基于纳米金探针的6-巯基嘌呤分析方法研究
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摘要
近年来,金纳米粒子因其具有很多宏观粒子所不具备的小尺寸效应、表面效应、光学效应以及特殊的生物亲和效应,已经广泛应用于临床医学、材料学等领域。而且在医学检验领域,金纳米粒子标记技术成为目前第四大标记技术。金纳米粒子所具备的特殊的物理与化学性质,在许多研究领域中表现出了潜在的应用价值。目前,金纳米粒子的分析应用主要是通过化学修饰后作为光谱探针与其它物质结合反应,但是用不经过任何化学改性的金纳米粒子作探针,结合光度法或RRS法测定药物的分析方法报道还很少见。研究的主要内容有：
综述了金纳米粒子的制备和表征方法、光谱性质及其在分析化学中的应用和抗癌药物6-巯基嘌呤的性质、作用及分析现状等。
利用金纳米粒子作探针,建立了一种分光光度法检测微量抗癌药物6-巯基嘌呤的新方法。6-巯基嘌呤自组装到金纳米粒子上以后,溶液的颜色由亮红色变为深蓝色,吸收光谱发生明显的变化。反应前,金纳米粒子在520 nm处出现了其典型的等离子共振吸收带,而6-巯基嘌呤在可见区无吸收峰；反应后,520 nm吸收峰处的吸光度降低,而在680 nm处出现一较强吸收峰,该吸收峰为结合产物的待征吸收峰。并且,在该波长下吸光度与6-巯基嘌呤的浓度呈良好线性关系,检出限为17μg/L。对浓度为0.25mg/L的6-巯基嘌呤进行了11次平行测定,RSD为2.9%。将该法用于检测人尿样中的6-巯基嘌呤含量,方法简便、灵敏度高、抗干扰性强,检测结果令人满意。
以金纳米粒子为探针,建立了一种共振瑞利散射光谱法检测6-巯基嘌呤的新方法。在pH为5.50的B-R缓冲溶液中,金纳米粒子与6-巯基嘌呤自组装成体积更大的结合产物,从而使体系的共振瑞利散射光谱强度急剧增强。在实验中,研究了其共振瑞利散射光谱特征,并且考察了影响反应的一些因素和共存物质的影响。结果表明：在最佳条件下,6-巯基嘌呤浓度在0.017～0.32 mg/L范围内与共振瑞利散射光谱强度成线性关系,方法的灵敏度较高,其检出限(3σ)为8μg/L,对浓度为0.12mg/L的6-巯基嘌呤进行了11次平行测定,RSD为5.1%。考察共存物质对6-MP测定的影响,发现该方法具有较好的选择性。根据所确定的最佳条件,建立了一种共振瑞利散射光谱法检测6-巯基嘌呤的新方法,并将该方法用于人尿样中6-巯基嘌呤含量的测定,获得较好的结果。
Gold nanoparticles had many special properties such as quantum size effects, surface effects, optical effects and compatibility effects. With these properties, gold nanoparticles were widely used in the fields of clinical medicine, materials and more. Moreover, in the field of medical tests, now gold nanoparticles labels had been the forth important markers. Gold nanoparticles show the potential applied value because of their special physical and chemical characteristics. At present, the analytical applications of gold nanoparticles are as optical probes or as electrochemical sensors after chemical modification. However, there are rarely reports that spectrophotometry or Resonance Rayleigh scattering method is used to the determination of pharmaceuticals with unmodified gold nanoparticle probes. Main content are listed as follow:
The preparation and characterization methods, spectroscopy characterization and analytical application of gold nanoparticles and the property, purpose and the present situation of its analysis of 6-mercaptopurine are revivwed here.
A novel spectrophotometry method has been developed for the determination of micro amounts of 6-mercaptopurine, employing gold nanoparticles as probes. The color of test solution changed to blue from bright red is significantly observed upon the gold nanoparticles self-assembled with 6-mercaptopurine and the absorption peak was obviously different. Before reaction, there is a plasmon band at 520 nm that is the characteristics of gold nanoparticles and 6-mercaptopurine has no absorption peak in rang of visible light region. After reaction, the peak of 520 nm is lower, in addition, there is a maximum absorption peak located at 680 nm, which is the characteristic peak of the binding product. The absorption at 680 nm of different concentrations of 6-mercaptopurine could be quantified with good linear, with a low detection limit of 17μg/L, under optimal conditions. The relative standard deviation (n=11) was 2.9%at 0.25 mg/L 6-mercaptopurine concentration level. The proposed method was successfully applied to determination of the drug spiked human urine. The results are satisfactory, which indicates that the method is not only sensitive, simply, but also reliable and suitable for practical application.
A novel Resonance Rayleigh scattering method was developed for the determination of micro amounts of 6-mecaptopurine, employing gold nanoparticles as probes. In pH 5.50 weak acid medium, the greatly enhanced Resonance Rayleigh scattering spectrum can be observed upon the gold nanoparticles self-assembled with 6-Mercaptopurine because both of them react with each other to form a binding product with bigger volume. In this work, The Resonance Rayleigh scattering spectral characteristics, optimum conditions for the reaction and the influencing factors were investigated. The relative Resonance Rayleigh scattering intensity (ΔIRRS) is directly proportional to the concentration of 6-Mercaptopurine in the range of 0.017-0.32 mg/L. The method has high sensitivity and the limit of detection for 6-Mercaptopurine is 8μg/L. The RSD (n=11) is 5.1% at 0.12 mg/L 6-mercaptopurine concentration level. The effect of coexisting substances on the determination of 6-mercaptopurine was studied and the results showed that this method had good selectivity. Therefore, the proposed method is successfully used to determine 6-mercaptopurine in human urine by Resonance Rayleigh scattering with gold nanoparticle probes.
