基于F68-Ce-Cu过氧化酶活性调控检测类烟产品中尼古丁  

Detection of Nicotine in Cigarette-like Products Based on the Regulation of Peroxidase-like Activity of F68-Ce-Cu

作　　者：陈丹 王春琼[1] 张轲 荣先芳 龙杰 李海燕 蔡洁云 陈志超 崔一凡 杨德志 杨亚玲 CHEN Dan;WANG Chun-qiong;ZHANG Ke;RONG Xian-fang;LONG Jie;LI Hai-yan;CAI Jie-yun;CHEN Zhi-chao;CUI Yi-fan;YANG De-zhi;YANG Ya-ling(Yunnan Institute of Tobacco Quality Inspection&Supervision,Kunming 650106,China;School of Materials Science and Engineering,Peking University,Beijing 100871,China;Research and Development Center,IMiracle(Shenzhen)Innovation Technology Co.,Ltd,Shenzhen 518126,China;Faculty of Life Science and Technology,Kunming University of Technology,Kunming 650500,China)

机构地区：[1]云南省烟草质量监督检测站,云南昆明650106 [2]北京大学材料科学与工程学院,北京100871 [3]爱奇迹(深圳)创新科技有限公司研发工程部,广东深圳518126 [4]昆明理工大学生命科学与技术学院,云南昆明650500

出　　处：《分析测试学报》2025年第3期445-453,共9页Journal of Instrumental Analysis

基　　金：云南省烟草质量监督检测站一般项目(20220001,20240002);中国烟草总公司云南省公司科技计划重点项目(2023530000241027)。

摘　　要：研究制备了具有过氧化物酶活性的铈、铜掺杂纳米酶(F68-Ce-Cu)用于“茶烟”、“花烟”等类烟产品中尼古丁的比色分析。在H_(2)O_(2)存在下,F68-Ce-Cu可以催化无色3,3',5,5'-四甲基联苯胺(TMB)转化为蓝色氧化型TMB(ox TMB)。研究发现尼古丁对F68-Ce-Cu过氧化物酶活性有显著增敏作用,且不会受到类烟产品中其它成分的干扰。米氏动力学研究结果发现,相较于单独Ce(K_(m)=2.04 mmol/L)或Cu(K_(m)=9.46 mmol/L)掺杂纳米酶,F68-Ce-Cu的米氏常数K_(m)显著降低(K_(m)=1.51 mmol/L),而在加入天然尼古丁之后,“F68-CeCu+尼古丁”的K_(m)为0.25 mmol/L,与底物的亲和性增强了6倍。Zeta电位测定结果表明,F68-Ce-Cu与尼古丁之间有较强的吸附作用,并能够促进产生更多的·OH和O_(2)^(·-),从而显著提高过氧化物酶活性。在0.017~0.52 mg/mL范围内,尼古丁质量浓度与654 nm处吸光度呈良好线性关系,检出限达到0.008 mg/mL。在实际类烟产品中,该方法的加标回收率为90.9%~103%,相对标准偏差为2.1%~4.6%,且与高效液相色谱法的检测结果几乎一致,表明该方法稳定性强、准确性高。Cerium and copper doped nanozyme(F68-Ce-Cu)with peroxidase-like activity was prepared for the colorimetric analysis of nicotine in“tea tobacco”and“flower tobacco”.In the presence of H2O2,F68-Ce-Cu could catalyze the conversion of colorless 3,3´,5,5´-tetramethylbenzidine(TMB)into blue oxidized TMB(oxTMB).It had been found that nicotine has a significant sensitizing effect on the peroxidase-like activity of F68-Ce-Cu without interference from other possible components in cigarette-like products.The results of the Michaelis-Menten kinetics study showed that compared to the Ce(K_(m)=2.04 mmol/L)or Cu(K_(m)=9.46 mmol/L)based-nanozyme alone,the Km of F68-Ce-Cu was significantly reduced(K_(m)=1.51 mmol/L),and the Km of“F68-Ce-Cu+nicotine”system was 0.25 mmol/L,which was 6 times more strongly bound to the substrate after the addition of natural nicotine.The results of Zeta potential determination showed that there was a strong adsorption between F68-Ce-Cu and nicotine,and it could promote the production of more·OH and O_(2)^(·-),which significantly increased the peroxidase-like activity.In the range of 0.017-0.52 mg/mL,nicotine concentration showed a good linear relationship with absorbance at 654 nm,and the detection limit reached 0.008 mg/mL.In the actual cigarette-like products,the recovery of the method was in the range of 90.9%-103%,and the relative standard deviation was in the range of 2.1%-4.6%,which was almost consistent with the detection results of high performance liquid chromatography,indicating that the method was stable and accurate.

关 键 词：尼古丁 类烟产品 纳米酶 过氧化物酶 比色检测 

分 类 号：O657.3[理学—分析化学] Q554.6[理学—化学]