高效液相色谱与化学发光联用技术测定神经递质的研究
|
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-K_3Fe(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C_(18)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~(-9)g／mL-1.0×10~(-7)g／mL，相关系数r=0.9995，检出限为7.6×10~(-10)g／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l
X 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%o
4.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。
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 Technology
A 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 Detection
The 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 Separation
A 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 Detection
The 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
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-K_3Fe(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C_(18)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~(-9)g／mL-1.0×10~(-7)g／mL，相关系数r=0.9995，检出限为7.6×10~(-10)g／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l
X 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%o
4.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。
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 Technology
A 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 Detection
The 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 Separation
A 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 Detection
The 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
引文
1．汤涛等．安徽医学．，1990，11(5)：9-11
2．章裳伦，等．分析化学．，2000，28(3)：293-295．
3. Wan X C, Yang T Z, Xu C T. Modernneurobiology[M]. Beijing: Beijing Medi-cal University & Chinese Xie he Medical University United Press, 1999:158-162,459-469
4. Ding R G, Asada H, Obata K. J.Brain.Res.,1998;800:105-113
5. Raggi M,Sabbioni C,Casamenti G,et al. J.Chromatogr.B., 1999,730:201-211
6. Wang Y, Fice D S,Yeung P K. J.Pharm.Biomed.Anal., 1999,21:519-525
7. Chi J D,Odontiadis J,Frankkin M. J. Chromatogr.B., 1999,731:361-367
8．黄新华，陈本美，陈新．湖南医科大学学报．，2002，27(5)：471-473
9. Raggi M A, Sabbioni C, Casamenti G,et al. J.Chromatogr.B., 1999, 730:2:201-211
10. Annika Vaarmann, Ants Kask,Uno Meorg. J.Chromatogr.B., 2002, 769 (1): 145-153
11. Wang H,Li J,Liu X,et al. J.Anal Biochem., 2000,281:15-20
12. Alberts G,Lameris T, Meiracker A H,et al. J.Chromatogr.B., 1999,730:213-219
13. TorsterJ P, Jurgen B, Klaus L. J. Clin. Chem., 1999,45:262-268
14．吴予明，等．中国卫生检验杂志，2000，10(5)：526-528
15. Umegae Yoshihiko, Nohta Hitoshi,Lee, Myungkoo,et al. J. Chem. Pharm. Bull 1990, 38: 8
16．王铎，汪竹平，赵丽．华西口腔医学杂志．，1998，16(2)：132-133
