Analysis Of Chemical And Nutritional Components And Safety Evaluation In Toxicity Of Peganum Harmala In Different Phenophases

Peganum harmala is a perennial herb of the Zygophyllaceae genus.It is widely distributed in the arid desert areas of Inner Mongolia,Xinjiang,Qinghai,Gansu,Shaanxi,Ningxia and other provinces.It has been used in ethnic minority areas since ancient times.As a medicinal plant,it can treat various diseases such as cough and asthma,rheumatism arthralgia,swelling toxin,skin pruritus,and rheumatoid arthritis.For a long time,China’s natural grassland has been over-utilized,the ecological environment has deteriorated,and edible pastures have decreased.However,poisonous and harmful plants such as St.chamaejasme,Ligularia and Oxytropis.The healthy breeding and ecological security of natural grassland grazing animals pose a potential threat.Different provinces have different opinions on whether Peganum harmala is a poisonous plant.At present,researches on the pharmacological effects of Peganum harmala are relatively rich and diverse,especially in its anti-cancer and anti-tumor activities.However,comparative studies on the chemical components of Peganum harmala in different phenological periods,especially the alkaloid components,nutritional components and toxicity are still blank.This study clarified the mature and wilt period by analyzing the chemical,alkaloid and nutrient components of the ground parts of Peganum harmala in different phenological periods（mature and wilt period）and the safety evaluation of acute and sub-chronic toxicity.The differences in the types of alkaloids contained and the difference in nutrients contained in the two phenological period of Peganum harmala is analyzed,and their toxicity and differences are compared,and the two different phenological period of mature and wilt are comprehensively evaluated.The nutritional value and toxicity of Peganum harmala provide theoretical basis for resource utilization and toxicity risk prevention of Peganum harmala in pastoral areas.The results of this test are as follows:1.Analysis of chemical components:The paper thin layer method and test tube method are used to detect the common chemical components of the ground parts of Peganum harmala in the mature and wilt periods,including alkaloids,alkaloids,volatile oil,triterpenoids,cardiac glycosides,organic acids,phenols,tannins,flavonoids,quinones and steroids and other compounds.2.Extraction,separation,analysis and identification of alkaloid components:The traditional extraction method of alkaloids is used to extract and separate the alkaloids from the ground parts of Peganum harmala,thin-layer chromatography analysis and GC-MS detection,and the results show:The extraction rate of the acidic dichloromethane part of Peganum harmala at the mature stage is 0.20%,the extraction rate of the alkaline dichloromethane part is 0.31%,and the extraction rate of the basic n-butanol part is 0.16%.TLC analysis shows 1,4 and 1 orange punctuations.After GC-MS detection and comparative analysis of existing databases,five alkaloids:Quinazoline,Deoxypeganine,Harmine,N-formyl-Cytisine,and Harmalacidine have been identified;the partial extraction rate was 0.27%,the partial extraction rate of alkaline dichloromethane was 0.45%,and the partial extraction rate of alkaline n-butanol was 0.19%.There were 1,5,and 1 orange punctuations in TLC analysis.After GC-MS detection,it was compared and analyzed with existing databases,and seven alkaloids,namely Quinazoline,Deoxypeganine,Leptaflorine,Harmaline,Harmine,N-formyl-Cytisine,and Harmalacidine,were identified.Among them,Leptaflorine and Harmaline were not found in mature period of Peganum harmala.3.Nutrient composition analysis:According to the national standard method,the nutrient components of the ground parts of the mature period and wilt period of Peganum Harmala were tested.The results showed that the crude protein,crude fat and crude fiber content of the ground parts of Peganum harmala in mature period were 13.5%respectively.,2.34%,25.97%;the major elements calcium,phosphorus,magnesium,and potassium content of the mineral elements are 10849.00 mg/kg,1275.50 mg/kg,6398.50 mg/kg,31376.75 mg/kg,trace elements iron,copper,and manganese,zinc,chromium and selenium content are 1099.70 mg/kg,4.78 mg/kg,54.02 mg/kg,12.67 mg/kg,9.71 mg/kg and 0.09mg/kg respectively;the ground part of Peganum harmala contains 17 kinds of amino acids,The total amount of amino acids is 9.83%.The crude protein,crude fat,and crude fiber content of the ground parts of Peganum harmala in wilt period were 11.15%,2.43%,and29.64%,respectively;the major mineral elements calcium,phosphorus,magnesium,and potassium were 11175.75 mg/kg,respectively.1105.75 mg/kg,6030.25 mg/kg,17571.50mg/kg,trace elements iron,copper,manganese,zinc,chromium and selenium content are1499.75 mg/kg,5.59 mg/kg,80.315 mg/kg,12.90 mg/kg,55.74 mg/kg and 0.11 mg/kg;Peganum harmala in wilt period contains 17 kinds of amino acids,and the total amount of amino acids is 7.04%.The results showed that both Peganum harmala in mature period and wilt period have certain nutritional value,rich in mineral elements,and a complete variety of amino acids.Especially in mature period of Peganum harmala,its crude protein content is up to 13%.4.Acute toxicity test and sub-chronic toxicity test in mice:Using mice as experimental animals,the results showed that:alcohol extract of Peganum harmala in mature period in mice of the oral LD₅₀ is 22.413±0.016 g/kg,and alcohol extract of Peganum harmala in wilt period in mice of the oral LD₅₀is 13.696±0.035 g/kg.Combined with the clinical symptoms and changes of necropsy in mice,the analysis showed that Peganum harmala in the mature and wilt periods have certain toxicity,and the toxicity of Peganum harmala in wilt period is obviously stronger than that in the mature period.The basic diet was supplemented with 11.21%(1/5 LD₅₀)and 2.80%(1/20 LD₅₀)powder of Peganum harmala in mature period,with 6.85%(1/5 LD₅₀)and 1.71%(1/20 LD₅₀)powder of Peganum harmala in wilt period.A 90-day sub-chronic toxicity test was carried out with observed the clinical manifestations of mice in each group,recorded feed intake and body weight changes,analyzed changes in blood physiological and biochemical indicators,and made organ tissue sections to observe pathological changes.The results showed that,compared with the control group,the clinical manifestations,blood physiological and biochemical indicators and tissues and organs of the heart,spleen,lung,brain,ovary,testis and other tissues and organs of the mature stage of Peganum harmala powder fed to mice at low and high doses,there was no obvious change and substantial damage,liver,kidneys and cerebellum have minor damage;and there were slight pathological changes in the liver and kidney;while the mice were fed with low and high doses of Peganum harmala in wilt period,and there were slight clinical manifestations,and the liver,kidneys,lungs and cerebellum damage of the high-dose group is more obvious.Conclusion:There are differences in the alkaloid composition,nutrient composition and toxicity of Peganum harmala in different phenological periods.The alkaloid composition of Peganum harmala in the wilt stage is more than that in the mature stage.The nutritional value of Peganum harmala both in the mature and the wilt stage has certain nutritional value.It contains rich mineral elements and a complete variety of amino acids,especially when it is mature.The crude protein content of Peganum harmala was as high as13%in the early stage,and the toxic Peganum harmala in the wilt stage was greater than that in the mature stage.After long-term feeding of mice,Peganum harmala in the wilt stage has more obvious effects on mice’s growth and development and damage to multiple organs.In general,Peganum harmala in the mature stage is safer than Peganum harmala in the wilt stage,and has the value of potential resource development and utilization.