The Effect Of Ethyl Acetate Fraction From Citrus Changshan-huyou Pupl On Glucose Consumption In 3T3-l1 Adipocytes

The Citrus changshan-huyou is hybrid rutanceae citrus plants grapefruit and orange,and it contains a lot of carbohydrates.The previous search has shown that huyou pulp ethyl acetate component has the effect of balancing blood sugar.This thesis is aimed to investigate the component composition further.The ethyl acetate fraction was separated by Sephadex LH-20,and the conditions for separation of mobile phase,the elution velocity,height diameter ratio of chromatography column and sample weight on the four factors were optimized.Then the ingredients separated which could increase the glucose uptake in 3T3-L1 adipocytes were selected.Finally,the high performance liquid chromatography-mass spectrometry was used to detect the main components.The results of the study are as follows:The ethyl acetate fraction was separated by Sephadex LH-20.The orthogonal experiment of four factors and three levels was designed basis of based the single factor experiments.The result showed that the best separating conditions were 40%methanol solution,the velocity of 0.2 ml/min,the height diameter ratio of chromatography column of 30:1,and the loading sample amount of 1%column volume.The degree of separation(?R)on the optimal conditions was 4.08 ± 0.16 and six components were separated.In these four factors,the impact on the degree of separation was as following:the velocity>mobile phase>height diameter ratio>the loading sample amount.The effect of the separated components of huyou pupl ethyl acetate fraction on 3T3-L1 adipocytes absorbing glucose were detected.The results showed that sample 1(1-25?g/ml),sample 2(1?100 pg/ml)and sample 3(1?5 ?g/ml)promoted significantly the uptake of glucose by the 3T3-L1 adipocytes(P<0.01),and the consumption of glucose decreased with the increase of concentration increased.Sample 4(1?100 ?g/ml)promoted significantly the uptake of glucose(P<0.01),and the consumption of glucose was balanced in a certain range;Sample 6(25?100 ?g/ml)also promoted significantly the uptake of glucose(P<0.01)with presenting dose dependent.After two two joint of these six components,excepting the sample 3 combined with sample 4 and sample 5 combined with sample6 having no external interaction,other combinations were presented antagonism effect,having the significance,and the effect of promoting the uptake ofglucose was weakened.Using liquid-mass spectrometry,the components structures of the components of the ethyl acetate fraction of huyou pulp were identified.By the information of two levels mass spectrometry,compared with the information of standards,five compounds were obtained:eriocitrin,neoeriocitrin,narirutin,naringin and neohesperidin.The huyou pulp balance blood sugar composition in the ethyl acetate fraction could provide a reference for huyou deep processing and utilization in this study.