17．赵永成．中草约．2001，32(6)：512-513
18. Raggi M A,,Sabbioni C,Gassmenti G,Gerra G.J.Chromatogr., 1997,730:201-211
19．彭贵华，唐银，曾立明，刘文恩．湖南医科大学学报．，2001，26(5)：485-487
20. Gooitzen Alberts, Thomas Lameris, Anton H. van den Meiracker, Arie J. Man in't Veld, Frans Boomsma. J.Chromatogr.B., 1999,730(2):213-219
21. Morier E,Ripa R.Chromatography in Biochemistry Medicine and Enviromental Researth.Elsevier Scientifec Publishing Company, Amsterdem, 1983:217-223
22. Ping Y L,Condtance S. J.Chromatogr.,1992,576-245
23. Opacka-Jufffry J,Tacconeel F, Coln C W. J Chromatogr., 1988,433:41-51
24．叶惟玲．色谱，19908(3)：159-162
25. Cheng F C,Kao J S,Shin Y, Lsi J S. J Chromatogr.,1997,730:201-211
26．徐桂芸，常溪燕，常理文．分析化学，2001，29(2)：199-201
27. Wen Zhang,Xuni Cao,Fangli,Wan,Song Zhang. J.Anal.Chim.Acta.,2002,472(1-2):27-35
28. Wen Zhang,Yunfeng Xie, Shiyun Ai, Fangli Wan,etc. J.Chromatogr.B.,2003,791(1-2):217-225
29. Cullison J K,et al.J.Pharm.Biomed.Anal.,1999,19(1-2):253-259
30. Dutton J,et al. J.Clin.Chem.,1999,Mar;45(3)394-399
31．朱爱芝，刘军，傅承光．分析测试学报．，1997，16(6)：47-49
32．辛梅华，李明春，徐金瑞，孙多先．化学通报．，2001，8：504-507
33. Juan Chen, Yan-Ping Shi, Jing-Yan Liu. J.Chromatogr.A., 2003, 1003(1-2)127-132
34．刘文第，齐伟，祁元明，郑惠良．兰州大学学报(自然科学版)，1999，35(2)111-114
35．杨士杰，张伟，岳海涛．白求恩医科大学学报．，1995，21(4)：434-436
36．张晓萍，王国民，邱宗荫等．重庆医科大学学报．，1997，22(3)194-197
37. Anne T Wood, Michael R Hall. J.Chromatogr.B: 2000, 744(1)221-225
38. se Marie Hansen, Jesper Kristiansen, Jeanet Lgsted Nielsen, et al. J. Talanta. 1999, 50:2:367-379
39. Sineon N, Myuers R, Bayle C,et al. J. Chromatogr.A.,2001,913(12):253 259
40. Maria A Fotopoulou,Penelope C Ioannou. J.Anal.Chim.Acta., 2002, 462:2:179-185
41．陈桂良，余琛，张慧．中国医药工业杂志，2001，32(7)：307-309
42．辛梅华，李明春，徐金瑞．分析化学．，2000，28(3)：387
43. Ci YX,Tie JK,Wang Q W et al.Anal Chim Acta, 1992;269:109
44. Ragab G H, Nohta H, Kai M, Ohkura Y. Anal.Chim.Acta.,1994,298:431
45. Gamal H. Ragab, Hitoshi Nohta, Kiyoshi Zaitsu. Anal.Chim.Acta.,2000, 4113(1-2):155-160
46. Chan E C, Ho P C. Raoid Commun Mass Spectrom., 2000,14(21):1959-1964
47. Acevedo L D, Xu Y, Zhang X, et al. J.Mass Spectrom., 1996,31(12):1399 1402
48．章裳伦，朱岩，郭丹等．分析化学．，2001，29(10)：1237
49. Magali Hay, Pierre Mormède. J.Chromatogr. B., 1997, 703(1-2)15-23
50. Guan CL, Ouyang J, Li Q L, et al. Talanta., 2000, 50:6:1197-1203
51．李金昶，石晶，邵长路，于惠芳．分析测试学报．，1994，13(1)：50-52
52．蒋银土，朱岩，王炯，郭莹莹．浙江大学学报(理学版)．，2001，28(4)：421-416
53．杨玉华，李崎．航天医学与医学工程．，1996，9(3)：217-221
54．庞志攻，汪宝琪．汪从莹．分析科学学报．，1994，10(1)：53-58
55. Milada Dolelowá, Magda Tkaczyko.,J.Pharm.Biomed.Anal. 1999, 19(3-4) 555-567
