Krüger S, Hüsken L, Fornasari R, Scainelli I, Morlock G E
Justus Liebig University Giessen, Institute of Nutritional Science, Chair of Food Science, Heinrich-Buff-Ring 26-32, 35392 Giessen, Germany.
On leave from University of Modena and Reggio Emilia, Via G. Campi 183, 41125 Modena, Italy.
J Chromatogr A. 2017 Dec 22;1529:93-106. doi: 10.1016/j.chroma.2017.10.068. Epub 2017 Oct 29.
Quantitative effect-directed profiles of 77 industrially and freshly extracted botanicals like herbs, spices, vegetables and fruits, widely used as food ingredients, dietary supplements or traditional medicine, gave relevant information on their quality. It allows the assessment of food, dietary supplements and phytomedicines with regard to potential health-promoting activities. In contrary to sum parameter assays and targeted analysis, chromatography combined with effect-directed analysis allows fast assignment of single active compounds and evaluation of their contribution to the overall activity, originating from a food or botanical sample. High-performance thin-layer chromatography was hyphenated with UV/Vis/FLD detection and effect-directed analysis, using the 2,2-diphenyl-1-picrylhydrazyl radical, Gram-negative Aliivibrio fischeri, Gram-positive Bacillus subtilis, acetylcholinesterase and tyrosinase assays. Bioactive compounds of interest were eluted using an elution head-based interface and further characterized by electrospray ionization (high-resolution) mass spectrometry. This highly streamlined workflow resulted in a hyphenated HPTLC-UV/Vis/FLD-EDA-ESI/ESI-(HR)MS method. The excellent quantification power of the method was shown on three compounds. For rosmarinic acid, contents ranged from 4.5mg/g (rooibos) to 32.6mg/g (rosemary), for kaempferol-3-glucoside from 0.6mg/g (caraway) to 4.4mg/g (wine leaves), and for quercetin-3-glucoside from 1.1mg/g (hawthorn leaves) to 17.7mg/g (thyme). Three mean repeatabilities (%RSD) over 18 quantifications for the three compounds were ≤2.2% and the mean intermediate precision over three different days (%RSD, n=3) was 5.2%.
77种工业提取和新鲜提取的植物性物质（如药草、香料、蔬菜和水果）常被用作食品成分、膳食补充剂或传统药物，对其进行的定量效应导向分析给出了有关其质量的相关信息。这有助于评估食品、膳食补充剂和植物药的潜在健康促进活性。与总和参数测定法和靶向分析相反，色谱法结合效应导向分析能够快速确定单一活性化合物，并评估其对源自食品或植物样本的整体活性的贡献。高效薄层色谱法与紫外/可见/荧光检测及效应导向分析联用，使用2,2-二苯基-1-苦基肼自由基、革兰氏阴性费氏弧菌、革兰氏阳性枯草芽孢杆菌、乙酰胆碱酯酶和酪氨酸酶测定法。使用基于洗脱头的接口洗脱感兴趣的生物活性化合物，并通过电喷雾电离（高分辨率）质谱进一步表征。这种高度简化的工作流程产生了一种联用的高效薄层色谱-紫外/可见/荧光-效应导向分析-电喷雾电离/电喷雾电离-（高分辨率）质谱方法。该方法对三种化合物显示出出色的定量能力。迷迭香酸含量范围为4.5mg/g（南非红叶茶）至32.6mg/g（迷迭香），山奈酚-3-葡萄糖苷含量范围为0.6mg/g（香菜）至4.4mg/g（葡萄叶），槲皮素-3-葡萄糖苷含量范围为1.1mg/g（山楂叶）至17.7mg/g（百里香）。这三种化合物在18次定量中的三个平均重复性（%RSD）≤2.2%，在三个不同日期的平均中间精密度（%RSD，n = 3）为5.2%。
