毛细管电泳在药物分析及物化常数测定中应用
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摘要
毛细管电泳是近年来发展最快的一种分离分析新技术。它具有高效、快速、
微量等特点。在化学、药学和生命科学等领域均有广泛的应用。
本文共包括三章。第一章是综述，介绍了毛细管电泳基本原理及在药物分析
中的应用，同时也简要叙述了毛细管电泳的发展方向。
第二章研究了毛细管胶束电动色谱法同时分离和测定夏枯草中齐墩果酸和
熊果酸异构体含量的新方法。以15mmol/L磷酸氢二钠（Na_2HPO_4）、15mmol/L硼
砂（Na_2B_4O_7）、10mmol/L十二烷基硫酸钠（SDS）、5％（v/v）乙醇的缓冲溶液作
为运行电解质溶液，在20分钟实现了齐墩果酸和熊果酸的基线分离。同时也研
究了SDS浓度和有机溶剂（乙醇）浓度对待测物质分离的影响。测得了不同提取
方法夏枯草中两种物质的含量。其次，采用上面优化的分离条件，测得了石楠叶
及凌霄花中齐墩果酸和熊果酸的含量。
第三章是毛细管电泳法在物化常数测定中的应用。第一部分在简单介绍基本
公式的基础上，对苯酚、对苯二酚、间苯二酚、间苯三酚、对羟基苯甲醇和天麻
素的pKa_1和pKa_2值进行了测定。结果与文献值吻合较好。第二部分系统的推导
出毛细管电泳法测定电离常数的完整理论公式（部分是新导的），并用此理论采
用非线性拟合方式首次测得了厚朴酚、和厚朴酚、槲皮素、姜黄素、己稀雌酚、
对乙酰氨基酚、丁香酚和丹皮酚的电离常数。
Capillary electrophoresis(CE) has been widely used as a powerful
separation and analytical technique in recent years because of its
advantages including high performance, high speed, low analytes and
sample consumptions. It has been applied to a wide range of research fields
such as chemistry, pharmaceutical analysis and life science et al.
This paper consists of three chapters. In the first chapter, the
elementary theory of CE and its application on the pharmaceutical analysis
and developing direction were introduced.
In the second chapter, a micelle electrokinetic capillary
chromatography (MECC) method to separate and determine oleanolic acid and
ursolic acid isomers simultaneously in Prune]]a vulgarisL. was developed.
The two compounds were baseline separated under the optimum buffer
condition of 15 znrnol/L disodium hydrogen phosphate + 15 mmol/L disodium
teterborate + 10 mmol/L sodium dodecyl sulphate(SDS) + 5%(v/v) alcohol
and their contents in Prunella vulgaris L. with different extracted
methods were quantitatively analyzed within 20 nun. In addition, the
effects of SDS concentration and organic modifier (alcohol) concentration
on the separation of the analytes were also investigated. In the second
section of chapter 2, the contents of the isomers in folium photiniae and
[los ca.mpsis were determined under the same optimum buffer condition as
above.
In the third chapter, the formulas for CE determination of pKa and pKa2
were introduced. The pKa and pKa2 values of phenol, hydroqninone,
resorcinol, phloroglucinol, pydroxybenzyl alcohol and gastrodin were
determined and were consistent with those in literatures. The pK, values
were achieved by noninear regression analysis by taking into
consideration the effect of activity coefficient completely. This was the
first report on determining the dissociation constants of magnolol,
honokiol, quercetin, curcumin, diethylsti lbestrol, 4--acetanidophenol,
ellgenol and paeonol.
