Study On Quality Equivalence And Safety Of Edible Semen Cassia From Two Species

Semen Cassia is the dried ripe seeds of Cassia obtusifolia L.or Cassia tora L.It is one of the best medicines in the Shen Nong's Herbal Classic and one of the 101types of Chinese medicine which are used as food and medicine,announced by National Health and Family Planning Commission.Semen Cassia's water adecoction which can replace tea as drink is popular in weight-losing,hypolipidemic,liver-protection and some other aspects.However some reports express that chinese medicines which contain anthraquinones have safely issues.It's important to do Effectiveness and Safety Evaluation of Semen Cassia including the time and dose of taking this medicine and toxic potential.Current study found differences in chemical compositions between two kinds of Semen Cassia,so we need further research to confirm if these two herbs both have reliable clinical effect.The essay including the following:A.Provides the basis for Cassia obtusifolia L.or Cassia tora L.can both be used as Semen Cassia by making a contrastive study of their therapeutic material bases.B.At the same time Semen Cassia's hypolipidemic effect as the research background,using small dose with health care products taking characteristics,long-term use of the way of reducing blood lipid in Semen Cassiae efficacy dose-response relationship of toxicity,safety,provide the basis for the rational use of Semen Cassiae.1.Contrastive study of two kinds of Semen Cassia1.1 AnthraquinonesAnthraquinones is the main therapeutic material bases of Semen Cassia.Current study found differences between two kinds of Semen Cassia's anthraquinones and there is little research about the content of anthraquinones of Cassia tora L.This experiment compared the content of anthraquinones between two kinds of Semen Cassia.Experimental results establish a method of“QASM”to detect 7 kinds of anthraquinones in Semen Cassia for the first time.The relative correction factors are determined for emodin:f_{Aurantio-obtusin/emodin}=1.403?f_{Aloe-emodin/emodin}=0.521?frhein/emodin=0.434?fchryso-obtusin/emodin=1.749?fchrysophanol/emodin=0.562?fphyscion/emodin=0.895.This experiment also adopted UV method to determine the total anthraquinone,total free anthraquinone,combined anthraquinone of Semen Cassia.The results have significant differences:the content of total anthraquinone in Cassia obtusifolia L.is more than Cassia tora L.The minimum deviation two multiplication?PLS?analysis of the main factors screening of anthraquinone content of the difference between the two,7 anthraquinones were determined by QASM as the independent variables,three kinds of content was determined with UV as the dependent variable,according to the grouping variable contribution value?VIP>1?,selected aurantio obtusifoli.n,emodin and chrysophanol.Aloe emodin as a marker,that the above 4 kinds of anthraquinone glycosides of anthraquinone or its corresponding factors exist is caused by different original anthraquinones content.1.2 NaphthopyroneThe Naphthopyrone of Semen Cassia have a pharmacological activity of protective effect on the liver and the anti hepatic virus.so the essay includes contrastive study on the content of the two origins.Firstly sample was refluxed with methanol in a water bath.Then we used HPLC method to determine the Naphthopyrone in Semen Cassia.The results showed that the contents of 10 batches of Cassia obtusifolia L.is between 2.48%⁴.42%,the average value is 3.12%,the highest one produced from Inner Mongolia;the contents of 10 batches of Cassia tora L.is between 2.38%⁴.37%,the average value is 2.92%,the highest one produced from Hebi,HeNan Province.There is no significant difference between two origins.1.3 Fatty AcidThe fatty acid of Semen Cassia is rich in linoleic acid and linolenic acid.It has the pharmacological activity of lowering lipid and cholesterol levels.At present,there are few reports on the content of linoleic acid and linolenic acid.Fatty acid was extracted and determined by petroleum ether reflux,results showed that There is no significant difference in fatty acid between two origins.The fatty acid component of different origins was analyzed qualitatively and quantitatively by using GC-MS-AMDIS combined with retention index to determine methyl esterified fatty acids.The results showed that there were more qualitative compounds than previous literature such as erucic acid,wood wax acid,nineteen alkane acid,wax acid.There are richer kinds of co mpounds and higher content of peanut acid methyl ester in Cassia obtusifolia L.Than Cassia tora L.The two origins both have Palmitic acid,seventeen acid,CIS-9-acid,linoleic acid,linolenic acid,linoleic acid,acid,twenty-two alkane acid.The sum of the relative contents of these 6 compounds