Comparative Analysis Of The Effective Components Of Fructus Aurantii Immaturus In Different Species And Place

Fructus Aurantii Immaturus(Zhishi),originated from "Shen Nong Ben Cao Jing",can dissipate stagnant qi,relieve food retention,eliminate phlegm and disintegrate masses. Zhishi is the dried immature fruits of Citrus aurantium L.and its cultivated variety,or Citrus sinensis Osbeck.It showed that distinct batches of Zhishi contain differences types of flavonoids which have different biological activities depending on their strcture.It is unknown whether there are differences in the types of alkaloid and volatile oil in Zhishi. Based on the species and quality of flavonoid,the quality of Synephrine,the species and quality of volatile oil,IR spectrum,the relation between the types and quality of effective components with species and place were studied.In this paper we provide an overview on the relevant study and development on Zhishi and developments at home and abroad in recent 50 years,and studies on the following aspects:The samples were collected from Sichuan,Hunan,Jiangxi and Guizhou,and the species were identified by specialist.The herbs were crosscut into two halves,dried by low temperature.By comparing the cross-sectional distinction among the various species,the feasibility of identifying Zhishi species from its characteristic was studied.The pulp of fruit percentage in the cross-sectional total area ranged from 0.57%to 15.0%in Citrus aurantium L.,13.2%to 19.9%in Citrus junos Sieb.ex Tanaka,20.1%to 44.2%in Citrus sinensis Osbeck and 25.4%to70.6%in Poncirus trifoliate Raf.,Thus we can discern the species of Zhishi by the pulp of fruit percentage of the cross-sectional ranged.RRLC method was used to determine and analyze flavonoids of Zhishi,the relations between the type and content of flavonoids in Zhishi to their species and places.The analysis of the relation of flavonoid types and species showed that the main flavonoids of Citrus aurantium L.,Citrus aurantium L.cv 'xiucheng',C.aurantium×P.trifoliata and Citrus junos Sieb.ex Tanaka are hesperidin,naringin,neohesperidin and narirutin.The content of naringin and neohesperidin in Citrus aurantium L.,Citrus aurantium L.cv'xiucheng',C.aurantium×P.trifoliata are higher than narirutin and hesperidin.Adversely,the content of narirutin and hesperidin in Citrus junos Sieb.ex Tanaka are higher than naringin and neohesperidin.The Citrus sinensis Osbeck.only have narirutin and hesperidin,and the content of hesperidin was high.In addition,the analysis of the relation of flavonoid types and places showed that the same species but various places have the same flavonoid types,and the same place but different species have differrent flavonoids.In conclusion,the type and content of flavonoids in Zhishi are related to their species but not their places.According to Pharmacopoeia of The People's Repubic of China(2005),the content of Synephrine in Zhishi of various species and places was determined by HPLC method.It showed that the contents of Synephrine have no significant differences in different species and places of Zhishi.To know if Citrus junos Sieb.ex Tanaka can belongs to Citrus aurantium L.,the fingerprints of Zhishi from different species and places were determined by RRLC,and were evaluated the differences of components in Zhishi of various species and places by Similarity Evaluation System software for Chromatographic Fingerprint of TCM and Hierarchical cluster software.The fingerprints of 30 samples could be divided into 3 classes:the first is Citrus aurantium L.(including Citrus aurantium L.cv 'xiucheng' and C.aurantium×P.trifoliata);the second is Citrus junos Sieb.ex Tanaka;the last is Citrus sinensis Osbeck. Referring to the reference spectrum,the similar value in same species is higher,the similar value of Citrus aurantium L.,Citrus aurantium L.cv 'xiucheng' and C.aurantium×P.trifoliata are higher than 0.9,but the similar valve of Citrus aurantium L.and Citrus junos Sieb.ex Tanaka are lower than 0.2.In hierarchical cluster tree,Citrus aurantium L.,Citrus junos Sieb.ex Tanaka and Citrus sinensis Osbeck can cluster to one class respectively. Conclusively,the classification results is same to the "Flora of China",and contribute to the conclusion of Citrus junos Sieb.ex Tanaka not belonging to Citrus aurantium L.The fragment characteristics of main flavonoids in Zhishi were analyzed by DAD and (±)ESI-MS~n detector.The summary of characteristics could know the MS split regularities, which can reflect the structural characteristics and differences.Compared with the literature data,UV,DAD and ESI-MS~n data are used to identify the major peaks in fingerprint.In conclusion,four components were identified in Citrus aurantium L.,include narirutin, naringin,hesperidin and neohesperidin;four components were identified in Citrus sinensis Osbeck,include lonicerin,narirutin,hesperidin and 3',4',5,7-tetrahydroxy-7-O-rutinose;six components were identified in Citrus junos Sieb.ex Tanaka.,include narirutin,naringin, hesperidin,neohesperidin and 3',4',5,7- tetrahydroxy-7-O-rutinose.Except for the known flavonoids,lonicerin was only finded in Citrus sinensis Osbeck.;3',4',5,7 -tetrahydroxy-7-O-rutinose was found both in Citrus aurantium L.and Citrus junos Sieb.ex Tanaka.The volatile oil of Zhishi from different species and places are analyzed by GC-MS to find the relation between species and place.The chemical constitutes of volatile oils are identified by Data Base Retrieval and the relative standard mass spectrum data.The total oil currents were analyzed by chemical pattern recognition to find the relation between the constitutes of volatile oils in Zhishi with the species and places.The cluster tree showed that Citrus sinensis Osbeck cluster to a class,Citrus aurantium L.cluster to a class,Citrus junos Sieb.ex Tanaka clustered to a small class which belongs to Citrus aurantium L.It is indicated that the components of volatile oil of Zhishi are related to the species.In addition,the distance of the same species in same place is smaller than the different places.Therefore,the components of volatile oil of Zhishi are not only related to species but also the place.FI-IR was used to measure the infrared spectrum of 10 batches Zhishi,which have differences in flavonoids.The relation studies of spectrum were analyzed by IR analysis software.Compared with the control medicinal material,the correlation coefficients of 3 batches of Citrus sinensis Osbeck are lower than 0.50;the correlation coefficients of 7 batches of Citrus aurantium L.and-Citrus junos Sieb.ex Tanaka are higher than 0.90.In conclusion,the FT-IR could be used to study the types differences of flavonoids in Fructus Aurantii Immaturus.