Analytical and Bioanalytical Chemistry

, Volume 397, Issue 2, pp 687–693 | Cite as

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

  • Maysa Faisal Suyagh
  • Godwill Iheagwaram
  • Prashant Laxman Kole
  • Jeff Millership
  • Paul Collier
  • Henry Halliday
  • James C. McElnay
Original Paper

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.

Keywords

Dried blood spots Guthrie card Metronidazole HPLC 

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Maysa Faisal Suyagh
    • 1
  • Godwill Iheagwaram
    • 1
  • Prashant Laxman Kole
    • 1
  • Jeff Millership
    • 1
  • Paul Collier
    • 1
  • Henry Halliday
    • 2
  • James C. McElnay
    • 1
  1. 1.Clinical and Practice Research Group, School of Pharmacy, Medical Biology CentreQueen’s University BelfastBelfastUK
  2. 2.Perinatal Medicine, Royal Maternity Hospital, and Department of Child HealthQueen’s University BelfastBelfastUK

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