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Development and validation of a dried blood spot–HPLC assay for the determination of metronidazole in neonatal whole blood samples

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Abstract

A selective and sensitive high-performance liquid chromatography method with UV detection for the determination of metronidazole in dried blood spots (DBS) has been developed and validated. DBS samples [spiked or patient samples] were prepared by applying blood (30 µL) to Guthrie cards. Discs (6 mm diameter) were punched from the cards and extracted using water containing the internal standard, tinidazole. The extracted sample was chromatographed without further treatment using a reversed phase system involving a Symmetry® C18 (5 µm, 3.9 × 150 mm) preceded by a Symmetry® guard column of matching chemistry and a detection wavelength of 317 nm. The mobile phase comprised acetonitrile/0.01 M phosphate solution (KH2PO4), pH 4.7, 15:85, v/v, with a flow rate of 1 mL/min. The calibration was linear over the range 2.5–50 mg/mL. The limits of detection and quantification were 0.6 and 1.8 µg/mL, respectively. The method has been applied to the determination of 203 DBS samples from neonatal patients for a phamacokinetic/pharmacodynamic study.

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References

  1. Paediatric Formulary Committee (2006) BNF for children. BMJ, London

    Google Scholar 

  2. Jager-Roman E, Doyle PE, Baird-Lambert J, Cvejic M, Buchanan N (1982) J Pediatr 100:651–654

    Article  CAS  Google Scholar 

  3. Hall P, Kaye CM, McIntosh N, Steele J (1983) Arch Dis Child 58:529–531

    Article  CAS  Google Scholar 

  4. Rubenson A, Rosetzsky A (1986) Eur J Clin Pharmacol 29:625–628

    Article  CAS  Google Scholar 

  5. Upadhyaya P, Bhatnagar V, Basu N (1988) J Pediatr Surg 23:263–265

    Article  CAS  Google Scholar 

  6. Sandall JM, Millership JS, Collier PS (2006) McElnay JC J Chromatogr B 839:36–44

    Article  CAS  Google Scholar 

  7. Smyth JM, Darwish M, Collier PS, Millership JS, Petersen S, Halliday HL, McElnay JC (2004) Br J Clin Pharmacol 58:249–258

    Article  CAS  Google Scholar 

  8. Wilcken B, Wiley V (2008) Pathology 40:104–115

    Article  Google Scholar 

  9. AbuRuz S, Millership JS, McElnay JC (2006) J Chromatogr B 832:202–207

    Article  CAS  Google Scholar 

  10. Millership JS, Collier PS, McElnay JC (2003) J Pharm Pharmacol 55:S65

    Google Scholar 

  11. Oliveira EJ, Watson DG, Morton NS (2004) J Pharm Biomed Anal 29:803–809

    Article  Google Scholar 

  12. Kaye CM, Sankey MG, Thomas LA (1980) Br J Clin Pharmacol 9:528–529

    CAS  Google Scholar 

  13. Gibson RA, Lattanzio L, McGee H (1984) Clin Chem 30:784–787

    CAS  Google Scholar 

  14. Okonkwo PO, Eta EI (1988) Life Sci 42:539–545

    Article  CAS  Google Scholar 

  15. Venkateshwaran TG, Stewart JT (1995) J Chromatogr B 672:300–304

    Article  CAS  Google Scholar 

  16. Jessa MJ, Barrett DA, Shaw PN, Spiller RC (1996) J Chromatogr B 677:374–379

    Article  Google Scholar 

  17. Yeung PK, Little R, Jiang Y, Buckley SJ, Pollak PT, Kapoor H, Veldhuyzen van Zanten SJ (1998) J Pharm Biomed Anal 17:1393–1398

    Article  CAS  Google Scholar 

  18. Menelaou A, Somogyi AA, Barclay ML, Bochner F (1999) J Chromatogr B 731:261–266

    Article  CAS  Google Scholar 

  19. Galmier MJ, Frasey AM, Bastide M, Beyssac E, Petit J, Aiache JM, Lartigue-Mattei C (1998) J Chromatogr B 720:239–243

    Article  CAS  Google Scholar 

  20. Stambaugh JE, Feo LG, Manthel RW (1968) J Pharmacol Exp Ther 161:373–381

    CAS  Google Scholar 

  21. Pahkla ER, Koppel T, Saag M, Pahkla R (2005) J Clin Peridontol 32:163–166

    Article  Google Scholar 

  22. do Nascimento TG, de Jesus Oliveira E, Macedo RO (2005) J Pharm Biomed Anal 37:777–783

    Article  Google Scholar 

  23. Mustapha KB, Odunola MT, Garba M, Obodozie O (2006) Afr J Biotechnol 5:1188

    CAS  Google Scholar 

  24. Emami J, Ghassami N, Hamishehkar H (2006) DARU 14:15–21

    CAS  Google Scholar 

  25. Sagan C, Salvador A, Dubreuil D, Poulet PP, Duffaut D, Brumpt I (2005) J Pharm Biomed Anal 38:298–306

    Article  CAS  Google Scholar 

  26. U.S. Department of Health and Human Services, Food and Drug Administration (2001) Guidance for industry: bioanalytical method validation. U.S. Department of Health and Human Services, Food and Drug Administration, Silver Spring, May

    Google Scholar 

  27. The European Agency for the Evaluation of Medicinal Products (1996) ICH harmonised tripartite guideline: validation of analytical procedures: methodology. ICH Topic Q2B (CPMP/ICH/281/95). The European Agency for the Evaluation of Medicinal Products, London.

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Authors and Affiliations

  1. Clinical and Practice Research Group, School of Pharmacy, Medical Biology Centre, Queen’s University Belfast, 97 Lisburn Road, Belfast, BT7 1NN, UK

    Maysa Faisal Suyagh, Godwill Iheagwaram, Prashant Laxman Kole, Jeff Millership, Paul Collier & James C. McElnay

  2. Perinatal Medicine, Royal Maternity Hospital, and Department of Child Health, Queen’s University Belfast, Belfast, BT7 1NN, UK

    Henry Halliday

Authors
  1. Maysa Faisal Suyagh
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  2. Godwill Iheagwaram
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  3. Prashant Laxman Kole
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  4. Jeff Millership
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  5. Paul Collier
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  6. Henry Halliday
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  7. James C. McElnay
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Correspondence to Jeff Millership.

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Suyagh, M.F., Iheagwaram, G., Kole, P.L. et al. Development and validation of a dried blood spot–HPLC assay for the determination of metronidazole in neonatal whole blood samples. Anal Bioanal Chem 397, 687–693 (2010). https://doi.org/10.1007/s00216-010-3571-5

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  • DOI: https://doi.org/10.1007/s00216-010-3571-5

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