阳离子铝酞菁红区荧光探针在纳克级RNA定量分析中的应用

发布时间：2017-12-27 08:24

本文关键词：阳离子铝酞菁红区荧光探针在纳克级RNA定量分析中的应用　出处：《光谱学与光谱分析》2016年03期 　论文类型：期刊论文

【摘要】：荷正电的四取代三甲基碘化铵铝酞菁(Tetra(trimethylammionio)aluminum phthalocyanine,TTMAAlPc)是新近出现的、很有应用潜力的新型红区荧光化合物。研究表明,弱碱性条件,RNA对TTMAAlPc的荧光具有高效猝灭作用,且猝灭程度与RNA含量呈线性正相关。据此发现建立了可测定纳克级RNA的高灵敏定量分析新方法,并对原理进行了讨论。考察了pH、缓冲体系、反应时间、温度、TTMAAlPc浓度的影响,确定了最佳反应条件(pH 8.0的广泛缓冲液、反应5 min、室温、浓度为2×10-6 mol·L-1的TTMAAlPc)。在最佳条件下,方法的线性回归方程为y=-15.0+0.51x,相关系数r=0.998 6,线性范围7.71~1 705.57ng·mL-1,检测限为1.55ng·mL-1。本法灵敏度高、线性范围宽、对RNA测定常见干扰物质包括阴、阳离子、表面活性剂和维生素等的抗干扰能力强,且操作简便。该法应用于实际样品RNA含量的测定,取得了满意的结果。还以参比法首次测定了不同pH下TTMAAlPc的荧光量子产率,结果显示TTMAAlPc具有较高的荧光量子产率,且对大范围的酸度稳定,表明TTMAAlPc是极具应用潜力的新型红色荧光探针,值得深入研究,开拓其应用。
[Abstract]:Positively charged four substituted three methyl ammonium iodide aluminum phthalocyanine (Tetra (trimethylammionio) aluminum phthalocyanine, TTMAAlPc) is a new and promising new red fluorescent compound. The results show that RNA has a high efficiency quenching effect on the fluorescence of TTMAAlPc, and the degree of quenching is linearly and positively correlated with the content of RNA. A new method of high sensitivity and quantitative analysis for the determination of NAC grade RNA is found, and the principle is discussed. The effects of pH, buffer system, reaction time, temperature and TTMAAlPc concentration were investigated. The best reaction conditions (pH 8 wide buffer, 5 min, room temperature and 2 L-1 10-6 mol L-1 TTMAAlPc) were determined. Under the optimal conditions, the linear regression equation of the method is y=-15.0+0.51x, the correlation coefficient r=0.998 6, the linear range 7.71~1 705.57ng. ML-1, and the detection limit of 1.55ng mL-1. This method has high sensitivity and wide linear range. It has strong anti-interference ability to RNA for the determination of common interfering substances, including anion, cationic, surfactant and vitamin, and it is easy to operate. The method is applied to the determination of the RNA content in the actual sample, and the satisfactory results have been obtained. The reference method for the first time the determination of fluorescence quantum yield of pH under different TTMAAlPc, the result shows that the fluorescence quantum yield of TTMAAlPc is higher, and the acidity of stability in a wide range, indicating that TTMAAlPc is a novel red fluorescent probe has great application potential, expand the application is worthy of further research.
【作者单位】：厦门大学医学院抗癌研究中心;
【基金】：国家自然科学基金重大研究计划项目(90206016) 福建省科技重点项目(2012Y0081) 省属公益类科研院所基本科研专项(2014R1102,2015R1101031-2) 厦门市科技计划项目(3502Z20143015)资助
【分类号】：O657.3;O629.74
【正文快照】： 引言随着生命科学、环境科学的快速发展,传统的荧光探针越来越显示出其局限性。因为生物、医学和环境样品通常组成复杂,在进行荧光分析时常受到背景荧光和散射光的显著干扰,客观上要求发展新一代荧光探针以解决这一问题。因此,长波荧光检测技术应运而生[1-2]。金属酞菁化合物

【相似文献】