A Non-Derivative Method For The Quantitative Analysis Of Isosteroidal Alkaloids From Fritillaria By High Performance Liquid Chromatography Combined With Charged Aerosol Detection

Fritillaria is a commonly used Traditional Chinese herbal medicine that has antitussive activity. The primary active ingredients of fritillaria are isosteroidal alkaloids – their analysis is critically important for the quality control of fritillaria or fritillaria-derived medicines. This can be challenging as the isosteroidal alkaloids do not possess a suitable chromophore for UV absorbance detection. To overcome this problem a non-derivative HPLC method was developed for the qualitative and quantitative analysis of isosteroidal alkaloids from Fritillaria thunbergii. During method development the performance of two universal detectors, the charged aerosol detector (CAD) and evaporative light scattering detector (ELSD), was evaluated. The CAD was found to be 30 to 55 times more sensitive than the ELSD enabling the measurement of low levels of reference compound impurities that could not be detected by the ELSD. For example, the peak area percent of the reference compound, peimisine, obtained by the CAD was only 50%, but 92% by the ELSD. The CAD showed good reproducibility with overall intra- and inter-day precision of less than 1.8% and 2.7%, respectively. The CAD had a dynamic range of up to 4 orders of magnitude (0.06 – 44 mg/L) for peimine and peiminine. The optimized method was used for the quantitative analysis of peimine and peiminine from Fritillaria thunbergii.