Chemical composition of Nypa palm (Nypa fruticans Wurmb.) vinegar and its effect on adipogenesis

Overweight and obesity are implications of excessive body fat that may give negative effects to health. Nowadays, overweight and obesity are known to be closely related to many types of chronic diseases such as type 2 diabetes, hypertension, coronary heart disease, stroke, and certain cancers. Anti-obesity drug therapy complemented with diet therapy and physical activity has been widely used to treat obesity. However, using the drug as a treatment of obesity could give harmful side effect to the obese patient. Therefore, the objective of this study was to evaluate the chemical composition and antiobesity effect of nypa palm vinegar (NPV) in vitro adipocyte-induced model in comparison to pioglitazone and orlistat as the positive controls. The present study began with the characterization of chemical composition in NPV including organic acids, phenolic compounds, sugar, and alcohol contents. Changes of chemical composition in NPV have been observed base on different fermentation periods and it might be due to naturally presence of microorganisms in Nypa fruticans Wurmb. sap. Organic acids and phenolic compounds of fresh nypa sap and its vinegar at different fermentation stages were determined using a cation exchange column of HPLC- DAD. Organic acids detected were acetic acid, lactic acid, succinic acid, tartaric acid, maleic acid, malic acid, quinic acid, oxalic acid, formic acid, and fumaric acid. The prominent organic acid of vinegar was acetic acid in the range of 5-10%. The concentration of acetic acid had significantly increased (p < 0.05) from 62.49 ± 0.55 mg/ 100 ml in fresh nypa sap to 2513.80 ± 10.24 mg/ 100 ml, 4510.07 ± 7.03 mg/ 100 ml, and 6036.32 ±5.56 mg/ 100 ml in 4 months, 8 months, and 60 months fermented sap, respectively. Nine phenolic compounds including gallic acid, p-coumaric, o-coumaric, protocatechuic, and chlorogenic acid, catechin, epicatechin, quercetin, and rutin were detected in the samples. The concentration of phenolic compounds significantly diverse (p < 0.05) in each sample depend on fermentation periods. Catechin was found the highest concentration in nypa sap (1014.36 ± 21.06 mg/100 ml) and 60 months fermented NPV (3249.40 ± 25.51 mg/ 100 ml) while protocatechuic acid showed the highest concentration in 4 months fermented NPV (1064.50 ± 24.35 mg/ 100 ml) and 8 months fermented NPV (1322.16 ± 7.14 mg/ 100 ml). HPLC-ELSD and GC-FID were employed to quantify sugar and alcohol, respectively. Sugar and alcohol showed significant decrement (p < 0.05) in fermented sap compared to non-fermented sap. DPPH, FRAP and Folin-Ciocalteu assays were used to assess antioxidant capacity in nypa sap and its vinegar. The results indicated that 60 months fermented NPV have the highest antioxidant activity compared to other samples. To assess the NPV effect on OP9 cells viability, MTT assay was carried out and IC50, 2.74% (v/v) of NPV was calculated. IC20 value also was determined to represent the non-toxic concentration of NPV for further analysis. Oil Red O (ORO) staining, triglycerides (TG), and glycerol-3-phosphate (G3P) assays were applied to evaluate antiadipogenesis properties of NPV. After ORO staining, relative lipid accumulation was calculated and NPV-supplementation decreased lipid accumulation in adipocyte up to 57 %. Regardless concentration of treatments, NPV- supplementation showed significant reduction (p < 0.05) of TG content compared to non-treated OP9 adipocytes (control). NPV-supplementation caused TG content reduction in the range of 48 - 70 %. Furthermore, 2.0 % (v/v) NPV-supplementation had also suppressed G3P concentration and comparable (p > 0.05) to positive control (orlistat), but significantly difference (p < 0.05) to pioglitazone. Collectively, the present study has determined bioactive compounds in nypa sap and its vinegar. Potential anti-obesity effects of NPV by in vitro study also has been elucidated.

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