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玄參藥材中高極性成分之分離及高效液相層析定量研究

Isolation and Determination by HPLC of Polar Constituents from Scrophularia ningpoensis

摘要


本研究選擇玄參藥材中高極性,可使用高效液相層析分析,在紫外線波長278nm具有吸收之成分,探討分離新方法。分離純化而得之2-(3-hydroxy-4-methoxyphenyl) ethyl 1-O-[α-L-arabinopyranosyl(1→6)]-feruloyl(1→4)]-α-L-rhamnopyranosyl(1→3)-β-D-glucopyranoside (SN-A),harpagoside(SN-B)及cinnamic acid(SN-C)三成分作為指標成分,利用高效液相層析儀探討其定量方法,並調查市售玄參藥材之指標成分含量。 高效液相層析法使用逆相層析管柱(Cosmosil 5C18-AR),移動相為乙腈及1.0%醋酸溶液,採線性梯度沖提[15:85→27:73 (44 min)],以SN-A(20~150μg/ml),B (5~50μg/ml)及C(20~200μg/ml)做為對照用標準品。其線性迴歸方程式及相關係數(r)分別為SN-A:Y=14.58X+11.25 (r=0.9907);SN-B:Y=2.26X-0.40 (r=0.9994);SN-C:Y=14.12X-29.28 (r=0.9981),均呈良好線性關係。三種指標成分之回收率,均在100±4%之內。同日內及異日間相對標準偏差試驗,其同日內為0.93~1.01%,異日間為0.28~1.97%,顯示高效液相層析法用於玄參指標成分之分析,具良好之再現性及準確性。 測定市售玄參藥材中三種指標成分之含量。結果SN-A之含量平均為0.30%(0.05%~1.71%),含量高低相差34倍;SN-B之含量平均為0.06%(未檢出~0.17%);SN-C之含量平均為025%(0.06%~0.53%),含量高低相差9倍。顯見各成分之含量分布範圍甚大,以SN-A相差34倍最高,而有一件藥材未檢測出SN-B成分。

並列摘要


Three polar constituents, 2-(3-hydroxy-4-methoxyphenyl) ethyl-1-O-[α-L-arabinopyranosyl (1→6)]-feruloyl(l→4)]-α-L-rhamnopyranosyl(1→3)-β-D-glucopyranoside(SN-A), harpagoside(SN-B) and cinnamic acid(SN-C) have been isolated from Scrophularia ningpoensis by a modified isolation method. These constituents were used as marker substances for quality analysis of Scrophulariae Radix. In this study we have also developed a high performance liquid chromatographic method for simultaneous determination of above marker substances in Scrophulariae Radix. This performance was done by using a gradient elution with a solution of acetonitrile in 1.0% acetic acid on a ODS column. The linear calibration ranges were SN-A 20-l50μg/ml (r=0.9907), SN-B 5-50μg/ml (r=0.9994) and SN-C 20-200μg/ml (r=0.9981). Recoveries were SN-A 100.47±3.23%, SN-B 98.49±2.62% and SN-C 96.15±4.39%. The relative standard deviations of three marker substances for intraday and interday analyses were 0.93-1.01% and 0.28-1.97%.. The contents of three marker substances in 24 crude drugs of Scrophulariae Radix were SN-A 0.05%~1.71%, SN-B non detected~0.17%, and SN-C 0.06%~0.53%, respectively.

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