Perpustakaan Digital ITB

Lactose is a disaccharide sugar derived from galactose and glucose that is found in milk. As an important source of carbohydrate and a factor of lactose intolerance, therefore analysis of this sugar content in milk is needed. The objective of this study was to verify the suitability and determination of sugar content in milk sample using HPLC. The milk sample is precipitated by acetonitrile with the volume of ratio of 1:1 and diluted with 100mL of distilled water in volumetric flask. Then, it is filtered through 0.45pm filter membrane. Lactose of milk sample were analyzed in HPLC with Refractive Index detector, NH2 polar bonded phase column, 10um (250mm x 4.6mm I.D.), room temperature, mobile phase of acetonitrile and water (65:35) and flow rate of 1.0mL/min. The method validation showed good linearity of equations for y=106434.5500x-77704.1667 with regression coefficients, r2 of 0.9997. The calculated regression function coefficients (Vx0) for lactose was 0.3763. The percentage recovery of lactose using standard addition method was found to be in a range of 99.11-108.64 and 97.076-102.94%. It is concluded that sugar contents in milk sample can be determined using verified HPLC method with Refractive Index Detector.