A sensitive LC-MS/MS methotrexate assay capable of assessing adherence to methotrexate therapy in rheumatoid arthritis
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Malcolm P. McTaggart
,
James Bluett
Abstract
Objectives
To develop a sensitive liquid chromatography-tandem mass spectrometry method capable of measuring serum methotrexate in patients with rheumatoid arthritis to assess adherence to drug treatment.
Methods
Isotopically labelled internal standard and deionised water were added to sample prior to solid phase extraction using a Waters Oasis Max ion-exchange 96-well plate. Following extraction, samples were analysed by LC-MS/MS on a TQS-micro mass spectrometer.
Results
Mean recovery was 107 % for four different concentrations of methotrexate spiked into seven patient samples, whilst post extraction spiking gave a mean recovery of 100 %. Between-batch and within-batch CVs were ≤6 % at three different concentrations of methotrexate in fresh frozen plasma. Mean bias was <5 % for between-batch and within batch analysis at three different weighed in concentrations of methotrexate certified reference material. The lower limit of quantification of the assay was 0.1 nmol/L with linearity up to approximately 100 nmol/L. Dilution linearity studies were used to validate the dilution of patient samples prior to analysis. There was no significant interference in the method from lipaemia, haemolysis or icterus.
Conclusions
A sensitive LC-MS/MS assay for methotrexate has been developed and validated. The method has been used to measure methotrexate adherence in patient samples from clinical trials and could be used in future research to assess the ability of the assay as a biofeedback intervention to improve adherence to methotrexate therapy.
Funding source: The MIRA study is funded by The Association of Physicians of Great Britain & Ireland and supported by Versus Arthritis (21754) and the NIHR Manchester Biomedical Research Centre (NIHR203308). N/A
Award Identifier / Grant number: N/A
Acknowledgments
We thank laboratory staff in biochemistry at Wythenshawe Hospital for providing a productive working environment allowing this work to be carried out. Additionally we thank UKNEQAS for providing samples free of charge.
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Research funding: The MIRA study is funded by The Association of Physicians of Great Britain & Ireland and supported by Versus Arthritis (21754) and the NIHR Manchester Biomedical Research Centre (NIHR203308).
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Author contributions: Professor Brian Keevil developed the method. Dr Malcolm McTaggart performed the other laboratory work including method validation and clinical trial work. Dr Malcolm Mctaggart produced the first draft of the manuscript. Dr James Bluett provided the clinical trial samples. All authors edited the manuscript and contributed to the final version.
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Competing interests: No conflict of interest.
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Informed consent: Informed consent was obtained from all individuals included in the MIRA study.
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Ethical approval: Ethical approval for MIRA was given by the North West – Haydock Research Ethics Committee (19/NW/0047) and all participants provided written informed consent. The method validation did not require ethical approval.
References
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2. National Institute for Health and Care Excellence. Rheumatoid arthritis in adults: management. NG100; 2018. https://www.nice.org.uk/guidance/ng100/resources/rheumatoid-arthritis-in-adults-management-pdf-66141531233989 [Accessed 1 Feb 2023].Search in Google Scholar
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Supplementary Material
This article contains supplementary material (https://doi.org/10.1515/cclm-2023-0350).
© 2023 Walter de Gruyter GmbH, Berlin/Boston
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Articles in the same Issue
- Frontmatter
- Editorial
- Biological management of diabetes mellitus, the laboratory medicine specialist and the patient
- Reviews
- Remote HbA1c testing via microsampling: fit for purpose?
- The effect of hormonal contraceptive therapy on clinical laboratory parameters: a literature review
- Opinion Paper
- Continuous glucose monitoring has an increasing role in pre-symptomatic type 1 diabetes: advantages, limitations, and comparisons with laboratory-based testing
- IFCC Paper
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- Determination of cortisone and cortisol in human scalp hair using an improved LC-MS/MS-based method
- Mild hypophosphatasia may be twice as prevalent as previously estimated: an effective clinical algorithm to detect undiagnosed cases
- Combined deficient response to polysaccharide-based and protein-based vaccines predicts a severe clinical phenotype
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- Letters to the Editor
- In-house diagnostic devices under the EU IVDR and unwanted side-effects of intentional transparency
- Use of thyroid function tests in urine: a position statement of the Belgian Thyroid Club
- State of the art of measurement uncertainty of serum ferritin
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