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Research On Proanthocyanidins From Sargentodoxae Caulis,Photinia Serrulata And Spatholobi Caulis Extracts

Posted on:2018-01-07Degree:MasterType:Thesis
Country:ChinaCandidate:Y LiFull Text:PDF
GTID:2310330515451174Subject:Botany
Abstract/Summary:PDF Full Text Request
Proanthocyanidins are oligomers or polymers of flavon-3-ol compounds in different ways which are synthesized through plant flavonoid biosynthetic pathway and widely distributed in the plant community.It is reported that proanthocyanidins have strong antioxidant ability to protect the skin and reproductive system,improve immunity inhibition,et al,which are so called the strongest natural antioxidant.In this study,the extraction of proanthocyanidins from Sargentodoxae caulis,Photinia serrulata,and Spatholobi caulis was carried out by ultrasonic-assisted organic solvent method and organic solvent method for continuous extraction,then the content of proanthocyanidins was determined by vanillin-HCl method and dimethyl amine cinnamic aldehyde method.And then proanthocyanidins were purified by macroporous adsorption resins;the antioxidant ability of proanthocyanidins was evaluated by DPPH free radical scavenging activity,ABTS free radical scavenging ability,iron ion reduction ability and copper ion reduction ability.Inhibitory effects of proanthocyanidins on ?-glucosidase,?-amylase,acetylcholinesterase and butyrylcholinesterase was also evaluated.Structure analysis was used by ultraviolet absorption spectrum,infrared absorption spectrum,reversed phase high-performance liquid chromatography(RP-HPLC),matrix-assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF MS)and 13 C nuclear magnetic resonance(13C-NMR).The main reserch and results are as follows:1?Proanthocyanidins were extracted from Sargentodoxae caulis extracts,and the crude extract was purified.The content was determined by an improved vanillin-HCl colorimetric method.Meanwhile,the antioxidant ability and enzyme inhibitory ability of each sample were measured.The results showed that the contents of proanthocyanidins in the crude extract,petroleum ether fraction,ethyl acetate insoluble fraction,ethyl acetate fraction,n-butanol fraction and water residue were(36.03 ± 0.40)%,(1.19 ± 0.11)%,(57.99 ± 0.22)%,(32.40 ± 0.07)%,(25.03 ± 0.08)% and(41.32 ± 0.37)%,respectively.Moreover,sample purified in Sephadex LH-20 was(98.35 ± 0.72)%,which was 2.73 times of the crude extract;the DPPH free radical scavenging activity was 1.12 times of L-ascorbic acid;the inhibitory ability against ?-glucosidase was 120.71 times of acarbose.It showed that proanthocyanidins from Sargentodoxae caulis was abundant and which had an obvious antioxidant ability and enzyme inhibitory ability in vitro.2?To investigate the relationship between antioxidant ability with proanthocyanidins(OPC),polyphenols(TP)and flavonoids(TF)of Photinia serrulata extracts,to inspect the influence of solvent against antioxidant ability,different parts were ultrasonic extracted using different solvent systems.The content of OPC,TF,TP were determined,the antioxidant ability was evaluated.Results indicated that: the antioxidant ability has a significant difference between different parts of Photinia serrulata,whereas branch phloem and infructescence stalk were more potent than the other parts due to their higher contents of OPC,TP,TF.Significant differences were found among different solvents,which showed that OPC were ranged from(10.77 ± 0.08)% to(29.31 ± 0.69)%,TF were ranged from(2.60 ± 0.02)% to(7.02 ± 0.01)%,TP were ranged from(3.15 ± 0.02)% to(6.42 ± 0.07)% for branch phloem,and 85 % methanol was best for OPC,70 % acetone was best for TP and TF.A significant linear correlation(P < 0.01)between OPC,TF,TP and antioxidant ability indicated that OPC,TP,TF were mostly cooperated responsible for their antioxidant ability.3?Proanthocyanidins from Spatholobi caulis extracts were purified by macroporous adsorption resins,the static adsorption and desorption capacity of 3 resins including D101,X-5 and AB-8 were compared and the best X-5 resin was chosen for purification.Afterwards,the optimum parameters of dynamic adsorption and desorption for proanthocyanidins purification were obtained as follows: the loading rate,loading amount,eluting rate and eluting amount were 2 BV/h,10 BV,1BV/h and 2 BV respectively,when the loading concentration was 6.00 mg/m L.The contents of proanthocyanidins in the purified products F70 obtained by elution with 70% ethanol were(97.53 ± 0.27)%,which was 2 times of the crude extract;the DPPH free radical scavenging activity was 1.26 times of L-ascorbic acid;the inhibitory ability against ?-glucosidase was 107.56 times of acarbose.It showed that proanthocyanidins from Spatholobi caulis was rich and which had an obvious antioxidant ability and ?-glucosidase inhibitory.4?The structures of proanthocyanidins from Spatholobi caulis have been analyzed with ultraviolet absorption spectrum,infrared absorption spectrum,reverse phase high-performance liquid chromatography,matrix-assisted laser desorption/ionization time of flight mass spectrometry and 13 C nuclear magnetic resonance.Results indicated that they were mainly composed of procyanidin and prodelphinidin,the terminal units were mainly composed with catechin,while epicatechin were the dominant of the extension units.Proanthocyanidins from trimers to fifteen tpolymers were detected by matrix-assisted laser desorption/ionization time of flight mass spectrometry,and together with galloyl substituents.
Keywords/Search Tags:Sargentodoxae caulis, Photinia serrulata, Spatholobi caulis, proanthocyanidins, antioxidant ability, enzyme inhibitory ability, structural analysis
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