| Objectives1.Sulfur fumigation for Fritillaria thunbergii influences of total alkaloid,total saponin and water solution pH value research.2.To establish the characteristic fingerprinting of the water soluble constituents of Fritillaria thunbergii by sulfur fumigation and fresh-cut,which its hierarchical clustering analysis and principal component analysis research.3.The study of sulfur fumigation for Fritillaria thunbergii influences the content of water soluble constituents.4.The survey of sulfur fumigation for Fritillaria thunbergii influences of volatile oil components.5.The survey of sulfur fumigation for Fritillaria thunbergii influences of antitussive,expectorant,anti-inflammatory action,and toxicity.6.Sulfur fumigation for Fritillaria thunbergii influences of Peimine and Peiminine in vivo pharmacokinetics research.Methods1.By using UV chromogenic method,gravimetric method and determination of the pH meter method investigate sulfur fumigation for Fritillaria thunbergii influences of the concentration of total alkaloid,total saponins,and in the water solution pH value.2.Samples from Fritillaria thunbergii by sulfur fumigation and fresh cut processing each 13,using the method of HPLC-DAD to establish the characteristic fingerprinting of the water soluble constituents of Fritillaria thunbergii by sulfur fumigation and fresh-cut processing,and by using the traditional Chinese medicine(TCM)chromatographic fingerprint similarity evaluation system for 2004 A edition similarity calculation,set up its joint mode;which by using the SPSS 19.0 software cluster analysis and by using the SIMCA-P 11.5 software principal component analysis the water soluble component fingerprint of Fritillaria thunbergii by sulfur fumigation and fresh cut processing.3.Samples from Fritillaria thunbergii by sulfur fumigation and fresh cut processing each batch of 13,using the method of HPLC-DAD investigates sulfur fumigation on the effect of the content of water-soluble constituents of Fritillaria thunbergii.4.The water vapor method is adopted to extract volatile oil components in Fritillaria thunbergii by sulfur fumigation and fresh cut processing,using the method of GC-MS investigates sulfur fumigation on the effect of volatile oil components in the Fritillaria thunbergii.5.Ammonia in mice led to coughing,phenol red in mice shedding,mouse ear swelling induced by xylene and acute toxicity experiment to investigate sulfur fumigation on the effect of antitussive,expectorant,anti-inflammatory and toxicity of Fritillaria thunbergii.6.Using the method of LC-MS investigates sulfur fumigation for Fritillaria thunbergii influences of peimine and peiminine in vivo pharmacokinetics.Results1.The average content of total alkaloid in Fritillariae thunbergii by sulfur fumigation and fresh-cut was respectively 0.0629%、0.0727%,and the content of total saponin was respectively 0.8567%、2.3157%,and its water solution pH value was respectively 4.14、5.92.2.The water soluble constituents of Fritillaria thunbergii by sulfur fumigation have 25 common chromatographic peak in fingerprint,and its No.7,No.9,No.13,No.16,No.20,No.24,No.25 peaks was respectively adenine,inosine,uracil,uridine,β-thymidine,adenosine,2-deoxy adenosine,and The water soluble constituents of Fritillaria thunbergii by fresh-cut have 25 common chromatographic peak in fingerprint,and its No.7,No.8,No.11,No.14,No.18,No.20,No.24,No.25 peaks was respectively adenine,cytosine nucleoside,inosine,uracil,uridine,β-thymidine,adenosine,2-deoxy adenosine.From different growing areas and batches of Fritillaria thunbergii,which the water soluble constituents fingerprint was compared,and its similarity reached 0.95 above.The cluster analysis and principal component analysis for the fingerprint,and distinguished the different of two types of the Fritillaria thunbergii by sulfur fumigation samples and fresh-cut samples.3.To qualitative compared the water soluble constituents fingerprint of Fritillaria thunbergii by sulfur fumigation and fresh-cut.In the same reservation time,and the chromatographic peak area has changed.The water soluble constituents chromatograms of Fritillaria thunbergii by sulfur fumigation,the peak area of adenine,cytosine nucleoside,inosine,uracil,adenosine,2-deoxy adenosine were reduced,but uridine and β-thymidine peak area were increased.The determination of the water soluble constituents content of Fritillaria thunbergii by sulfur fumigation and fresh-cut from different growing areas and batch,which the average content of adenine was respectively range of 0.0050%and 