Development and Validation of the HPLC Method for Simultaneous Quantification of Glucose and Xylose in Sugar Cane...

A validated rapid high-performance liquid chromatography (HPLC)-refractive index (RI) method was developed for the identification and quantification of glucose and xylose in hydrolyzed bagasse extract. The separation of compounds was achieved on Eurokat® H column (300 × 8 mm, 10 μm) at 75°C, using 0.01 N sulfuric acid solution as mobile phase and 0.6 mL/min as flow rate. The method was validated based on accuracy, precision, linearity, robustness, uncertainty, limit of detection (LOD) and limit of quantification (LOQ). Total chromatographic analysis time per sample was ~11 min. Calibration plots were linear over the concentration ranges 11-100 μg/100 μL for glucose and xylose. The LOD was 0.8 ppm and LOQ was 2.5 ppm. The high recovery and low relative standard deviation confirm the suitability of the method for determination of glucose and xylose in bagasse extract. The proposed HPLC-RI method was accurate, fast and robust and required less run time due to less analytes retention times and allowed optimal energy consumption owing to lower column oven temperature.