微量等特点。在化学、药学和生命科学等领域均有广泛的应用。
本文共包括三章。第一章是综述，介绍了毛细管电泳基本原理及在药物分析
中的应用，同时也简要叙述了毛细管电泳的发展方向。
第二章研究了毛细管胶束电动色谱法同时分离和测定夏枯草中齐墩果酸和
熊果酸异构体含量的新方法。以15mmol/L磷酸氢二钠（Na_2HPO_4）、15mmol/L硼
砂（Na_2B_4O_7）、10mmol/L十二烷基硫酸钠（SDS）、5％（v/v）乙醇的缓冲溶液作
为运行电解质溶液，在20分钟实现了齐墩果酸和熊果酸的基线分离。同时也研
究了SDS浓度和有机溶剂（乙醇）浓度对待测物质分离的影响。测得了不同提取
方法夏枯草中两种物质的含量。其次，采用上面优化的分离条件，测得了石楠叶
及凌霄花中齐墩果酸和熊果酸的含量。
第三章是毛细管电泳法在物化常数测定中的应用。第一部分在简单介绍基本
公式的基础上，对苯酚、对苯二酚、间苯二酚、间苯三酚、对羟基苯甲醇和天麻
素的pKa_1和pKa_2值进行了测定。结果与文献值吻合较好。第二部分系统的推导
出毛细管电泳法测定电离常数的完整理论公式（部分是新导的），并用此理论采
用非线性拟合方式首次测得了厚朴酚、和厚朴酚、槲皮素、姜黄素、己稀雌酚、
对乙酰氨基酚、丁香酚和丹皮酚的电离常数。
Capillary electrophoresis(CE) has been widely used as a powerful
separation and analytical technique in recent years because of its
advantages including high performance, high speed, low analytes and
sample consumptions. It has been applied to a wide range of research fields
such as chemistry, pharmaceutical analysis and life science et al.
This paper consists of three chapters. In the first chapter, the
elementary theory of CE and its application on the pharmaceutical analysis
and developing direction were introduced.
In the second chapter, a micelle electrokinetic capillary
chromatography (MECC) method to separate and determine oleanolic acid and
ursolic acid isomers simultaneously in Prune]]a vulgarisL. was developed.
The two compounds were baseline separated under the optimum buffer
condition of 15 znrnol/L disodium hydrogen phosphate + 15 mmol/L disodium
teterborate + 10 mmol/L sodium dodecyl sulphate(SDS) + 5%(v/v) alcohol
and their contents in Prunella vulgaris L. with different extracted
methods were quantitatively analyzed within 20 nun. In addition, the
effects of SDS concentration and organic modifier (alcohol) concentration
on the separation of the analytes were also investigated. In the second
section of chapter 2, the contents of the isomers in folium photiniae and
[los ca.mpsis were determined under the same optimum buffer condition as
above.
In the third chapter, the formulas for CE determination of pKa and pKa2
were introduced. The pKa and pKa2 values of phenol, hydroqninone,
resorcinol, phloroglucinol, pydroxybenzyl alcohol and gastrodin were
determined and were consistent with those in literatures. The pK, values
were achieved by noninear regression analysis by taking into
consideration the effect of activity coefficient completely. This was the
first report on determining the dissociation constants of magnolol,
honokiol, quercetin, curcumin, diethylsti lbestrol, 4--acetanidophenol,
ellgenol and paeonol.
引文
1． Hjerten S. Chromatogr Rev. 1967, 9: 122
2． Everaerts F M et al. Sci Tools. 1970, 17: 25
3． Mikkers F E P, Everaerts F M, Verheggen Th P E M. J Chromatogr. 1979, 169: 1&11
4． Jorgenson J W, Lukacs K D. Anal Chem. 1981, 52: 1298
5． Hjerten S. J Chromatogr. 1983, 270: 1
6． Hjerten S, Zhu M D. J Chromatogr. 1985, 346: 265
7． Terabe S et al. Anal Chem. 1984, 56: 111
8． Lauer H H, Mcmanigill D. Anal Chem. 1986, 58: 166