0.0218%,the average content of cytosine nucleoside was respectively not detected and 0.1222%,the average content of inosine was respectively range of 0.0913%and 0.2610%,the average content of uracil respectively was range of 0.0046%and 0.0282%,the average content of uridine respectively was range of 0.0576%and 0.0242%,the average content ofβ-thymidine was respectively range of 0.1398%and 0.1001%,the average content of adenosine was respectively range of 0.2381%and 0.3593%,the average content of 2-deoxy adenosine was respectively range of 0.0360%and 0.0786%.4.The volatile oil components extracted from fritillaria thunbergii by sulfur fumigation was identified 13 kinds of components,and main component relative percentage content was L-(+)-Asorbic acid 2,6-dihexadecanoute(27.06%),3-Methyleneandrostan-17-ol(20.50%),Kaurene,(8.beta.,13.beta.)(10.76%)and 9,12-Octadecadienoic acid(10.07%).The volatile oil components extracted from fritillaria thunbergii by fresh-cut was identified 15 kinds of components,and main component relative percentage content was Linoleic acid ethyl ester(36.93%),Kaur-16-ene(22.85%),Hexadecanoic acid,ethyl ester(10.67%)and 3-Methyleneandrostan-17-ol(10.59%).5.The Pharmacology experiment results show that compared with negative control group,water decoction of Fritillaria thunbergii by sulfur fumigation in 1.5 g/kg,3 g/kg doses have obvious antitussive and expectorant effect(P<0.05,P<0.01),0.75 g/kg,0.75 g/kg,3 g/kg doses has obvious anti-inflammatory effects(P<0.01);Water decoction of Fritillaria thunbergii by fresh-cut in 0.75 g/kg,1.5 g/kg,the 3 g/kg dose group have obvious antitussive,expectorant and anti-inflammatory effects(P<0.05,P<0.01).Water decoction group of Fritillaria thunbergii by sulfur fumigation compared with water decoction group of Fritillaria thunbergii by fresh-cut in 0.75 g/kg,1.5 g/kg,3 g/kg doses the antitussive,expectorant and anti-inflammatory effects have no obvious difference(P>0.05).The extract of Fritillaria thunbergii by sulfur fumigation and fresh-cut was not obtained a median lethal dose.The extract of Fritillaria thunbergii by sulfur fumigation in mice maximum tolerance was 68.00(crude drug)g/kg,the extracts of Fritillaria thunbergii by fresh cut in mice maximum tolerance was 51.37(crude drug)g/kg;The weight of mice in Fritillaria thunbergii by sulfur fumigation and fresh cut extract group compared with negative control group have no significant difference(P>0.05),To naked eye observe the anatomical heart,liver,spleen,lung,kidney and other organs have found no obvious abnormalities.6.Rats gavage after the extract of Fritillaria thunbergii by sulfur fumigated and fresh cut,its main pharmacokinetics parameters showed that a maximum blood drug concentration(Cmax)of Peimine was respectively 66.40±4.65 ng/mL,186.37±18.82 ng/mL,the medication in the area under the curve(AUC)of Peimine was respectively 181.79±7.85(ng/mL)h,197.70±118.69(ng/mL)h;which a maximum blood drug concentration(Cmax)of Peiminine was respectively 146.72±10.88 ng/mL,227.65±7.01 ng/mL ng/mL,the medication in the area under the curve(AUC)of Peiminine was respectively457.38±58.81(ng/mL)h,566.16±41.55(ng/mL)h.Conclusion1.Sulfur fumigated for Fritillaria thunbergii reduced the content of total alkaloid,total saponins,and water solution pH value was more partial acid to a certain extent.2.To establish water soluble constituents fingerprint study methods and hierarchical cluster analysis of Fritillaria thunbergii by sulfur fumigated and fresh cut,which can quality control and identify Fritillaria thunbergii by sulfur fumigated and fresh-cut samples.3.The sulfur fumigated have different influence on the water soluble constituents content in the Fritillaria thunbergii,the content of adenine,cytosine nucleoside,inosine,uracil,adenosine,2-deoxy adenosine was reduced,but the content of uridine,β-thymidine was increased.4.The sulfur fumigated changed the composition of volatile oil components in Fritillaria thunbergii and their relative percentage content.5.Fritillaria thunbergii by sulfur fumigated and fresh-cut,which antitussive,expectorant and anti-inflammatory pharmacological effect is the difference;The water soluble extract of Fritillaria thunbergii by sulfur fumigated has low toxicity,and the sulfur fumigated improved the water soluble extract of Fritillaria thunbergii maximum tolerance dose.6.Sulfur fumigated reduced the Peimine and Peiminine of Fritillaria thunbergii in rat blood drug concentration(Cmax)and largest medication in the area under the curve(AUC). |
PDF Full Text Request