Investigation on chemiluminescence detection of vitamin B12 with capillary electrophoresis
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摘要: 基于维生素B12对鲁米诺-双氧水体系良好的化学发光(CL)催化活性, 研究确立了一种新型毛细管电泳---化学发光(CE-CL)检测方法, 用于这一重要营养物质的准确CL分析. 预处理阶段, 连二亚硫酸钠(Na$_{2}$S$_{2}$O$_{4}$)被确定为将VB12(Co(III))转化成VB12(Co(II))的适当还原剂, 以使VB12在luminol-H$_{2}$O$_{2}$发光体系中获得良好的催化信号. 这一过程产生的CL干扰组分, 则通过毛细管电泳(CE)加以分离除去. 实验中, VB12可被温和还原并由催化所得CL信号精确测定. 采用实验室自行组建的CE-CL检测装置, 该测试过程可在20 min内完成;检测下限(LOD)2$\times $10$^{-7}$ mol/L($S$/$N$=3), 线性范围6$\times $10$^{-7}\!\sim $6$\times $10$^{-4}$ mol/L($r^{2}$=0.968), 相对标准偏差(RSD)2.4{\%}($n$=9). 对药品中VB12含量进行加标回收实验, 回收率97{\%}$\sim $106{\%}. 相较于已有关于VB12的CL分析研究, 本文首次考察了预还原处理中产生的干扰组分及其分离过程, 以便捷操作获得更为可靠的检测数据.Abstract: Based on the chemiluminescence (CL) catalysis of vitamin B12 (VB12 in luminol-H$_{2}$O$_{2}$ system, a novel capillary electrophoresis-chemiluminescence (CE-CL) method was developed for the accurate determination of this important nutrient substance. First of all, sodium hydrosulfite (Na$_{2}$S$_{2}$O$_{4})$ was found to be used as an competent reduction reagent to pre-reduce VB12(Co(III)) into VB12(Co(II)), which produced strong CL emission in the CL reaction of luminol-H$_{2}$O$_{2}$. In this process, some interfering CL noise was observed and capillary electrophoresis (CE) was taken to remove it by efficient separation. In this way, VB12 could be reduced gently and determined accurately by the catalyzed CL signal with our lab-constructed CE-CL system. The proposed CE-CL determination could be finished in 20 min with a detection limit (LOD) of 2$\times $10$^{-7}$ mol/L ($S$/$N$=3), a linear range from 6$\times $10$^{-7}$ to 6$\times$10$^{-4}$ mol/L ($r^{2}$=0.968) and a relative standard deviation (RSD) of 2.4{\%} ($n$=9). Utilizing this method, satisfactory results were obtained by testing VB12 tablets with a recovery of 97{\%}$\sim $106{\%}. Compared to previously reported CL analysis of VB12, we dealt with the interference components in pre-reducing solution for the first time to gain more reliable data with convenient operations.
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Key words:
- vitamin B12 /
- chemiluminescence /
- capillary electrophoresis /
- sodium hydrosulfite
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