| Ligularia naryensis,Ligularia virgaurea,Ligularia sagitta,and Ligularia discoidea for ligularia perennial herbaceous plants,the compositae in xinjiang,qinghai,gansu,sichuan and Tibet grassland pastoral area of natural grassland has formed dominant population,as a kind of poisonous plant breeding for grassland livestock health and ecological safety there is a potential hazard.At present,the researches on Ligularia naryensis,Ligularia virgaurea,Ligularia sagitta,and Ligularia discoidea are mainly focused on the biological characteristics,allelopathic effects and the prevention.This paper collected from xinjiang,qinghai,gansu and Tibet in natural grassland of Ligularia naryensis,Ligularia virgaurea,Ligularia sagitta,and Ligularia discoidea the ground part of the whole grass alkaloid composition and nutrient analysis,and the toxicity of in vivo and in vitro studies the extraction parts,preliminary clarify four ligularia contained alkaloids types of aerial part and the main toxic ingredients,analysis and evaluation of the nutritional value of high and low,not only enrich the research content,the plants of the genus ligularia for plants of the genus ligularia toxicology research and its development and utilization to provide basic support,the test results are as follows:1.Chemical composition system research: The paper thin layer method and test tube method were used to detect 4 kinds of ligularia,and the chemical constituents of 4 kinds of ligularia were preliminarily determined,including volatile oil and grease,alkaloid,tannin,organic acid,amino acid,peptide and protein,carbohydrate,flavonoids,coumarin,terpenolactone and phenolic compounds.In addition,Ligularia naryensis also contains quinones;There are also steroids,triterpenes,cardiac glycosides and quinones in Ligularia sagitta.There are also steroids and triterpenoids in Ligularia virgaurea.Ligularia discoidea also contains steroids,triterpenes and quinones.2.Isolation and identification of alkaloids: In this experiment,the traditional extraction method of phytochemical components was used to extract and separate the above-ground alkaloids of 4 kinds of ligularia,and then the TLC analysis and GC-MS detection were carried out.The results showed that the extraction rates of acidic dichloromethane,alkaline dichloromethane and alkaline n-butanol of Ligularia naryensis were 2.00%,0.05% and 0.76%.There were 1,5 and 1 alkaloid speckles in TLC analysis.GC-MS analysis identified five alkaloid components,namely 2,4-dimethylpyrrole,Heliotridine,Caulophylline,Sophoranol and Sophoramine,from the site of alkaline dichloromethane.The extraction rates of acidic dichloromethane,alkaline dichloromethane and alkaline n-butanol of Ligularia sagitta were 0.80%,0.16% and 2.54%.There were 1,4 and 0 alkaloid speckles in TLC analysis.Two alkaloids,2-Pyrrolidinone and 1-Pyrrolidinecarbaldehyde,were identified from the site of alkaline dichloromethane by GC-MS analysis.The extraction rates of acidic dichloromethane,alkaline dichloromethane and alkaline n-butanol of Ligularia virgaurea were 1.80%,0.02% and 3.20% respectively.There were 0,5 and 1 alkaloid speckles in TLC analysis.Three kinds of alkaloids were identified by GC-MS analysis from the site of alkaline dichloromethane,that is,Senecionine,Integerrimine and Platyphylline.The extraction rates of acidic dichloromethane,alkaline dichloromethane and alkaline n-butanol of Ligularia discoidea were 4.00%,0.08% and 3.20%.There were 0,5 and 0 alkaloid speckles in TLC analysis.Six alkaloids were identified by GC-MS analysis from the site of alkaline dichloromethane,namely 2-Pyrrolidinone,Benzothiazole,Benzothiazolone,Senecionine,Integerrimine and Platyphylline.3.Acute toxicity test in mice: The toxicity of aboveground