Development and validation of simultaneous HPLC-PDA analysis method for quality control of Hwang-ryeon-hae-dok-tang: An analytical quality by design approach

Abstract

This study introduces an advanced analytical method for quality control of botanical drugs, focusing on Hwang-ryeon-hae-dok-tang (HRHDT), a traditional herbal prescription utilized for inflammation, gastritis, fever, and hypertension. Botanical drugs pose a challenge due to their complex composition, requiring reliable and robust analytical techniques. Our step-by-step procedure included defining the analytical target profile, risk assessment, screening and optimizing design of experiment, measuring the design space, and Monte Carlo simulation. The established HPLC-PDA method enables the simultaneous analysis of four major compounds (geniposide, baicalin, palmatine, and berberine) present in HRHDT. The optimized conditions involve a gradient elution with acetonitrile and water, both containing 0.1% trifluoroacetic acid, a 32 °C column temperature, 254 nm detection wavelength, and 1.0 mL/min flow rate. A successful validation demonstrated excellent linearity (R²: 0.9984–0.9989) and accuracy (recovery: 100.61%–102.44%), with precise repeatability and intermediate precision (%RSD: 0.39%–0.83%). These results further confirm the reliability and accuracy of the analytical method. Overall, our design-based approach not only facilitated the development of robust analytical methods but also allowed for flexible operation ranges. Therefore, this methodology holds significant potential for ensuring the quality of botanical drug products within the pharmaceutical industry.

Keywords:

• Simultaneous quantitation
• quality control
• analytical quality by design (AQbD)
• HPLC-PDA analysis
• hwang-ryeon-hae-dok-tang (HRHDT)

Acknowledgments

This study was funded by a grant (21173MFDS561) from the Ministry of Food and Drug Safety in Korea.

Authors’ contributions

Geonha Park: conceptualization, project administration, resources, softwares, validation, writing—original draft, writing—review and editing. Seung Hyeon Go: conceptualization, data curation, formal analysis, investigation, methodology, resources, software, visualization, writing—original draft. Sejin Ku: investigation, resources, validation. min kyoung kim: conceptualization, visualization. Young Pyo Jang: conceptualization, supervision, project administration, writing—review and editing, funding acquisition.

Disclosure statement

The authors declare no conflict of interest.

Additional information

Funding

Ministry of Food and Drug Safety in Korea (21173MFDS561).