LC–MS Based Workflows for Qualitative and Quantitative Analysis for Homeopathic Preparation of Hydrastis canadensis

Hydrastis canadensis L. is one of the important herbal medicinal plants, which is used for homeopathic preparations. It contains berberine, hydrastine as major and canadine, hydrastinine, berberastine and canadaline as minor bioactive compounds. The aim was to develop (I) a sensitive mass spectrometric-based quantitative/qualitative method for berberine in homeopathic dilutions of H. canadensis, (II) with identification of berberine and its structurally related compound present in the homeopathic preparation of H. canadensis using predictive MRM workflow, (III) confirmatory analysis for the structurally related compound in homeopathic dilutions of H. canadensis using high-resolution mass spectrometer. Different mass spectrometric-based workflows (MRM to EPI; pMRM to EPI and IDA-DBS) were used to develop the method to analyze the homeopathic dilutions of H. canadensis. A high-throughput liquid chromatography-tandem mass spectrometry method using MRM to EPI workflow was developed to analyze berberine in homeopathic preparation of H. canadensis. Multiple reaction monitoring transitions of 336.0/292.08 and 336.0/277.9 were selected for developing the quantitation method of berberine. The method was linear from 5.0 to 2000.0 pg/mL. The validated method has been successfully applied for the analysis of different potencies of homeopathic preparation of H. canadensis. Predictive MRM to enhanced product ion workflows was used to identify and confirm the structurally related molecules of berberine. Non targeted information dependent acquisition-DBS method was developed to identify the compounds in H. canadensis mother tincture using high-resolution mass spectrometry. The established method using different mass spectrometric workflows facilitates the convenient and fast quality control analysis for the herbal preparation of H. canadensis.