partial alcohol extract of four kinds of ligularia was determined by intragastric administration of mice,and the results showed that: Mice with oral LD50 of Ligularia naryensis,Ligularia sagitta,Ligularia virgaurea and Ligularia discoidea alcohol extract were respectively 4.50 g/kg(95% confidence limit of 3.65-5.47 g/kg),8.30 g/kg(95% confidence limit of 7.75-8.89 g/kg),9.28 g/kg(95% confidence limit of 8.49-10.15 g/kg)and 9.28 g/kg(95% confidence limit of 8.03-10.72 g/kg).Combined with the clinical symptoms of mice and the changes of autopsy,it was concluded that Ligularia naryensis,Ligularia sagitta,Ligularia virgaurea and Ligularia discoidea all had toxicity,and the toxicity sequence was that Ligularia naryensis was greater than Ligularia sagitta than Ligularia discoidea than Ligularia virgaurea.4.Cytotoxicity evaluation: CCK 8 method is used to detect the four kinds of ligularia the ground part of petroleum ether part,methylene chloride and n-butyl alcohol and alkaline methylene chloride and alkaline n-butyl alcohol extract of BRL-3A and TCMK-1 cell toxic effects,the results showed that four kinds of ligularia are alkaline methylene chloride parts for BRL-3A and cell toxicity strongest TCMK-1,followed by methylene chloride are parts in ligularia(Ligularia naryensis),petroleum ether part of Ligularia virgaurea,n-butanol part Ligularia sagitta,petroleum ether and alkaline n-butyl alcohol parts(Ligularia discoidea).The results showed that different chemical sites had different toxicity to the two kinds of cells,among which the site of alkaline dichloromethane was the most toxic,proving that the site of alkaline dichloromethane was the main toxic site of the four species of ligularia.5.Analysis of nutritional composition: In accordance with the national standard method,the nutritional components of the aboveground part of 4 kinds of ligularia were tested.The results showed that the contents of crude protein,crude fat and crude fiber in Ligularia naryensis were 0.87%,3.26% and 43.63% respectively.The contents of calcium,phosphorus,magnesium,zinc,selenium,iron,copper and manganese were 7622 mg/kg,1960 mg/kg,9383 mg/kg,38.11 mg/kg,0.09 mg/kg,744.50 mg/kg,9.07 mg/kg and 6.34 mg/kg,respectively.It contains 17 kinds of amino acids,the total content is 7.43%.The contents of crude protein,crude fat and crude fiber in Ligularia virgaurea were 6.09%,3.77% and 26.40%,respectively.The mineral elements of calcium,phosphorus,magnesium,zinc,selenium,iron,copper and manganese were 10500 mg/kg,936.67 mg/kg,2635 mg/kg,18.80 mg/kg,0.005 mg/kg,188 mg/kg,7.43 mg/kg and 35.20 mg/kg,respectively.With 17 kinds of amino acids,the total content was 4.85%.The contents of crude protein,crude fat and crude fiber in Ligularia sagitta were 7.40%,2.10% and 24.67%,respectively.The mineral elements of calcium,phosphorus,magnesium,zinc,selenium,iron,copper and manganese were 11700 mg/kg,1413.33 mg/kg,2895 mg/kg,19.25 mg/kg,0.022 mg/kg,428 mg/kg,6.77 mg/kg and 40.15 mg/kg,respectively.The total amino acid content was 5.06%.The contents of crude protein,crude fat and crude fiber in Ligularia discoidea were 10.80%,2.70% and 25.50%,respectively.The mineral elements of calcium,phosphorus,magnesium,zinc,selenium,iron,copper and manganese are 14150 mg/kg,1265 mg/kg,2730 mg/kg,37.30 mg/kg,0.0025 mg/kg,165 mg/kg,6.39 mg/kg and 77.55 mg/kg respectively.The total amino acid content was 8.84%.The results showed that the four species of ligularia had certain nutritional value,abundant mineral elements and complete amino acids,especially the crude proteins of the three species of Ligularia virgaurea,Ligularia sagitta,and Ligularia discoidea were higher than 5%,and the four species had the potential of forage utilization. |
PDF Full Text Request