核磁共振电信号内标法在人体尿液定量分析中的应用  

Human Urinary Metabolite Quantification in Nuclear Magnetic Resonance-based Metabonomics Using Electronic Internal Reference

作　　者：杨亮[1,2] 茹阁英[1,2] 唐惠儒[1] 刘朝阳[1] 

机构地区：[1]波谱与原子分子物理国家重点实验室,武汉磁共振中心,中国科学院武汉物理与数学研究所,武汉430071 [2]中国科学院研究生院,北京100039

出　　处：《分析化学》2010年第6期789-794,共6页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(Nos.10675162;20825520);国家科技支撑计划(No.2006BAK03A22);国家重点基础研究发展计划(No.2007CB914701)资助

摘　　要：研究了Varian谱仪核磁共振电信号内标法在人体尿液代谢物浓度测定上的应用,通过实验证明了该方法进行定量分析的可靠性。NMR电信号内标法原理上是通过Varian谱仪去耦通道在常规一维谱图上产生一个参考信号,并利用谱仪软件程序来调整该信号在频谱上的强度、频率、衰减速率等参数。避免了代谢组学中NMR定量实验需要添加已知浓度物质(例如TSP)作为内标而引起的内标物与样品相互作用、谱峰重叠、内标物不溶及弛豫时间太长等问题。研究结果表明,Varian谱仪去耦通道产生的电信号稳定可靠(标准偏差0.36%),能够用于定量分析;当样品浓度大于20mmol/L或小于2mmol/L时,该方法测定的相对误差分别为1%和5%。通过配制低浓度的尿液模型样品,验证了电信号内标法测量人体尿液代谢物的浓度的可行性,最后使用该方法测量真实的人体尿液中常见代谢物的浓度,测定结果与医院常用生化分析仪器的测定结果相符。We report a complete assessment for urinary quantitative analysis by using an electronic signal as an internal reference, which was generated via decoupling channel on a Varian NMR spectrometer with its amplitude, frequency and transverse relaxation time digitally programmable according to a decoupling shape file. The electronic signal with excellent stability （standard deviation 0.4%） avoided possible problems such as insolubility, interaction and spectral overlap for a traditional internal reference. Concentration determination using the electronic signal proved to be reliable by a series of quantitative experiments. The relative error of quantification was about 1% and 5% when the analyte concentration was above 20 mmol/L and below 2 mmol/L respectively. We further measured the concentration of human urinary metabolites in a simultaneous fashion using this electronic internal reference and obtained consistent results with the values measured by the biochemical analysis instrument used in hospitals.

关 键 词：核磁共振 电信号内标 人体尿液 定量分析 

分 类 号：R446.1[医药卫生—诊断学] R445.2[医药卫生—临床医学]