Authentication of Cassia seeds on the basis of two-wavelength HPLC fingerprinting with the use of chemometrics

High performance liquid chromatographic(HPLC) fingerprints of Cassia seed,a traditional Chinese medicine(TCM),were developed by means of the chromatograms at two wavelengths of 238 and 282 nm.Then,the two data sets were combined into one matrix.The application of principal component analysis(PCA) for this data matrix showed that the samples were clustered into four groups in accordance with the plant sources and preparation procedures.Furthermore,partial least squares(PLS),back propagation artificial neural...

Box-Behnken response surface method combined with fingerprint to optimize the extraction process of total anthraquinone from Cassia seeds

Objective:The Box-Behnken response surface method combined with fingerprints was used to optimize the extraction process of total anthraquinone from Cassia seeds.Methods:A three-factor,three-level response surface test was conducted based on the single-factor test with comprehensive evaluation as the measurement index.The comprehensive evaluation indexes included the extraction rate of total anthraquinone of Cassia seeds or the equivalent amount of herbs per gram of total anthraquinone of Cassia seeds,the normalized value of peak areas of 5 index components such as aurantio obtusin in the fingerprint of each sample to 16 shared peaks and the similarity of fingerprints(the reference fingerprint was established by the extraction solvent for the determination of Cassia seeds content in the Chinese Pharmacopoeia 2020 edition)with the weights of 0.2,0.5 and 0.3,respectively.Results:The best extraction process was obtained:the liquid-to-material ratio was 20:1(mL·g-1),the extraction solvent was mixture of 60%ethanol-ethyl acetate(2:1),and the extraction time was 15.12 min.The results of five sets of validation experiments showed that the overall evaluation of total anthraquinone of cassia seeds by the best process was 0.528(RSD=0.45%),and the prediction result of response surface method model was 0.531,and the relative error with the prediction result was 0.531.The relative error of the predicted results was 0.56%,and the best extraction process was consistent with the model prediction,and the obtained best process could be used for the extraction of total anthraquinone from Cassia seeds.Conclusion:The Box-Behnken response surface method combined with the fingerprint technique can be used to find the best reaction conditions and examine the interactions among the factors in a comprehensive and accurate manner,which can provide reference for the optimization and evaluation of the extraction process of Chinese medicine.

Determination of Emodin and Chrysophanol Contents in Callus of Cassia tora L. Leaf

[ Objective] Through inductive culture, emodin and chrysophanol contents in cassia seed were increased. [ Method ] MS culture medium was used to bourgeon seedlings of cassia seed and to induce cotyledon callus of cassia seed while HPLC method was adopted to determine emodin and ehrysophanol eontents.[ Result] Emedin and chrysophanol contents in cotyledon callus were 0. 099% and 0. 312%, respectively, while they were 0.029% and 0. 190% respectively in cassia seed. [ Conclusion] The method of inducing cotyledon callus was helpful for increasing emodin coment in cassia seed.

Ethanol extract of cassia seed alleviates metabolic dysfunction-associated steatotic liver disease by acting on multiple lipid metabolism-related pathways

Background and objective:In northern China's cold regions,the prevalence of metabolic dysfunction-associated steatotic liver disease(MASLD)exceeds 50%,significantly higher than the national and global rates.MASLD is an important risk factor for cardiovascular and cerebrovascular diseases,including coronary heart disease,stroke,and tumors,with no specific therapeutic drugs currently available.The ethanol extract of cassia seed(CSEE)has shown promise in lowering blood lipids and improving hepatic steatosis,but its mechanism in treating MASLD remains underexplored.This study aims to investigate the therapeutic effects and mechanisms of CSEE.Methods:MASLD models were established in male Wistar rats and golden hamsters using a high fat diet(HFD).CSEE(10,50,250 mg/kg)was administered via gavage for six weeks.Serum levels of total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),aspartate aminotransferase(AST),and alanine aminotransferase(ALT),as well as liver TC and TG,were measured using biochemical kits.Histopathological changes in the liver were evaluated using Oil Red O staining,Hematoxylin-eosin(H&E)staining,and transmission electron microscopy(TEM).HepG2 cell viability was assessed using the cell counting kit-8(CCK8)and Calcein-AM/PI staining.Network pharmacology was used to analyze drug-disease targets,and western blotting was used to confirm these predictions.Results:CSEE treatment significantly reduced serum levels of TC,TG,LDL-C,ALT,and AST,and improved liver weight,liver index,and hepatic lipid deposition in rats and golden hamsters.In addition,CSEE alleviated free fatty acid(FFA)-induced lipid deposition in HepG2 cells.Molecular biology experiments demonstrated that CSEE increased the protein levels of p-AMPK,p-ACC,PPARα,CPT1A,PI3K P110 and p-AKT,while decreasing the protein levels of SREBP1,FASN,C/EBPα,and PPARγ,thus improving hepatic lipid metabolism and reducing lipid deposition.The beneficial effects of CSEE were reversed by small molecule inhibitors of the signaling pathways in vitro.Conclusion:CSEE improves liver lipid metabolism and reduces lipid droplet deposition in Wistar rats and golden hamsters with MASLD by activating hepatic AMPK,PPARα,and PI3K/AKT signaling pathways.