PENGARUH PENYIAPAN SAMPEL PADA PENGEMBANGAN METODE ANALISISLAKTOSA DALAM SUSU FORMULA SECARA KROMATOGRAFI CAIR KINERJATINGGI

• Main Author: Mardiana
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/79057
• Institution: Institut Teknologi Bandung

Lactose is considered to provide beneficial effects for intestinal physiology, including prebiotic effects, and enhancement of water, sodium and calcium absorption. Therefore, the IEG considers it prudent to include lactose in infant milk formula. High Performance Liquid Chromatography (HPLC) with refractive index detector is a technique for the determination of lactose. Sample preparation before injecting to the chromatograph is a critical step in analysis with HPLC. This research performed three different sample preparation procedures based on procedures had been used by Ferreira et al., Chavez-Servin et al. and Merck, and Coppa et al.. The aims of this research was to test the performance of the sample preparation procedures to the determination of lactose content in infant milk formula accurately, and to developed and validated the method of lactose content determination in infant milk formula using HPLC with index refraction detector. Lactose concentration was determined using HPLC technique under following optimum conditions: refractive index detector, NH2 column Inertsil 5 µm, 250 x 4.6 mm i.d, mobile phase ACN-H2O (65:35) with the flow rate 1 mL/minute. Under this conditions, the system suitability criteria parameters were as follow: resolution factor (Rs) = 5.38, tailing factor (?) = 1.11, and selectivity factor (?) = 3.57. Coefficient variance (CV) of injection repeatability for retention time, Area Under Curve, and height of chromatogram were 0.09%, 1.63%, and 1.23%, respectively. The method gave calibration curve of y = iv 364048.15x-17235.22 with r = 0,9998; regression coefficient variance (Vxo) = 1.25%; limit of detection (LOD) = 0.12 mg/mL; and limit of quantitation (LOQ) = 0.35 mg/mL. Precision, stated as CV, of sample preparation procedure I, II, and III were 15.07%, 3.06%, 1.93%, respectively. Recoveries of lactose in sample preparation procedure I, II, and III were 65.9633%, 96.29%, and 101.66%, respectively. Based on this result, it was concluded that the sample procedure preparation III could be used to the determination of lactose in infant formula by HPLC technique. Applying this method, the concentration of lactose in infant formula sample of commercially available were found to be at infant formula sample A 53.71 g/100 g and infant formula sample B 60.13 g/100 g.