56. Martin Jeffrey T, Barchas Jack D, Faull Kym F. Anal.Chem., 1982, 54:11 1806-1811
57. Kuhlenbeck D L, et al. J.Chromatogr.B.,2000,738(2):319-330
58. guriS.J.Chromatogr.B.,2000,747:1
59. Katariina Vuorensola, Heli Sirén. J.Chromatogr.B., 2000, 895(1-2)317-327
60．于遐，罗振，宋承木，Robert AL．山东医科大学学报．，1994，32：347
61．林玲，等．福建医科大学学报，2000，34(4)：403-404
62. Dabek-Zlotorzynska E,Maruszak W.J.Chromatogr.B.,1998,714:77
63. Nouadji G, Nertz M, VerdeguerP H,Couderc F. J.Chromatogr.A.,1995,717:335
64. Oguri S, Watanabe S, Abe S. J.Chromatogr.A.,1997,790:177
65．张东明，付敏，马万云，陈瓞延．分析科学学报．，2002，18(4)：265-268
66. Chang, Huan-Tsung; Yeung, Edward S. J.Anal.Chem. ,1995, 67:6 1079-1083
67. Lu Xiaoning, Chen Yi. J.Chromatogr.A., 2002,955:133
68. Chen F T, Tusak A, Steinberg J. J.Chromatogr.A.,1993,652:355
69. Yarmoluk S M, Kostenko A M, Dubey I Y, Bioorganic Med. Chem.Lett.,2000,10:2201
70. Southwick P L.Cytometr.,1990,11:418
71. Gostkowski, Michael L.; Shear, Jason B., J.,Amer.Chem.Soc. 1998, 120:49 12966-12967
72. Wallingford R A, Ewing A G. Anal.Chem.,1987,59:1762
73. Lan Zhang, Guonan Chen, Qin Hu,et al. J. Anal.Chim.Acta.,2001, 431:2:287-292
74．杨冰仪，莫金垣，杨晓云，王立世．中山大学学报(自然科学版)．，1999，38(4)：36-39
75．金文睿，汪乃兴，赵昕．生物电分析化学[M]．济南：山东大学出版社，1994．78
76．胡深，庞代文，王宗礼，程介克．分析化学．，1998，26(6)：752-756
77. Kristenson H K,Lau Y Y, Ewing A G. J.Neurosci.Methods., 1994,51:183-187
78．王立世，杨晓云，莫金垣．色谱．，1999，17(5)：435-437
79．吴友谊，周明嘉．分析化学，2001，29(2)：138-141
80．朱英，陈义．分析化学．，2001，29(6)：661-663
81. Zlatuse D Peterson, Milton L Lee, Steven W. Graves, et al. J. Chromatogr.B., 2002, 776(2) 221-229
82. Zhang Y, Cheng J K, Huang B, al. Analytica Chimica Acta., 1998,363(2-3):157-163
83. Hui-chuan Tsai, Chen-wen Whang. Electrophresis., 1999,20(12):2533-2538
84. Bobbitt D R, Jackson W A, Hendrickson H P. Talanta., 1998,46(4):565-572
85 Weiping Yang, Zhujun Zhang, Wei Deng. Talanta., 2003,59(5):951-958
86. Takahiro Umeno, Yasuaki Okamito, Terufumi Fujiwara. 分析化学．，2003,52(12):1199-1202
87. ArleighHartkopf, etal.AnalLett, 1974,7(1):78-88
88．武竟存，章竹君，吕九如．分析化学，1994；22(4)：396
89．章竹君，武竟存．陕西师范大学学报，1989(分析化学专辑)：1
90．武竟存，耿征，吕九如．化学试剂，1994；16(1)：35
91. Worsfold P J, In: Stanley P E, Kricka L Jeds. Biolu-minescence and chemiluminescence: currentstatus,Chichester:John Wiley, UK; 1991:371
92. de Jong G J, Kwakman P J M. J.Chromatogr.,1989;492:319
93. Nieman T A, In:Birks J Wed. Chemiluminescence and photochemical lreaction detection in chromatogra-phy,NewYork:VCH,1989:99
94. Nieman T A.J.Res.Natl.Bur.Stand.,1988;93(3):501.
95. Imai K.Chromatogr Sci.,1990;8(DetectOrientedDe-rivTecbLiqChromatgr):359
96．张国清．中国药科大学学报．，1987；18(4)：309
