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-metlioxypheiiyl) ethyl l-0-[ot-L-ara-binopyranosyl(1^6)]-feruloyl(l~>4)]-a-L-rhamnopyranosyl(1^3)-P-D-glucopyranoside (SN-A) > harpagoside(SN-B)及cinnamic acid(SN-C)三成分作爲指操成分’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)，移動相爲乙月青及1.0%醋酸溶液， 採線性梯度沖提[15:85—27:73 ( 44 1^11)]，以3队八(20~150问/1111)，；6(5~50叫/1111)及(：(20~ 20(Hlg/ml)做爲對照用標準品。其線性迺歸方程式及相關係數(r)分別爲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 0- 0.9981)，均呈良好 線性關係。三種指標成分之回收率，均在1〇〇±4%之内。同日内及異日間相對標準偏差試 驗，其同日内爲0.93~1,01%，異日間爲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之含量平均爲0.30%(0.05%〜 1.71%)，含量高低相差34倍；SN-B之含量平均爲0.06%(未檢出〜0.17%) ; SN-C之含量平均爲 0.25%(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。

Three polar constituents, 2-(3-hydroxy-4- methoxyphenyl) ethyl 1-0-[o:-L-arabinopyranosyl (1--+6)]-feruloyl(1--+4)]-o:-L-rhamnopyranosy I ( 1--+ 3 )-~- 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- 150J.!g/ml ( r=0.9907 ), SN-B 5-50J.!g/ml ( r=0.9994 ) and SN-C 20-200J.!g/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.