Determination of Triterpene Glycosides in Cimicifuga racemosa (Black Cohosh) by HPLC-CAD

Significance of the Topic

Black cohosh is widely used to alleviate menopausal symptoms and its active compounds are triterpene glycosides that lack strong UV chromophores

Study Objectives and Overview

This application study compares charged aerosol detection with traditional evaporative light scattering detection for quantifying triterpene glycosides in black cohosh extract

Methodology and Instrumentation

Sample Preparation

• Standard solution of 27-deoxyactein prepared in methanol with serial dilutions from 10 to 100 micrograms per milliliter
• Sample extract dissolved in methanol by sonication, filtered through 0.22 micron PTFE filters

Chromatographic Conditions

• Column Fused Core C18 4.6 x 150 mm 2.7 micron
• Mobile phase A 0.1 percent formic acid in water B acetonitrile gradient from 30 to 60 percent B over 36 minutes
• Flow rate 1.0 milliliter per minute, column temperature 35 degrees Celsius, injection volume 10 microliters

Instrumentation

• HPLC system with quaternary pump and autosampler
• Charged aerosol detector for improved sensitivity and linearity
• Evaporative light scattering detector operated for comparison

Results and Discussion

Calibration Performance

• Charged aerosol detection produced a linear calibration curve for 27-deoxyactein with high correlation
• ELSD required a logarithmic calibration and showed greater variance at low and high concentrations
• Signal to noise improved by approximately tenfold with charged aerosol detection

Sample Analysis

• Eleven triterpene glycosides quantified using relative retention times against the 27-deoxyactein standard
• Total glycoside content in the extract measured at 3.27 percent

Benefits and Practical Applications

Charged aerosol detection offers mass based response nearly independent of analyte structure and enables consistent quantitation of nonchromophoric glycosides in quality control of nutraceuticals

Future Trends and Opportunities

Expansion of charged aerosol detection to other natural product classes and adoption of inverse gradient techniques to correct for solvent composition effects will further improve robustness and throughput in industry and research labs

Conclusion

Charged aerosol detection outperforms ELSD for triterpene glycoside analysis by delivering linear calibration, enhanced sensitivity and reproducibility making it a superior choice for QC applications

References

1. NSF International Black Cohosh Assay by ELSD INA Method 113.001