Analysis of Ginsenoside and Notoginsenoside in Panax Notoginseng

Importance of Topic

Panax Notoginseng is valued for its bioactive dammarane type saponins, notably ginsenosides Rb1 and Rg1 and notoginsenoside R1. Accurate quantitation of these compounds is essential for quality control in herbal products, pharmacokinetic studies, and evaluation of neuroprotective and antioxidant activity.

Study Objectives and Overview

This study demonstrates a robust reversed phase HPLC method using a Shim-pack GISS C18 column to separate and quantify ginsenoside Rb1, ginsenoside Rg1, and notoginsenoside R1 in extracts of Panax Notoginseng. A standard mix was analyzed to establish retention times and calibration conditions before applying the procedure to sample extracts.

Methodology and Instrumentation

• Sample preparation:
  1. Commercial Panax Notoginseng powder sieved and 0.6 g extracted in 50 mL methanol with overnight soak and 2-hour reflux at 80°C.
  2. Post-extraction weight adjustment with methanol and filtration into HPLC vials.
• Chromatographic conditions:
  1. Column Shim-pack GISS C18 (250×4.6 mm, 5 μm).
  2. Mobile phase water (A) and acetonitrile (B) with gradient from 19% B to 36% B over 60 min, returning to 19% B by 75 min.
  3. Flow rate 1.0 mL/min, column temperature 25°C, injection volume 10 μL.
  4. Detection UV absorbance at 203 nm; LabTotal vials for LC/LCMS used.

Main Results and Discussion

The method achieved baseline separation of notoginsenoside R1, ginsenoside Rg1, and ginsenoside Rb1, with retention times suitable for quantitative analysis. The standard mix containing 0.4 mg/mL of R1 and Rg1 and 0.1 mg/mL of Rb1 produced well-resolved peaks. The gradient profile provided consistent elution reproducibility, and UV detection at 203 nm exhibited sufficient sensitivity for routine quality control applications.

Benefits and Practical Applications

• Provides reliable quantitation of key ginsenosides in herbal formulations and raw materials.
• Applicable to quality assurance in nutraceutical and pharmaceutical production.
• Supports pharmacological research into antioxidant and neuroprotective properties.

Future Trends and Potential Applications

• Integration with mass spectrometric detection to expand structural characterization of minor saponins.
• Method adaptation for high throughput screening using UHPLC or core shell columns to reduce run times.
• Development of standardized reference materials for harmonization across laboratories.

Conclusion

This reversed phase HPLC approach using Shim-pack GISS C18 reliably separates and quantifies major ginsenosides in Panax Notoginseng extracts. Its straightforward sample preparation and gradient program make it well suited for routine analysis in both research and industrial settings.

Reference

Shimadzu Corporation Application note ERAS-1000-0030 First Edition March 2023.