不同聚合度板栗壳原花青素的体外抗氧化活性、抑菌活性及其组成分析

Antioxidant,Antimicrobial Activity and Structure Analysis of Procyanidins with Different Polymerization Degree from Chinese Chestnut Shell

为了充分利用板栗壳资源,本文以新鲜板栗壳为原料,分别提取四种不同聚合度的板栗壳原花青素样品F1(70%乙醇粗提物)、F2(AB-8大孔树脂纯化物)、F3(乙酸乙酯萃取物)及F4(水相萃取物),对其进行平均聚合度的测定,分析其体外抗氧化活性和抑菌活性,并采用高效液相色谱和高分辨质谱技术对活性最高级分进行组成分析。结果表明:样品F1、F2、F3及F4的平均聚合度分别为9.42±0.41、4.14±1.06、3.32±0.34、5.07±0.75,最小聚合度样品F3具有最大的抗氧化活性,其对ABTS自由基、羟基自由基与超氧阴离子自由基清除率最大,IC50值分别为11.24、1.18和93.98μg/m L。抑菌圈试验初筛结果显示,F1对金黄色葡萄球菌表现出抑制作用,对沙门氏菌和大肠杆菌抑制作用不明显。以金黄色葡萄球菌为指示菌,研究四种不同聚合度板栗壳原花青素抑菌效果,结果显示,最小聚合度样品F3对金黄色葡萄球菌抑制作用最大,最小抑菌浓度(MIC)为0.31 mg/m L。高效液相色谱和高分辨质谱结果显示,样品F3中含有没食子酸、儿茶素/表儿茶素和原花青素B型二聚体。实验结果表明,板栗壳原花青素具有较好的抗氧化活性和抑菌活性,且低聚合度原花青素的抗氧化和抑菌效果更为显著,可作为天然抗氧化剂和抑菌剂的原料。

In order to make full use of the chestnut shell resources,four procyanidins sample with different polymerization degree F1( 70% ethanol extract),F2( AB-8 macroporous resin purification),F3( ethyle acetate extract) and F4( water-soluble fraction) were extracted from fresh chestnut shell were measured for mean degree of polymerization. In vitro antioxidative and antibacterial activity of procyanidins in chestnut shell was analyzed. The compositions of the part with highest activity were analyzed by high performance liquid chromatography( HPLC) and high resolution mass spectrometry( HRMS). The results showed that the mean degrees of polymerization of F1,F2,F3 and F4 were 9.42 ± 0.41,4.14 ± 1.06,3.32 ± 0.34,5.07 ± 0.75,F3 with the smallest degree of polymerization had the highest antioxidant activity. It had the best scavenging rate against ABTS radical,hydroxyl radical and superoxide anion radical with the IC50 values of 11.24,1.18 and 93.98 μg/m L,respectively. The preliminary screening of the inhibition zone experiment showed that F1 had inhibitory effect on Staphylococcus aureus,the inhibition on Escherichia coli and Salmonella were not obvious.Staphylococcus aureus was the indicator,antimicrobial activities of four procyanidins with different polymerization degree were measured. The results of measurement showed that F3 with the smallest degree of polymerization had significant bacteriostatic effect on Staphylococcus aureus,and the minimal inhibitory concentration( MIC) was 0.31 mg/m L. The results of HPLC and HRMS showed that gallic acid,catechin/epicatechin and B-type procyanidin were present in F3.In conclusion,chestnut shell proanthocyanidins have good antioxidant and bacteriostatic activity,and the antioxidant and bacteriostatic effects of low-polymerization proanthocyanidins were more significant.They could be used as raw materials for natural antioxidants and bacteriostatic.