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Simultaneous Analysis of Bioactive Metabolites from Rehmannia glutinosa by HPLC-DAD-MS/MS  

Won, Tae-Hyung (Natural Products Research Institute, College of Pharmacy, Seoul National University)
Ryu, Sung-Kwang (Natural Products Research Institute, College of Pharmacy, Seoul National University)
Kang, Sam-Sik (Natural Products Research Institute, College of Pharmacy, Seoul National University)
Shin, Jong-Heon (Natural Products Research Institute, College of Pharmacy, Seoul National University)
Publication Information
Natural Product Sciences / v.16, no.2, 2010 , pp. 116-122 More about this Journal
Abstract
A high-performance liquid chromatography (HPLC) with diode array detector (DAD) and electrospray ionization mass spectrometry (ESI-MS) was established for the simultaneous determination of five representative metabolites of the iridoid and phenolic classes from Rehmannia glutinosa. The optimal chromatographic conditions were obtained on an ODS column (5 mm, $4.6{\times}250\;mm$) with the column temperature at $25^{\circ}C$. The mobile phase was composed of water and acetonitrile using a gradient elution with the flow rate 0.3 mL/min. Detection wavelength was set at 205 nm. All calibration curves showed good linear regression ($r^2$ > 0.997) within test ranges. Limits of detection (LOD) and quantitation (LOQ) values were lower than 0.123 and $0.373\;{\mu}g/mL$, respectively. The developed method provided satisfactory precision and accuracy with overall intra-day and inter-day variations of 0.09 - 0.76% and 0.16 - 1.41%, respectively, and the overall recoveries of 99.03 - 102.67% for all of the compounds analyzed. In addition, effectiveness of diverse extraction methods was compared to each other for the development of standard analytic method. The verified method was successfully applied to the quantitative determination of five representative metabolites in twenty-one commercial Rehmannia glutinosa samples from different markets in Korea and China. The analytical results showed that the contents of the five analytes vary significantly with sources.
Keywords
HPLC-DAD-MS; Rehmannia glutinosa; iridoids; phenolic compounds;
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