Studies on the unknown ninhydrin-positive substance, appearing in the developing fruits (pericarp and seeds) of tea plants (Camellia sinensis L.)

对茶树（Camellia sinensis L.）正在发育的果实（果皮和种子）中出现的未知茚三酮阳性物质的研究

• 批准号: 07660082
• 负责人: SUZUKI Takeo
• 金额: $ 1.28万
• 依托单位: Kyoto Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07660082/
• 关键词: tea (Camellia sinensis); pipecolic acid; cyclic imino acid; proline analogue; fruit development; seed formation; 種子形成; チャ(Comellia sinensis); 果実成育; Camellia sinensis

An unknown ninhydrin-positive substance, especially appearing developing fruits (pericap and seeds) of tea plants, was detected, analyzed and identified as pipecolic acid by two-dimensional thin-layr chromatography (TLC) and by an HPLC system using fluoresence detector.(a) Free amino acids were separated on TLC.The unknown compound and pipecolic acid, a cyclic amino acid, on the plate reacted with nihydrin sprays, giving distinctive color (blue). They also produced a greenish-blue color with isatin.(b) Standard solution containing amino acids, theanine, and pipecolic acid, or fresh extracts from developing tea fruits (pericarp or seeds) were separated on a Shimpack Amino-Li coloumn using a linear gradient and detected by a post-column derivative method using fluoresence detector (OPA method). In this method, more than 20 free amino acids, including theanine and pipecolic acid, were detected in tea extracts. The main free amino acid was pipecolic acid (70-80%) but not theanine in fresh unripe fruit pericarp and seeds, dissapearing in mature seeds. Lysine and *-aminoadipic acid were also detected in these extracts, suggesting that lysine-pipecolic acid pathway play an important rol in fruit development and seed formation in tea plants.(c) Pipecoli acid was also detected in flush tea leaves (5-15%), indicating that pipecolic acid is regarded as one of important free amino acids in manufactured tea leaves.

采用二维薄层色谱（TLC）和荧光检测器高效液相色谱（HPLC）系统对一种未知的茚三酮阳性物质进行了检测、分析和鉴定，特别是在茶树发育的果实（果皮和种子）中出现。(a)薄层色谱分离游离氨基酸。未知的化合物和pipecolic酸（一种环氨基酸）在平板上与nihydrin喷雾剂反应，产生独特的颜色（蓝色）。他们还用isatin生产出了一种蓝绿色。(b)含有氨基酸、茶氨酸和果酸的标准溶液，或发育中的茶果（果皮或种子）的新鲜提取物，在Shimpack amino - li柱上用线性梯度分离，并用荧光检测器（OPA法）柱后导数法检测。在该方法中，检测了茶氨酸、茶果酸等20多种游离氨基酸。新鲜未成熟果皮和种子中游离氨基酸以果酸为主（70-80%），茶氨酸次之，成熟种子中游离氨基酸消失。在这些提取物中还检测到赖氨酸和*-氨基己二酸，表明赖氨酸-管果酸途径在茶树果实发育和种子形成中起重要作用。(c)冲茶茶叶中也检测到果酸（5-15%），表明果酸被认为是成品茶叶中重要的游离氨基酸之一。