葡萄科植物中 resveratrol 衍生物之高效液相層析定量研究   全文下載

Simultaneous Determination of Resveratrol Derivatives in Vitis thunbergii Plant by High Performance Liquid Chromatography

小本山葡萄（Vitis thunbergii Sieb. & Zucc.，葡萄科）又名細本葡萄，其根部（VtR）具清熱利濕、強肝明目、舒筋活絡之效，為民間常用之藥材。我們之前的研究發現此藥材富含具生物活性的resveratrol衍生物。基於市面上充斥非真品的切片藥材，本研究乃以自行分離的ampelopsin C 、(+)-vitisin A 、(-)-vitisin B 、(+)-ε-viniferin四化合物與另一購買的resveratrol作為標準品，建立一種簡單、快速、靈敏且高穩定性的HPLC分析方法，除了測定五個化合物在VtR中的含量，並探討其在市售切片藥材中的含量。結果顯示前三個化合物為VtR的主要成分，含量分別為3.57、2.99與1.69 mg/g；resveratrol則表現0.33 mg/g的低含量。檢測五樣市售的切片藥材（Vt-1-Vt-5），三個主要成分的含量分別為ND-0.35、ND-1.10與ND-1.20 mg/g，明顯低於其在VtR中的含量。此外本研究亦探討同科另六種本土植物－－葡萄（V. vinifera）、基隆葡萄（V. kelungensis）、毛山葡萄（Ampelopsis brevipedunculata）、翼莖粉藤（Cissus pteroclada）、粉藤（C. repens）的根部及小葉葡萄（V. thunbergii var. taiwaniana）莖部中是否含上述化合物，結果顯示五個化合物均存在於前三種植物中，但在翼莖粉藤中均未被偵測到，小葉葡萄與粉藤二者則只偵測到resveratrol化合物。

The roots of Vitis thunbergii Sieb. Zucc. (Vt-R) are well-known bone-protective medicinal material and rich in active resveratrol derivatives, as revealed in our previous studies. The present study used five compounds, including four isolated ones, ampelopsin C, (+)-vitisin A, (-)-vitisin B, and (+)-e-viniferin, and one purchased resveratrol, as standards to establish an HPLC method in order to simultaneously determine these contents in the sample materials. The analyses method was validated and was performed by using a reverse-phase column (Cosmosil 5C18-AR-II) with absorbance detection at 280 nm. The results showed that ampelopsin C, (+)-vitisin A and (-)-vitisin B were the major three constituents in Vt-R and their contents were 3.57, 2.99, and 1.69 mg/g, respectively. Meanwhile, resveratrol gave a level of 0.33 mg/g, which was much lower than those three major constituents. To determined the contents of five standards in the stems and leaves of this plant, except for (+)-e-viniferin, the levels of ampelopsin C, (+)-vitisin A, (-)-vitisin B, and resveratrol were 1.13, 0.27, 0.57, and 0.14 mg/g, respectively, in the stems, which were much less than those in root part. However, five compounds were not detected in leaves. This analytic method was also employed to investigate the qualities of five piece-packed raw materials (Vt-1-Vt-5) of root of V. thunbergii collected from the herb market. The levels of ampelopsin C, (+)-vitisin A, and (-)-vitisin B were in the range of ND-0.35, ND-1.10, and ND-1.20 mg/g, respectively, which were markedly less than those in Vt-R. These five compounds in other six plants of Vitaceae were also investigated, including V. vinifera L., V. kelungensis Moriyama, Ampelopsis brevi-pedunculata (Maxim.) Traut., V. thunbergii var. taiwaniana Lu, Cissus pteroclada Hayata, and C. repens Lam. As a result, the former three plants were found to contain all of five standards. Except for resveratrol, other four compounds were not detected in V. thunbergii var. taiwaniana and C. repens. None of five compounds was found in C. pteroclada.