TLC-Bioautography is a fast and effective method for assessing the inhibitory effect of compounds present in plant extracts against microbial species. However, this method has a hidden, currently underutilized potential for evaluating the presence of inhibitory compounds against selected enzymes. The aim of this work was to design a functional TLC-Bioautography method for the evaluation of protease inhibitors present in plant extracts. The method is based on the hydrolysis of Nα-benzoyl-DL-arginine-p-nitroanilide hydrochloride (BApNA) by α-chymotrypsin as a representative serine protease to produce coloured para-nitroaniline (pNA). Derivatization of pNA with both sodium nitrite and N-(1-naphthyl) ethylenediamine (NPED) leads to the formation of a pink azo dye. This step improves the resolution of active compounds on the chromatogram, which appear as light spots on a pink background. The developed method was tested for the analysis of protease inhibitors in different plant materials such as grape pomace from Vitis vinifera, Picea abies bark, Hippophae rhamnoides berries, Hordeum sativum bran, Triticum aestivum bran and Avena sativa bran. Plant extracts, which could not be analyzed by a commonly used spectrophotometric method due to interference, were assessed by this method.

中文翻译：

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TLC-Bioautography是一种快速廉价的筛选方法，可用于检测粗植物提取物中的α-胰凝乳蛋白酶抑制剂。

TLC-Bioautography是一种快速有效的方法，用于评估植物提取物中存在的化合物对微生物物种的抑制作用。但是，该方法具有潜在的，未充分利用的潜力，可用于评估针对所选酶的抑制性化合物的存在。这项工作的目的是设计一种功能性TLC-生物自显影法，用于评估植物提取物中存在的蛋白酶抑制剂。该方法基于α-胰凝乳蛋白酶作为代表的丝氨酸蛋白酶水解Nα-苯甲酰基-DL-精氨酸-对硝基苯胺盐酸盐（BApNA），以产生有色对硝基苯胺（pNA）。用亚硝酸钠和N-（1-萘基）乙二胺（NPED）衍生化pNA会导致形成粉红色偶氮染料。此步骤可提高色谱图中活性化合物的分离度，在粉红色的背景上显示为亮点。测试了所开发的方法用于分析不同植物材料中的蛋白酶抑制剂，例如葡萄中的葡萄渣，云杉云杉，沙棘浆果，大麦麸，普通小麦和燕麦。通过这种方法评估了由于干扰而无法通过常用分光光度法进行分析的植物提取物。