毛细管电泳—化学发光法分离检测金属离子的新体系
摘要
毛细管电泳以其独特的分离效率高,进样量小,分析速度快等优越性能。已在分离科学的众多研究领域和实际应用中得到了迅速发展,并不断开拓新的研究领域和方法,成为分析科学的前沿领域。而化学发光分析法具有很高的灵敏度,但选择性差限制了其应用,将毛细管电泳的高分离效率和化学发光检测的高灵敏度相结合,具有广泛的发展和应用前景。本文主要进行了金属离子的分离及检测研究:
金属离子的分离及检测在环境和生物领域有着极其重要的价值。传统的方法或是仪器昂贵或是常常需要进行衍生处理,其过程繁琐、耗时。目前,毛细管电泳-化学发光法用于金属离子分离检测所采用的发光体系基本为鲁米诺-过氧化氢体系,但是该体系最大的缺陷在于:过氧化氢不稳定,容易产生气泡引起毛细管堵塞,从而影响仪器的运行。而鲁米诺-铁氰化钾化学发光体系具有稳定、反应快速、分析灵敏度高等特点,但是,该体系用于金属离子的分离检测还未见报道。本研究基于某些金属离子对鲁米诺-铁氰化钾体系有催化化学发光反应的特性,采用毛细管电泳-化学发光法成功实现了对Co(Ⅱ)和Cu(Ⅱ)的分离检测,并对化学发光条件和电泳条件进行了优化:电泳分离时所用的介质是10 mmol/L醋酸钠溶液(含0.8 mmol/L的鲁米诺,2 mmol/L HIBA,用醋酸溶液调节pH为4.8);柱后试剂:2.0 mmol/L铁氰化钾(用氢氧化钠溶液调节pH值为13.0)。在优化条件下,Co(Ⅱ)和Cu(Ⅱ)的检出限(S/N=3)分别为7.5×10-11 mol/L和7.5×10-9 mol/L,线性范围分别为7.5×10-9 mol/L～1.0×10-6 mol/L和7.5×10-8 mol/L～5.0×10-5 mol/L,迁移时间和峰高的相对标准偏差(n=5)分别为2.2%、2.9%(Co(Ⅱ) );4.2%、4.7%(Cu(Ⅱ) )。
In the past years, capillary electrophoresis(CE) has gained significant acceptance in the modern analytical laboratory. This technique yields highly efficient separations while requiring only amounts of sample. The constraints of on-column detection generally result in poor detection limits and have reduced the overall application of CE.
Chemiluminescence(CL) detection has been shown to be a highly sensitive detection method. Due to its simple optical system and low background nature, CL is expected to be an ideal detection method for CE. CE-CL technique has received significant attention over the past several years. The research results obtained in the thesis are listed as follows: Separation and detection of metal ions are important in both environmental and biological applications. The main disadvantage of conventional methods are high costs, and some require tedious pre-column treatments and require large amount of analyte. In the previously used interface for CE-CL detection, the presence of gas bubbles in the reaction capillary due to decomposed hydrogen peroxide in alkaline solution would affect the stability of the current and background, therefore ,would disturb the detection. CL(luminol with potassium ferricyanide ) detection with CE for metal ions analysis has not yet been reported. Based on the fact that some metal ions can catalyze the chemiluminescence (CL) reaction of luminol with potassium ferricyanide, a novel capillary electrophoresis CL method was developed for the determination of Co(Ⅱ)and Cu(Ⅱ). The separation was carried out with a 10mmol/L sodium acetate solution containing 0.8 mmol /L luminol and 2.0 mmol/L HIBA (adjusted to pH 4.8 by HAc solution). The post-capillary reagent was 2.0 mmol/L K3Fe(CN)6 which was adjusted to pH 13.0 by NaOH solution. Under the optimum conditions, the detection limits (S/N=3) for Co(Ⅱ) and Cu(Ⅱ) were 7.5×10-11 mol/L and 7.5×10-9 mol/L, with the linear range of 7.5×10-9 mol/L~1.0×10-6 mol/L and 7.5×10-8 mol/L~5.0×10-5 mol/L, respectively. The RSDs (n=5) for migration time and peak height were 2.2%、2.9% for Co(Ⅱ) ,and 4.2%、4.7% for Cu(Ⅱ),respectively.
