| Rationales:Ziziphi Spinosae Semen(ZSS)is the dried mature seeds of Ziziphus jujube Mill.var.spinosa(Bunge)Hu ex H.F.Chou,a plant of the genus Rhamnaceae.ZSS is the first choice for nourishing the heart and calming the nerves in Chinese medicine clinically.Its clinical application involves single medicine,compound medicine,food and beverage,etc.Over the years,a large number of studies were carried out,but most of them focused on the analysis of its production,processing and storage,mildew and oil loss according to thethe previous study.It was found that the processing method of ZSS in the market was extensive.As agriculture products,the processing mode of ZSS follows the traditional open,small workshops style.In addition,the study on the effect of storage time on the quality of ZSS is still a blank.Therefore,it is important to study the processing method and storage time of ZSS to improve the quality of ZSS.Objective:To compare the contents of seven compounds in ZSS under long-term stability conditions and accelerated stability conditions.The content of chemical components is changed to comparing the color is correlated with the intrinsic content index value representing the quality,and the correlation between the content of chemical components and the color and the influence of antioxidant capacity are explored,which provides a reference for the quality evaluation of the commonly used traditional Chinese medicine ZSS.Methods:1.The effect of two different pulping methods on moisture content and aflatoxin content of ZSS were compared.The contents of total alkaloids,total flavonoids and total saponions of ZSS in different pulping methods were determined by ultraviolet spectrophotometry.The effects of two pulping methods on the contents of seven chemical components in ZSS were determined by HPLC-UV-ELSD.2.Primary metabolites in ZSS were analyzed by ~1H-NMR and secondary metabolites were analyzed by UHPLC-Q-Orbitrap-MS,and principal component analysis(PCA)was performed,partial least squares discriminant analysis(PLS-DA),orthogonal partial least squares discriminant(OPLS-DA)and other multivariate statistical methods were used to find the difference of primary and secondary metabolites in the samples of ZSS before and after storage.3.By measuring the color difference value and content of seven chemical components of ZSS at different acceleration times,the appearance index representing color was correlated with the internal content index representing quality,and the correlation between the content of chemical components and color as well as the influence of its oxidation resistance of ZSS were discussed.The ZSS was stored in plastic self-sealed bag of drug stability test chamber with temperature of 40±2?and relative humidity of 75±5%.The effect of different acceleration time on the color of ZSS samples were analyzed with the contents of seven chemical components and the color of the samples as the inspection indexes.DPPH·,ABTS·+and FRAP were used to evaluate the antioxidant activity of ZSS in vitro and the accelerated stability test was carried out for six months.Results:1.After the fresh jujube samples were harvested,compare the hand-breaking of the inner pulp by drying in a 60?oven,smashing the seed husks(dry pulping)and soaking for a week until the pulp is soft,then the pulp is moved and the remaining pulp is washed and dried.The two methods of natural drying(wet pulping)showed that the content of total alkaloids,total flavonoids and total saponins were higher than of wet-pulping.2.In total,thirty primary medabolites were identified by ~1H-NMR and seven metabolites were considered as different components by OPLS-DA.Compare to ZSS at 0 year,the relative content of cysteine,betaine and sucrose decreased significantly.A total of seventeen differential secondary metabolites were identified by LC-MS combined with multivariate stasistical analysis and the content of seventeen secondary metabolites decreased significantly after storage for one year.HPLC-UV-ELSD method was used to compare the contents of seven chemical constituents temporarily stored in room temperature and cold storage for three months.It was found that the chemical constituents were relatively stable in the two storage environments for three months.3.With the extension of the acceleration time,the contents of the seven chenmical components have all decreased and the antioxidant capacity has also decreased.The results of the six months accelerated stability test showed that the L*value of the sample decreased significantly,the a*value increased and the b*value of remained basically unchanged.The results of antioxidation experiments showed that the scavenging ability of ZSS samples with different storage time gradually increased in a dose-dependent manner.When the conrentrations were 6.25 mg/m Land12.5 mg/m L,compared with 0 months,the clearance rate of accelerated 6-month samples was significantly reduces(p<0.001).It showed that the antioxidant capacity of ZSS was significantly reduced after 6 months of acceleration.Conclusion:1.By comparing the effects of different pulping methods on the chemical composition,moisture and aflatoxin of ZSS and considering the environment pollution of wet pulping methods and increasing the risk of aflatoxin pollution,it is recommend to use dry pupling method in the production and processing links of enterprises.2.After storage in the cold storage for one year,the contents of pharmacopoeia index of ZSS decreased significantly,the relative contents of seven primary defferential metabolites showed different trends of increasing and decreasing.The contents of seventeen secondary differential metabolites decreased significantly.There was no significant change in the contents of seventeen chemical components at room temperature and in cold storage for six months.3.With the extension of accelerated storage time,the contents of seven chemical components decreased and the antioxidant capacity also decreased.Compared with PLS analysis of seven chemical components and antioxidant activity in ZSS samples at different acceleration times,it was concluded that magnoflorine and spinosin contributed the most to the antioxidant activity.To provide experimental basis for the production and processing of ZSS and the research of storage validity. |
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