| Sphallerocarpus gracilis(S.gracilis)is a perennial member of the Apiaceae family.According to ancient records,the entire S.gracilis plant was widely used to treat rheumatic arthritis,tetanus and ulcers in traditional Chinese medicine.The chemical composition and functionality of essential oil and phenolic compounds in S.gracilis seed were studied in this paper,which has a very important practical significance for the development of natural antibacterial agents and antioxidants,utilization and develpoment of S.gracilis and promotion of the local economics.The main research results of this paper are as follows:1.The chemical composition,yield,color,state and flavor of the essential oil from S.gracilis seed varied greatly with the extraction methods.The essential oil obtained by steam distillation was colorless,pungent with a lower yield,and its main constituents were p-cymene,y-terpinene and a-asarone.The essential oil extracted by supercritical carbon dioxide was light yellow,soften with a higher yield and its major components were p-cymene,?-terpinene and a-asarone.The main constituents of the essential oil extracted by solid-phase microextraction were ?-Pinene,p-cymene and ?-terpinene.In addition,the ultrastructure of the S.gracilis seed varied greatly with the different extraction methods.The ultrastructure of the S.gracilis seed treated with steam distillation and supercritical carbon dioxide extraction was greatly damaged.Whereas,no significant destroy was observed of the S.gracilis seed treated with solid-phase microextraction.2.The antibacterial activity of the essential oil from S.gracilis seed varied greatly with the extraction methods.Among the 13 tested microorganisms,no inhibitory effect was found against L.monocytogenes of the essential oil obtained by steam distillation,whereas no inhibitory effects were observed against L.monocytogenes and M.luteus of the essential oil obtained by supercritical carbon dioxide.As far as the A.niger was concerned,the essential oil extracted by steam distillation and supercritical carbon dioxide showed no inhitory effect.As a whole,the antibacterial activity of the essential oil obtained by steam distillation was stronger than the the essential oil extracted by supercritical carbon dioxide.Among all the tested microorganisms,the essential oil from S.gracilis seed showed the strongest inhibitory effect against K.pneumoniae.3.The components of the essential oil from S.gracilis seed effected the growth cruve of microorganisms and inhibited the germination of microorganisms.Physical damage and considerable morphological alteration were observed to the tested microorganisms treated with the essential oil.The cells present deformity in the cell walls,leakage of intracelluar constituents,surface roughness,fragmentation and lysis.Additionally,the cellular ultrastructure were also damaged by the essential oil from S.gracilis seed.The cell losted its whole structure and cell wall was destoried.Additionally,it was difficult to distinguish cell membrane and cell wall and the cytoplasm was not homogeneous.In some cells,the cytoplasm showed a serious collapse and vacuoles structure was observed.4.The total phenolic content in S.gracilis root,stem and seed was different.Higher levels of phenolic content was detected in seed,followed by stem and root;The content of free,soluble glycoside,conjugated and insoluble cell-wall-bound phenolics was different in S.gracilis root,stem and seed.In addition,the phenolic content of the S.gracilis root varied greatly with the harvest time and higher amounts of phenolic content was found in root harvested in spring.5.Chlorogenic acid,caffeic acid,peonidin-diglucoside,luteolin-glucoside,di-caffeoylquinic acid glucoside and catechin glucoside were identified in S.gracilis seed and their contents in free,soluble glycoside,conjugated and insoluble cell-wall-bound phenolic extracts were different.6.Significant ferric reducing antioxidant power,inhibitory effects on site and non-site hydroxyl radical mediated 2-deoxy-D-ribose degradation and scavenging capacity of DPPH and ABTS radicals were found of the free,soluble glycoside,conjugated and insoluble cell-wall-bound phenolic extracts from S.gracilis seed.Furthermore,the antioxidant activities of different phenolic extracts in different antioxidant systerms were various.7.The data demonstrated that significant correlation was found between phenolic content and DPPH radicals scavenging capacity,FRAP and TEAC.Conversely,no significant correlation was observed between phenolic content and inhibitory effects on site and non-site hydroxyl radical mediated 2-deoxy-D-ribose degradation.Among the methods used for quantifying antioxidant activity,significant correlation was found between DPPH radicals scavenging capacity,FRAP and TEAC,and inhibitory effects on site and non-site hydroxyl radical mediated 2-deoxy-D-ribose degradation.In contrast,there was no significant correlation between inhibitory effects on hydroxyl radical mediated 2-deoxy-D-ribose degradation and DPPH radicals scavenging capacity,FRAP and TEAC.8.Significant-OH and ROO· mediated DNA damage protective effects were observed of free,soluble glycoside,conjugated and insoluble cell-wall-bound phenolic extracts from S.gracilis seed.Moreover,-OH mediated DNA damage protective effects of the phenolic extracts higher than the ROO-mediated DNA damage.Results of the correlation analysis indicated that ·OH and ROO· mediated DNA damage protective effects of the phenolic extracts from S.gracilis seed not only depended on the phenolic content but also the type of phenolic constituent present in each fraction.9.Significant inhibitory effects were found of free,soluble glycoside,conjugated and insoluble cell-wall-bound phenolic extracts from S.gracilis seed on the Human Colon Carcinoma Lovo Cells.Furthermore,the inhibition rate gradually enhanced followed by the phenolic concentrations increasing.Based on the morphological observation,the morphology of the cells treated with the phenolic extracts changed greatly.Cell shrinkage and nuclear condensation were observed,which suggested that phenolic extracts from S.gracilis seed can induce the Human Colon Carcinoma Lovo Cells apoptosis.10.Results indicated that the free phenolic extract from S.gracilis seed showed preventive effect on liver injury induced by CCl4.The phenolic extract from S.gracilis seed remarkably inhibited the increase of liver index,spleen index,the activities of serum ALT,AST,AKP and liver TG,TC,MDA content,and elevated activities of SOD?CAT and GSH-PX induced by CCl4.Moreover,the middle dose group is the most advantage.The mice hepatic cells treated with middle dose were slightly damaged with a little intense coagulation necrosis,which further showed the hepatoprotective effects of the phenolic extract against carbon tetrachloride induced liver injury in rats. |
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