综述了金纳米粒子的制备和表征方法、光谱性质及其在分析化学中的应用和抗癌药物6-巯基嘌呤的性质、作用及分析现状等。
利用金纳米粒子作探针,建立了一种分光光度法检测微量抗癌药物6-巯基嘌呤的新方法。6-巯基嘌呤自组装到金纳米粒子上以后,溶液的颜色由亮红色变为深蓝色,吸收光谱发生明显的变化。反应前,金纳米粒子在520 nm处出现了其典型的等离子共振吸收带,而6-巯基嘌呤在可见区无吸收峰；反应后,520 nm吸收峰处的吸光度降低,而在680 nm处出现一较强吸收峰,该吸收峰为结合产物的待征吸收峰。并且,在该波长下吸光度与6-巯基嘌呤的浓度呈良好线性关系,检出限为17μg/L。对浓度为0.25mg/L的6-巯基嘌呤进行了11次平行测定,RSD为2.9%。将该法用于检测人尿样中的6-巯基嘌呤含量,方法简便、灵敏度高、抗干扰性强,检测结果令人满意。
以金纳米粒子为探针,建立了一种共振瑞利散射光谱法检测6-巯基嘌呤的新方法。在pH为5.50的B-R缓冲溶液中,金纳米粒子与6-巯基嘌呤自组装成体积更大的结合产物,从而使体系的共振瑞利散射光谱强度急剧增强。在实验中,研究了其共振瑞利散射光谱特征,并且考察了影响反应的一些因素和共存物质的影响。结果表明：在最佳条件下,6-巯基嘌呤浓度在0.017～0.32 mg/L范围内与共振瑞利散射光谱强度成线性关系,方法的灵敏度较高,其检出限(3σ)为8μg/L,对浓度为0.12mg/L的6-巯基嘌呤进行了11次平行测定,RSD为5.1%。考察共存物质对6-MP测定的影响,发现该方法具有较好的选择性。根据所确定的最佳条件,建立了一种共振瑞利散射光谱法检测6-巯基嘌呤的新方法,并将该方法用于人尿样中6-巯基嘌呤含量的测定,获得较好的结果。
Gold nanoparticles had many special properties such as quantum size effects, surface effects, optical effects and compatibility effects. With these properties, gold nanoparticles were widely used in the fields of clinical medicine, materials and more. Moreover, in the field of medical tests, now gold nanoparticles labels had been the forth important markers. Gold nanoparticles show the potential applied value because of their special physical and chemical characteristics. At present, the analytical applications of gold nanoparticles are as optical probes or as electrochemical sensors after chemical modification. However, there are rarely reports that spectrophotometry or Resonance Rayleigh scattering method is used to the determination of pharmaceuticals with unmodified gold nanoparticle probes. Main content are listed as follow:
The preparation and characterization methods, spectroscopy characterization and analytical application of gold nanoparticles and the property, purpose and the present situation of its analysis of 6-mercaptopurine are revivwed here.
A novel spectrophotometry method has been developed for the determination of micro amounts of 6-mercaptopurine, employing gold nanoparticles as probes. The color of test solution changed to blue from bright red is significantly observed upon the gold nanoparticles self-assembled with 6-mercaptopurine and the absorption peak was obviously different. Before reaction, there is a plasmon band at 520 nm that is the characteristics of gold nanoparticles and 6-mercaptopurine has no absorption peak in rang of visible light region. After reaction, the peak of 520 nm is lower, in addition, there is a maximum absorption peak located at 680 nm, which is the characteristic peak of the binding product. The absorption at 680 nm of different concentrations of 6-mercaptopurine could be quantified with good linear, with a low detection limit of 17μg/L, under optimal conditions. The relative standard deviation (n=11) was 2.9%at 0.25 mg/L 6-mercaptopurine concentration level. The proposed method was successfully applied to determination of the drug spiked human urine. The results are satisfactory, which indicates that the method is not only sensitive, simply, but also reliable and suitable for practical application.
A novel Resonance Rayleigh scattering method was developed for the determination of micro amounts of 6-mecaptopurine, employing gold nanoparticles as probes. In pH 5.50 weak acid medium, the greatly enhanced Resonance Rayleigh scattering spectrum can be observed upon the gold nanoparticles self-assembled with 6-Mercaptopurine because both of them react with each other to form a binding product with bigger volume. In this work, The Resonance Rayleigh scattering spectral characteristics, optimum conditions for the reaction and the influencing factors were investigated. The relative Resonance Rayleigh scattering intensity (ΔIRRS) is directly proportional to the concentration of 6-Mercaptopurine in the range of 0.017-0.32 mg/L. The method has high sensitivity and the limit of detection for 6-Mercaptopurine is 8μg/L. The RSD (n=11) is 5.1% at 0.12 mg/L 6-mercaptopurine concentration level. The effect of coexisting substances on the determination of 6-mercaptopurine was studied and the results showed that this method had good selectivity. Therefore, the proposed method is successfully used to determine 6-mercaptopurine in human urine by Resonance Rayleigh scattering with gold nanoparticle probes.
引文
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