应用薄层扫描光密度法检测南极放线菌N-1中的α-葡萄糖苷酶抑制剂的含量  

ATLC-Scanning Quantitative Method for α-Glucoside Inhibition from Antarctic Pole Actinomyces N-1

作　　者：倪孟祥[1] 张扬[1] 葛羚超 魏爽[1] 

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京210009

出　　处：《药物生物技术》2014年第2期139-143,共5页Pharmaceutical Biotechnology

摘　　要：以比色法(PNPG法)对10株海洋微生物经摇床的发酵液进行α-葡萄糖苷酶抑制活性检测,发现南极放线菌N-1的代谢产物对α-葡萄糖苷酶抑制活性最强。文章旨在建立稳定的南极放线菌N-1产α-葡萄糖苷酶抑制剂的分析方法,为α-葡萄糖苷酶抑制剂的含量测定和纯化做前期工作。以乙酸乙酯∶乙醇=27∶1为展开剂,将南极放线菌N-1的发酵液在GF254硅胶板上展开后,以紫外灯254 nm波长下显影观察得到两个组份,组份A的抑制率为10.03%,纯度达到81.6%,组份B的抑制率为2.91%,纯度达到71.32%,组份A可作为α-葡萄糖苷酶抑制剂的相对工作标准品。分别用高效液相和薄层扫描光密度法绘制其标准曲线,然后对已经分离纯化的成品进行纯度分析。用高效液相检测成品的含量是88%,薄层扫描光密度法测定成品的含量是91.6%,比较接近。结果表明薄层扫描光密度法可以作为检测α-葡萄糖苷酶抑制剂含量的一种新方法。Ten marine microorganisms were screened by the method of PNPG. It showed that the metabolite of the Antarctic Pole Actinomyces N-1 had the stronger α-glucoside inhibition activity than others. The purpose of the study was to establish a stable analysis method of α-glucoside inhibition from Antarctic Pole ActinomycesN-1, preparing for the determination and purification of α- glycosidase inhibitor. The mobile phase was a mixture of EA： EtOH = 27： 1. Fluorescent was displayed under UV at GF254 silica gel. There were two components. The inhibitory rate of the component A was 10.03% and the purity reached to 81.6%. The inhibitory rate of the component B was 2.91% and the purity reached to 71.32%. The component A can be used as standard relative works of α-glycosidase inhibitors. The standard curves were drawn by HPLC and TLC scanning optical density method. The results showed that the content of purified product was 88% using high performance liquid chromatography and the content of purified product was 91.6% using TLC-scanning ,which was quite approaching. This indicates that the TLC-Scanning method can be used as a new method for the determination of a-glycosidase inhibitor.

关 键 词：南极放线菌N-1 Α-葡萄糖苷酶 Α-葡萄糖苷酶抑制剂 分离纯化 薄层扫描光密度法 高效液相 GF254硅胶板 

分 类 号：O658.64[理学—分析化学]