Analysis of Ginsenosides and Notoginsenoside in Herbal medicine

Significance of the topic

The accurate quantification of ginsenosides and notoginsenosides in Panax-based herbal preparations is essential for quality control, efficacy assessment and regulatory compliance. Reversed-phase high-performance liquid chromatography (RP-HPLC) offers a reliable platform to resolve these structurally related triterpene saponins, ensuring consistent dosing and safety in botanical products.

Objectives and study overview

This study aimed to establish and validate an RP-HPLC method using Shimadzu’s Shim-pack GISS C18 column to separate and quantify four key analytes: notoginsenoside R1, ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1. The protocol was applied to both standard mixtures and a commercially available herbal medicine to demonstrate its practicality in routine analysis.

Methodology and instrumentation

Sample Preparation:

• Weigh 0.5 g of finely powdered herbal product into a stoppered flask.
• Add 50 mL of 70 % methanol–water, seal and record weight.
• Ultrasonicate at 250 W, 33 kHz for 30 minutes.
• Allow cooling, adjust solvent to original weight with 70 % methanol–water, mix and filter into an LC vial.

Chromatographic Conditions:

• Column: Shim-pack GISS C18, 250 mm × 4.6 mm i.d., 5 µm.
• Mobile phase: (A) water; (B) acetonitrile with linear gradient from 19 % B (0–35 min) to 29 % B (55–70 min) and 40 % B (100 min).
• Flow rate: 1.0 mL/min; Column temperature: 30 °C; Injection volume: 20 µL.
• Detection: UV absorption at 203 nm; Vial: LabTotal™ vial for LC/LCMS.

Main results and discussion

The method achieved baseline separation of notoginsenoside R1, ginsenosides Rg1, Re and Rb1 with distinct retention times. Calibration standards at concentrations between 0.05 and 0.20 mg/mL exhibited linear responses, demonstrating good sensitivity. Analysis of the herbal medicine sample showed clear peak identification and consistent quantitation, highlighting the robustness of the procedure.

Benefits and practical applications

This RP-HPLC protocol provides:

• High resolution of structurally similar saponins.
• Reproducible retention times and quantitation suitable for quality assurance.
• Applicability to routine testing in research, manufacturing QC and regulatory labs.

Future trends and applications

Advancements may include coupling with mass spectrometry for enhanced selectivity, adoption of green solvent systems to reduce environmental impact, and implementation of shorter columns or elevated temperatures for faster throughput. Integration into automated workflows will further streamline quality control of botanical products.

Conclusion

The developed RP-HPLC method on Shimadzu’s Shim-pack GISS C18 column offers a straightforward, reliable approach for the simultaneous determination of key Panax saponins in herbal formulations, supporting comprehensive quality assessment and regulatory compliance.

Instrumentation used

• Shim-pack GISS C18 column (250 mm × 4.6 mm i.d., 5 µm).
• Shimadzu HPLC system with UV detector at 203 nm.
• Ultrasonic bath (250 W, 33 kHz).
• LabTotal™ LC/LCMS vials.