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.

用二维薄层色谱法和高效液相色谱荧光检测器检测、分析和鉴定了一种茚三酮阳性物质，特别是在茶树果实（果皮和种子）发育过程中出现的茚三酮阳性物质。(a)薄层层析分离游离氨基酸，未知化合物与环氨基酸哌啶甲酸在薄层板上与尼醇喷雾反应，显蓝色。他们还与靛红产生了蓝绿色。(b)含有氨基酸、茶氨酸和哌啶酸的标准溶液，或来自发育中的茶果实（果皮或种子）的新鲜提取物，在Shimpack Amino-Li柱上使用线性梯度分离，并通过使用荧光检测器的柱后衍生方法（OPA方法）检测。在该方法中，超过20种游离氨基酸，包括茶氨酸和哌啶酸，茶叶提取物中检测。新鲜未成熟果皮和种子中游离氨基酸主要为哌啶酸（70-80%），而茶氨酸不存在，成熟种子中消失。赖氨酸和 *-氨基己二酸也在这些提取物中检测到，表明赖氨酸-哌啶酸途径在茶树果实发育和种子形成中起重要作用。(c)在冲茶中也检测到哌可酸（5-15%），表明哌可酸被认为是成品茶叶中重要的游离氨基酸之一。