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Study on Separation Conditions of Alkaloids in Peels of Carya Cathayensis Sarg, and Influence of Isoliensinine on Activity of CYP3A in Rats

Author: NanLi
Tutor: XuanGuiDa
School: Zhejiang University
Course: Pharmacy
Keywords: Carya cathayensis Sarg. Alkaloids Separation Isoliensinine CYP3A
CLC: R285
Type: Master's thesis
Year: 2012
Downloads: 62
Quote: 0
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Abstract


In the first part of this paper, we have studied on the separation conditions of alkaloids in the peels of Carya cathayensis Sarg. to provide a scientific basis for the further exploitation and utilization.First of all, a mixture extracts were obtained by ultrasound aided extraction. Then alkaloids were seperated from the mixture extracts by acid sinking alkali formulation method. After that, system extraction of alkaloids methods were used to divided the alkaloids into four parts according to their polarity. Fianlly We investigated the separation conditions of alkaloids in the peels of Carya cathayensis Sarg. by silica gel column chromatography and thin layer chromatography(TLC). The results show that: the extraction yield of mixture extracts, weakly alkaline, tertiary alkaloids, chloroform extract and ethyl acetate extract(C) were 6.09%, 0.22%, 0.018%, 0.15% and 1.69% respectively. For part C, use chloroform-methanol(30 ’ 1—>2 ? 1; V/V) as washing solvent, gradient elution, got three parts: E, F, G. The same separation conditions was used to part E,then got two parts:H,I.For partⅠ,the optimal separation conditions was using petroleum ether- ethyl acette(3,:1→1:1;Ⅴ/Ⅴ)as washingslvent,gradient elutionIn the second part of this paper,we have studied the influence that Isoliensinine (IL)to CYP3A enzymes in rats,to know the possible interactions when use IL and the substrates of CYP3A enzymes together.In the second part of this paper,we have investigated the influence of IsolienSinine (IL) upon CYP3A enzymes in rats,to demonstrate the possible interactions between IL and the substrates of CYP3A enzymes.First of all, a mixture metabolic system of liver microsome enzymes was established in vitro.Then different concentrations of IL and a certain amount of testosterone were incubated together in above mixture metabolic system,respectively. After that,the reduction amount of testosterone was measured to explore the suitable incubation conditions.Here,testosterone was used as a probe to test the metabolic activity of CYP3A by HPLC method.The results show that:In mixture metabolic system of liver microsome enzymes,the best incubation concentration of testosterone was 200μmol/L and the best incubation time was 210 min.However,IC50>1000μmol/L,indicating IL has little inhibition on CYP3A.IL could be used with substrates of CYP 3A enzymes.

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