Development and validation of a dried blood spot-HPLC assay for the determination of metronidazole in neonatal whole blood samples.
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 microL) 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(R) C18 (5 microm, 3.9 x 150 mm) preceded by a Symmetry(R) guard column of matching chemistry and a detection wavelength of 317 nm. The mobile phase comprised acetonitrile/0.01 M phosphate solution (KH(2)PO(4)), 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 microg/mL, respectively. The method has been applied to the determination of 203 DBS samples from neonatal patients for a phamacokinetic/pharmacodynamic study.
Clinical and Practice Research Group, School of Pharmacy, Medical Biology Centre, Queen's University Belfast, 97 Lisburn Road, Belfast, BT7 1NN, UK.
Maysa Faisal
MF
Suyagh
Godwill
G
Iheagwaram
Prashant Laxman
PL
Kole
Jeff
J
Millership
Paul
P
Collier
Henry
H
Halliday
James C
JC
McElnay
D000890:Anti-Infective Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D007231:Infant, Newborn; D057230:Limit of Detection; D008795:Metronidazole / Q000097:blood*
D000890:Anti-Infective Agents; D008795:Metronidazole