Research On Quality Evaluation Of The Process Of The Harvest And Processing Of Paeoniae Radix Alba Based On Quality Marker

Objective:Paeoniae Radix Alba（PRA）is the dried root of Paeonia lactiflora Pall.It has the effect of nourishing blood and regulating menstruation,collecting Yin and antiperspirant,softening liver and relieving pain,and suppressing liver Yang.Bo PRA is also one of the Ten Anhui medicines,there are mainly“XT”,“PB”,“XPB”and other varieties,harvesting years and months are different.In this study,by studying the specific chromatogram and the changes of active components in the transmission process of PRA materia medica-decoction piece-standard decoction,the traceable quality marker components of the whole process of PRA production were selected,and a Q-Marker multi-evaluation quality evaluation system was established to optimize the harvesting period and processing technology of PRA.To further improve the clinical efficacy of PRA decoction pieces and quality evaluation of PRA.Methods:1.Study on quality markers of PRA based on the transfer process of medicinal materials-decoction pieces-standard decoctions:Collecting 10 batches of PRA in Bozhou City and preparing them as medicinal materials,decoction pieces and standard decoctions,HPLC is adopted to establish the specific chromatogram of medicinal materials,decoction pieces and standard decoctions of PRA,combined with the similarity evaluation and OPLS-DA,the content and transfer rate of the preliminarily determined indicator components in the process of medicinal materials,decoction pieces and standard decoctions were analyzed,the quality transfer law in the production process of PRA is explored,and the Q-Marker of PRA is comprehensively screened.2.A multi-evaluation quality evaluation system of PRA was established based on Q-Markers:Using paeoniflorin as internal standard,the relative correction factor f_i/svalues of oxypaeoniflorin,Paeoniflorin,gallic paeoniflorin and benzoyl Paeoniflorin were calculated.The relative correction factors of Agilent 1260 and Waters e2695HPLC were investigated respectively.Agilent ZORBAX SB-C18,SHIMADZU Shim-pack GISC18,Welch Ultimate Plus C18 column;25℃,30℃,35℃column temperature;0.8m L/min,1.0m L/min,1.2m L/min flow rates,0.01%phosphoric acid solution,0.05%phosphoric acid solution,0.1%phosphoric acid solution mobile phase and 228nm,230nm,232nm detection wavelength durability.The content of paeoniflorin and other five chemical components in 10 batches of samples from PRA were compared with that of QAMS method to verify the accuracy and feasibility of QAMS method.3.Study on the optimum harvesting period of PRA based on Q-Marker:Samples of four-year-old“XT”,“PB”and“XPB”PRA were collected at fixed sampling points every month from June to December 2021 to investigate different harvesting months.In September 2021,samples of“XT”aged 3-5 years and“PB”aged 3-4 years from Bozhou PRA were collected to investigate different growth years.The fresh weight and dry weight of samples were determined and the drying rate was calculated.The yield and quality of PRA in each harvesting month and different growth years were analyzed combined with Q-Marker content,and the weights of five components were determined by entropy weight method.The comprehensive score of content was calculated.According to the maximum yield and the best quality,the highest total score of weighted score was indeed the best harvest period.4.Study on processing methods of PRA based on Q-Marker:Fresh medicinal materials of PRA in the best harvest period were collected.Two kinds of medicinal materials processed from different origin after boiling/steaming and scraping and drying,and three kinds of decoction pieces processed by soaking softening,steaming softening,origin processing and processing integration were investigated.Entropy weight method was used to optimize the best processing method.Through single factor and orthogonal experiment,the technological parameters of boiling process,steaming process,drying temperature and drying time were investigated,and the optimum technological parameters of processing method of PRA decoction pieces were determined.Results:1.Study on quality markers of PRA based on the transfer process of medicinal materials-decoction pieces-standard decoctions:There were 9 co-peaks in the HPLC specific chromatogram of decoction pieces and standard decoctions of PRA and the intragroup similarity was high.OPLS-DA results showed that No.9 co-peak cannot be transmitted stably.In the process of medicinal materials-decoction pieces,the transfer rate of gallic acid,oxypaeoniflorin,catechin,albiflorin,paeoniflorin,gallophilyl paeoniflorin,1,2,3,4,6-O-pentagalloglucose,benzoylpaeoniflorin was more than 70%,they can transfer stability.In the process of decoction pieces-standard decoctions,the the transfer rate of oxypaeoniflorin,albiflorin,paeoniflorin,gallophilyl paeoniflorin,benzoylpaeoniflorin mainly monoterpenes and their glycosides was stable and the transfer was more than 60%,they can transfer stability;the transfer rate of gallic acid was more than 200%,the transmission of gallic acid in the decoction pieces of PRA was not fully reflected.The contents of catechin,1,2,3,4,6-O-pentagalloglucose were unstable and the transfer rate was less than 25%.2.A multi-evaluation quality evaluation system of PRA was established based on Q-Markers:Different detection wavelength（230±2）nm had significant effects on the correction factors（RSDS>5.0%）,while other factors had little effect on the relative correction factors（RSDS<5.0%）.The relative correction factor had good repeatability and durability（RSDS<5.0%）.Relative correction factors f_{oxide paeoniflorin/Paeoniflorin}=4.911,falbiflorin/Paeoniflorin=1.232,fgallic Paeoniflorin/Paeoniflorin=1.011,fbenzoyl Paeoniflorin/Paeoniflorin=0.704.The relative deviation between QAMS content values of 5 chemical components in 10batches of decoction pieces and measured values of external label was less than 5.0%,and there was no significant difference.3.Study on the optimum harvesting period of PRA based on Q-Marker:The results of the growth of PRA in different harvesting months showed that the yield of“XT”and“PB”reached the maximum in June,and that of“XPB”reached the maximum in August;The results of entropy weight method showed that the content of“XT”,“PB”and“XPB”reached the maximum in November,September and June,respectively.The results of weighted scoring method showed that the best harvest time of“XT”,“PB”and“XPB”were October,September and June,respectively.The results of different growth years and different years of the same plant showed that the best growth years of“XT”and“PB”were both 4 years.4.Study on processing methods of PRA based on Q-Marker:The T-test results of boiling and steaming processing methods from different producing areas showed that there was no significant difference in the other four components except albiflorin.The results of entropy weight method showed that steaming processing method was superior to boiling processing method.The results of ANOVA showed that the content of albiflorin and paeoniflorin integrated by steaming softening and origin processing and processing integration was significantly higher than that of traditional soaking softening,and the result of entropy weight method showed that the processing method origin processing and processing integration was the best.Conclusion:comprehensive screening to determine oxypaeoniflorin,albiflorin,paeoniflorin,gallophilyl paeoniflorin,benzoylpaeoniflorin five components can be used as Q-Marker of PRA.This method has good repeatability and accurate results,which can provide reference for quality control of Radix Paeoniae Radix decoction pieces.The best harvesting month for“XT”of PRA was October,the best harvesting month for“PB”was September,and the best harvesting month for“XPB”was June.The best growth years of“XT”,“PB”and“XPB”PRA were 4 years.The best processing method of PRA decoction pieces is integration of origin processing and decoction piece processing,the optimal process parameters of PRA decoction pieces were as follows:take fresh PRA,steam in boiling water for 5 min,remove and scrape the skin,bake at 60℃for 8h,cut2mm slices,and dry at 60℃.