97. McGownLB,WarnerIM.AnalChem., 1990;62(12):255R
98．章竹君，吕九如，张新荣．化学试剂．，1987；9(3)：149
99．武竟存，章竹君，吕九如．化学试剂．，1992；14(2)：95
100. Ishida J,Horike N,Yamaguchi M.Anal.Chim.Acta., 1995;302:61
101. Rauhut M M, Bollyky L J, Roberts B G, etal. J.Amer.Chem.Soc.,1967;89:6515
102. Kewis S W, Worsfold P J.Anal.Proc., 1992;29:10
103. Kwakman P J M, BrinkmanUA T h. Anal.Chim.Ac-ta.,1992;266:175
104. Imai K.Trendsin. Anal.Chem., 1985;4(7):170
105. Givens R S, Schowen R L. In:Birks J Wed. Chemi-luminescence and photochemical reaction detection in chromatography.,NewYork:VCH,1989:125
106. Imai K, Nishitani A, Akitomo H, etal. J.Biolumin.Chemilumin.,1989;4:500
107. Lee W Y, Nieman T A. J.Chromatogr.A.,1994;659:111
108. Imai K.Chromatogr Sci, 1990;48(DetectOrientedDerivTechLiqChromatogr):359
109. Imai K,Tsu kamotoY.Rinsho.Byori,,1988;6(4):402
110. Yamazaki S, Hara K, et al. J.High Resolut Chromatogr.,2000,23:127
111. Knight A W, Greenway G M. Analyst。,1994;119:879
112．于建国，叶水英．分析测试通报．，1988；7(1)：67
113. Hill E, Humphreys E, Malcolme-Lawes D J. J.Chro-matogr.,1986;370:427
114. Hill E, Humphreys E, Malcolme-Lawes D J. J.Chro-matogr.,1988;441:394
115. Hill E, Humphreys E.Laboratory Microcomput.,1988;7(4):132
116. Wang Y, Yeung E S. Anal.Chim.Acta., 1992;266:295
117．崔华，贺彩霞，赵贵文．高等学校化学学报，2000，21(6)：876-878
118．周光明，杨光明，齐东梅等．色谱，2002，20(6)：550-553
119. Hua Cui, Caixia He and Guiwen Zhao. J.Chromatogr.A.,1999,855(1):171-179
120. Ayako Ogawa, Hisako Arai, Hisayuki Tanizawa,et al. Anal.Chim.Acta., 1999, 383(3):221-230
121. Wei Zhang, Neil D.Danielson. Anal.Chim.Acta.,2003,493:2:167-177
122. Neary M P, Seitz W R. Anal, Lett.,1974,7(8&9):583.
123. Jones P, William T.EbdonL.Anal.Chim.Acta.,1989,217(1):157
124. Burguera J L, Burguera M, Townshend A. Anal.Chim.Acta, 1981,127(1): 199
125. Willams T, Jones P,Ebdon L. J.Chromatogr., 1989,482(2):361
126. Poulscn J R, Birks J W.J.Chromatogr., 1986,360(3):371
127. Kawasa k T, Maed M, Tsuji A.J.Chromatogr.,1985,328(1):121
128. Jansen I I, Frei R W. Jchromatogr., 1998,440(1):217
129. Hara T, Toriyama M, Euehi T. Bull.Chem.Soc.Jpn.,1985,58:109
130. Maltesev V G. J.Chromatogr.,1987,416(1):45
131. Kawasaki H,Maeda N, YukiH.J.Chro-matogr.,1990,516(2):450.
132. Kather H. Anal.Biochem., 1987,163(1):45
133. Frei B.Anal.Biochem.,1988,175(1):120
134. Candelman M S, Birks J W.J.Chromatogr.,1982,242(1):21.
135．朱龙，封满良，万秀琴，吕九如．高等学校化学学报．，1996，17(1)：1693-1696
136．王亦群，杨小峰，封满良，章竹君．西北药学杂志．，2001，16(1)：6-7
137．傅承光，徐宏达，王志．分析测试学报．，1994，13(1)：28-32
138．吴迎春，刘谦光，王亦群等．分析实验室．，2000，19(6)：30-32
139．段复利，中岛宪一郎，秋山修三，黑田直敬．中国药学杂志．，1996，31(1)：673-676