金属离子的分离及检测在环境和生物领域有着极其重要的价值。传统的方法或是仪器昂贵或是常常需要进行衍生处理,其过程繁琐、耗时。目前,毛细管电泳-化学发光法用于金属离子分离检测所采用的发光体系基本为鲁米诺-过氧化氢体系,但是该体系最大的缺陷在于:过氧化氢不稳定,容易产生气泡引起毛细管堵塞,从而影响仪器的运行。而鲁米诺-铁氰化钾化学发光体系具有稳定、反应快速、分析灵敏度高等特点,但是,该体系用于金属离子的分离检测还未见报道。本研究基于某些金属离子对鲁米诺-铁氰化钾体系有催化化学发光反应的特性,采用毛细管电泳-化学发光法成功实现了对Co(Ⅱ)和Cu(Ⅱ)的分离检测,并对化学发光条件和电泳条件进行了优化:电泳分离时所用的介质是10 mmol/L醋酸钠溶液(含0.8 mmol/L的鲁米诺,2 mmol/L HIBA,用醋酸溶液调节pH为4.8);柱后试剂:2.0 mmol/L铁氰化钾(用氢氧化钠溶液调节pH值为13.0)。在优化条件下,Co(Ⅱ)和Cu(Ⅱ)的检出限(S/N=3)分别为7.5×10-11 mol/L和7.5×10-9 mol/L,线性范围分别为7.5×10-9 mol/L～1.0×10-6 mol/L和7.5×10-8 mol/L～5.0×10-5 mol/L,迁移时间和峰高的相对标准偏差(n=5)分别为2.2%、2.9%(Co(Ⅱ) );4.2%、4.7%(Cu(Ⅱ) )。
In the past years, capillary electrophoresis(CE) has gained significant acceptance in the modern analytical laboratory. This technique yields highly efficient separations while requiring only amounts of sample. The constraints of on-column detection generally result in poor detection limits and have reduced the overall application of CE.
Chemiluminescence(CL) detection has been shown to be a highly sensitive detection method. Due to its simple optical system and low background nature, CL is expected to be an ideal detection method for CE. CE-CL technique has received significant attention over the past several years. The research results obtained in the thesis are listed as follows: Separation and detection of metal ions are important in both environmental and biological applications. The main disadvantage of conventional methods are high costs, and some require tedious pre-column treatments and require large amount of analyte. In the previously used interface for CE-CL detection, the presence of gas bubbles in the reaction capillary due to decomposed hydrogen peroxide in alkaline solution would affect the stability of the current and background, therefore ,would disturb the detection. CL(luminol with potassium ferricyanide ) detection with CE for metal ions analysis has not yet been reported. Based on the fact that some metal ions can catalyze the chemiluminescence (CL) reaction of luminol with potassium ferricyanide, a novel capillary electrophoresis CL method was developed for the determination of Co(Ⅱ)and Cu(Ⅱ). The separation was carried out with a 10mmol/L sodium acetate solution containing 0.8 mmol /L luminol and 2.0 mmol/L HIBA (adjusted to pH 4.8 by HAc solution). The post-capillary reagent was 2.0 mmol/L K3Fe(CN)6 which was adjusted to pH 13.0 by NaOH solution. Under the optimum conditions, the detection limits (S/N=3) for Co(Ⅱ) and Cu(Ⅱ) were 7.5×10-11 mol/L and 7.5×10-9 mol/L, with the linear range of 7.5×10-9 mol/L~1.0×10-6 mol/L and 7.5×10-8 mol/L~5.0×10-5 mol/L, respectively. The RSDs (n=5) for migration time and peak height were 2.2%、2.9% for Co(Ⅱ) ,and 4.2%、4.7% for Cu(Ⅱ),respectively.
引文
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