Objective To establish the HPLC fingerprint of rhubarb and determine the effect of processing time on the quality of rhubarb based on the variation of peak area of the main components.Methods The chromatographic column was Eclipse Plus C18(4.6 mm×200 mm,5.0 μm).Gradient elution was performed with methanol-0.1％phosphoric acid.The flow rate was 1.0 mL·min-1.The detection wavelength was 280 nm and the column temperature was 35 ℃.The chromatographic fingerprints of rhubarb samples from different origins were processed based on the Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System(2012 edition)of National Pharmacopoeia Committee.The HPLC chromatograms of rhubarb charcoal samples with different processing time were compared.Results Totally 18 common peaks were identified in the established HPLC fingerprint of rhubarb.By comparing with the reference substance,8 components were identified,namely gallic acid,catechin,aloe emodin,rhein,emodin,chrysophanol,physcion,and 5-hydroxymethylfurfural.Five new components were found,and the content of each component changed with processing time.Conclusion The established rhubarb HPLC fingerprint method is reproducible and stable,which provides reference for the quality control and evaluation of rhubarb charcoal.
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