| Polygonum Multiflorum is the dried tuberous root of Polygonum multiflorum Thunb.The Chinese pharmacopoeia 2015 edition contains a collection of processed Polygonum Multiflorum slices,including stewed black bean juice pieces,steamed black bean juicepieces,steamed pieces.This three kinds of prepared Polygonum Multiflorum pieces have the same quality standards,so the lack of specific processing technology and quality standards leads to the inaccurate processing technology and difficult quality control,resulting in uneven quality of the decoction pieces,which seriously affects the clinical efficacy and drug safety.Therefore,on the basis of the 2015 edition of Chinese Pharmacopoeia,the processing technology,quality standard and QTOF-LC/MS qualitative analysis of main active components of Polygonum Multiflorum stewed with black bean juice were studied in this paper,which provided a reference basis for establishing the standard processing technology and the scientific and rational quality standard of Polygonum Multiflorum stewed with black bean juice.The principal research substance are summarized as follows:1 Study on processing technology of Polygonum Multiflorum stewed with black bean juiceDifferent process points in accordance with the traditional empirical judgment criteria of the Chinese pharmacopoeia 2015 edition were selected as the processing end point.The appearance characters,stilbene glycosides and anthraquinone contents of Polygonum Multiflorum stewed black bean juice for 8 hours to 96 hours were analyzed and studied.The content of stilbene glycoside decreased gradually with the increase of stewing time,compared with 8 h,the content decreased by 79% at 64 h.The content of emodin-8-O-β-D-glucoside and emodin methyl ether-8-O-β-D-glucoside increased first and then decreased,reaching the highest value at 24 h,and then decreased to the level similar to the content of 8 h after 40 h,then fluctuated slightly.The content of emodin and emodin methyl ether first increased and then decreased,reached the maximum when stewed for 32 h,then slowly decreased and tended to be stable.This indicated that the ingredients of processed Polygonum Multiflorum changed significantly with different stewing time,and the changing trend was different.The traditional empirical judgment standard led to too broad processing end point,which may be the chief reason for the uneven quality of processed Polygonum Multiflorum.2 Qualitative analysis of stewed Polygonum Multiflorum with black bean juice by HPLC-QTOF-MSIn order to further comprehensively characterize the differences of chemical components in processed Polygonum Multiflorum,this section selected the time points of 24 to 48 hours,which has a great difference in the ratio of chemical components,and performs a qualitative and quantitative analysis to determine the best processing end point of processed Polygonum Multiflorum stewed with black bean juice.HPLC-QTOF-MS was used to identify and analyze the multiple active components in Polygonum Multiflorum stewed with black bean juice.A total of 69 compounds were identified by the established QTOF-LC/MS qualitative analysis method,including 19 distyrene glycosides,11 quinones,11 flavonoids and 28 other compounds.The isoflavones of daidzin,daidzein,glycitein and genistein were the components of black beans.The unsupervised mode PCA analysis and supervised mode PLS-DA analysis of the peak areas of the marked compounds confirmed that the chemical compositions of different processing time Polygonum Multiflorum changed significantly,among which stilbene and emodin 8-O-β-D-glucoside is the most significant chemical constituents in Polygonum Multiflorum stewed in black bean juice at different processing time points.Consequently,stilbene glycoside is more reasonable as an index of quality control in Pharmacopoeia.Based on the results of qualitative analysis and quantitative analysis,the optimal processing time was determined to be 24 h.3 Quality standard of Polygonum Multiflorum stewed with black bean juiceAccording to the optimal processing time point to prepare processed Polygonum Multiflorum samples,the scientific and reasonable quality standards were formulated.The research on the quality control of Polygonum Multiflorum stewed with black bean juice is mainly focused on stilbene glycosides and anthraquinone glycosides,most of these ingredients reduced after processing,which were inconsistent with the efficacy trend of supplementary enhancement after processing.So there are some limitations in using them as index components for the processing quality of processed Polygonum Multiflorum.Therefore,the addition of sugars as the quality control index of Polygonum Multiflorum stewed with black bean juice can effectively avoid theoccurrence of toxic and side effects caused by inadequate processing.TLC method was used for qualitative identification.The characteristic chromatograms and the contents of stilbene glycosides and anthraquinones were determined by HPLC-DAD.The content of polysaccharide was determined by UV spectrophotometry,and the contents of D-fructose,D-glucose anhydrous and sucrose were determined by HPLC-ELSD.The final quality regulation of Polygonum Multiflorum pieces stewed with black bean juice were as follows: The consequence of The thin-layer chromatography of Polygonum Multiflorum stewed with black bean juice were clear and showed 4 color bands.The characteristic maps detected 9relatively stable chromatographic peaks as the indicator peak for qualitative identification.The content of stilbene glycoside should not be less than 0.70%in dry products.The total amount of emodin and emodin methyl ether should not be lower than 0.15%.The total contents of emodin-8-O-β-D-glucoside and emodin methyl ether-8-O-β-D-glucoside should not be below 0.01%.The content of polysaccharide was no less than 20%.The total amount of monosaccharide and disaccharides which were measured by D-fructose,D-glucose anhydrous and sucrose should not be less than 15%. |
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