| Highland barley belongs to genus Hordeum,gramineous crop.It has been used intensively as a foodstuff,food material,brewing material and feed material in highland areas,and other varieties of barley also has been widely cultivated and applied all over the world.At present,the highland barley is mainly distributed in Tibet,Qinghai,Yunnan,Sichuan,Guizhou and other provinces in China.As a nutritious food with special characteristics in the plateau region,different kernel colors have different nutritional qualities.Because highland barley is mainly used as whole grain food,there are few studies on intensive processing and utilization.This study mainly uses different extraction methods to study vitamin E content and main functions in barley from different places,and treat those different highland barley by fluorescent labeling microsatellite technology.A comparative study of kinship provides a reference for possible further research.The main findings include the following:1?The experiment firstly optimizes the HPLC method.Extracts dissolve in n-hexane were injected into a forward Venusil XBP Silica column with an isocratic phase of n-hexane: isopropanol=95:5(v/ v)and flow rate of 1.0 mL/min,the column temperature was regulated at 30 ° C,injection volume 10 ?L.Through the single factor experiment of saponification,the KOH addition amount,soap time,saponification temperature and antioxidant addition amount were used as indicators,and on the basis of single factor,the Box-Benhnken response surface was used to optimize the experimental factors level to extract the vitamin E from the highland barley.Vitamin E concentration was highest in the extraction when conditions are: 1.98 ml of KOH,34.15 min of saponification time,87.3 °C of saponification temperature,and 5.25 ml of antioxidant.Under this condition,the maximum extraction content of vitamin E is 370.236 ?g/g.Ultrasonic method was used to extract highland barley from different habitats.The effects of various factors on the extraction of vitamin E from different habitats were studied by single factor experiment with liquid-solid ratio,ultrasonic time and ultrasonic temperature as factors.At the same time,the Box-Benhnken response surface was used to optimize the experimental factors.Vitamin E concentration was highest in the extraction when conditions are: liquid-solid ratio 17.27:1,ultrasonic temperature 59.33 °C,ultrasonic time 56.24 min.Under this condition,the maximum vitamin E extraction was obtained.The content was 1982.74 ?g / g.When use subcritical extraction method to extract the highland barley from different habitats,the effects of extraction temperature,liquid-solid ratio and extraction time on the vitamin E of sub-critical extraction of highland barley from different places were investigated by single factor experiment.The process optimization of subcritical extraction was carried out by the Box-Benhnken response surface experiment.The optimal process conditions for subcritical extraction were as follows: extraction temperature 62.33 ° C,ratio of liquid to material 25.92:1,extraction time 67.19 min,total vitamin E extraction content under this condition was 2113.72 ?g/g.Verification and comparison of different extraction methods,the actual value of vitamin E extracted from saponification method compared with the theoretical value is within 2%;the actual value of the vitamin E obtained by ultrasonic extraction and subcritical extraction method is compared with the theoretical value are within 1%.At the meantime,compared the extraction content of vitamin E in different field,the extraction amount of vitamin E obtained by ultrasonic extraction was 5 times that of the saponification method.2?The TBARS method was used to investigate the effect of different vitamin E extracts on the inhibition rate of lipid peroxidation.The results showed that the lipid peroxidation inhibition rate increased with the concentration of vitamin E extract.At a concentration of 350 ?g/mL,the inhibition curves of each producing area reached their apex,and Yunnan barley>Sichuan barley>Qinghai barley>Tibet barley.The POV value test method was used to explore the antioxidant activity of different vitamin E extracts against palm oil from highland barley.The results showed that the vitamin E extract of Yunnan barley was more active than the other three producing areas.The POV values were 32.95-0.15 meq/kg and 39.91-0.21 meq/kg during the 6th-8th day,respectively,which were significantly lower than the other three producing areas.The experimental results showed that the most suitable addition amount of the vitamin E extract was the value of 36 ?g/g that antioxidant activity of palm oil was the maximum.3?The 21 pairs of microsatellite primers randomly distributed on the 7 chromosomes of highland barley were fluorescently labeled,and then the relative genetic relationship between the 4 farmer cultivars used in the previous study and the identified 9 barley varieties was compared and established 11 varieties' fingerprints.The results showed that 17 pairs of primers detected polymorphic loci,accounting for 80.95% of the selected primers.A total of 82 alleles were detected in 17 pairs of primers,with an average of 4.82 per locus and a fragment length between 143 and 388 bp..Among them,the Bmag206 primer located on chromosome 7H detected 13 alleles of them.The Shannon's index is between 0.5481 and 2.3593,the polymorphic information content varies from 0.2389 to 0.8702,and 9 of the 17 markers are highly polymorphic,accounting for 52.94% of the total marker.The results of clustering analysis showed that the genetic similarity coefficient was between 0.6206 and 0.9706.When the genetic similarity coefficient was 0.6206,the materials were divided into two categories.The farmland species Y1,which was produced in Kunming,Yunnan,was divided into category ?.The rest of the materials were classified as category ?.17 pairs of microsatellite primers can separate the each variety from different producing areas,and the phylogenetic relationships of different sources are different.The cultivars from different producing areas may also have closer genetic relationship. |
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