【作者】 张迎雪；

【导师】 章竹君；

【作者基本信息】 西南师范大学 ， 分析化学， 2004， 硕士

【摘要】 化学发光（Chemiluminescence，简称CL）是化学反应的反应物或生成物吸收了反应释放的化学能，电子由基态跃迁至激发态，再由激发态返回基态时所产生的光辐射。根据化学反应在某一时刻的发光强度来确定物质组成含量的分析方法叫化学发光分析法。化学发光分析法具有灵敏度高、仪器设备简单、线性范围宽、分析速度快以及不需外加光源和单色器，没有散射光等干扰，容易实现自动化等优点，但选择性差。液相色谱，特别是高效液相色谱法（HPLC），以其高效、高速和高分辨能力等特征给人们留下了深刻的印象。高灵敏度的化学发光检测手段与高分辨力的高效液相色谱法（HPLC）结合于一体，成为一种理想的分离分析方法，两种方法能取长补短，充分发挥各自优势，非常适合复杂生物样品的分析。可以说，HPLC-CL联用技术为现代痕量分析带来勃勃生机，并且具有广阔应有前景 神经递质（又名神经介质）在神经信息的传导中发挥着重要作用，它们广泛参与了机体内一些重要的生理机能活动。 本文包括两个部分。第一部分是综述，主要介绍神经递质的分类和常用的分析方法，重点评述高效液相色谱法、化学发光法及二者联用技术的应用。 第二部分是研究报告。主要报告了基于儿茶酚胺类神经递质对鲁米诺-铁氰化钾、鲁米诺-过氧化氢化学发光体系的增敏作用，利用高效液相色谱与化学发光联用技术检测生物样品中的儿茶酚胺类神经递质，希望为在线活体分析奠定基础。 1．HPLC与微超滤技术联用化学发光检测人体血清中的盐酸多巴酚丁胺 研究发现，盐酸多巴酚丁胺在碱性条件下能大大增强Luminol-K₃Fe（CN）₆化学发光强度，基于此，并结合HPLC分离技术，建立起盐酸多巴酚丁胺的HPLC-CL分析新方法。并且在C₁₈反相键合相色谱柱，乙腈为流动相的条件下，实现了对人体血清中盐酸多巴酚丁胺的分离与测定，方法的线性范围为1.0×10^-9g／mL-1.0×10^-7g／mL，相关系数r=0.9995，检出限为7.6×10^-10g／mL，相对标准偏差为2.9％（n=11）。，1，文摘要2.高效液相色谱与化学发光联用测定人体血清及尿样中的盐酸多巴胺 铁氰化钾在碱性条件下能够氧化鲁米诺产生化学发光，盐酸多巴胺的存在可以大大增强其发光强度。基于此，结合高效液相色谱分离技术，在色谱柱为C!。反向柱，流动相为邻苯二甲酸氢钾（0 .01M）:甲醇等于92:8（体积比），pH值等于4.5的条件下，实现了对人体血清及尿样中的盐酸多巴胺的分离与测定，方法的线性范围为5.0火10-0一1 .0 xlo一gimL，检出限为8 xlJ’“留mL。对6.0 X10’8gimL的盐酸多巴胺连续测定11次的相对标准偏差为3.2%。.3.高效液相色谱与化学发光检测人体血清及尿样中的左旋多巴 基于左旋多巴对Luminol·K3Fe（CN）6化学发光体系的增敏作用，在C，。反相色谱柱，邻苯二甲酸氢钾（0 .OIM）甲醇（8%）（磷酸氢二钠调至PH=4 .5）为流动相的条件下，实现了对人体血清及尿样中的左旋多巴的分离与测定，方法的线性范围为1 X10月g/mL一lX 10币g/mL，相关系数r=o.9995，检出限为3x10一99/mL，对于浓度为5x10月g/mL的左旋多巴平行测定n次，其相对标准偏差为2.8%o4.高效液相色谱化学发光法测定人体血清中的重酒石酸去甲肾上腺素 本文研究发现，铁氰化钾与碱性鲁米诺溶液混合产生化学发光，重酒石酸去甲肾上腺素的存在可以大大增强其化学发光强度，结合高效液相色谱，在C::反相柱上，在统动相为邻苯二甲酸氢钾O.Olmo功、甲醇（v:二92%:8%），pH值等于9.6的条件下，实现了对人体血清中重酒石酸去甲肾上腺素的分离与测定，方法的线性范围为5x10母咖l一10一6留mL，检出限为1.0xl0一9留mL。对sxlo一8留mL的重酒石酸去甲肾上腺素连续进行11次平行测定的相对标准偏差为2.6%。5.高效液相色谱化学发光检测人体血清及尿样中的盐酸肾上腺素 本文研究发现，在介质NaC仇一Na0II中，铁氰化钾与鲁米诺溶液混合产生化学发光，盐酸肾上腺素的存在可以大大增强其化学发光强度，结合高效液相色谱，在cl.反向柱上，在流动相为邻苯二甲酸氢钾0. olm。1/1、甲醇（v:v二92%:8%），PH值等于9.8的条件下，实现了对人体血清及尿样中盐酸肾上腺素的分离与测定，方法的线性范圈为10飞/m1一5 x lrg/mL，检出限为4.ox10书g/mL。对sxlo一g/mL的盐酸肾上腺素连续进行H次平行测定的相对标准偏差为3.0%。6.流动注射化学发光法测定吗丁琳 2中文摘要 在碱性条件下，过氧化氢氧化鲁米诺，产生化学发光，吗丁琳对该体系的化学发光具有显著的增敏作用。基于此，建立了一种直接测定吗丁琳的流动注射化学发光分析法。该方法的线性范围在1.ox 10一6一3.0x10一日g/mL，检测限（30）为5.0X10一g/mL，相对标准偏差为2.9%（下11），方法已用于吗丁琳片剂及血清中吗丁琳的分析。更多还原

