Quantitative Clinical Diagnostic Analysis of Acetone in Human Blood by HPLC: A Metabolomic Search for Acetone as Indicator.
Using high-performance liquid chromatography (HPLC) and 2,4-dinitrophenylhydrazine (2,4-DNPH) as a derivatizing reagent, an analytical method was developed for the quantitative determination of acetone in human blood. The determination was carried out at 365 nm using an ultraviolet-visible (UV-Vis) diode array detector (DAD). For acetone as its 2,4-dinitrophenylhydrazone derivative, a good separation was achieved with a ThermoAcclaim C18 column (15 cm × 4.6 mm × 3 μm) at retention time (t R) 12.10 min and flowrate of 1 mL min(-1) using a (methanol/acetonitrile) water elution gradient. The methodology is simple, rapid, sensitive, and of low cost, exhibits good reproducibility, and allows the analysis of acetone in biological fluids. A calibration curve was obtained for acetone using its standard solutions in acetonitrile. Quantitative analysis of acetone in human blood was successfully carried out using this calibration graph. The applied method was validated in parameters of linearity, limit of detection and quantification, accuracy, and precision. We also present acetone as a useful tool for the HPLC-based metabolomic investigation of endogenous metabolism and quantitative clinical diagnostic analysis.
Department of Forensic Medicine, Faculty of Medicine, Canakkale Onsekiz Mart University, Terzioglu Campus, 17020 Canakkale, Turkey.
Esin
E
Akgul Kalkan
Department of Leather Engineering, Faculty of Engineering, Ege University, Bornova, 35100 İzmir, Turkey.
Mehtap
M
Sahiner
Department of Clinical Biochemistry, Faculty of Medicine, Canakkale Onsekiz Mart University, Terzioglu Campus, 17020 Canakkale, Turkey.
Dilek
D
Ulker Cakir
Department of Chemical Engineering, Faculty of Engineering, Yuzuncu Yil University, 65080 Van, Turkey.
Duygu
D
Alpaslan
Department of Chemistry, Faculty of Sciences and Arts, Canakkale Onsekiz Mart University, Terzioglu Campus, 17020 Canakkale, Turkey.
Selehattin
S
Yilmaz