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Article Title

Isolation and Determination by HPLC of Polar Constituents from Scrophularia ningpoensis

Abstract

Three polar constituents, 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) 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 20-150μg/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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