The Authenticity Detection Of Walnut Milk Beverage And Bioactivity Detection Of Walnut Green Husk' Extract

Walnut(Julans regia L),also called walnut or walnut kernel,is a member plant of walnut genera in the walnut family.Walnut have a very high nutritional value and health effects.It is ranked the first in the world's four dried fruit which the other three are hazelnut,cashew and almond.Kernels are the most widely used in the walnut family because the rich oil and protein are good for brain and can enhance memory function.Walnut tree and its branches,leaves,flowers have great application in some aspects such as food,medicine and agriculture.The exploitation and utilization of walnut resources have been a key problem to the development of walnut industry.At the same time,due to the high price of walnut,the quality of its products has always haunted consumers and been a problem of quality prosecution.As a byproduct of walnut industry,a large number of walnut green husk had been discarded,it is a waste of resources.We should give full play to the utilization value of walnut green husk and realize the value of it.This study take walnut milk and walnut green husk as research objects,establish a detection of walnut milk drinks(dew)doped with other plant components and the method of study on development and application of extracts from walnut green husk.This research includes three parts.1.Walnut milk put walnut kernel as a main raw material and it's popular in many consumers because of its good health care function and rich nutrient.Illegal businessmen use shoddy raw materials such as soybean or peanut to infringe on the legitimate rights and interests of consumers.To ensure the quality of walnut milk(dew)drinks and protect the rights and interests of consumers,it is necessary to establish detection of other plant compounds in walnut milk(dew).This research described a rapid PCR method for the detection of soybean and peanut materials in walnut milk(drew).The primer pairs Bd 28K-F/R and Arah 1-F/R were based on the soybean Gly m Bd28 K gene and peanut Arah 1 gene respectively.The results showed that the two designed primers had strong specificities,the detection limitation was 1%for peanut and 0.1%for soybean respectively.It can be an effective method for the detection of soybean,peanut materials in walnut milk(drew)and some other walnut products such as walnut oil and walnut powder's adulteration,and also provide some reference for authenticity of other foods' related study.2.Using walnut green husk as the test object,the active site was extracted by soaking in the ethanol and petroleum ether,chloroform,ethyl acetate,n-butanol extraction method.The inhibitory activities of 4 kinds of plant pathogenic fungi were test by mycelia growth method and the results showed that the inhibition of ethyl acetate extract is the best with some concentration,the inhibiting rate of Fusarium graminearum could be 90.24%and the EC50 values is 7.3538mg/ml.Turbidimetric method and the minimum inhibition concentration(MIC)experiment were used to test the inhibitory activity of seven kinds of common pathogenic bacteria.The results showed that the inhibition of ethyl acetate extract is the best:diameter of the inhabited ring is 22.99mm to Staphylococcus aureus,MIC value and minimal bactericidal concentration(MBC)value are 0.781mg/ml and 1.563 mg/ml respectively.The different culture contions such as culture medium and solution pH.temperature,NaCl concentration,sucrose concentration,ultraviolet irradiation were changeded in antimicrobial tests.The results showed that temperature and ultraviolet(UV)have no effects on antibacterial activity,NaCl and sucrose has little influence on it,but it is greatly influenced by the pH of culture medium and solution.Transmission electron microscope(TEM)was used to observe whether the ethyl acetate extract has impact on the cell membrane or cell wall structure of the bacteria,and its influence on the growth of bacteria,conductivity and macromolecular dissolution such as DNA and RNA of bacteria culture supernatant.The preliminary study on the antibacterial mechanism showed that the bacteria cell membrane and cell wall and the contents were damaged,this lead to changes in growth curve,macromolecular dissolution and electrical conductivity's content.3.Extraction method of active substances has been changed on the basis of the first part.Extracts from walnut green husk were selected by MTT assay against human cervical cancer Hela cells and human liver cancer Huh7 cells.In order to observe the morphological changes,we use a kind of dye named Heochst33342.The study results show that with the increase of concentration of all the extraction,their inhibitory effect on cell is stronger.The antitumor effect of petroleum ether extraction is the most obvious,the IC50 is 45.33mg/ml,it can induce Huh7 cell's apoptosis.