Laboratory of Advanced Research on Foods and Drugs, Department of Food Science and Technology, Faculty of Pharmacy, University of Concepción, Chile.
Laboratory of Biotechnology and Genetic of the Foods, Department of Food Science and Technology, Faculty of Pharmacy, University of Concepcion, Chile; Center for Biotechnology, University of Concepcion, Chile.
J Chromatogr A. 2019 Dec 20;1608:460415. doi: 10.1016/j.chroma.2019.460415. Epub 2019 Jul 31.
α-glucosidase inhibitors (AGIs) are very attractive bioactive compounds due to their therapeutic profile that includes beneficial effects over glycemic control in type 2 diabetes mellitus and viral infections. Its detection and identification in plants and fruits has gained growing attention, and certainly requires efficient screening methodologies. The objective of the present work was to develop a fast methodology to detect and identify AGIs in cherimoya fruit (Annona cherimola Mill.) applying effect-directed analysis via high-performance thin layer-chromatography (HPTLC) linked with bioassay and mass spectrometry (MS). Both, HPTLC and bioassay conditions, were optimized accomplishing 50% and 83% reduction on enzyme concentration and incubation time respectively, compared to the original method. Additionally, the contrast between inhibitory bands and purple background was also enhanced by enzyme substrate impregnation on HPTLC plate. The optimized detection conditions established were the following: 5.0 U mL of enzyme solution, 1.0 mg mL of 2-naphthyl-α-D-glucopyranoside substrate, 1.0 mg mL of Fast Blue B salt solution and 10 min as incubation time. Applying this methodology, coupled to HPTLC-MS and ultra-high-performance liquid chromatography (UHPLC)-diode array detector (DAD)-MS/MS, it was possible for the first time to detect and identify three AGIs in cherimoya peel and seeds. Compounds were tentatively assigned as phenolamides (phenylethyl cinnamides): N-trans-feruloyl tyramine (m/z 314 [M+H]; UV λ 293 and 316 nm), N-trans-p-coumaroyl tyramine (m/z 284 [M+H]; UV λ 296 nm) and N-trans-feruloyl phenethylamine (m/z 298 [M+H]; UV λ 288 nm). To the best of our knowledge, the presence of latter compound is reported for the first time in cherimoya.
α-葡萄糖苷酶抑制剂（AGIs）因其治疗谱而成为非常有吸引力的生物活性化合物，包括对 2 型糖尿病和病毒感染的血糖控制有益的作用。其在植物和水果中的检测和鉴定越来越受到关注，当然需要有效的筛选方法。本工作的目的是开发一种快速检测和鉴定番荔枝（Annona cherimola Mill.）中 AGIs 的方法，应用高效薄层色谱（HPTLC）结合生物测定和质谱（MS）进行定向分析。与原始方法相比，HPTLC 和生物测定条件分别实现了 50%和 83%的酶浓度和孵育时间减少。此外，通过在 HPTLC 板上浸渍酶底物，还增强了抑制带与紫色背景之间的对比度。优化的检测条件如下：酶溶液 5.0 U mL、2-萘基-α-D-吡喃葡萄糖苷底物 1.0 mg mL、Fast Blue B 盐溶液 1.0 mg mL 和 10 min 的孵育时间。应用该方法，结合 HPTLC-MS 和超高效液相色谱（UHPLC）-二极管阵列检测器（DAD）-MS/MS，首次能够在番荔枝果皮和种子中检测和鉴定三种 AGIs。化合物被初步鉴定为酚酰胺（苯乙基肉桂酰胺）：N-反式-阿魏酰酪胺（m/z 314 [M+H]+；UV λ 293 和 316nm）、N-反式-对香豆酰酪胺（m/z 284 [M+H]+；UV λ 296nm）和 N-反式-阿魏酰苯乙胺（m/z 298 [M+H]+；UV λ 288nm）。据我们所知，后一种化合物在番荔枝中是首次报道。