140．杨维平，张琰图，章竹君．化学学报．，2003，61(2)：303-306
141．朱龙，封满良，万秀琴．吕九如．分析化学．，1996，24(1)：1295-1297
142. Yen Sun, Mitsuhiro Wada, Osama Al-Dirbashi, et al. J.Chromatogr,B., 2000, 749(1):49-56
143. Manuel Cobo, Manuel Silva. J.Chromatogr.A., 1999, 848(1-2): 105-115
144. Michael E Bolden, Neil D Danielson. J.Chromatogr.A., 1998, 828(1-2):421-430
145. Matthew R Oates, William Clarke, Alden Zimlich,et al. Anal.Chim.Acta., 2002, 470(1): 37650
146. Kazuhiko Tsukagoshi, Kazuhide Miyamoto, Riichiro Nakajima, et al. J.Chromatogr.A., 201,919(2):331-337
147. Masaaki Kai, Hiromi Kinoshita, Kazuko Ohta, et al. J.Pharm.Biomed.Anal., 2003, 30(6): 1765-1771
148. Hideyuki Yoshida, Keiko Hidaka, Junichi Ishida. Anal.Chim.Acta., 2000, 413(1-2): 137-145
149. Eva Orejuela, Manuel Silva. J. Chromatogr.A., 2003, 1007(1-2): 197-201
150. Mitsuhiro Wada, Keiyu Inoue, Ayuko Ihara, et al. J. Chromatogr.A., 2003, 987(1-2):189-195
2．章裳伦，等．分析化学．，2000，28(3)：293-295．
3. Wan X C, Yang T Z, Xu C T. Modernneurobiology[M]. Beijing: Beijing Medi-cal University & Chinese Xie he Medical University United Press, 1999:158-162,459-469
4. Ding R G, Asada H, Obata K. J.Brain.Res.,1998;800:105-113
5. Raggi M,Sabbioni C,Casamenti G,et al. J.Chromatogr.B., 1999,730:201-211
6. Wang Y, Fice D S,Yeung P K. J.Pharm.Biomed.Anal., 1999,21:519-525
7. Chi J D,Odontiadis J,Frankkin M. J. Chromatogr.B., 1999,731:361-367
8．黄新华，陈本美，陈新．湖南医科大学学报．，2002，27(5)：471-473
9. Raggi M A, Sabbioni C, Casamenti G,et al. J.Chromatogr.B., 1999, 730:2:201-211
10. Annika Vaarmann, Ants Kask,Uno Meorg. J.Chromatogr.B., 2002, 769 (1): 145-153
11. Wang H,Li J,Liu X,et al. J.Anal Biochem., 2000,281:15-20
12. Alberts G,Lameris T, Meiracker A H,et al. J.Chromatogr.B., 1999,730:213-219
13. TorsterJ P, Jurgen B, Klaus L. J. Clin. Chem., 1999,45:262-268
14．吴予明，等．中国卫生检验杂志，2000，10(5)：526-528
15. Umegae Yoshihiko, Nohta Hitoshi,Lee, Myungkoo,et al. J. Chem. Pharm. Bull 1990, 38: 8
16．王铎，汪竹平，赵丽．华西口腔医学杂志．，1998，16(2)：132-133
17．赵永成．中草约．2001，32(6)：512-513
18. Raggi M A,,Sabbioni C,Gassmenti G,Gerra G.J.Chromatogr., 1997,730:201-211
19．彭贵华，唐银，曾立明，刘文恩．湖南医科大学学报．，2001，26(5)：485-487
20. Gooitzen Alberts, Thomas Lameris, Anton H. van den Meiracker, Arie J. Man in't Veld, Frans Boomsma. J.Chromatogr.B., 1999,730(2):213-219
21. Morier E,Ripa R.Chromatography in Biochemistry Medicine and Enviromental Researth.Elsevier Scientifec Publishing Company, Amsterdem, 1983:217-223
22. Ping Y L,Condtance S. J.Chromatogr.,1992,576-245
23. Opacka-Jufffry J,Tacconeel F, Coln C W. J Chromatogr., 1988,433:41-51
