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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。
The colorimetric method (PNPG method) was used to detect the α-glucosidase inhibitory activity of 10 strains of marine microorganisms in the fermentation broth. It was found that the metabolites of Antarctic actinomycetes N-1 had the strongest α-glucosidase inhibitory activity . This article aims to establish a stable Antarctic actinomycetes N-1 α-glucosidase inhibitor analysis method for the α-glucosidase inhibitor determination and purification of the preliminary work. With ethyl acetate: ethanol = 27: 1 as developing solvent, the fermentation broth of Antarctic actinomycetes N-1 was developed on GF254 silica gel plate and observed under UV light at a wavelength of 254 nm. Two components were obtained. The inhibitory rate of A was 10.03%, the purity was 81.6%, the inhibitory rate of component B was 2.91% and the purity was 71.32%. Component A could be used as relative working standard of α-glucosidase inhibitor. Respectively, using high-performance liquid and thin-layer scanning densitometry drawing its standard curve, and then purified the finished product purity analysis. With high-performance liquid detection of the finished product content is 88%, TLC scanning optical density of the finished product content is 91.6%, relatively close. The results show that TLCS can be used as a new method to detect the content of α-glucosidase inhibitors.