【Abstract】 Chemiluminescence(CL) is defined as the emission of light from an electronically excited state species which is produced during the course of a chemical reaction, and is observed when the electronically excited product or intermediate formed decays to the ground state with a photon emitted. There are many merits on CL such as high sensitivity, wide linear range, rapidity, simplicity and so on. Due to the power ability of seperation, high performance liquid chromatography coupled with chemiluminescene has been widely applied in analysis on complicated, biologic samples and has become the most powerful technique in analytical chemistry.The thesis consists two parts. The first part is a review concering the definiton, the classification and the analytical methods of neurotransmitters. Emphasis was placed on the development of technique of high performance liquid chromatography coupled with chemiluminescene detection in the recent ten years.The other is research reports which is composed of 6 parts as follows. There are two chemiluminescence systems was described concluding Luminol-K.3Fe(CN)6 and luminol-H2O2 systems. Neurotransmitters were separated and determined by HPLC-CL, and satisfied results were obtained.1.Chemiluminescence Determination of Dobutamine Hydrochloride in Human Serum by HPLC Separation coupled with Ultrafiltration TechnologyA novel chemiluminescence determination of dobutamine hydrochloride in human serum by HPLC separation connected with ultrafiltration technology is described in this paper. The signal produced by the chemiluminescent reaction(CL) between luminol and ferricyanide was greatly increased in the presence of dobutamine which was eluted by HPLC. The linear range of dobutamine concentration was 1.0 X 10-9gmL-1 - 1.0 X 10-7gmL-1 (r=0.9995)and the detection limit was 7.6X 10-10gmL-1.2.Determination of Dopamine Hydrochloride in Human Serum and Urine Samples by High Performance Liquid Chromatography with Chemiluminescence DetectionThe separation and determination of dopamine hydrochloride in human serum and urine samples by high performance liquid chromatography with Luminol-K3Fe(CN)6 chemiluminescence detection were investigated first time in this paper.The separation was carried out on C18 colum using the moble phase of 0.01mol/L potassium hydrogen phthalste solution and methyl alcohol(V1:V2=92%:8%).Under the optimum conditions of 2 X 10-4mol/L Luminol(NaHCO3/NaOH solution ,pH=9.8), 2X10-5mol/L K3Fe(CN)6,the linear range and detection limit for dopamine hydrochloride were 5~1000ng/mL and 8.0 X 10-10g/mL, respectively. The relative standard derivation for 100ng/mL dopamine hydrochloride was 3.2%(n=11).3.Chemiluminescence Determination of Levodopa in Human Serum and Urine SaBipies by HPLC SeparationA flow injection chemiluminescence method for the determination of levodopa in human serum by HPLC separation connected with ultrafitration was described in this paper.The method is based on the chemiluminescence produced during the oxidation of luminol by potassium ferricyanide in an alkaline solution.The emission intensity is greatly enhanced in the presense of levodopa which was eluted by potassium hydrogen phthalate solution.Under the optimal conditions ,the CL intensity was linear to the concentration in the range of 1 .0X 10-8g/mL-1.0 X 10-6g/mL(r=0.9995),With a detection limit of 3.0 X 10-9g/mL.Relative standard deviation was 2.8%.The proposed method was simple,quick and sensitive. 4. Determination of Noradrenaline Bitartrate in Human Serum by HPLC Separatien with Chemiluminescence DetectionThe separation and determination of noradrenaline nitartrate in human serum by high performance liquid chromatography with Luminol-K.3Fe(CN)6 chemiluminescence detection were investigated first time in this paper.The separation was carried out on C18 colum using the moble phase of 0.01mol/L potassium hydrogen phthalate solution and methyl alcohol(V1:V2=92%:8%).Under the optimum conditions of 2 X 10-4mol/L Luminol (NaHCO3/NaOH solution ,pH=9.8), 3X 10-5mol/L K3Fe(CN)6,the更多还原