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为对烟碱代谢中间产物进行安全性和生理功能评价,以烟碱为底物,利用节杆菌Z3发酵法合成烟碱代谢中间产物L-6-羟基烟碱。采用HPLC监测发现发酵反应24 h,目标产物积累量达到最大。以V(乙酸乙酯)∶V(甲醇)∶V(氨水)=6∶1∶0.01的混合溶液为洗脱剂,用柱层析对发酵粗产物进行分离纯化,目标产物利用HPLC检测其纯度为99.00%,经IR、1HNMR、13CNMR和MS进行了结构表征,证实了烟碱代谢中间产物L-6-羟基烟碱。
In order to evaluate the safety and physiological functions of the intermediates of nicotine metabolism, Nicotine was used as a substrate to synthesize nicotine intermediate metabolite L-6-hydroxynicotine using Arthrobacter Z3 fermentation. The results of HPLC showed that the fermentation product reached the maximum 24 h after fermentation. The crude fermentation product was separated and purified by column chromatography with a mixed solvent of V (methanol): V (methanol): V (ammonia) = 6:1:0.01 as eluant. The purity of the target product was determined by HPLC Was 99.00%. Structural characterization by IR, 1HNMR, 13CNMR and MS confirmed the nicotine metabolism intermediate L-6-hydroxynicotine.