24．叶惟玲．色谱，19908(3)：159-162
25. Cheng F C,Kao J S,Shin Y, Lsi J S. J Chromatogr.,1997,730:201-211
26．徐桂芸，常溪燕，常理文．分析化学，2001，29(2)：199-201
27. Wen Zhang,Xuni Cao,Fangli,Wan,Song Zhang. J.Anal.Chim.Acta.,2002,472(1-2):27-35
28. Wen Zhang,Yunfeng Xie, Shiyun Ai, Fangli Wan,etc. J.Chromatogr.B.,2003,791(1-2):217-225
29. Cullison J K,et al.J.Pharm.Biomed.Anal.,1999,19(1-2):253-259
30. Dutton J,et al. J.Clin.Chem.,1999,Mar;45(3)394-399
31．朱爱芝，刘军，傅承光．分析测试学报．，1997，16(6)：47-49
32．辛梅华，李明春，徐金瑞，孙多先．化学通报．，2001，8：504-507
33. Juan Chen, Yan-Ping Shi, Jing-Yan Liu. J.Chromatogr.A., 2003, 1003(1-2)127-132
34．刘文第，齐伟，祁元明，郑惠良．兰州大学学报(自然科学版)，1999，35(2)111-114
35．杨士杰，张伟，岳海涛．白求恩医科大学学报．，1995，21(4)：434-436
36．张晓萍，王国民，邱宗荫等．重庆医科大学学报．，1997，22(3)194-197
37. Anne T Wood, Michael R Hall. J.Chromatogr.B: 2000, 744(1)221-225
38. se Marie Hansen, Jesper Kristiansen, Jeanet Lgsted Nielsen, et al. J. Talanta. 1999, 50:2:367-379
39. Sineon N, Myuers R, Bayle C,et al. J. Chromatogr.A.,2001,913(12):253 259
40. Maria A Fotopoulou,Penelope C Ioannou. J.Anal.Chim.Acta., 2002, 462:2:179-185
41．陈桂良，余琛，张慧．中国医药工业杂志，2001，32(7)：307-309
42．辛梅华，李明春，徐金瑞．分析化学．，2000，28(3)：387
43. Ci YX,Tie JK,Wang Q W et al.Anal Chim Acta, 1992;269:109
44. Ragab G H, Nohta H, Kai M, Ohkura Y. Anal.Chim.Acta.,1994,298:431
45. Gamal H. Ragab, Hitoshi Nohta, Kiyoshi Zaitsu. Anal.Chim.Acta.,2000, 4113(1-2):155-160
46. Chan E C, Ho P C. Raoid Commun Mass Spectrom., 2000,14(21):1959-1964
47. Acevedo L D, Xu Y, Zhang X, et al. J.Mass Spectrom., 1996,31(12):1399 1402
48．章裳伦，朱岩，郭丹等．分析化学．，2001，29(10)：1237
49. Magali Hay, Pierre Mormède. J.Chromatogr. B., 1997, 703(1-2)15-23
50. Guan CL, Ouyang J, Li Q L, et al. Talanta., 2000, 50:6:1197-1203
51．李金昶，石晶，邵长路，于惠芳．分析测试学报．，1994，13(1)：50-52
52．蒋银土，朱岩，王炯，郭莹莹．浙江大学学报(理学版)．，2001，28(4)：421-416
53．杨玉华，李崎．航天医学与医学工程．，1996，9(3)：217-221
54．庞志攻，汪宝琪．汪从莹．分析科学学报．，1994，10(1)：53-58
55. Milada Dolelowá, Magda Tkaczyko.,J.Pharm.Biomed.Anal. 1999, 19(3-4) 555-567
56. Martin Jeffrey T, Barchas Jack D, Faull Kym F. Anal.Chem., 1982, 54:11 1806-1811
57. Kuhlenbeck D L, et al. J.Chromatogr.B.,2000,738(2):319-330
58. guriS.J.Chromatogr.B.,2000,747:1
59. Katariina Vuorensola, Heli Sirén. J.Chromatogr.B., 2000, 895(1-2)317-327
60．于遐，罗振，宋承木，Robert AL．山东医科大学学报．，1994，32：347
61．林玲，等．福建医科大学学报，2000，34(4)：403-404
62. Dabek-Zlotorzynska E,Maruszak W.J.Chromatogr.B.,1998,714:77
63. Nouadji G, Nertz M, VerdeguerP H,Couderc F. J.Chromatogr.A.,1995,717:335
64. Oguri S, Watanabe S, Abe S. J.Chromatogr.A.,1997,790:177
65．张东明，付敏，马万云，陈瓞延．分析科学学报．，2002，18(4)：265-268