is between 65%⁸0%which means there is a high similarity between the two components of unsaturated fatty acids.1.4 Comparative study on UPLC fingerprintAt present,there is nothing but fingerprints of anthraquinones of Semen Cassia.This operation is tedious and have no specificity.In this paper,we established the UPLC fingerprint method for the determination of the components Naphthopyrone and anthraquinones in Semen Cassiae for the first time,and compared the chemical composition of the two components.The results of the two origins in the chromatogram has similarities and differences,the components are basically the same,differences only exist in the relative content of individual components.The difference is mainly reflected in the 3,5,8peak:A.The relative ratio of 3 peak and marker peak of Cassia tora L.is larger than Cassia obtusifolia L.B.The relative ratio of 8 peak and marker peak of Cassia tora L.is smaller than Cassia obtusifolia L.C.The relative ratio of 4 peak and 5 peak of Cassia tora L.is larger than Cassia obtusifolia L.D.Chromatogram of Cassia obtusifolia L.is richer than Cassia tora L.and the total peak area is lager than Cassia tora L.in the chromatographic peaks after 25min which means the contents of small-polar free anthraquinones in Cassia obtusifolia L.is higher.Therefore,the chromatographic peak after 25min can be used as the fingerprint identification area of two species.In summary,this paper made contrastive studies of the two origins from the above 3 categories.The results showed that except for the anthraquinones content of Cassia obtusifolia L.is higher and the relative contents of their common components exist difference there is no obvious diffe rence between the two origins.We need further study to confirm whether the difference has an effect on the pharmacological effects of the two drugs.2.Study on safety and effectiveness2.1 Study on dose-response relationship of hypolipidemicThe effect of different doses of Semen Cassiae Decoction on blood lipid in rats with hyperlipidemia were dynamically observed.The model was established and dividedinto:model group,positive group Xuezhikang,0.2 g·kg-1)and5cassiae semen water decoction group?with crude drug?0.315?0.945?1.575?3.15?6.3g·kg^-1.Continuous gavage for 90d,weight,TG,TC,HDL-C,LDL-C,AI were respectively measured on 14,35,56,77 and 90d.TC?TG?MDA?T-SOD in the liver tissue is measured after 90d.Compared with the model group,At the 56d after gavage,the levels of TC,TG,LDL-C and AI were significantly decreased?P<0.01,P<0.05?,while the levels of HDL-C were significantly increased?P<0.01,P<0.05?in the positive group and two cassiae semen water decoction groups?3.15,6.3 g·kg-1?.The level of TG,LDL-C and AI were significantly decreased?P<0.01,P<0.05?in the medication groups of 1.575g·kg-1 at the 77d after gavage.There was no significant difference in serum lipid level in 0.315 and 0.945 g·kg-1 groups?P>0.05?,The levels of TC and TG in positive and 3.15,6.30 g·kg-1 groups were significantly decreased?P<0.01?.The activities of T-SOD in liver tissue were significantly increased?P<0.01?,and had no effect on MDA?P>0.05?in medication groups.There was no statistical significance between medication groups and model group?P>0.05?.There was no statistical significance between medication groups and model group?P>0.05?.The results suggest that Semen Cassia has hypolipidemic effect on hyperlipemia model of rats and has a dose-effect relationship.After 56d of treatment,the high dose?1.575g·kg-1?group had a stable and sustained lipid-lowering effect.2.2 Safety evaluation of Semen Cassia based on Hypolipidemic EffectThere is no report on the potential toxicity of low dose of Semen Cassia.Based on the study of the effect of reducing blood lipid of Semen Cassiae,this paper discusses the toxic and side effects.During administration period,each group appeared the colour yellow,dark yellow urine,the spirit of low-spirited,loose stools,diarrhea symptoms.The results of pathological examination showed that there were toxic and side effects in each dose group and it was positively correlated with the amount of Semen Cassia.Liver tissue can be seen:Hepatocyte vacuole like degeneration,swelling.Liver nucleus pycnosis,necrosis.Hepatic sinus inflammatory cells infiltration.Kidney tissue can be seen:Renal tubule epithelial cell swell,degeneration and exfoliate.Renal interstitial inflammatory cell infiltration.The detection results of serum related index showed that the levels of aspartate aminotransferase?AST?and alanine aminotransferase?ALT?of Semen Cassia administration group were significantly different between model groups?P<0.01,P<0.05?,and the results were consistent with those of the liver tissues.So there is a certain toxic effects for long-term use of Semen Cassiae water decoction.