Biological activity and quantification of bioative compounds in yerba mate extract and its application in fish hamburger

Abstract The yerba mate has a high content of biologically active compounds with industrial application. Caffeine, rutin and chlorogenic acid present in this product have antioxidant activity and the latter one is also an antimicrobial agent. This \paper aimed to assess the biological activity of yerba mate extract and quantify its bioactive compounds, as well as apply the extract in fish hamburger to determine its preservative activity. The antioxidant power was evaluated by DPPH (2,2-diphenyl-1-picrylhydrazyl) and FRAP (ferric reducing antioxidant power). Total phenolic compounds and flavonoids were quantified by spectrophotometry, and caffeine, rutin and chlorogenic acid by ultra-performance liquid chromatography. The antimicrobial activity was assessed by Minimum Inhibitory Concentration (MIC). Associations of extract with the synthetic preservative butyl hydroxyanisole (BHA) were tested. Fish hamburgers incorporated with yerba mate were prepared; centesimal composition, lipid and microbiological oxidation were carried out during 28 days. Results: DPPH IC50 of 7.91 µg mL-1 in the extract, FRAP of 4922.67 µmol for Fe (II) g-1, 104.54 EAG g-1 of total phenolics, 23.11 EQ g-1 of flavonoids, and 49.6, 34.5 and 23.1 mg g-1 of chlorogenic acid, caffeine and rutin, respectively. The extract presented a CIM of 10, 5 and 10 mg mL-1 against Escherichia coli, Staphylococcus aureus and Salmonella enterica, respectively. The combination of extract and BHA reduced the amount of synthetic preservative for the same antioxidant activity and presented an additive effect to S. aureus. The fish hamburgers with 1% of extract or with BHA showed statistical difference in mesophiles counting from the control group. Thus, the yerba mate extract presented biological activity and the growth of mesophiles was inhibited in fish hamburguers incorporated with BHA or 1% of extract, indicating that the quantified phenolic compounds are likely to be preservative agents.

摘要 马黛茶（yerba mate）富含具有工业应用价值的生物活性成分。该产品中含有的咖啡因、芦丁和绿原酸均具备抗氧化活性，其中绿原酸同时兼具抗菌性能。本研究旨在评估马黛茶提取物的生物活性并定量分析其生物活性成分，同时将该提取物应用于鱼肉汉堡以探究其防腐效能。本研究采用DPPH（2,2-二苯基-1-苦基肼基）法与FRAP（铁还原抗氧化能力）法评估提取物的抗氧化能力；采用分光光度法对总酚类物质与总黄酮进行定量，采用超高效液相色谱法对咖啡因、芦丁及绿原酸进行定量；采用最低抑菌浓度（Minimum Inhibitory Concentration, MIC）法评估其抗菌活性。此外，还测试了提取物与合成防腐剂丁基羟基茴香醚（butyl hydroxyanisole, BHA）的复配效果。本研究制备了添加马黛茶提取物的鱼肉汉堡，并在28天的储存周期内对其进行了百分营养组成、脂质氧化与微生物氧化检测。结果显示，该提取物的DPPH半抑制浓度（IC₅₀）为7.91 µg·mL⁻¹，FRAP值为4922.67 µmol Fe(II)·g⁻¹，总酚含量为104.54 没食子酸当量（EAG）·g⁻¹，总黄酮含量为23.11 槲皮素当量（EQ）·g⁻¹；绿原酸、咖啡因与芦丁的含量分别为49.6、34.5与23.1 mg·g⁻¹。该提取物对大肠埃希氏菌（Escherichia coli）、金黄色葡萄球菌（Staphylococcus aureus）与肠炎沙门氏菌（Salmonella enterica）的最低抑菌浓度分别为10、5与10 mg·mL⁻¹。提取物与BHA复配可在达到同等抗氧化活性的前提下降低合成防腐剂的使用量，且对金黄色葡萄球菌表现出协同增效作用。添加1%提取物或BHA的鱼肉汉堡，其需氧菌总数与对照组存在显著统计学差异。综上，马黛茶提取物具备良好的生物活性，添加BHA或1%提取物的鱼肉汉堡可抑制需氧菌的生长，表明本研究定量得到的酚类物质或可作为天然防腐成分加以应用。