred over plasma levels due to binding of hydroxychloroquine to red blood cells and its lysosomal sequestration in leukocytes (28).”
From Association of Higher Hydroxychloroquine Blood Levels With Reduced Thrombosis Risk in Systemic Lupus Erythematosus (2021) · cites this paper as [28]
The Conclusions (page unavailable) identify whole blood as the preferred matrix for hydroxychloroquine measurement. The Discussion (page unavailable) attributes its advantage to better reproducibility in patient samples; it does not establish red-blood-cell binding or lysosomal sequestration in leukocytes as the reasons for that preference. (LLM, conf=0.88)
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Petri, M., Konig, M. F., Li, J., & Goldman, D. W. (2021) · Arthritis & Rheumatology · doi:10.1002/art.41621 · Cited by 65 · see citing works
- Authors
- Carlsson, H., Hjorton, K., Abujrais, S., Rönnblom, L., Åkerfeldt, T., & Kultima, K.
- Affiliations
- Uppsala University (SE)
- Corresponding author
- Kultima, Kim
- Venue
- Arthritis Research & Therapy
- Published
- 2020-06-01
- Volume
- 22
- Issue
- 1
- Pages
- 125-125
- ISSN
- 1478-6354 (ISSN-L)
- 1478-6362
- PMC
- PMC7261520
- OpenAlex
- W3026022611
- Cited by
- 48
- References
- 38
- Citations by year
- 2026: 1 · 2025: 4 · 2024: 7 · 2023: 9 · 2022: 11 · 2021: 14 · 2020: 1 · 2018: 1
- FWCI
- 4.53
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Arthritis Research & Therapy (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Uppsala University Publications (Uppsala University) (submittedVersion, other-oa)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Region Uppsala (n.a.)
- the Swedish Rheumatism Association
- King Gustaf V's 80-years Foundation
- the Swedish Society of Medicine
- Topics
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
- Health Sciences › Medicine › Rheumatology › Systemic Lupus Erythematosus Research
- Life Sciences › Immunology and Microbiology › Immunology › Biosimilars and Bioanalytical Methods
- Keywords
- hydroxychloroquine, systemic lupus erythematosus, therapeutic drug monitoring, whole blood, plasma, serum, sample matrix comparison, LC-HRMS, assay validation
- MeSH terms
- Betacoronavirus, COVID-19, SARS-CoV-2, Adult, Aged, Chromatography, Liquid, Female, Humans, Hydroxychloroquine, Lupus Erythematosus, Systemic, Male, Middle Aged, Plasma, Pneumonia, Viral, Mass Spectrometry, Drug Monitoring, Coronavirus Infections, Serum, Young Adult, Pandemics
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