Multi-wavelength fusion spectral extraction-assisted HPLC for simultaneous quantification of multiple secondary metabolites in herbal matrices

Abstract

Abstract The existing HPLC methods for the assaying of multiple markers in standardized herbal extracts use a single wavelength for quantification, which usually leads to lower assay than actual claims. It was because screening multiple phytochemical standards irrespective to their absorption maxima which drive decrease in peak are response which ultimately impact the assay. To overcome this issue, we have applied a methodology, called Mixed Standard Multi-Signal (MSMS) simultaneous detection, where multiple reference standards get extracted each with their respective λ max and quantification will be done at the same λ max in a single sequence data setup. This method was standardized using Single Signal Multi-Standards, Single Signal Single Standard, Multi-Signal Single Standard, for all individual reference standards, mixed reference standards, plant extracts, and Ayurvedic formulations. The results of our study demonstrate that higher assay reported total actives 5.04% by conventional detection and for current proposed method 13.81% with shorter span of analysis time and lesser solvent consumption. It helps to decide the actual standardization label claim on product which drives better commercial costings, and dosage regimen claims. Graphical Abstract

Keywords

• HPLC
• DAD
• MSMS
• Simultaneous detection
• Bioactives
• Food