9． Cox H C, Hessels J K C, Teven J M. J Chromatogr. 1972, 66: 19
10． Ghowsi K Foley J P, Gale R J. Anal Chem. 1990, 62: 2714
11． Terabe S, Otsuka K, Ando T. Anal Chem. 1989, 61: 251
12． Chu I, Bodnar J A, White E L, et al. Quantification of vincristine and vinblastine in catharanthus roseus plants by CZE. J Chromatogr A, 1996, 755: 281
13． Sun S W, Lee S S, Chen L Y, et al. Separation of aporphine alkaloids by MEKC. J Chromatogr A, 1997, 767: 277
14． Sun S W, Wa A C. Optimizationof the capillary electrophoretic separation of bisbenzylisoquinoline alkaloids by an overlopping resolution mapping scheme. J Chromatogr A, 1998, 814: 223
15． Liebich H M, Lehmann R, Stefano C D, et al. Analysis of tradition Chinese anticancer drugs by CE. J Chromatogr A, 1998, 795: 388
16． Trenerry V C, Wells R J, Robertson J. Determination of morphine and related alkaloids in crude morphine, poppy straw and opium preparations by MEKC. J Chromatogr A, 1995, 718: 217
17．王实强，首弟武。高效毛细管电泳测定罂栗壳中生物碱的含量。色谱，1997，15(5)：438
18.Unger M, Stockigt J. Improved detection of alkaloids in crude extracts applying CE with field amplified sample injection. J Chromatogr A, 1997, 791:323
19.Liang H R, Siren H, Riekkola M L, et al. Optimized separation of pharmacologically active flavonoids from epimedium species by CE.J Chromatogr A, 1996, 746:123
20.Liang H R, et al.J Chromatogr Sci, 1997, 35(3): 117
21.Gil M I, Ferreres F, Tomas-Barberan F A. MEKC of methylated flavone aglycones.J Liq Chromatogr A, 1995, 18(15): 3007
22.Wu H K, Sheu S J. CE determinotion of the constituents of paeoniae radix. J Chromatogr A, 1996, 753:139
23.Moodley V E, Mulholland D A, Raynor M W. MEKC of limonoid glucosides from citrus seeds. J Chromatogr A,1995, 718:187
24.Stuppner H, Egger R. Application of CZE to the analysis betalains from beta vulgaris. J Chromatogr A, 1996, 735:409
25.Hiermann A, Badl B. Analysis of aromatic plant acids by CZE. J Chromatogr A, 1998, 803:311
26.Summanen J, et al. J Chromatogr Sci., 1995, 33(12): 704
27．张凤云，毛富春。用毛细管区带电泳技术测定杜仲叶中绿原酸含量的研究。分析测试通报，1996，15(3)：66
28．杨更亮，王德先，宋秀荣，等。高效毛细管电泳鉴别常规中草药。中国化学会论文集，2001：521
29.Chen C T, Sheu S J. Separation of coumarins by MEKC. J Chromatogr A, 1995, 710:323
30.Ochocka R J, Rajzer D, Kowalski P, et al. Determinationof coumanns from chrysanthemum segetum L. by CE.J Chromatogr A, 1995, 709:197
31.Zhang H Y, Hu Z D, Yang G L, et al. Separation and determination of magnolol and honokiol in crude and burn-fried magnolia officinalis.Anal Lett.,1997,30(13):2327
32.Oehrle S A.AnalysiS of ginkolides and biobalides by CE.J Liq Chromatogr,1995,18(14):2855
33．柴逸峰，等。中国药学杂志，1997，32(增刊)：80
34.Hu Z D,et al. J Liq Chromatogr & Relat Technol,1997,20(1):63
35．朱萱萱，陈福林，等。毛细管电泳法测定麻黄中生物碱。中国中药杂志，1996，21(10)：587
36．赵霞，陆阳，陈泽乃，等。毛细管电泳法定量分析贝母中几种生物碱。中草药，2001，32(2)：116
37．袁炜，吕建德，傅小芸，等。槟榔中槟榔碱和槟榔次碱的毛细管电泳法分析。分析化学，2000，28(6)：749
38．杨更亮，宋秀荣，张红医，等。胶束毛细管电泳法测定牡丹皮及芍药中的牡丹酚和芍药甙。分析化学，1999，27(1)：1
