Optimisation for simultaneous determination of iridoid glycosides and oligosaccharides in Radix Rehmannia by microwave assisted extraction and HILIC‐UHPLC‐TQ‐MS/MS. Issue 3 (3rd January 2020) - Wang, Xin
Abstract: Hydrophilic iridoid glycosides and oligosaccharides are the major active ingredients of Radix Rehmanniae. Analysis of oligosaccharides is a challenging task because they are highly hydrophilic, with similar chemical structures and absence of chromophore and fluorophore groups. The difficulty for simultaneous analysis of iridoid glycosides and oligosaccharides in Radix Rehmanniae is increased due to the polarity difference between the two types of ingredients. Objective: To develop a method for simultaneous determination of iridoid glycosides (ajugol, catalpol) and oligosaccharides (sucrose, melibiose, raffinose, mannotriose and stachyose) in Radix Rehmanniae. Methodology: Microwave‐assisted extraction (MAE) was established to extract target analytes from Radix Rehmanniae samples using methanol–water (60:40, v / v ) as the extraction solvent. Fast separation of seven analytes was achieved by hydrophilic interaction liquid chromatography (HILIC) using an Accucore‐150‐Amide‐HILIC column. Sensitive and selective detection of the analytes was performed by triple quadrupole tandem mass spectrometry (TQ‐MS/MS) using multiple reaction monitoring in positive electrospray ionisation mode. Results: Good linearities were achieved for all the analytes with the correlation coefficients above 0.9991. The precisions resulted in deviations of less than 5.0% and the recoveries ranged from 93.8% to 105.5%. The established method was successfully applied to the analysis of iridoid glycosides and oligosaccharides in 12 samples of crude and processed Radix Rehmanniae. Conclusion: A simple, rapid and sensitive method based on MAE combined with HILIC‐UHPLC‐TQ‐MS/MS was developed for simultaneous determination of iridoid glycosides and oligosaccharides in Radix Rehmanniae for the first time. The method exhibited excellent performance with simple sample preparation, short analysis time, high selectivity and sensitivity. Abstract : A simple, rapid and sensitive method based on microwave‐assisted extraction (MAE) combined with hydrophilic interaction liquid chromatography ultra‐high‐performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry (HILIC‐UHPLC‐TQ‐MS/MS) was developed for simultaneous determination of iridoid glycosides and oligosaccharides in Radix Rehmanniae for the first time. The developed method was successfully applied to determine two iridoid glycosides and five oligosaccharides in crude and processed Radix Rehmanniae. The method exhibited excellent performance with simple sample preparation, short analysis time, high selectivity and sensitivity.