HPLC Method Development for the Separation and Quantitative Determination of Rg1 and Re Ginsenosides from Tissue Culture
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Abstract
Ginsenosides are triterpenoid saponins found in Panax ginseng. They are well known for their health benefits, including antioxidant, anticancer, and antidiabetic effects, along with their support for cardiovascular and neurological function. Our previous study showed that cultivating Panax ginseng in tissue culture successfully led to its rapid growth. Several active compounds, including Rg1 and Re ginsenosides, were identified in the powdered extract of cultivated Panax ginseng. As part of a continuous study, an analytical method was developed to ensure the consistency of yield composition with the required quality standards. The analytical method optimization involved selecting the mobile phase composition and adjusting the flow rate in both isocratic and gradient systems to achieve optimal separation. The gradient elution system, with a constant flow rate of 1.0 mL/min, was more effective than the isocratic system for separating ginsenosides. The optimized method was then validated, showing acceptable validation parameters. The validated method was applied to the ginsenosides assay in the powdered extract of the Panax ginseng sample. Eight out of fourteen ginsenosides were identified, including Rg1 and Re, which were quantified at 0.16% and 0.45%, respectively. A baseline resolution of about 5.16 between Rg1 and Re ginsenosides was achieved. The analytical method developed using HPLC in this study effectively separated ginsenoside compounds simultaneously. These findings proposed an analytical method to evaluate the quality standards of ginsenosides in Panax ginseng.
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