Study Of Detection Of Buckwheat Based On HPLC-FLD

D-fagomine,that also called 1,2-double deoxynojirimycin,is a kind of six membered ring iminocyclitols,for the first time discovered and isolated from from buckwheat,and also exists in other plants such as mulberry and Chinese wolfberry.D-fagomine is a carbohydrate analogue that has the same spatial configuration of hydroxyl as sugar(such as glucose),the difference is that nitrogen replaces the internal epoxy of carbohydrates.Recent study shows that D-fagomine plays a dual role in the intestinal:first,D-fagomine is an inhibitor of intestinal glycosidase,so the decomposition of sucrose and starch can be effectively delayed by inhibiting glycosidase;secondly,D-fagomine selectively agglutinate Enterobacteriaceae,such as Escherichia coli,Salmonella typhimurium,and promote probiotic adhesion to the intestinal mucosa.D-fagomine as a kind of iminosugars to reduce postprandial blood glucose,improve the level of blood glucose in patients with diabetes has a good effect,but because of the absence of the chromophore and the simple spectrophotometric method can not for the determination of the content of D-fagomine,while that gas chromatography(GC)and Evaporative light scattering detector(ELSD)are more troublesome for detecting D-fagomine?Mass spectrometry(MS)is expensive.Therefore,this study using pre-column derivatization with fluorene methoxy chloride reagent,endow D-fagomine with fluorescent chromophore,and then use the high performance liquid chromatography with fluorescence detector(HPLC-FLD)to detect the content,establish a simple and quick method for detection of D-fagomine.The purpose of this study is based on the existing technology problems and defects,to establish a detection method for D-fagomine by high performance liquid chromatography with fluorescence detector,and the stability of derivatives,the precision,the accuracy and the sensitivity of the method were studied.In addition,this study discussed existing form of D-fagomine and analysed the content of D-fagomine in different parts and different varieties of buckwheat buckwheat by high performance liquid chromatography with fluorescence detector derived.The results are as follows:(1)The standard curve equation was Y=6×10~6X+19514,R~2=0.999;(2)The results show that the derivative can be stored for 3 days in the refrigerator at a temperature of 4?,the RSD of the precision is 0.73%,the RSD of repeatability is 3.67%,the accuracy(the average recovery rate)is 106.38%,the sensitivity of the method(the LOD and LOQ)are respectively 14.5?g/Kg and 43.9?g/Kg;(3)The results show that there are significant differences in different parts of buckwheat,the content of buckwheat flour powder was the highest,with an average of 3.61mg/Kg,followed by buckwheat bran powder,with an average of 2.60 mg/Kg,while the content of the shell powder was the least,with an average of 2.39 mg/Kg;(4)The results show that there are significant differences in different varieties of buckwheat,the contents of D-fagomine in Ding tian NO.1(12.79±0.18 mg/Kg)?Long tian NO.1(11.83±0.42 mg/Kg)are higher;Yu 6-21(4.39±0.09 mg/Kg)was the lowest;Analysis showed that the content of D-fagomine in buckwheat was significantly higher than that in Tartary buckwheat.