The Survey Of Chemical Components And Pharmacological Effects In Citrus Changshan-huyou Y.B.Chang. And To Compared With Fructus Aurantii

Objectives(1)To compare the pharmacognostic characteristics;(2)To establish the HPLC fingerprint of C.changshan-huyou and compare with Citrus aurantium L.;(3)To analyze and compare the chemical components among the above TCMs with reference control method and LC/MS method;(4)To establish an HPLC method for the determination of multiple components in C.changshan-huyou,and compare with Citrus aurantium L.;(5)To establish TLC identification between the above 2 TCMs;(6)To compare the pharmacological effects between the above 2 TCMs.Methods(1)Compared the differences through medicinal properties and microscopic characteristics.(2)Optimized chromatographic conditions,and established HPLC fingerprint of C.changshan-huyou,compared the similarities between the above 2 TCMs,and compared with other variants to investigate the specificity of fingerprints.(3)Used reference substance and LC/MS to analyze the chromatographic peaks in side the fingerprint,made clear their chemical components,and used LC/MS technique to confirm their similarity.(4)Established an HPLC method for the determination of multiple components in C.changshan-huyou,and compare with Citrus aurantium L..(5)Used TLC to compare the differences between the above 2 TCMs.(6)Carried out the comparison experiments of normal intestinal carbon propelling and tracheal phenol red excretion in mice.Results(1)The pharmacognosy characteristics of C.changshan-huyou and Citrus aurantium L.were similar.(2)Established the fingerprint to wards C.changshan-huyou's characteristic components for the first time,the similarity measurement results showed that C.changshan-huyouand Citrus aurantium L.had high similarity,with the similarity as?0.90,while obviously different from Citrus maxima,Citrus reticulate 'Daidai'and Citrus wilsonii Tanaka Citrus maxima and so on;the cluster analysis classified C.changshan-huyou and Citrus aurantium L.into one group,while Citrus aurantium‘Chuluan' was significantly different from Citrus aurantium L.and its variants Citrus aurantium 'Daidai',Citrus aurantium 'Huangpi',(3)Reference control and LC/MS identified 10 components from C.changshan-huyou's fingerprint,namely rutinnaringin,naringin,hesperidin,neohesperidin,merancin hydrate,luteolin,merancin,nobiletin,hesperetin,aurapten,the component comparison revealed that these 2 TCMs had all 10 components.(4)established the HPLC method for the simultaneous determination of 8 comp-onents in C.changshan-huyou for the first time,rutinnaringin,naringin,hesperidin,neohes-peridin,merancin hydrate,luteolin,nobiletin,hesperetin,exhibited good linear relationship in 23.30?1164.80?g(r=1.000),170.84?8541.91 ?g(r=1.000),12.22?861.20 ?g(r=0.9995),156.17?7808.64 ?g(r=1.000),0.91?45.40 ?g(r=1.000),3.13?156.48 ?g(r=0.9995),0.90?45.23 ?g(r=1.000),0.85?42.27 ?g(r=0.9995),respectively,with the recovery ratesas 98.8%(RSD=1.7%),101.3%(RSD=3.0%),98.3%(RSD=3.3%),96.8%(RSD=1.7%),101.8%(RSD=4.9%),101.7%(RSD=2.2%),108.9%(RSD=3.4%).The component determination results,combined with principal component analysis,further revealed that them in components and their contents were similar between the above 2 TCMs.(5)TLC showed C.changshan-huyouand Fructus aurantii both contain synephrine,and TLC exhibited the similar results.(6)mice were orally administrated the above TCMs,compared with the control group,C.changshan-huyou and Citrus aurantium L.with different administration doses could significantly increase the normal in test in a carbon propelling rate(P<0.05),which there was no significant difference among different dose groups(P>0.05);compared with the control group,the above TCMs with the same administration dose could significantly increase the amount of tracheal phenol red excretion in mice(P<0.05),and there was no significant difference among different dose groups(P>0.05).Conclusions(1)Appearance,microscopic characteristics,and synephrine TLC comparison suggested that C.changshan-huyou and Citrus aurantium L.were basically similar.(2)The fingerprint established had strong specificity,and could be used to effectively identify C.changshan-huyou from Citrus maxima,Citrus reticulate Blanco cv.Suavissia and Citrus wilsonii Tanaka,Citrus maxima and so on,etc.The similarity measurement showed high similarity between C.changshan-huyou and Citrus aurantium L.;the component analysis results showed that the main component peaks were the same between them.(3)RP-HPLC method was simple and accurate,and could provide the basis for the comprehensive evaluation between the above 2 TCMs.(4)The pharmacological effects of C.changshan-huyou and Citrus aurantium L.were basically the same.The above results provided references for C.changshan-huyou to be used as the new resource of Citrus aurantium L..