Characterization of Hemicellulose by High-Performance Liquid Chromatography

The method of characterization by high-performance liquid chromatography [HPLC] is as follows 25 mg of hemicellulose was put into 50 mL volumetric flask and pure water was added to the mark. The mixture was shaken and then filtered [first few milliliters of filtrate was discarded]. The solution was then filtered through a membrane filter of 0.2 pm cellulose nitrate. Then about 100 mL solution was injected into the HPLC system via a loop injector with a 20 mL, using an isocratic elution system with distilled water with a mobile phase, flow rate 0.8 mL/min. The detection was made using a UV detector at a wavelength of 280 nm [34]. 

The chromatogram recorded and performed a qualitative analysis of the area, peak height, and retention time corn cobs hemicellulose. 

From Fig. 8.1 it can be seen that the yield of hemicellulose is a hydrophilic compound as it is soluble in the distilled water. The HPLC testing using aquabidest as the mobile phase, using a C18 column, with a flow rate of 0.8 cm /min and ultraviolet light detector at a wavelength of 280 nm produces retention time of 1.8, peak height of 5.11558 and symmetrical area of 0.82 [34]. 

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