Ultrasonication Extraction and Solid Phase Extraction Clean-up for Quantification of Aristolochic Acid I in the Thai Herbal Antitussive 3 Formula by High Performance Liquid Chromatography with Photodiode Array Detection
Keywords:
Aristolochic acid; Aristolochia tagala Cham.; Thai Herbal Antitussive 3 Formula; High Performance Liquid Chromatography (HPLC)Abstract
Objective: To establish a reliable High Performance Liquid Chromatography method with photodiode array detection (HPLC-PDA) and a suitable extraction method for identification and quantification of Aristolochic acid I (AA-I) in 3 different sample types, i.e. Aristolochia tagala Cham.(ATC), Thai Herbal Antitussive 3 Formula (ATF3), and Thai Herbal Antitussive 3 Formula containing ATC (ATF3+ATC).
Methods: Two extraction methods, ultrasonication extraction (UE) and solid phase extraction (SPE), were used in the present study. Reversed phase separations were performed with a SunFire C18 column and HPLC-PDA.
Mobile phase was performed by 1 mL/min infusion of mixture of 0.1% o-phosphoric acid and methanol (35:65).
Results: Calibration curve showed a good linearity with R2 equal to 0.9986. Every sample type has been analyzed. While the concentration of AA-I in ATF3 cannot be determined due to the absence of the AA-I peak, the AA-I concentrations in ATC and ATF3+ATC were 7.50-11.34 and 6.78-11.77 mg/mL respectively. In comparison with UE, SPE is more effective in eradication of the interfering compounds and improves the absorbance spectrum of the peak and increases extraction efficiency.
Conclusion: A reliable HPLC-PDA and SPE methods for identification and quantification of AA-I in ATF3+ATC, ATF3, and ATC were established in the present study. The SPE was more effective than UE in the pretreatment step.
Downloads
Published
How to Cite
Issue
Section
License
Users are free to share, copy, and redistribute all articles published in the Siriraj Medical Journal (SMJ) in any medium or format as long as you follow the following terms:
- Attribution — You must give appropriate credit, provide a link to the material, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the publisher endorses you or your use.
- NonCommercial — You may not use the material for commercial purposes.
- NoDerivatives — If you remix, transform, or build upon the material, you may not distribute the modified material.
- No additional restrictions — You may not apply legal terms or technological measures that legally restrict others from doing anything the license permits.