Determination of metformin and its prodrugs in human and rat blood by hydrophilic interaction liquid chromatography.
Simple and specific hydrophilic interaction liquid chromatography (HILIC) method with ultraviolet (UV) detection was developed for the simultaneous determination of highly water-soluble metformin and its more lipophilic prodrugs in human and rat blood samples. The sample preparation was accomplished by precipitating proteins with acetonitrile, which enabled the direct injection of supernatants to the HPLC. Chromatographic separation was performed on an analytical normal phase silica column using a mixture of 0.01 M ammonium acetate pH 5.0 and acetonitrile (40:60, v/v) as a mobile phase at flow rate of 1 ml/min and at the wavelength of 235 nm. The method was validated in terms of specificity, linearity, accuracy, precision, recovery, and analyte stability. The UV-HILIC method was suitable for detecting both metformin and one of its more lipophilic prodrugs simultaneously in human and rat blood samples.
Huttunen
Kristiina M
KM
Department of Pharmaceutical Chemistry, University of Kuopio, P.O. Box 1627, FI-70211 Kuopio, Finland. Kristiina.Huttunen@uku.fi
Rautio
Jarkko
J
Leppänen
Jukka
J
Vepsäläinen
Jouko
J
Keski-Rahkonen
Pekka
P
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D007004:Hypoglycemic Agents / Q000097:blood* / Q000737:chemistry; D008297:Male; D008687:Metformin / Q000097:blood* / Q000737:chemistry; D011355:Prodrugs / Q000493:pharmacokinetics*; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results
D007004:Hypoglycemic Agents; D011355:Prodrugs; D008687:Metformin