| Objective:To optimize the extraction process of total anthraquinone from cassia seed.with total anthraquinone extract as the principal agents,suitable excipients were selected to prepare cassia seed dispersible tablets.The total anthraquinone extract from cassia seed was used as the principal agents and suitable excipients were screened to prepare cassia seed dispersible tablets.To establish the quality standards of cassia seed dispersible tablets.To study the protective effect of cassia seed dispersible tablets on acute liver injury in rats and its spectrum-effect relationship.Methods:1)Determination of the extraction process of total anthraquinone from cassia seed.Taking the yield of dry paste and the content of total anthraquinones as evaluation indexes,the optimum material liquid ratio,ethanol concentration,reflux time and extraction times were selected,and the extraction process of total anthraquinones from cassia seed was also optimized by using orthogonal test.The dry paste yield and the total anthraquinone content were used as evaluation indicators,the optimum material to liquid ratio,ethanol concentration,reflux time and number of extractions were selected,and the extraction process of total anthraquinone from cassia seed was also optimized by using orthogonal test.2)Preparation of cassia seed dispersible tablets.Cassia seed dispersible tablets were prepared by taking the total anthraquinone extract of cassia seed as the principal agents,using appearance,hardness and disintegration time as evaluation indicators to screen out suitable fillers,binders or liquid binders,disintegrat and lubricant.On this basis,The formulation of cassia seed dispersible tablets was optimized by central composite design-response surface method.Preparation of cassia seed dispersible tablets using total anthraquinone extract of cassia seed as the main drug.The appearance,hardness and disintegration time of the dispersible tablets were used as evaluation indicators,to screen for suitable fillers,binders or wetting agents,disintegrants and lubricants.On this basis,the central composite design was used to optimize the prescription of cassia seed dispersible tablets.3)Establishment of quality standards for cassia seed dispersible tablets.Referring to Chinese Pharmacopoeia(2020 edition)and related literature,the identification method of cassia seed dispersible tablets was established;The inspection indexes such as dispersion uniformity,dissolution and fingerprint similarity were defined;The content of total anthraquinone in dispersible tablets was determined by UV Vis;The contents of 6 anthraquinones in dispersible tablets were determined by HPLC,and the fingerprint of cassia seed dispersible tablets was established.Referring to the Chinese Pharmacopoeia(2020 Edition)and related literature,to establish the identification of cassia seed dispersible tablets;determine the inspection indicators such as dispersion uniformity,dissolution rate,fingerprint profiles similarity and so on;determination of total anthraquinone in cassia seed dispersible tablets by UV-Vis;determination of six anthraquinones in cassia seed dispersible tablets by HPLC,and establishment of fingerprint profiles for cassia seed dispersible tablets.4)The cassia seed dispersible tablets were administered to rats by gavage at high,medium and low doses(6,3 and lg/kg)for consecutive 7 days.After the last administration for 10 hours,models were made with carbon tetrachloride olive oil solution(carbon tetrachloride:olive oil,V/V=1:1,2mL/kg).24 hours later,the liver was taken out and the appearance was observed.Pathological sections were made,and the contents of AST and ALT in serum were determined.Combined with the HPLC fingerprint of cassia seed dispersible tablets,the spectrum-effect relationship of the protective effect of cassia seed dispersible tablets on acute liver injury in rats was studied by grey correlation method.Cassia seed dispersible tablets were administered to rats by gavage at three doses:high,medium and low(6,3 and 1 g/kg),continuous administration for 7 days,10h after the last administration,modeling with carbon tetrachloride olive oil solution(carbon tetrachloride:olive oil,V/V=1:1,2mL/kg).After 24h,the liver was removed,the appearance was observed,pathological sections were made and serum levels of AST and ALT were measured.Establishment of fingerprint profiles for cassia seed dispersible tablets,and to study the protective effect of cassia seed dispersible tablets on acute liver injury in rats and its spectrum-effect relationship.Results:1)The optimum extraction process of total anthraquinones from cassia seed is as follows:the ratio of material to liquid is 1:16(g:mL),the concentration of ethanol is 75%,the reflux time is 2 hours and the extraction times are 2 times.2)The best prescription for cassia seed dispersible tablets is as follows:150g of total anthraquinone extract of cassia seed,283g of corn starch,20g of L-HPC,21g of PVPP,11g of CCMC-Na,50g of micropowder silica gel,50%ethanol in the right amount and 5g of talcum powder to prepare 1000 tablets.The best prescription for cassia seed dispersible tablets is 150g of cassia seed extract,283g of corn starch,20g of L-HPC,21g of PVPP,11g of CCMC-Na,50g of colloidal silicon dioxide,50%ethanol in the right amount and 5g of talcum powder to prepare 1000 