66. Chang, Huan-Tsung; Yeung, Edward S. J.Anal.Chem. ,1995, 67:6 1079-1083
67. Lu Xiaoning, Chen Yi. J.Chromatogr.A., 2002,955:133
68. Chen F T, Tusak A, Steinberg J. J.Chromatogr.A.,1993,652:355
69. Yarmoluk S M, Kostenko A M, Dubey I Y, Bioorganic Med. Chem.Lett.,2000,10:2201
70. Southwick P L.Cytometr.,1990,11:418
71. Gostkowski, Michael L.; Shear, Jason B., J.,Amer.Chem.Soc. 1998, 120:49 12966-12967
72. Wallingford R A, Ewing A G. Anal.Chem.,1987,59:1762
73. Lan Zhang, Guonan Chen, Qin Hu,et al. J. Anal.Chim.Acta.,2001, 431:2:287-292
74．杨冰仪，莫金垣，杨晓云，王立世．中山大学学报(自然科学版)．，1999，38(4)：36-39
75．金文睿，汪乃兴，赵昕．生物电分析化学[M]．济南：山东大学出版社，1994．78
76．胡深，庞代文，王宗礼，程介克．分析化学．，1998，26(6)：752-756
77. Kristenson H K,Lau Y Y, Ewing A G. J.Neurosci.Methods., 1994,51:183-187
78．王立世，杨晓云，莫金垣．色谱．，1999，17(5)：435-437
79．吴友谊，周明嘉．分析化学，2001，29(2)：138-141
80．朱英，陈义．分析化学．，2001，29(6)：661-663
81. Zlatuse D Peterson, Milton L Lee, Steven W. Graves, et al. J. Chromatogr.B., 2002, 776(2) 221-229
82. Zhang Y, Cheng J K, Huang B, al. Analytica Chimica Acta., 1998,363(2-3):157-163
83. Hui-chuan Tsai, Chen-wen Whang. Electrophresis., 1999,20(12):2533-2538
84. Bobbitt D R, Jackson W A, Hendrickson H P. Talanta., 1998,46(4):565-572
85 Weiping Yang, Zhujun Zhang, Wei Deng. Talanta., 2003,59(5):951-958
86. Takahiro Umeno, Yasuaki Okamito, Terufumi Fujiwara. 分析化学．，2003,52(12):1199-1202
87. ArleighHartkopf, etal.AnalLett, 1974,7(1):78-88
88．武竟存，章竹君，吕九如．分析化学，1994；22(4)：396
89．章竹君，武竟存．陕西师范大学学报，1989(分析化学专辑)：1
90．武竟存，耿征，吕九如．化学试剂，1994；16(1)：35
91. Worsfold P J, In: Stanley P E, Kricka L Jeds. Biolu-minescence and chemiluminescence: currentstatus,Chichester:John Wiley, UK; 1991:371
92. de Jong G J, Kwakman P J M. J.Chromatogr.,1989;492:319
93. Nieman T A, In:Birks J Wed. Chemiluminescence and photochemical lreaction detection in chromatogra-phy,NewYork:VCH,1989:99
94. Nieman T A.J.Res.Natl.Bur.Stand.,1988;93(3):501.
95. Imai K.Chromatogr Sci.,1990;8(DetectOrientedDe-rivTecbLiqChromatgr):359
96．张国清．中国药科大学学报．，1987；18(4)：309
97. McGownLB,WarnerIM.AnalChem., 1990;62(12):255R
98．章竹君，吕九如，张新荣．化学试剂．，1987；9(3)：149
99．武竟存，章竹君，吕九如．化学试剂．，1992；14(2)：95
100. Ishida J,Horike N,Yamaguchi M.Anal.Chim.Acta., 1995;302:61
101. Rauhut M M, Bollyky L J, Roberts B G, etal. J.Amer.Chem.Soc.,1967;89:6515
102. Kewis S W, Worsfold P J.Anal.Proc., 1992;29:10
103. Kwakman P J M, BrinkmanUA T h. Anal.Chim.Ac-ta.,1992;266:175
104. Imai K.Trendsin. Anal.Chem., 1985;4(7):170
105. Givens R S, Schowen R L. In:Birks J Wed. Chemi-luminescence and photochemical reaction detection in chromatography.,NewYork:VCH,1989:125
