SIMULTANEOUS QUANTIFICATION OF TWO MAJOR ACTIVE COMPONENTS IN RADIX ANGELICA SINENSIS BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH AN INTERNAL STANDARD CORRECTION METHOD

Abstract

Ligustilide and ferulic acid are the major active components in Angelica sinensis (Oliv.) Diels (AS). However, no simple and practical method to determine AS is available because the reference substance of ligustilide is difficult to prepare, unstable, and expensive. Recently, the Internal Standard Correction Method (ISCM) by high-performance liquid chromatography was developed to determine ligustilide and ferulic acid in AS simultaneously. In the present article, ferulic acid is used as the internal reference standard, and its content is determined via an external standard method. The relative correction factor (RCF) of ligustilide to ferulic acid is calculated. Ligustilide content is determined by the ISCM, which is calculated using ferulic acid content and its RCF. The accuracy of the new method is evaluated by comparing the calculated contents with the determined contents in 22 batches of samples collected from different areas. Upon validation of the analysis procedures, the calculated and determined contents in 22 batches of AS show no significant difference. The calculated RCF has a confidence value of 0.2209. The relative retention time of ligustilide to ferulic acid is 16.918 min. The proposed method was developed on the premise that it could determine ligustilide and ferulic acid simultaneously without the reference substance of ligustilide. Hence, it is a simple, sensitive, and reliable method for the quality control of AS and Chinese herbal preparations containing AS.

Keywords:

• Angelica sinensis
• high-performance liquid chromatography
• internal standard correction method
• related correction factor

ACKNOWLEDGMENT

The Cultivation Fund of Scientific and Technical Research of Traditional Chinese Medicine of Gansu Province (GZK-2010-57) and Cultivation Fund of the Scientific and Technical Innovation Project of Gansu province (1104FKCA122) financially supported this research work.

Notes

^a x, amount of substance (µg); y, peak area.

^b LOD, limit of detection (S/N = 3).

^c LOQ, limit of quantification (S/N = 10).

^a RSD (%) = (SD/mean) × 100.

^b Three different concentrations of the reference solution containing two reference standard compounds.

^a External standard method.

^b ISCM method.