tablets.3)Cassia seed dispersible tablets are dark brown,fragrant and slightly bitter.With petroleum ether(60-90℃)-acetone(2:1)as the developing agent,autrantio-obtusin and chrysophanol in cassia seed dispersible tablets can be separated well;when placed in ammonia vapor,the spots change color,in which bright yellow is aurantio-obtusin and pink is chrysophanol.Cassia seed dispersible tablets can also react with magnesium acetate for colour development.Cassia seed dispersible tablets can disintegrate within 3min;the cumulative dissolution at 5min was more than 70%;other routine inspection items of tablets meet the requirements.Each cassia seed dispersible tablet contains 2.8mg of total anthraquinone.Cassia seed dispersible tablets are dark brown in colour,aromatic and slightly bitter in taste.With petroleum ether(60-90℃)-acetone(2:1)as the unfolding agent,aurantio-obtusin and chrysophanol which are in cassia seed dispersible tablets were well separated,when placed in ammonia vapour,the spots change colour,bright yellow for aurantio-obtusin and pink for chrysophanol.Cassia seed dispersible tablets can react with magnesium acetate for colour development.Cassia seed dispersible tablets can disintegrate and leak off the screen within 3min;the cumulative dissolution rate at 5min is more than 70%;other tablets routine checks are in accordance with the regulations.Each tablet of cassia seed dispersion contains 2.8mg of total anthraquinone,1501.78μg of aurantio-obtusin,4.07μg of aloe-emodin,470.64μg of rhein,116.62μg of emodin,353.84μg of chrysophanol and 84.43μg of physcion.4)Cassia obtusifolia dispersible tablets significantly reduced the liver patholo gical changes and significantly lowered serum AST and ALT levels in rats.Ther e were 63 common peaks in 10 batches of cassia seed dispersible tablets.With t he administered dose increased from lg/kg to 6g/kg,the correlation between the common peaks of cassia seed dispersible tablets and AST gradually increased,bu t the correlation between the common peaks and ALT first increased and then d ecreased.At different doses,the trend of common peak correlation was basically the same,but the correlation degree between peak 29 and AST first increases a nd then decreases,and the correlation degree with ALT gradually decreases.The correlation order of high,medium and low doses(6,3 and lg/kg)was inconsis tent,and the correlation order of AST was(top 15 chromatographic peaks):54>32>40>29(aurantio-obtusin)>42>37>56>62>33>60(chrysophanol)>24>27>52>36(rhe in)>63(physcion);29>32>40>54>42>37>56>62>33>60>24>52>27>36>63;29>54>32>40>42>37>56>62>33>60>24>27>52>36>63.The correlation order of ALT was(to p 15 chromatographic peaks):32>42>40>37>54>56>62>29>33>60>24>52>27>36>63;29>54>32>40>42>37>56>62>33>60>24>27>52>36>63;29>32>54>40>42>37>56>62>33>60>24>52>27>36>63.Cassia seed dispersible tablets significantly reduced liver lesions and significantly lowered serum AST and ALT levels in rats.Ther e were 63 shared peaks in 10 batches of cassia dispersible tablets.As the admin istered dose increased from lg/kg to 6g/kg,the correlation between the shared p eaks of cassia seed dispersible tablets and AST gradually increased,but the corre lation with ALT first increased and then decreased.The trend of correlation with shared peaks was basically the same for the different doses administered,but p eak 29 showed an increasing and then decreasing correlation with AST and a de creasing correlation with ALT.The order of association among the 3 doses of hi gh,medium,and low(6,3 and lg/kg)was inconsistent,the association orders of AST were(top 15 chromatographic peaks)54>32>40>29(aurantio-obtusin)>42>37>56>62>33>60(chrysophanol)>24>27>52>36(rhein)>63(physcion);29>32>40>54>42>37>56>62>33>60>24>52>27>36>63;29>54>32>40>42>37>56>62>33>60>24>27>52>36>63.The association orders of ALT were(top 15 chromatographic peaks)32>42>40>37>54>56>62>29>33>60>24>52>27>36>63;29>54>32>40>42>37>56>62>33>60>24>27>52>36>63;29>32>54>40>42>37>56>62>33>60>24>52>27>36>63.Conclusion:The orthogonal test method can better optimize the extraction of cassia seed,and the process is simple and reliable.The prescription of cassia seed dispersible tablets is reasonable and has fast dispersibility.The cassia seed dispersible tablets can meet the requirements of its quality standard(draft).The dispersible tablets have protective effect on acute liver injury induced by CCl4 in rats.he protective effect comes from the interaction between various active ingredients related to the dosage.With the different dosage,the correlation order of the active chemical components is not the same,but the main components of the 3 dose groups are relatively consistent.Cassia seed dispersible tablets are well prescribed and designed to have a fast dispersion.The cassia seed dispersible tablets are able to meet the requirements of their quality standard(draft).The protective effect of cassia seed dispersible tablets against acute liver injury caused by CCl4 in rats,and the protective effect results from the interaction between multiple active ingredients in relation to the dose administered.With different doses administered,the chemical components that play a role are not in the same order of association,but the main components that exerted their effects were more consistent across the 3 dose groups. |
PDF Full Text Request