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PHILIPP AGRIC SCIENTIST Vol. 96 No 3, 308–313 September 2013 ISSN 0031-7454 Antioxidant Properties of Bignay [Antidesma bunius (L.) Spreng] Wine at Different Stages of Processing Ma. Desiree Belina-Aldemita1,*, Veronica C. Sabularse1, Erlinda I Dizon2, Wilma A Hurtada3 and Mary Ann O. Torio1 1 Institute of Chemistry, College of Arts and Sciences; 2Institute of Food Science and Technology, College of Agriculture; 3Institute of Human Nutrition and Food, College of Human Ecology; University of the Philippines Los Baños, College, Laguna, Philippines * Author for correspondence; e-mail:; Tel: +63 49 5362220, Tel/Fax: +63 49 5362241, Mobile: +63 932 8689348 Bignay [Antidesma bunius (L.) Spreng] fruits were used in the preparation of red wine Must and wine samples at different stages of processing were analyzed for total antioxidant activity (TAA), total phenolics (TP), total flavonoids (TF) and total anthocyanins (TA). Ranges in values obtained were as follows:

53.37 ± 057 to 7835 ± 212% for TAA, 65139 ± 309 to 93007 ± 2618 mg L -1 gallic acid equivalents for TP, 371.62 ± 584 to 70367 ± 1280 mg L-1 catechin equivalents for TF, and 9697 ± 472 to 372.62 ± 477 mg L-1 cyanidin-3,5-diglucoside for TA Results indicated that changes in the TAA of the must/wine samples varied during the different stages in the processing of bignay wine. These changes could not be completely attributed to a certain group of phenolic compounds as the changes in TP, TF and TA also varied from one stage to another. Key words: anthocyanins, Antidesma bunius (L.) Spreng, antioxidant, bignay, flavonoids, must, phenolics, wine Abbrevations: CE – catechin equivalents, CydGE – cyanidin-3,5-diglucoside equivalents, GAE – gallic acid equivalents, TA – total anthocyanins, TAA – total antioxidant activity, TF – total flavonoids, TP – total phenolics INTRODUCTION Red wine has considerably been strongly associated with cardioprotection and chemoprotection

than other alcoholic beverages (Rimm et al. 1996) It was hypothesized that some phenolic compounds in red wine such as polyphenols, flavonoids and anthocyanins play an important role in the prevention of coronary heart diseases. In the Philippines, a variety of tropical fruit wines are produced either for home consumption or commercial purposes. However, there is lack of information on the polyphenolic composition of these locally manufactured wines and their antioxidant capacity, which is related to polyphenolic content, as these have not yet been examined. This study was conducted to determine the antioxidant properties, specifically the total antioxidant activity, total phenolics, total flavonoids and total anthocyanins of bignay [Antidesma bunius (L.) Spreng] wine during processing. 308 Bignay is native to the Philippines and often grows in mountainous areas with a tropical climate. It belongs to the Euphorbiaceae family and has ovoid-shaped fruits clustered together in a bunch

of 30–40 small fruits. The fruit is green, and turns red to black as it ripens. It has a sour sweet taste when ripe and is commonly used to make jam and wine. There has been no report in the literature about the changes in the antioxidant properties at every stage of bignay wine processing. This is the first study to investigate these changes and to explain their biochemical aspects. A good knowledge of the changes in the antioxidant properties of wine during processing would contribute to the development of high-quality wines produced locally. Research on the antioxidant activity and phenolics present in locally manufactured wines would be beneficial to local wine manufacturers. Inclusion of this information on the bottle label will orient the consumer about the quality and health benefits of the purchased wine. It may also promote consumption of locally made wines over imported wines and other alcoholic beverages. The Philippine Agricultural Scientist Vol. 96 No 3 (September 2013)