39．宋秀荣，杨更亮，李杰，等。毛细管电泳法测定桑叶中的有效成分。中成药，2000，22(2)：158
40．王德先，杨更亮，王琳颖，等。毛细管区带电泳测定芦荟中的有效成分。色谱，2000，18(5)：423
41.Wang Dexian,Yang Gengliang,H.Engelhardt,Liu HaiXing.Chromatographia,2001,53(3／4):185
42．定天明，田颂九，张正行，等。胶束电动毛细管色谱法测定水飞蓟素及其制剂中主要有效成分含量。药学学报，2000，35(10)：778
43．丁岗，陆蕴如，冀春茹，等。诃子及其易混淆品中二种丹宁成分的高效毛细管电泳分析。药学学报，2001，36(4)：292
44．陈刚，李利军，叶建农，等。黄芩及其制剂中黄芩素和黄芩甙的毛细管区带电泳—电化学检测。分析测试学报，2001，20(1)：26
45．王德先，赵敬湘，杨更亮，等。毛细管区带电泳法测定中药金银花中绿原酸的含量。中草药，2000，31(6)：432
46．范国荣，胡晋红，李博华。四倍体板蓝根中五种有机酸成分的毛细管电泳分离分析。中国药科大学学报，2000，31(2)：111
47．陈勇，程智勇，韩凤梅，等。高效毛细管区带电泳法测定生药当归中阿魏酸的含量。分析化学，1999，27(12)：1424
48.Seu S J, Chen H R. Determination of five major anthraquinoids in Chinese herbal preparations by MEKC. Anal Chim Acta, 1995, 309:361
49.Seu S J, Chen H R. Simultaneous detertion of twelve constituents of I-tzu-tang, a Chinese herbal preparation, by HPLC and CE. J Chromatogr A, 1995, 704:141
50.Seu S J, et al.J High Resol Chromatogr, 1995, 18(4): 269
51．张国华，王延仲，张永久，等。高效毛细管电泳法测定黄连及成药小檗碱型生物碱的含量。色谱，1995，13(4)：247
52.Huang H Y, Kuo K L, Heieh Y Z.Determination of cinnamaldehyde,cinnamic acid, paeoniflorin, glycyrrhizin and [6]-gingerol in the traditional Chinese medicinal preparation kuei-chih-lang by cyclodextrin-modified micellar electrokinetic chromatography.J Chromatogr A, 1997, 771:267
53.Wang C Y, Hang H Y, Kuo K L, et al. Analysis of puerariae radix and its medicinal preparations by CE.J Chromatogr A, 1998, 802:225
54.Ku Y R, et al.Chin Pharm J, 1998, 50(3): 157
55．陈勇，程智勇，韩凤梅，等。毛细管区带电泳法测定十全大补丸中的川芎嗪和芍药甙。分析化学，2000，282(2)：186
56．杨惠芳，丁天惠，等。毛细管胶束电动色谱法测定复方茶碱片的有效含量。分析仪器，1999，2：44
57．李前峰，张红医，王怀文，等。毛细管电泳法测定甲基苄胺嘧啶和磺胺甲基异恶唑。分析化学，2000，28(7)：793
58．刘勤，周永新，汤立合，等。毛细管区带电泳法检测神经性毒剂降解产物。分析化学，2000，28(7)：810
59．王杰，陈海峰，赵岚峰，等。胶束电动色谱分离5种二氢吡啶类钙拮抗剂。分析化学，1999，27(1)：82
60．林秀丽，李关宾，主沉浮，等。以L—白氨酸为手性选择剂用毛细管电泳拆分12种手性药物。色谱，2001，19(2)：109
61．杨冰仪，莫金桓，杨晓天等。用高效毛细管电泳安培法检测扑热息痛及其水解物对氨基酚。分析测试学报，2000，19(1)：13
62．贾丽，张君倩。氧氟沙星片剂与针剂中的有效成分。分析测试学报，2000，19(6)：78
2． Everaerts F M et al. Sci Tools. 1970, 17: 25
3． Mikkers F E P, Everaerts F M, Verheggen Th P E M. J Chromatogr. 1979, 169: 1&11
4． Jorgenson J W, Lukacs K D. Anal Chem. 1981, 52: 1298
5． Hjerten S. J Chromatogr. 1983, 270: 1
6． Hjerten S, Zhu M D. J Chromatogr. 1985, 346: 265
7． Terabe S et al. Anal Chem. 1984, 56: 111
8． Lauer H H, Mcmanigill D. Anal Chem. 1986, 58: 166
9． Cox H C, Hessels J K C, Teven J M. J Chromatogr. 1972, 66: 19
10． Ghowsi K Foley J P, Gale R J. Anal Chem. 1990, 62: 2714
11． Terabe S, Otsuka K, Ando T. Anal Chem. 1989, 61: 251
12． Chu I, Bodnar J A, White E L, et al. Quantification of vincristine and vinblastine in catharanthus roseus plants by CZE. J Chromatogr A, 1996, 755: 281
13． Sun S W, Lee S S, Chen L Y, et al. Separation of aporphine alkaloids by MEKC. J Chromatogr A, 1997, 767: 277
14． Sun S W, Wa A C. Optimizationof the capillary electrophoretic separation of bisbenzylisoquinoline alkaloids by an overlopping resolution mapping scheme. J Chromatogr A, 1998, 814: 223
