A novel dried blood spot-LCMS method for the quantification of methotrexate polyglutamates as a potential marker for methotrexate use in children

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Hawwa, Ahmed F. ; Albawab, Abdelqader ; Rooney, Madeleine ; Wedderburn, Lucy R. ; Beresford, Michael W. ; McElnay, James C.
  • Journal: (vol: 9)
  • Related identifiers: doi: 10.1371/journal.pone.0089908, pmc: PMC3934981
  • Subject: Chemistry | Research Article | Biology | Diagnostic Medicine | Test Evaluation | Rheumatology | Medicine | Analytical Chemistry | Pediatric Epidemiology | Epidemiology | Epidemiological Methods | Population Biology | Pediatrics | JUVENILE IDIOPATHIC ARTHRITIS, ACUTE LYMPHOBLASTIC-LEUKEMIA, RHEUMATOID-ARTHRITIS, LIQUID-CHROMATOGRAPHY, HPLC DETERMINATION, PHARMACOKINETICS, THERAPY, DERMATOMYOSITIS, SPECTROMETRY, IMMUNOASSAY

Objective: Development and validation of a selective and sensitive LCMS method for the determination of methotrexate polyglutamates in dried blood spots (DBS). <br/><br/>Methods: DBS samples [spiked or patient samples] were prepared by applying blood to Guthrie cards which was then dried at room temperature. The method utilised 6-mm disks punched from the DBS samples (equivalent to approximately 12 μl of whole blood). The simple treatment procedure was based on protein precipitation using perchloric acid followed by solid phase extraction using MAX cartridges. The extracted sample was chromatographed using a reversed phase system involving an Atlantis T3-C18 column (3 μm, 2.1x150 mm) preceded by Atlantis guard column of matching chemistry. Analytes were subjected to LCMS analysis using positive electrospray ionization. <br/><br/>Key Results: The method was linear over the range 5-400 nmol/L. The limits of detection and quantification were 1.6 and 5 nmol/L for individual polyglutamates and 1.5 and 4.5 nmol/L for total polyglutamates, respectively. The method has been applied successfully to the determination of DBS finger-prick samples from 47 paediatric patients and results confirmed with concentrations measured in matched RBC samples using conventional HPLC-UV technique. <br/><br/>Conclusions and Clinical Relevance: The methodology has a potential for application in a range of clinical studies (e.g. pharmacokinetic evaluations or medication adherence assessment) since it is minimally invasive and easy to perform, potentially allowing parents to take blood samples at home. The feasibility of using DBS sampling can be of major value for future clinical trials or clinical care in paediatric rheumatology. © 2014 Hawwa et al.
  • References (28)
    28 references, page 1 of 3

    1. Ravelli A, Martini A (2007) Juvenile idiopathic arthritis. Lancet 369 (9563):767- 778.

    2. Ramanan AV, Campbell-Webster N, Ota S, Parker S, Tran D, et al. (2005) The effectiveness of treating juvenile dermatomyositis with methotrexate and aggressively tapered corticosteroids. Arthritis and rheumatism 52 (11):3570- 3578.

    3. Murray KJ, Lovell DJ (2002) Advanced therapy for juvenile arthritis. Best practice & research Clinical rheumatology 16 (3):361-378.

    4. Choy EH, Isenberg DA (2002) Treatment of dermatomyositis and polymyositis. Rheumatology 41 (1):7-13.

    5. Pilkington CA, Wedderburn LR (2005) Paediatric idiopathic inflammatory muscle disease: recognition and management. Drugs 65 (10):1355-1365.

    6. Cutolo M, Sulli A, Pizzorni C, Seriolo B, Straub RH (2001) Anti-inflammatory mechanisms of methotrexate in rheumatoid arthritis. Annals of the rheumatic diseases 60 (8):729-735.

    7. Bannwarth B, Pehourcq F, Schaeverbeke T, Dehais J (1996) Clinical pharmacokinetics of low-dose pulse methotrexate in rheumatoid arthritis. Clinical pharmacokinetics 30 (3):194-210.

    8. Dalrymple JM, Stamp LK, O'Donnell JL, Chapman PT, Zhang M, et al. (2008) Pharmacokinetics of oral methotrexate in patients with rheumatoid arthritis. Arthritis and rheumatism 58 (11):3299-3308.

    9. Schmiegelow K, Bjork O, Glomstein A, Gustafsson G, Keiding N, et al. (2003) Intensification of mercaptopurine/methotrexate maintenance chemotherapy may increase the risk of relapse for some children with acute lymphoblastic leukemia. Journal of clinical oncology: official journal of the American Society of Clinical Oncology 21 (7):1332-1339.

    10. Dervieux T, Hancock M, Evans W, Pui CH, Relling MV (2002) Effect of methotrexate polyglutamates on thioguanine nucleotide concentrations during continuation therapy of acute lymphoblastic leukemia with mercaptopurine. Leukemia 16 (2):209-212.

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