| Objective:In this experiment,by comparing the anti-inflammatory and analgesic eff ects of water huangqi tang and its different extracts and different concentrations of ethan ol extract,the preparation method of supply huangqi tang was determined.Establish the H PLC fingerprint of Huang Qi huangqi tang and single herb,determine the main chemical composition of body-em and its flavor of a drug;Establish the HPLC fingerprint of serum body-em and single herbs in studies huangqi tang,and clarify the source of each compone nt in studies huangqi tang;Through uplc detection of the chemical composition and serum chemical composition of studies huangqi tang,the blood transitional composition of studi es Yellow was determined,and the pharmacodynamic material basis of its effect was preli minarily clarified.Methods:1.In this experiment,water and 20 %,50 %,75 % and 95 % ethanol were used as the extraction solvent,and the water extract was extracted to different parts of petroleum ether,ethyl acetate and n-butanol to compare their anti-inflammatory and analgesic effects.Taking dashuzu swelling degree as anti-inflammatory evaluation index,hot plate method and acetic acid torsion body method time as the analgesic evaluation index,the spss19.0software package was used for statistical analysis to determine the preparation method of Huang Qi huangqi tang.2.By using HPLC method,the HPLC fingerprint of mobile phase and single herbs was established to determine the number of common peaks,the " correlation coefficient" and " included angle cosine" were used to calculate the similarity of fingerprint similarity,precision,stability and repeatability were studied.the chemical constituents of studies Huang Qi tang were determined.3.Through the preparation method of serum sample,serum treatment method,blood sampling time and days,the serum treatment method is: SD rats,gavage to 4.2g/mL(2 mg /mL),two times a day,the last time after 1h abdominal aorta blood,centrifuged serum.The protein was added with three times of acetonitrile,and the supernatant was purged with NBS-I,and then the flow phase was furong.The HPLC fingerprint of body-em Sum andsingle herbs Tang tang was established,and the content of into the blood in the serum of rats was analyzed by the fingerprint atlas.The HPLC chromatogram of the extracts,blank serum and containing serum were compared,and the chemical constituents of body-em were initially determined and their flavor of a drug were assigned.4.Using UPLC-ms techniques to prevent huangqi tang on serum medicinal chemistry research,extraction in the m/z 282-284,416-418,418-416 of positive ions,m/z in 313-313,345-487,609-619,619-609,845-850,850-985 anion,find out the blood transitional comp onents and metabolites,clarify the studies of astragalus root soup into the composition of blood and the efficacy of anti-arthritic efficacy material base.Results:1.Anti-inflammatory experiments showed:75%,95%ethanol extract on rats foot swel Ling swelling degree effect after 30 min inflammatory inhibition effect is better.Analgesic experiments show: the water extract group,50% and 50% ethanol extracts of caused by hot plate in mice licking foot response is best,and the water extract of 75% ethanol extract of twisting reaction induced by injection of acetic acid is slightly better.To sum up: 75% etha nol extracts from the best effect.2.Established acutum huangqi tang of the whole party and single herbs extract HPLC-DAD fingerprint,identified 28 common peak,using the method of correlation coefficient and the included Angle cosine method "to calculate the fingerprint similarity,above results were greater than 0.90,and the common peak,5,6,and 7(calycosinseparately),10,12,14(Mans handle flower glucoside),17(calycosin separately),18,21(awn,the handle),24,peak from astragalus membranaceus,2,8,9(licorice glycosides),11,13,15,liquoricegly cosides),16(licorice root element),19,20,22,23(glycyrrhizic acid)peak from licorice,25,27(f alkali Lin),28(such as powder out the alkali)is derived from the studies.1,3,4,26,has not found its attribution,may produce a component for the new.Atractylodes single herbs fingerprint chromatography of fengfeng shape is better,the studies astragalus root soup all extract fingerprint chromatographic peak is not obvious.3.Built out the astragalus root soup drug-containing serum of the whole party and single herbs HPLC-DAD fingerprint,detected out the fingerprint of 19 common peak of astragalus root soup among them 1,2,5,6(calycosin separately glycosidase),9 from theblank serum,7,8,10,12,13 peaks from astragalus membranaceus,3,4,14,15 peaks from licorice,17,18(f alkali Lin),19(powder out the alkali)is derived from the studies,11,16 not seen in the single medicinal serum,probably for metabolites.4.Using UPLC-ms techniques to prevent huangqi tang on serum medicinal chemi stry research.Detected so far,the studies in huangqi tang extract 30 chemical composition,16 and identify the chemical composition,respectivelyKaempferol-3-O-rutinoside、Magn ocurarine、Liquiritin、Calycosin-7-O-D-glucoside、phellodendrine、Rutin、Wogonin qisu、Fangchinoline、Ononin、Isoliquiritin、Tetrandrine、9,10-Dimethoxyptercarpan-3-O-β-D-glucopyranoside、Calycosin、LicoRicesaponine A3、Glycyrrhizic acid、Astragaloside II.The drug-containing serum of rats in the detected 21 chemical composition,11 of the m are from the astragalus root soup,and identify the chemical composition,10,respectivel y,Kaempferol-3-O-rutinoside、Liquiritin、Calycosin-7-O-D-glucoside、phellodendrine Wogonin qisu、Fangchinoline、Isoliquiritin、Tetrandrine、LicoRicesaponine A3、Astrag aloside II.other10 possible for metabolites.Conclusion:1.This study established the acutum huangqi tang of Chinese herbs were tested prepa ration method of 75% ethanol reflux extraction 2 times,each time 2 hours.2.Set up out the huangqi tang of the whole party and single herbs fingerprint,the met hod repeatability,stable and reliable;And the common peaks of the fingerprint of 28,10 of them are from astragalus membranaceus,11 from licorice,three from the studies,four new ingredients.3.Set up out the astragalus membranaceus soup all parties and single herbs serum of HPLC fingerprint,detected out the astragalus root soup drug-containing serum has 19 com mon peak,five from the blank serum,five from astragalus membranaceus,four from licor ice,three from the studies.4.Using UPLC-ms techniques to prevent huangqi tang on serum medicinal chemistry research.Detected out the extract of astragalus membranaceus soup 30 chemical composi tion,and identified 16 chemical composition;Medicated serum of rats detected 21 in che mical composition,11 of them are from the astragalus root soup,10 and identify the chemi cal composition.In serum,Kaempferol-3-O-rutinoside、Liquiritin、Calycosin-7-O-D-glucoside、phellodendrine、Wogonin qisu、Fangchinoline、Isoliquiritin、Tetrandrine、Lico Ricesaponine A3、Astragaloside II From out the astragalus root soup,Magnocurarine、Rutin、Ononin9,10-Dimethoxyptercarpan-3-O-β-D-glucopyranoside、Calycosin、Gly cyrrhizic acid is not into the blood stream.other 10 possible for metabolites. |
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