Analysis of Ginsenoside and Notoginsenoside in Panax Notoginseng

Significance of the Topic

Panax notoginseng is widely used in traditional medicine for its neuroprotective and antioxidant properties. The bioactivity is mainly attributed to dammarane-type saponins called ginsenosides. Reliable analytical methods to quantify key ginsenosides such as Notoginsenoside R1, Ginsenoside Rg1 and Ginsenoside Rb1 are essential for quality control, standardization and pharmacological studies.

Objectives and Study Overview

This application note describes the development of a reversed-phase liquid chromatography method to separate and analyze three major ginsenosides in Panax notoginseng. The goal is to achieve baseline resolution in a single run for both standard mixtures and commercial samples.

Methodology and Sample Preparation

The workflow consists of two main parts:

• Sample Pretreatment
  1. Grind and sieve commercial Panax notoginseng to uniform particle size.
  2. Weigh 0.6 g and add 50 mL methanol in a stoppered flask.
  3. Let stand overnight, then reflux at 80 °C for 2 hours.
  4. After cooling, adjust the extract volume back to original weight with methanol and filter into vials.
• Chromatographic Conditions
  1. Column: Shim-pack GIS C18 (250 × 4.6 mm, 5 µm).
  2. Mobile Phase: A) Water, B) Acetonitrile; gradient from 19 % B at 0 min to 36 % B at 60 min, then return to 19 % B by 60.1 min.
  3. Flow Rate: 1.0 mL/min; Column Temperature: 20 °C; Injection Volume: 10 µL.
  4. Detection: UV at 203 nm.

Used Instrumentation

• Shimadzu HPLC system equipped with Shim-pack GIS C18 column (P/N 227-30106-08).
• LabTotal™ Vial for LC/LCMS (P/N 227-34001-01).
• UV detector set at 203 nm.

Main Results and Discussion

The method achieves clear separation of Notoginsenoside R1, Ginsenoside Rg1 and Ginsenoside Rb1 in both standard mixes and real samples. Retention order is R1 first, followed by Rg1 and then Rb1. Chromatograms show sharp peaks, good resolution and reproducible retention times, confirming method robustness.

Benefits and Practical Applications

This LC method offers:

• Accurate quantification for quality control of herbal products.
• Applicability to routine analysis in research and industrial laboratories.
• Compatibility with other detection techniques for expanded profiling.

Future Trends and Opportunities

Emerging directions include:

• Coupling with mass spectrometry for structural elucidation of minor saponins.
• Green extraction methods to reduce solvent consumption.
• High-throughput platforms for large-scale screening of Panax species.

Conclusion

The presented reversed-phase HPLC method on Shim-pack GIS C18 delivers reliable separation of key ginsenosides in Panax notoginseng. Its precision and reproducibility make it a valuable tool for quality assurance and research on bioactive compounds.