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以银杏种仁为材料,采用乙醇浸提、石油醚萃取的方法提取酚酸,研究银杏种仁酚酸硅胶柱层析纯化工艺。采用液相色谱(HPLC)及液-质联用(LC-MS)分析银杏种仁中酚酸的组成及含量。以2种细菌及4种真菌为供试菌,比较银杏种仁酚酸的抑菌活性,并确定其对各菌种的最小抑菌浓度(MIC)。结果表明,以流速为1.2 m L/min的石油醚-乙醚-甲醇(89∶11∶1,V/V/V)为洗脱液,对银杏酚酸粗提物进行硅胶柱层析纯化,取得较好的纯化效果。HPLC及LC-MS分析表明:银杏酚酸主要由C13∶0,C15∶1,C17∶2,C15∶0和C17∶1等5种酚酸组分组成。以C13∶0,C15∶1,C17∶1计,银杏种仁中的酚酸含量为0.11 g/kg脱脂干粉,其中C13∶0,C15∶1,C17∶1的含量分别为0.013 g/kg脱脂干粉、0.042 g/kg脱脂干粉、0.055 g/kg脱脂干粉。抑菌试验表明,银杏酚酸对供试的2种细菌(大肠杆菌和枯草芽孢杆菌)以及4种真菌(青霉、产紫青霉、沙门柏干酪青霉、黑曲霉)有抑制作用。银杏酚酸对大肠杆菌、枯草芽孢杆菌及青霉菌有较好的抑菌效果,其MIC分别为7.5,15和25 mg/m L。
Using ginkgo seed as material, phenolic acid was extracted by ethanol extraction and petroleum ether extraction, and the purification technology of phenolic acid silica gel by ginkgo seed was studied. The composition and content of phenolic acids in ginkgo seeds were analyzed by liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS). Two kinds of bacteria and four kinds of fungi were used as test bacteria to compare the antibacterial activities of phenolic acids of Ginkgo biloba seeds and to determine their minimum inhibitory concentration (MIC) against each strain. The results showed that the crude extract of ginkgolic acids was purified by silica gel column chromatography with petroleum ether-diethyl ether-methanol (89:11:1, V / V / V) at a flow rate of 1.2 m L / Get better purification effect. HPLC and LC-MS analysis showed that ginkgolic acids mainly consisted of five phenolic acids such as C13: 0, C15: 1, C17: 2, C15: 0 and C17: 1. The content of phenolic acids in ginkgo kernels was 0.11 g / kg in terms of C13: 0, C15: 1 and C17: 1, and the contents of C13: 0, C15: 1 and C17: 1 were 0.013 g / kg Non-fat dry powder, 0.042 g / kg dry powder, 0.055 g / kg dry powder. Bacteriostasis test showed that ginkgolic acids inhibited the two kinds of bacteria (Escherichia coli and Bacillus subtilis) and four fungi (Penicillium, Penicillium purpurogenum, Penicillium salmonella, Aspergillus niger) tested. Ginkgolic acids had good antibacterial activity against Escherichia coli, Bacillus subtilis and Penicillium sp., With MICs of 7.5, 15 and 25 mg / mL respectively.