106. Imai K, Nishitani A, Akitomo H, etal. J.Biolumin.Chemilumin.,1989;4:500
107. Lee W Y, Nieman T A. J.Chromatogr.A.,1994;659:111
108. Imai K.Chromatogr Sci, 1990;48(DetectOrientedDerivTechLiqChromatogr):359
109. Imai K,Tsu kamotoY.Rinsho.Byori,,1988;6(4):402
110. Yamazaki S, Hara K, et al. J.High Resolut Chromatogr.,2000,23:127
111. Knight A W, Greenway G M. Analyst。,1994;119:879
112．于建国，叶水英．分析测试通报．，1988；7(1)：67
113. Hill E, Humphreys E, Malcolme-Lawes D J. J.Chro-matogr.,1986;370:427
114. Hill E, Humphreys E, Malcolme-Lawes D J. J.Chro-matogr.,1988;441:394
115. Hill E, Humphreys E.Laboratory Microcomput.,1988;7(4):132
116. Wang Y, Yeung E S. Anal.Chim.Acta., 1992;266:295
117．崔华，贺彩霞，赵贵文．高等学校化学学报，2000，21(6)：876-878
118．周光明，杨光明，齐东梅等．色谱，2002，20(6)：550-553
119. Hua Cui, Caixia He and Guiwen Zhao. J.Chromatogr.A.,1999,855(1):171-179
120. Ayako Ogawa, Hisako Arai, Hisayuki Tanizawa,et al. Anal.Chim.Acta., 1999, 383(3):221-230
121. Wei Zhang, Neil D.Danielson. Anal.Chim.Acta.,2003,493:2:167-177
122. Neary M P, Seitz W R. Anal, Lett.,1974,7(8&9):583.
123. Jones P, William T.EbdonL.Anal.Chim.Acta.,1989,217(1):157
124. Burguera J L, Burguera M, Townshend A. Anal.Chim.Acta, 1981,127(1): 199
125. Willams T, Jones P,Ebdon L. J.Chromatogr., 1989,482(2):361
126. Poulscn J R, Birks J W.J.Chromatogr., 1986,360(3):371
127. Kawasa k T, Maed M, Tsuji A.J.Chromatogr.,1985,328(1):121
128. Jansen I I, Frei R W. Jchromatogr., 1998,440(1):217
129. Hara T, Toriyama M, Euehi T. Bull.Chem.Soc.Jpn.,1985,58:109
130. Maltesev V G. J.Chromatogr.,1987,416(1):45
131. Kawasaki H,Maeda N, YukiH.J.Chro-matogr.,1990,516(2):450.
132. Kather H. Anal.Biochem., 1987,163(1):45
133. Frei B.Anal.Biochem.,1988,175(1):120
134. Candelman M S, Birks J W.J.Chromatogr.,1982,242(1):21.
135．朱龙，封满良，万秀琴，吕九如．高等学校化学学报．，1996，17(1)：1693-1696
136．王亦群，杨小峰，封满良，章竹君．西北药学杂志．，2001，16(1)：6-7
137．傅承光，徐宏达，王志．分析测试学报．，1994，13(1)：28-32
138．吴迎春，刘谦光，王亦群等．分析实验室．，2000，19(6)：30-32
139．段复利，中岛宪一郎，秋山修三，黑田直敬．中国药学杂志．，1996，31(1)：673-676
140．杨维平，张琰图，章竹君．化学学报．，2003，61(2)：303-306
141．朱龙，封满良，万秀琴．吕九如．分析化学．，1996，24(1)：1295-1297
142. Yen Sun, Mitsuhiro Wada, Osama Al-Dirbashi, et al. J.Chromatogr,B., 2000, 749(1):49-56
143. Manuel Cobo, Manuel Silva. J.Chromatogr.A., 1999, 848(1-2): 105-115
144. Michael E Bolden, Neil D Danielson. J.Chromatogr.A., 1998, 828(1-2):421-430
145. Matthew R Oates, William Clarke, Alden Zimlich,et al. Anal.Chim.Acta., 2002, 470(1): 37650
146. Kazuhiko Tsukagoshi, Kazuhide Miyamoto, Riichiro Nakajima, et al. J.Chromatogr.A., 201,919(2):331-337
147. Masaaki Kai, Hiromi Kinoshita, Kazuko Ohta, et al. J.Pharm.Biomed.Anal., 2003, 30(6): 1765-1771
148. Hideyuki Yoshida, Keiko Hidaka, Junichi Ishida. Anal.Chim.Acta., 2000, 413(1-2): 137-145
149. Eva Orejuela, Manuel Silva. J. Chromatogr.A., 2003, 1007(1-2): 197-201
150. Mitsuhiro Wada, Keiyu Inoue, Ayuko Ihara, et al. J. Chromatogr.A., 2003, 987(1-2):189-195