| Objective:Gas purge microsyringe extraction (GP-MSE) was used for extracting the volatile compounds of Asarum heterotropoid.es Fr.Schmidt val. Mandshuricum (Maxim.) Kitag. The compounds were analyzed by gas chromatographic-mass spectrometric (GC-MS) and HPLC for establish GP-MSE-GC-MS and HPLC fingerprints.HPLC fingerprints of Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.)Kitag extract and serum of rats after taking the extract were established with serum pharmacochemistry method.These were distinct,and the basis experimental evidence is provided for the rationality of the clinical application and the establishment of quality standard.Methods:1. Study on Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.)Kitag by GP-MSE-GC-MS DB-5MS capillary column (30m×0.25mm×0.25mm) was used. Helium of high-purity was used as the carrier gas at a flowrate of1.0mL/min. Splitless injection was conducted and the injection volume is2μL.The injector temperature was280℃. The GC oven temperature was programmed as follows:the initial oven temperature was45℃and held for1min, and then to250℃at a rate of4℃/min and finally to280℃at a rate of6℃/min and held for5min. El for the ion source and the temperature is200℃, electron bombardment energy is70eV.2. Study on Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.)Kitag by HPLC HPLC chromatographic conditions:Kroasil C18column(4.6mm×250mm,5μm), using methanol-water solution as mobile phase gradient elution. The flow rate was1.0mL.min-1with detection wavelength of290nm. Column temperature was25℃. The injection volume is20μL3. Preliminary Study on Serum Pharmacochemistry of Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.)Kitag Methanol extraction solvent was medicinal materials of Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.)Kitag.SD Rats to the dose of5.4g/kg, twice a day, fill the stomach of rats for5days, on the last day to fill the stomach after eyeball blood make in an hour. According to the above method for HPLC chromatographic analysisResults:1. Study on Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.^Kitag by GP-MSE-GC-MSGP-MSE to get the kinds of volatile oil in the Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.)Kitag is obvious more than SD.38peaks were isolated, identified30peaks, identified components accounted for98.42of total volatile oil by SD.96peaks were isolated, identified46components which accounted for94.23of total volatile oil by GP-MSE.69components were separated but39were identified in leaves,79components were separated but34were identified in petioles and84components were separated but33were identified in rhizome.which identified components accounted for the percentage of total volatile components were89.78%81.99%and89.20%respectively. Twenty-one common components were identified in them. Relative content is more than1%for aldehyde sugar、sugar alcohol、Methyleugenol、hexadecane acid And pentadecane. The largest percentage content in the rhizome is Methyleugenol, followed by N-Isobutyl-(2E,4Z,8Z,10E)dodecatetraenamide. The largest percentage content in the leaves is linoleic acid content, followed by Methyleugenol. The largest percentage content in the petioles is hexadecanoic acid content,followed by2,3-dihydro coumarone.10kinds of special components were Identified in the leaves which were2,3-dihydrocouumaro、Humulene、Viridiflorol、1,2-Dimethyl-1-cyclooctene Pentadecanal、Hexahydrofarnesyl、acetone Phytol、Eicosanoic acid、1-Heptacosanol and Lignoceryl alcohol.2,5-dihydroxy acetophenone was exist in leaves. Corylon and alpha.-Bulnesene were exist in rhizome. Using GP-MSE-GC-MS for10batches of different origin of Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.)Kitag samples for analysis,37common peaks were identified and the fingerprint is established. Through principal component analysis get different batches Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.)Kitag which can classified as a class very well 2. Study on Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.)Kitag byHPLC Using HPLC for10batches of different origin of Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.)Kitag samples for analysis,13common peaks were identified and the fingerprint is established.3. Preliminary Study on Serum Pharmacochemistry of Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.)Kitag.,10rats serum was established HPLC fingerprint of medicinal chemistry,6peaks appeared after lavage,3peaks in the form of original drug into rats and3peaks may be metabolites of medicinal materials in certain ingredients and dosage of rats after stress and material.Conclusion:1. GP-MSE is a method of high sensitivity and specificity method than SD for Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.^Kitag.,2. The kinds and content of the volatile components from different parts varies a lot in the Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.,)Kitag.3. The GC-MS fingerprint and HPLC fingerprint Can be used in the quality evaluation and quality control of Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.)Kitag.4. The chemical composition of serum pharmacochemistry of Asarum heterotropoides Fr.Schmidt val. Mandshuricum(Maxim.)Kitag mainly comes from prototype components and metabolites. |
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