离体条件下鱼对灭幼脲的代谢

IN VITRO METABOLISM OF CCU IN FISH

  • 摘要: 实验选用甘氨酸-氢氧化钠(pH9±0.01)缓冲溶液,草鱼或鲤鱼肝脏匀浆液经低速离心后取上清液作为酶,反应系统中加入CCU后在25℃保温5h,用醋酸乙酯萃取代谢产物,用TLC对代谢产物进行分离纯化,用HPLC方法对代谢产物进行定性定量测定,证明有对氯苯基脲素CPU产生.为进一步证实代谢产物为CPU,用能给出绝对光谱定性数据的紫外检测器在一定的波长范围内,对标样及斑点中测定的组分进行扫描,从光谱学上证明了产物是CPU.当加入专一性酯酶抑制剂后,酶活性被抑制89-99%,证明酯酶在CCU的降解中起重要作用.

     

    Abstract: The reaction system includes 0.1 M GIy-NaoH buffer and the supernatant ofthe homogenate of the grass carp and common carp liver representing the enzymesolution. After addition of CCU,the reaction mixture was incubated at 25℃for 5h. The metabolite was extracted with ethy1 acetate,separated by TLC and identifiedbv HPLC,It was demonstrated that CPU was produced in the mixture,That theproduct was CPU was further verified by spectroscopy,using a specia1 UV detectorwhich could give absolute spectral evidence for the identification of CCU metabolite,and comparing the UV scanning chromatograph for CPU standard with that of themetabolite spot at the certain range of wavelengths. When specific inhibitor of es- terase was applied to the reaction mixture the activity of enzyme was decreased by 89~99 percent.It shows the key role of esterase in the production of CPU.

     

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