| Objective:To establish HPLC fingerprints of different grades of Corydalis decumbens medicinal materials,combined with chemical pattern recognition methods for processing and analysis,with a view to providing feasibility reference data for better selection of high-quality Corydalis decumbens medicinal materials;stir-frying with vinegar of Corydalis decumbens processing technology was preferred to develop Corydalis decumbens new Processed products provide relevant process parameters and technical references;evaluate the safety of Corydalis decumbens and its new processed products through acute toxicity,and provide references for later clinical safety medication;compare the analgesic effects of Corydalis decumbens and its vinegar products,and explore The liver-protecting effect of stir-frying with vinegar of Corydalis decumbens provides,to provide a scientific theoretical basis for the later development of Corydalis decumbens artillery products。Method:1.Based on the HPLC method,14 batches of 3 grades of Corydalis decumbens medicinal materials were measured,and the similarity was calculated using the 2012A version of"Chinese Medicine Fingerprint Similarity Evaluation Software",and the content of 6 alkaloids was determined.PCA and Orthogonal Partial Least Squares-DiscriminantAnalysis（OPLS-DA）for pattern recognition of samples。2.The key influencing factors and scope in the processing process were determined by single factor test.Based on response surface methodology,four factors including vinegar dosage,soaking time,frying temperature and frying time were investigated.The contents of water,ash,leachate,alkaloids（protopine,palmatine hydrochloride,tetrahydropalmatine,bicuculline）are taken as measurement indexes.Using CRITIC method combined with G1 method to calculate the weight coefficient,and through comprehensive score to optimize the processing technology of stir-frying with vinegar of Corydalis decumbens。3.Bliss method was used to calculate the 50%lethality（LD50）of raw products and stir-frying with vinegar of Corydalis decumbensr anhydrous extracts.After 14 consecutive days of observation,anatomy was performed to calculate the mouse heart,liver,spleen,lung,renal organ index,and blood Observation of biochemical indicators ALT,AST,BUN,CK,LDH,CA and HE staining pathological sections of various organs。4.Analgesic evaluation was performed on glutamate-induced writhing reaction in mice with Corydalis decumbens and its vinegar products。5.The mice were randomly divided into a normal group,a model group,a positive group（bifendate）（1.2 g·kg-1）,and high,medium,and low（1.8,1.5,0.9 g·kg-1）dose groups of vinegar,After 7 days of continuous intragastric administration,except for mice in the normal group,the mice were injected intraperitoneally with edible oil at a concentration of 0.1g/ml,and 0.4%CCl4 edible oil solution was injected intraperitoneally to establish an acute liver injury model.After 24h of modeling,blood was collected from the eyeballs,and serum and liver tissue were collected by centrifugation.Measure serum alanine aminotransferase（ALT）and aspartate aminotransferase（AST）activities,activity of superoxide dismutase（SOD）,glutathione peroxidase（GSH-Px）in liver homogenate,and malondialdehyde（MDA）Content,and expression levels of tumor necrosis factor-α（TNF-α）and interleukin-6（IL-6）,calculate liver index and observe pathological changes in liver tissue。Results:1.The content determination conditions were verified by methodology,precision,linearity,accuracy,stability and reproducibility were good.HPLC fingerprints of alkaloids in Corydalis decumbens were established,and the similarities were all greater than0.94.It shows that there is no difference in composition in the Corydalis decumbens of different producing areas,12 peaks are calibrated,and 6characteristic peaks are designated.The orthogonal partial least squares-discriminant analysis（OPLS-DA）can better distinguish the different grades and can judge the chemical components with strong correlation with Corydalis decumbens.The content of the six alkaloids still differed in the Corydalis decumbens in different producing areas.2.The optimal processing technology of vinegar-baked Corydalis Decumbentis was 30%vinegar dosage,soaking time 1 h,frying temperature170-190℃,frying time 5 min,OD value 95.64.Three batches of samples were prepared in parallel under the obtained conditions,and the calculated OD values were 95.05、96.51and 95.36 respectively.RSD was0.80%。3.The 50%lethality of the original medicines in the Corydalis decumbens and stir-frying with vinegar of Corydalis decumbens is 48g/kg,54g/kg;the difference in acute toxicity is not obvious,which is equivalent to 240 and 270 times the safe dosage of the human body（60 kg）;Compared with the normal group,except for AST（P<0.05）,there was no significant difference in the levels of other ALT,CK,LDH,UREA,and CREA（P<0.05）。4.Compared with the model group,the Corydalis decumbens group can significantly reduce the number of writhing in mice（P<0.05）.the results of the stir-frying with vinegar of Corydalis decumbens group is moreObvious。5.Compared with the model group,the stir-frying with vinegar of Corydalis decumbens group can significantly reduce the levels of ALT and AST in the serum of mice（P<0.01）,enhance the activity of SOD and GSH-PX in liver tissue（P<0.01）,and reduce the content of MDA in liver And the levels of TNF-αand IL-6（P<0.01）,the difference was statistically significant.Especially the effect of the stir-frying with vinegar of Corydalis decumbens high-dose group was more significant;In addition,it can reduce the pathological damage and swelling of CCl4on the liver。Conclusion:The high-performance liquid chromatography-based Corydalis decumbens component fingerprints are characteristic and easy to use.Combined with multi-component chemical pattern recognition technology,the quality can be better controlled,which has guiding significance and reference value for Corydalis decumbens’s level classification;The predicted value of stir-frying with vinegar of Corydalis Decumbens process regression model is close to the measured value,with good repeatability,good prediction effect,rationality and feasibility.To provide reference for standardizing the processing technology of Rhizoma Corydalis Decumbentis,At the same time,the method for determining the content of four alkaloids in Corydalis decumbens is fast and reliable,and it can be used for quality control of Corydalis decumbens medicine herbs;The extract water of Corydalis Decumbentis and its vinegar products LD50>10g/kg native medicine,which is practically non-toxic.The toxic effect of the extract water of Corydalis Decumbentis is slightly greater than the extract of stir-frying with vinegar of Corydalis Decumbens,indicating that after stir-frying with vinegar can reduce a slight attenuation toxicity;The analgesia effect of stir-frying with vinegar of Corydalis Decumbens is slightly better than that of raw products.It provides a reference for the basic theory of Chinese medicine that stir-frying with vinegar into the liver and has an analgesic effect.Oxidation is related,providing scientific basis for later development and utilization。... |
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