利用化學成分分析技術建立山藥品系與指標活性成分關係

Isolation and analytical diosgenin characterisation for the application of simultaneous extraction and hydrolysis from Yam in Taiwan

林旺慶(Lin wang-ching)

指導教授 ： 鄧正賢

亞洲大學/健康學院/保健營養生技學系碩士班/碩士(2011年)

摘要

本研究建立薯蕷皂配基 (diosgenin) 萃取、酸水解、提取三步驟合一的雙相酸水解法，以簡單方便快速的測定山藥中薯蕷皂配基 (diosgenin) 的含量，可作為篩檢不同品系山藥優劣的參考方法。
此方法採用採用雙相酸水解法確定山藥中薯蕷皂配基的含量，其步驟為取樣品凍乾粉末加入含 2.5 mol/L 鹽酸的 60％ 甲醇水溶液和石油醚各 100 mL，於沸水浴中加熱回流提取，溶劑輸送系統為 Waters 600E Gradient Pump，附自動注射器 (Autosampler 717+)，連接 Waters 486 紫外光/可見光偵測器，管柱為 Merck 公司 Lichrospher 100 RP-18e ( 5 m, 4.0 I.D.×250 mm；注射濾膜為 Millipore (PVDF) 13 mm Syringe Filter 0.45 m；分析山藥皂苷元 (diosgenin) 指標成分時，注射量為 20 μL，移動相為 CH3CN / H2O (90: 10, v/v)，流速為 1.0 mL/min，偵測波長為 203 nm，管柱壓力為 1060±50 psi。利用上述分析技術進行對照標準品薯蕷皂配基分析，薯蕷皂配基在濃度 100-1000 g/mL，得線性迴歸方程式 (Y=mX+b) 及相關係數(r)為 Y=0.0515X-9.277 (r=0.9916)，顯示良好線性關係。
薯蕷皂配基同日內及異日間相對標準偏差 1.39-3.54% 及 2.05-4.77 %，顯示再現性可以接受。山藥添加回收率為 64.2-72.0%，以薯蕷皂配基需由薯蕷皂苷酸水解去除醣基及液液萃取而言，尚在可接受的範圍；以三倍於指標成分和雜訊之波峰高度比的最小濃度視為儀器的偵測極限，薯蕷皂配基的偵測極限為 35 g/mL。
台灣地區基隆山藥(D. japonica Thunb. var. pseudojaponica (Hayata) Yamamoto)、花蓮 3 號 (D. batatas Dence.) (壽豐山藥)、恆春山藥 (D. doryophora Hance) 檢品的檢測，其中基隆山藥中山薯蕷皂配基含量為皮部 23.1±10.3 mg / 5 g 凍乾粉末、肉部 4.5±1.3 mg / 5 g 凍乾粉末；恆春山藥皮部 1.04±0.13 mg / 5 g 凍乾粉末，其餘則未檢出。

關鍵字

薯蕷皂配基(diosgenin) ； 雙相酸水解法 ； 液相層析

並列摘要

To establish an RP-HPLC method for determination of diosgenin in Dioscoreae spp. for quality assessment index in Sub-project 3. The process of simultaneous ultrasonic extraction and hydrolysis for the recovery of diosgenin from Yam was optimized by different pre-processing tests. To accurately determinating the diosgenin, the glycosyl groups on the saponins, both spirostanol and furostanol glycosides, shall be removed by acid hydrolysis and converted to diosgenin (aglycone) before HPLC. The samples were analyzed by HPLC on a Lichrospher 100 RP-18e column and detected at 203 nm with acetonitrile water (90: 10, v/v) as the mobile phase at a flow rate of 1.0 mL/min. The regression equations of diosgenin was Y = 0.0515X-9.277 (r = 0.9916). The intraday and interday relative standard deviations of diosgenin were at the levels of 1.39-3.54% and 2.05-4.77%, respectively. The recoveries of diosgenin added in yam were between 64.2% and 72.0%. The method was applied to three yams grown in Taiwan (D. japonica Thunb. var. pseudojaponica (Hayata) Yamamoto, D. batatas Dence. and D. doryophora Hance), diosgenin was detected in two samples, measurable at 1.04-23.1 mg/g. It was found that the cortex of D. japonica yam tuber contained 4.6±2.1 mg/g which was the highest amount comparison with different parts of yam.

並列關鍵字

Yam ； Dioscorea spp ； diosgenin

參考文獻

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