METHOD DEVELOPMENT FOR SIMULTANEOUSLY DETERMINATION OF BHA, BHT, PROPYL GALLATE, AND TBHQ IN MARGARINE WITH HIGH PERFORMANCE LIQUID CHROMATOGRAPY

• Main Author: Christinawaty
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/45941
• Institution: Institut Teknologi Bandung

Background: Antioxidants are food additives in oil and fat products usually to decrease the process of rancidity by preventing the oxidation such as in margarine. BHA, BHT, propyl gallate, and TBHQ are commonly used antioxidants. Combinations of antioxidants are generally used in the food product. Therefore, analytical method which is capable to measure the antioxidants simultaneously, is required. This study was aimed to obtain analytical method for determination of BHA, BHT, propyl gallate, and TBHQ using High Performance Liquid Chromatography (HPLC) in margarine samples. Methods: The analysis was using Waters column Xbridge C18 (4.6 x 150 mm, 5?m) and 280 nm ultraviolet detector. The mobile phase was a mixture of 1% acetic acid in acetonitrile and 1% acetic acid (3:7). The flow rate was set at 1.7 ml/min and the column temperature was 32 ºC. Samples were prepared using vortex and centrifugation. Further, they were homogenized and filtered with a membrane filter of 0.45 ?m. Results: The calibration curve of BHA with equation regression y = 9385.2x –10263 was linear in the concentration range from 9.98245 to 89.84202 µg/ml and correlation coefficient r = 0.9999, with limit of detection and limit of quantitation were 2.6217 and 7.9445 g/ml, respectively. The calibration curve of BHT with equation regression y = 5970.1x - 31248 linear in the concentration range from 5.66546 to 101.97835 g/ml and correlation coefficient r = 0.9996, with limit of detection and limit of quantitation were 4.1837 and 12.6777 g/ml, respectively. The calibration curve of propyl gallate with the regression equation y = 29851x + 50616 was linear in the concentration range from 5.38025 to 96.84451 ?g/ml and correlation coefficient r = 0.9998, with limit of detection and limit of quantitation were 2.494 and 7.559 g/ml, respectively. The calibration curve of TBHQ with the regression equation y = 7793.1x + 27100 was linear in the concentration range from 11.89258 to 107.03326 g/ml and correlation coefficient r = 0.9998, with limit of detection and limit of quantitation were 2.1272 and 6.4461 µg/ml, respectively. Inter day precision of the method was expressed as RSD with result; BHA, BHT, propyl gallate, and TBHQ were 0.1553; 0.4576; 0.0237; 4.8106 %; respectively whereas recovery using standard addition method of BHA, BHT, propyl gallate, and TBHQ in margarine samples were 105.38; 107.87; 86.50; 83.64 % respectively. Based on the obtained results, it can be concluded that this method has been successfully developed and validated for the determination of antioxidants BHA, BHT, propyl gallate, and TBHQ in margarine samples.