This study describes the successful cleanup of alkaloids (hydrastine and berberine) in goldenseal using the Agilent Bond Elut Plexa solid phase extraction sorbent. HPLC separation was achieved in less than 6 minutes using an Agilent RRHD ZORBAX Eclipse Plus C18 column (4.6 x 75 mm, 3.5 µm) at 35 °C with a gradient elution of 0.1% phosphoric acid/methanol mobile phase. The calibration curves of both alkaloids showed a good correlation coefficient of 0.9994. The recoveries of hydrastine ranged from 76-102% while those of berberine ranged from 99-104% with a relative standard deviation (RSD) % of less than 5.
This document details an analytical method for determining the alkaloids hydrastine and berberine in goldenseal dietary supplements. The method utilizes Agilent Bond Elut Plexa solid phase extraction (SPE) sorbent to clean up complex sample matrices before analysis.
The study validates the SPE procedure using high-performance liquid chromatography with a diode array detector (HPLC-DAD). Results indicate that the method provides accurate and reproducible quantification, with recoveries ranging from 76-102% for hydrastine and 99-104% for berberine, and relative standard deviations (RSD) of less than 5%.
| Column dimensions | 4.6 x 75 mm, 3.5 µm |
|---|---|
| Column temperature | 35 °C |
| Flow rate | 1.00 mL/min |
| Run time | 6 min |
| Hydrastine recovery range | 76-102% |
| Berberine recovery range | 99-104% |
| Hydrastine LOD | 0.50 pg/mL |
| Berberine LOD | 0.47 pg/mL |
The recovery range for hydrastine is 76-102%.
The recovery range for berberine is 99-104%.
The limit of detection for hydrastine is 0.50 pg/mL.