| Anthocyanin is a kind of flavonoid widely found in plants.Anthocyanin is popular in medicine and food industry,and become one of the hot topics for scholars to study because of its antioxidant,anti-tumor,anti-aging,anti-inflammatory,and cardiovascular and cerebrovascular diseases.The current research on anthocyanin concentrates on one plant or bioactivity,especially the antioxidant activity of fruit extracts such as mulberry and blueberry,which provides a certain reference basis for the development of anthocyanin products,and many products have been saling on the market.There are differences in the types and properties of anthocyanins between different plants,and external factors such as different research platforms and operators will cause some errors in the results of the study.Under the same platform,the comparative study of multiple plants at the same angle can eliminate the errors caused by humans as much as possible.However,there are fewer studies conducted in this multi-dimensional and multi-angle manner.Therefore,thesix different anthocyanin-rich fruits and vegetables were selected to construct characteristic chromatogram,determine the anthocyanin content and biological activity,and conduct comparative studies under the same platform.Then providing theoretical basis for the precise development and industry of subsequent related products.The study was conducted by purchasing six kinds of anthocyanin-rich fruits and vegetables（blueberries,purple cabbage,onions,purple grapes,red plums,and blackberries）in at least 10 different provinces and cities.Then put them on the same conditions,52%ethanol was used to extracted by microwave and made into extract,and analyzed by high performance liquid chromatography（HPLC）to systematically establish the HPLC characteristic chromatogram of six kinds of fruits and vegetables.Spectrophotometric（SP）was used to determinate anthocyanin content and antioxidant（DPPH and ABTS）,inhibitory activity of acetylcholinesterase（AChE）andα-amylase to compare the differences of six batches of fruit and vegetable extracts in different batches.Furthermore,an HPLC method was established to test inhibitory activity of AChE.Common characteristic peaks,anthocyanin contents and biological activities such as antioxidant,inhibitory activity of AChE andα-amylase as factors to analysis of anthocyanins by using Grey Relational Analysis（GRA）,Partial Least Squares（PLS）,and Pearson Correlation Analysis（PCA）.Preliminary expoler the relationship between composition and biological activity,and the results are as follows:（1）The content of anthocyanin in six batches of fruits and vegetables was determined by the hydrochloric acid-methanol method.The results showed that the content of the anthocyanins varied greatly among batches,purple cabbage with the highest,followed by blueberry and blackberry,and grape,red plum and onion were lower,among which the anthocyanin content of Jiangxi purple cabbage was the highest with 43.58 mg·g-1 in extract.（2）The results of evaluating antioxidant activity using DPPH and ABTS were similar.Most of one sample in different batches had similar antioxidant at the same concentration,and few batches would appear extremely low or extremely high.Among them,1 mg·mL-1 of red plum,blackberry,blueberry,grape and purple cabbage extracts all had good antioxidant activity,and blackberry was the best.The antioxidant activities of onion extracts in 10 mg·mL-11 were all above 50%.The six kinds of fruit and vegetable anthocyanin extracts in different batches had inhibitory activity of AChE but with difference,of which the best one was blackberry,and the inhibition rate was in the range of 29.4-40%.Grape,blueberry,onion,and red cabbage in different regions exhibited good inhibitory effects onα-amylase activity,but blackberry and red plum not showed significant inhibition onα-amylase.（3）In addition,a method for determinating AChE inhibitory activity by HPLC was established in the study.Five factors affecting on the enzymatic reaction which are reaction temperature,pH,AChE concentration,ATCI concentration and time were optimized for HPLC method,and the optimal reaction conditions were pH with value of 7.0 to 8.0,AChE concentration of 0.064 U·mL-1,ATCI concentration of 12.5 mM,a reaction time of 30 min,and reaction temperature of 37°C.Then huperzine A and galantamine hydrobromide were used to verify the specificity,and cyanidin-3-O-glucoside,blueberry and purple potato extracts was used to validate feasibility.The results confirmed that the HPLC method for testing inhibitory activity of AChE was feasible.Comparing with the traditional SP at the inhibitor concentration,HPLC method has a wider response range and higher sensitivity.The AChE inhibitory activity of low concentration of inhibitors showed great difference,but the high concentration was opposite,and the inhibition of AChE of low concentration of cyanidin-3-O-glucoside presented a dose-effect relationship.（4）The results of PCA,PLS and GRA showed DPPH and ABTS methods were used to evaluate the antioxidant activity of the six fruits and vegetables extracts were significantly related.Then three analyses were acted on common characteristic peaks with DPPH,ABTS,anthocyanin content,α-amylase and AChE inhibitory activity,the coefficients of GRA were mostly between 0.60 and 0.88,it belongs strong correlation.The results of comprehensive PLS and PCA showed that the characteristic chromatogram of purple cabbage 1th and 18th peaks were the main substance of anthocyanins and antioxidants,13th peak wasα-amylase inhibitory activity,and 5th was AChE inhibitory activity.For the characteristic chromatogram of blueberry exracts,its 8 common peaks were significantly associated with antioxidant activity and anthocyanin content,and 5th,7th and 8th peaks had inhibitory effects onα-amylase,5th,6th,7th 8th,9th and 10th peaks had a greater contribution and correlation to AChE inhibitory activity.For the blackberry,the four common peaks hadnotobviouscorrelationwithantioxidantactivity,1thpeak（cyanidin-3-O-glucoside）was the main component of anthocyanin,and 3th had effect on inhibitory activity of AChE.For onion,its 6 common peaks were significantly related to antioxidant activity except for 12th,and all were bound up content of anthocyanin except for 3th.The 1th,12th,5th,and 6th contributed toα-amylase inhibitory activity,and 3th,1th and 2th peaks played a major role in AChE.For red plum,2 common peaks were not obviously correlated in antioxidants,inhibition ofα-amylase and AChE activity.For grape,it had 5 common peaks,50th and 21th peaks were mainly acted on antioxidant activity,peaks of 12th,50th and 51th were related to the anthocyanin content,peaks of 9th,21th and 55th hadα-amylase inhibitory activity,and 12th,21th,and 50th peaks were significantly associated with AChE inhibitory activity.Most of the characteristic peaks which were significantly correlated with antioxidant activity by obtaining from the combination of three analyses were also related to to AChE andα-amylase inhibitory activity,but still had differences.It indicating that the structure of anthocyanin related to biological activity. |
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