Detection of 2-Deoxyglucose by Pre-column Derivation Reversed Phase High Performance Liquid Chromatography
A method was developed for the determination of 2-deoxyglucose(2-DG)by reversed phase high performance liquid chromatography(RP-HPLC) after pre-column derivated with p-aminobenzoic ethyl ester(ABEE).The results showed that the optimum derivation conditions were as follows: reaction temperature was 85 ℃,reaction time was 1.5 h and molar ratio of derivative reagent(ABEE) to monosaccharide(2-DG/Glu) was 25∶1.The derivative was analyzed on a Hypersil ODS 2(250 mm×4.6 mm,5 μ) column with a mixture of acetonitrile to 0.02% trifluoroacetic acid(TFA)(2∶80,V/V) as the mobile phase,at the flow rate of 1.0 mL/min and the detection wavelength at 307 nm.This method has a good linearity at the concentration from 1 mg/L to 700 mg/L(r=0.9999).The limit of detection was 100 μg/L while the quantitative limit was 450 μg/L.The average recovery of 2-DG was 101.7% and the relative standard deviation(RSD) was 0.7%.The inter-day and intra-day precision were both less than 0.7%,This method is of high sensitivity,precision and repetition,indicating that it is suitable for precise quantitative determination of 2-DG.