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OBJECTIVE:To optimize the extraction technology for flavonoids in leaves and seeds of Hippophae rhamnoides. METHODS:Using the total extraction rate of 6 flavonoid aglycones(catechins,rutin,myricetin,quercetin,kaempferol,isorham-netin)as index,ethanol volume fraction,extraction time,extraction times,material-lipid ratio as investigation indexes,L9(34)or-thogonal test was designed to optimize the extraction technology of flavonoids in leaves and seeds of H. rhamnoides,and verifica-tion test was carried out. RESULTS:The optimum extraction technology for flavonoids in leaves of H. rhamnoides was ethanol vol-ume fraction of 70%,extracting for 3 times with material-lipid ratio of 1:16,and 2.0 h each time;and that of seeds was ethanol volume fraction of 50%,extracting for 3 times with material-lipid ratio of 1:24,and 1.5 h each time. In verification test,the total extraction rate of 6 flavonoid aglycones was 56.4 mg/g in the leaves (RSD=1.4%,n=3) and 15.4 mg/g in the seeds (RSD=3.4%,n=3). CONCLUSIONS:Optimized extraction technology is simple,stable,feasible,and can be used for extracting the fla-vonoids in leaves and seeds of H. rhamnoides.
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To establish a rapid determination method of six flavonoids:catechin,rutin,myricetin,quercetin,kaempferol and isorhamnetin,from seabuckthorn leaves by RP-HPLC-DAD.The seabuckthorn leaves were first degreased by petroleum ether,extracted by ethanol,and determined by RP-HPLC-DAD.The six flavonoids were separated and eluted by a Shimadzu C18(150 mm ×2.1 mm,5 μm) column with methanol-water (0.1％ phos phoric acid) (60∶ 40) at a flow rate of 1.0 mL/min.The detection wavelength were as follow:catechin 208 nm,rutin 257 nm,myricetin 373 nm,quercetin 371 nm,kaempferol 367 nm,and isorhamnetin 371 nm,respectively.The injection volume was 20 μL.The contents of the six flavonoids were in the range of 0.47 to 30.00 μ,g/mL with good linearity.The validation of the method,including precision,stability and recovery rate,was acceptable.The established method can be used for fast determination of the content of six flavonoids in seabuckthorn leaves.
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@#In this study, normal isolated rat heart model was adopted to investigate the inotropic and electrocardiac effect of AF-HF001 with Lanfendorff technique on the left ventricular systolic pressure(LVSP), left ventricular end diastolic pressure(LVEDP), peak rate of rise of left ventricular pressure(+dp/dtmax)and peak rate of fall of left ventricular pressure(-dp/dtmax). The results indicated that AF-HF001 could significantly enhance LVSP, LVEDP and ±dp/dtmax in a dose-dependent manner and exerted a positive inotropic effect on normal isolated rat hearts. This study also analyzed the electrocardiac effect of AF-HF001 with the electrodes located on tip of heart, ventricular free wall and aortic root. The results exhibited that AF-HF001 did not change PR, QRS, QT or HR at various concentrations, which means that AF-HF001 could safely increase cardiac contractility. This research lays a good foundation for the further study on AF-HF001 and research on structure-activity relationship of its analogs.