15． Liebich H M, Lehmann R, Stefano C D, et al. Analysis of tradition Chinese anticancer drugs by CE. J Chromatogr A, 1998, 795: 388
16． Trenerry V C, Wells R J, Robertson J. Determination of morphine and related alkaloids in crude morphine, poppy straw and opium preparations by MEKC. J Chromatogr A, 1995, 718: 217
17．王实强，首弟武。高效毛细管电泳测定罂栗壳中生物碱的含量。色谱，1997，15(5)：438
18.Unger M, Stockigt J. Improved detection of alkaloids in crude extracts applying CE with field amplified sample injection. J Chromatogr A, 1997, 791:323
19.Liang H R, Siren H, Riekkola M L, et al. Optimized separation of pharmacologically active flavonoids from epimedium species by CE.J Chromatogr A, 1996, 746:123
20.Liang H R, et al.J Chromatogr Sci, 1997, 35(3): 117
21.Gil M I, Ferreres F, Tomas-Barberan F A. MEKC of methylated flavone aglycones.J Liq Chromatogr A, 1995, 18(15): 3007
22.Wu H K, Sheu S J. CE determinotion of the constituents of paeoniae radix. J Chromatogr A, 1996, 753:139
23.Moodley V E, Mulholland D A, Raynor M W. MEKC of limonoid glucosides from citrus seeds. J Chromatogr A,1995, 718:187
24.Stuppner H, Egger R. Application of CZE to the analysis betalains from beta vulgaris. J Chromatogr A, 1996, 735:409
25.Hiermann A, Badl B. Analysis of aromatic plant acids by CZE. J Chromatogr A, 1998, 803:311
26.Summanen J, et al. J Chromatogr Sci., 1995, 33(12): 704
27．张凤云，毛富春。用毛细管区带电泳技术测定杜仲叶中绿原酸含量的研究。分析测试通报，1996，15(3)：66
28．杨更亮，王德先，宋秀荣，等。高效毛细管电泳鉴别常规中草药。中国化学会论文集，2001：521
29.Chen C T, Sheu S J. Separation of coumarins by MEKC. J Chromatogr A, 1995, 710:323
30.Ochocka R J, Rajzer D, Kowalski P, et al. Determinationof coumanns from chrysanthemum segetum L. by CE.J Chromatogr A, 1995, 709:197
31.Zhang H Y, Hu Z D, Yang G L, et al. Separation and determination of magnolol and honokiol in crude and burn-fried magnolia officinalis.Anal Lett.,1997,30(13):2327
32.Oehrle S A.AnalysiS of ginkolides and biobalides by CE.J Liq Chromatogr,1995,18(14):2855
33．柴逸峰，等。中国药学杂志，1997，32(增刊)：80
34.Hu Z D,et al. J Liq Chromatogr & Relat Technol,1997,20(1):63
35．朱萱萱，陈福林，等。毛细管电泳法测定麻黄中生物碱。中国中药杂志，1996，21(10)：587
36．赵霞，陆阳，陈泽乃，等。毛细管电泳法定量分析贝母中几种生物碱。中草药，2001，32(2)：116
37．袁炜，吕建德，傅小芸，等。槟榔中槟榔碱和槟榔次碱的毛细管电泳法分析。分析化学，2000，28(6)：749
38．杨更亮，宋秀荣，张红医，等。胶束毛细管电泳法测定牡丹皮及芍药中的牡丹酚和芍药甙。分析化学，1999，27(1)：1
39．宋秀荣，杨更亮，李杰，等。毛细管电泳法测定桑叶中的有效成分。中成药，2000，22(2)：158
40．王德先，杨更亮，王琳颖，等。毛细管区带电泳测定芦荟中的有效成分。色谱，2000，18(5)：423
41.Wang Dexian,Yang Gengliang,H.Engelhardt,Liu HaiXing.Chromatographia,2001,53(3／4):185
42．定天明，田颂九，张正行，等。胶束电动毛细管色谱法测定水飞蓟素及其制剂中主要有效成分含量。药学学报，2000，35(10)：778
43．丁岗，陆蕴如，冀春茹，等。诃子及其易混淆品中二种丹宁成分的高效毛细管电泳分析。药学学报，2001，36(4)：292
44．陈刚，李利军，叶建农，等。黄芩及其制剂中黄芩素和黄芩甙的毛细管区带电泳—电化学检测。分析测试学报，2001，20(1)：26
45．王德先，赵敬湘，杨更亮，等。毛细管区带电泳法测定中药金银花中绿原酸的含量。中草药，2000，31(6)：432
46．范国荣，胡晋红，李博华。四倍体板蓝根中五种有机酸成分的毛细管电泳分离分析。中国药科大学学报，2000，31(2)：111
47．陈勇，程智勇，韩凤梅，等。高效毛细管区带电泳法测定生药当归中阿魏酸的含量。分析化学，1999，27(12)：1424
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