DETERMINATION OF 5-HYDROXYMETHYLFURFURAL IN HONEY BY HIGH PERFORMANCE CHROMATOGRAPHYL METHOD (HPLC-DAD)

Abstract

In this research, high performance liquid chromatography coupled with photodiode array detector (HPLC-DAD) has been reported to quantify 5-hydroxymethyfurfural (HMF) compounds in honey. This method has been validated according to the Association of Official Analytical Chemists (AOAC) and the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines and shown to be specificity, precise, accurate, lower limit of detection (LOD) and lower limit of quantitation (LOQ). The separation was developed by selecting Gemini NX C18 (250 mm × 4.6 mm, 5 µm) column as stationary phase and acetonitril:H2O = 85:15 (v/v) as mobile phase, flow rate of mobile phase was maintained at 1 mL/min at room temperature throughout the experiment, injection volume 50 µL. Quantification was achieved with photodiod arrays (DAD) detection at 284 nm. A good linearity was observed in the concentration range of 2-100 µg/mL with a correlation coefficient (R2 = 0.9996). LOD and LOQ on the sample were 0.25 mg/mL and 0.75 mg/mL, respectively and the recovery of method from 87.3 to 105.8%. Results of analysis of HMF content in 12 samples of honey in the market valued from 47.8 to 522.5 mg/kg. However, the results revealed that 75% of the samples were failing to meet the Codex Alimentarius standards because HMF content in most of samples was above the limit of 80 mg/kg. 

Keywords: Hydroxymethylfurfural (HMF), honey, liquid chromatography (HPLC), determination, process validation.