Establishment Of CE-ECL Technique For Detection Of Spermine And The Relationship Between Endogenous Spermine And Blueberry Fruit Quality

Capillary electrophoresis electrochemiluminescence(CE-ECL)technology is characterized by high efficiency(resolution,speed of analysis,sensitivity),good selectivity,less sample consumption,simple and miniaturization instrument,low cost and wide range of applications.These features make CE-ECL detection technology in drug analysis,biological analysis and other fields more and more widely used.Spermine,a kind of polyamines,is concerned by researchers because of its widespread presence in the organism and its important promoting effect on its own.Blueberry anthocyanins are also highly favored for their rich pharmacological and health benefits.Based on CE-ECL technology’s unique advantages in the detection of amines containing nitrogen groups,this article has established an analytical method for detection of spermine in fruits and vegetables.At the same time,the effects of spermine on some indexes and functional components during the growth and development of blueberry and post-harvest storage were studied.The purpose is to provide a theoretical basis for the accumulation of blueberry anthocyanins and the quality of blueberry fruit.The main experimental results are as follows:(1)To establish a capillary electrophoresis with electrochemiluminescence(CE-ECL)method for determination of spermine in fruit.The content of spermine in fruit was determined by CE-ECL.Under the optimized condition: The ECL intensity was linear with the concentrations of spermine over the ranges from 2×10-4 ~ 3×10-1 mmol/L.The linear equation was I =1114.2 c + 99.41,correlation coefficient r = 0.9996(n = 5)with detection limit(S/N = 3)of 9.9×10-5 mmol/L.Spermine was found in mulberry,apple and strawberry samples,adding standard recoveries were 95.1% ~ 101.2%,99.0% ~ 100.8% and 100.6% ~ 106.0%,respectively.This method has the characteristics of high separation efficiency,short time,simple operation,high sensitivity,low cost and trace consumption.(2)The dynamic changes of endogenous spermine,its fresh-keeping quality and physiological and biochemical indexes during the growth and development of blueberry were studied,and their correlation analysis was also conducted.The fresh-keeping,physiological and biochemical indexes and the content of endogenous spermine were measured with the blueberry fruit of "Zuozhibaoshi" at different growth stages.The results show that: The variation of the endogenous spermine content in blueberry fruits during the development period was found for the first time.The endogenous spermine of blueberry fruit was significantly positively correlated with hardness and titratable acid and negatively correlated with soluble sugar.The endogenous spermine of blueberry fruit was significantly positively correlated with SOD,PPO,POD activity,total phenolic and flavonoids,and significantly negatively correlated with anthocyanin.It can be seen that the relationship is very close between endogenous spermine and these indicators.However,the effect of anthocyanin is contradictory to previous studies and needs further study.The directions for subsequent genome and metabolome studies are indicated.(3)The effects of exogenous spermine on endogenous spermine,anthocyanin,total phenols and flavonoid in blueberry fruit at different storage temperatures were studied."Jietu blueberry,Yuanlan blueberry,Lansha blueberry,Hongbaoshi blueberry" four varieties of blueberry as the research object,0.3 mmol/L,0.9 mmol/L,1.5 mmol/L,3 concentrations of exogenous spermine and 3 storage temperatures at 0℃,4℃ and 20℃ were used to detect anthocyanins,total phenols and flavonoid in blueberry fruits under different conditions.The results showed: With the extension of the storage cycle,the content of endogenous spermine decreased as a whole and rapidly declined to normal level in the first cycle.With the increase of storage temperature,the endogenous spermine content decays slowly.At the late stage of storage,the endogenous spermine content of blueberry was significantly higher in the 0.9 mmol/L exogenous spermine group than in the other treatment groups,which effectively slowed the decrease of endogenous spermine in blueberries.The variations of the other three varieties(Yuanlan,Lansha,Hongbaoshi)are similar to those of Jietu blueberry.It can be seen that the effect of exogenous spermine on endogenous spermine is very clear.The effect of exogenous spermine on anthocyanins,total phenolics,and flavonoids of blueberry is closely related to its species and storage temperature.It is speculated that the effect of spermine on the anthocyanins,total phenolics,and flavonoids of blueberry is not a direct relationship and may be indirectly affected by glycometabolism.These studies provide direction and theoretical basis for the follow-up study of specific mechanisms of action.