Characterization and Quantitation of Anthocyanins of the Pigmented Tea Cultivar TRI 2043 (Camellia sinensis L.) from Sri Lanka

Philipp Hopfstock, Pitumpe Appuhamilage Nimal Punyasiri, Mats Kiene, Jeevan Dananjava Kottawa-Arachchi, Recep Gök, Peter Winterhalter
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Abstract

Tea leaves are rich in diverse bioactive compounds. The tea accession TRI 2043 is unique due to its pigmented leaves caused by anthocyanins, clonal origin, resistance to blister blight disease, and high pubescence density. Because of its peculiarity, TRI 2043 is used to produce high-quality silver tip tea, a premium type of tea that commands high prices. This study was carried out to clarify and elucidate the types of anthocyanins in this particular accession. Four different anthocyanin species were identified and quantitated as cyanidin-3-O-β-d-galactoside and delphinidin-3-O-β-d-galactoside equivalents for leaf blades and stems of the cultivar TRI 2043. The characterization was performed by comparison with commercially available reference substances and further confirmed using ion mobility high-resolution time-of-flight-mass spectrometry (IMS-HRTOF-MS). Quantitation was carried out using ultra-high-performance liquid chromatography ultraviolet–visible detection (UHPLC-UV-vis) with cyanidin-3-O-β-d-glucoside as an internal standard. E- and Z-geometric isomers of 6-p-coumaroyl derivates of delphinidin and cyanidin-3-O-β-d-galactopyranosides were observed, and collision cross section (CCS) values were determined for all four different anthocyanidin species. The content of anthocyanins in leaf blades of cultivar TRI 2043 was 856.32 ± 41.56 µg/g dry weight, with cyanidin being the more abundant anthocyanin (69.8%). Conversely, the stem material contained an anthocyanin amount of 459.5 ± 44.7 µg/g dry weight, with a higher content of delphinidin (69.6%). In summary, an enrichment strategy using analytical membrane chromatography was established to fully elucidate and quantify the anthocyanin profile of plant samples such as the special tea variety TRI 2043.
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斯里兰卡色素茶栽培品种 TRI 2043(Camellia sinensis L.)花青素的特性和定量分析
茶叶富含多种生物活性化合物。茶叶品种 TRI 2043 因其花青素引起的叶片色素沉着、克隆原产地、对水泡病的抗性和较高的短柔毛密度而独树一帜。由于其特殊性,TRI 2043 被用来生产高品质的银尖茶,这是一种价格高昂的优质茶叶。本研究旨在澄清和阐明这一特殊品种的花青素类型。在 TRI 2043 栽培品种的叶片和茎中,以氰苷-3-O-β-d-半乳糖苷和鹅膏素-3-O-β-d-半乳糖苷当量的形式对四种不同的花青素进行了鉴定和定量。通过与市售参考物质进行比较,并使用离子迁移高分辨飞行时间质谱法(IMS-HRTOF-MS)进一步确认了其特征。以氰苷-3-O-β-d-葡萄糖苷为内标,采用超高效液相色谱紫外可见检测法(UHPLC-UV-vis)进行定量。观察到了花翠素和花青素-3-O-β-d-吡喃半乳糖苷的 6-对香豆酰衍生物的 E- 和 Z-几何异构体,并测定了所有四种不同花色苷的碰撞截面(CCS)值。TRI 2043 栽培品种叶片中的花青素含量为 856.32 ± 41.56 µg/g 干重,其中花青素含量最高(69.8%)。相反,茎干材料中的花青素含量为 459.5 ± 44.7 µg/g 干重,其中花翠素含量较高(69.6%)。总之,利用分析膜色谱法建立了一种富集策略,以全面阐明和量化植物样品(如特殊茶叶品种 TRI 2043)的花青素特征。
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