Antioxidant Properties of Bignay Wine MATERIALS AND METHODS Wine processing was conducted at the Food Microbiology Laboratory (FML), Institute of Food Science and Technology (IFST), College of Agriculture (CA), University of the Philippines Los Baños (UPLB), College, Laguna. The analysis of antioxidant properties was done at the Institute of Chemistry, College of Arts and Sciences, UPLB. Processing of Bignay Wine The bignay wine was prepared using the method of Dizon (2009). Fully ripe fruits were obtained from the Institute of Plant Breeding, UPLB, College, Laguna. The materials were transported to the FML, IFST, CA, UPLB, College, Laguna. Must preparation. The bignay fruits were sorted manually in the laboratory to obtain fully ripe and undamaged fruits. The fruits were removed from the stems, washed and drained. Water (75 L) was added to 2.5 kg of fruits and then macerated using a blender to produce the must. The sugar content of the must was adjusted from 1.4 to 20 °Brix by

addition of 225 kg of refined cane sugar. The preparation was mixed thoroughly and about 10% of the must was separated for starter preparation. Starter preparation. Saccharomyces bayanus was obtained from the FML, IFST, CA, UPLB, College, Laguna. The cultures were transferred into potato dextrose agar slants and incubated at 28–30 °C for 48–36 h. Ten percent of the total volume of the must was placed in an Erlenmeyer flask and plugged with cotton. This volume was pasteurized in boiling water for 30 min, cooled to 40–45 °C and inoculated with S. bayanus Fermentation was allowed to take place for 24 h at room temperature. The fermented mixture served as the starter for wine making. Treatment of the must. Five milliliters of 10% sodium metabisulfite was added per 3.79 L of the prepared must to destroy spoilage microorganisms. The must was distributed in gallon jars where space was allotted for the addition of starter and the rise of the must during active fermentation. The gallon

jars were plugged with clean cotton and allowed to stand for 24 h at room temperature. Fermentation. The previously prepared starter was added to the treated must. Aerobic fermentation was allowed to take place for 3 d. The cotton plugs were then replaced with fermentation locks and anaerobic fermentation was allowed to proceed for 3 wk. The Philippine Agricultural Scientist Vol. 96 No 3 (September 2013) Ma. Desiree Belina-Aldemita et al Harvest, storage and aging. The wines were filtered through clean cheesecloth and placed in sterilized gallon jars. Five milliliters of 10% sodium metabisulfite was added per 3.79 L of wine The gallon jars were covered and stored at room temperature (20–22 °C), in a dry place for aging. Analysis of Antioxidant Properties The antioxidant properties of the following samples were analyzed: must upon dilution with water, must after adjustment of sugar content, must before addition of wine yeast, must before aerobic fermentation, must during aerobic

fermentation (after 1 and 3 d), must during anaerobic fermentation (end of 1st and 2nd wk), raw wine, and aged wine (1st, 2nd and 6th mo). All the assays were done in nine replicates. Total antioxidant activity (TAA). TAA was determined using the β-carotene bleaching method as described by Marco (1968) with some modifications. In a 50-mL beaker, 5 mg of β-carotene was dissolved in 5 mL of dichloromethane. A 15-mL volume of Tween 40 was added to the solution. The dichloromethane was evaporated under the hood for 5 min. Linoleic acid (030 mL) was added and the remaining solvent was removed by evaporation for another 5 min. Distilled water (20 mL) was added to the emulsion, and stirred thoroughly during the addition. The mixture was transferred to a 1-L Erlenmeyer flask and distilled water was added to obtain a final volume of approximately 650 mL. The mixture was constantly swirled for 5 min prior to use. The assay mixture, containing 25 mL β-carotene emulsion and 1 mL methanolic

extract, was incubated in a constant temperature water bath at 50 °C. The reference sample (1 mg mL-1 vitamin E) and control (distilled water) were also assayed along with the samples. The decrease in the absorbance of β-carotene was monitored using a Shimadzu UV-mini 1240 spectrophotometer set at 470 nm for 100 min at 20-min intervals. The TAA was expressed as percent of inhibition relative to the control at 100 min using the formula (Suja et al. 2005): TAA (% inhibition) = degradation rate of control – degradation rate of sample degradation rate of control x 100 where: degradation rate = absorbance (470 nm) at 0 min absorbance (470 nm) at 100 min ÷ 100 min 309