| Objective:To optimize the extract method of jujubosides from Semen Zizyphi Sponosae(SZS).To establish fingerprint and methods for qulity control of jujubosides.To investigate the antidepressive mechanmism of jujubosides in chronic unpredictable stress induced depression model in mice.Method:1.LAccording to the speciality of jujubosides,optimized the extract method.2.To establish the fingerprint of jujubosides by high performance liquid chromatography-diode array detection(HPLC-DAD).A method for simultaneously determining jujuboside A,jujuboside B,betulinic and betulin in jujubosides has been established by using HPLC-DAD.3.The chronic unpredictable mild sress(CUMS)animal model was used to determine the antidepressant-like effects of jujubosides in mice.The forced swimming test(FST),tail suspension test(TST),open field test(OFT),body weight of mice and sugar water consumption test were observed.Mice immobility duration in FST and TST,spontaneous activity frequency in OFT,the body weight of mice and sugar water consumption were observed as evaluate index to judge the antidepressant effect of jujubosides.The mechanisms were examined by measuring monoamine oxidase in mice brain.Result:1.The content of total sapoin in jujubosides was obtained by the new method was higher than the systemic extract method.2.The fingerprint of jujubosides was established by HPLC-DAD.Chromatographic condition:Chromatographic column:Agilent TC-C18(4.6 mm×250 mm,5?m);Mobile phase:acetonitrile(A)-water(B),gradient elute;Time:0 min,15%A;0?10 min,15?25%A;10?20 min,25?45%A;20?30 min,45?60%A;30?45 min,65?70%A;45?55 min,70%A;55?70 min,70?80%A;70?90 min,80?100%A;90?120 min,100%A;Flow rate:1 mlˇmin-1;Column temperature:35 ?;Sample size:20 ?l;Determine wavelength:204 nm.The content of jujuboside A,jujuboside B,betulinic and betulin in jujubosides was simultaneously determined by HPLC-DAD.The jujuboside A concentration was linear relationship in the range of 1.00?16.00 ?g.The equation of linear regression was Y=31992X-86180,r=0.9993.The average recovery was 99.18%and RSD was 1.11%.The jujuboside B concentration was linear relationship in the range of 0.50?12.00 ?g.The equation of linear regression was Y=30824X-23938,r=0.9995.The average recovery was 99.48%and RSD was 1.40%.The betulinic concentration was linear relationship in the range of 2.50?40.00 ?g.The equation of linear regression was Y=77095X-6×106,r=0.9990.The average recovery was 100.2%and RSD was 1.59%.The betulin concentration was linear relationship in the range of 0.25?4.00 ?g.The equation of linear regression was Y=69369X-5×106,r=0.9993.The average recovery was 99.50%and RSD was 1.14%.The result shows that the method is simple,accurate,highly sensitive and well rugged.It's feasible for quality control of jujubosides.3.Compared with the model group,jujubosides treatment group led to a reduction in the immobility period could in FST and TST.There were no significant differences on mice spontaneous activity in OFT.And jujubosides significantly inhibited monoamine oxidase-A(MAO-A)and monoamine oxidase-B(MAO-B)after 21-day CUMS exposure.Conclusion:According to the speciality of jujubosides,the optimized extract method was established.The method is simple,feasible and suitable for industry production.By using HPLC-DAD,the fingerprint of jujubosides was established.The determination of jujuboside A,jujuboside B,betulinic and betulin was simple and accurate and could be used as quality standards of jujubosides.The pharmacodynamic result showed jujubosides produced an antidepressant-like effect.In addition,the mechanism of action involves the monoamine oxidase enzyme systems. |
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