An Economical Method for Preparative Purification of Five Alkaloids from Coptis Chinensis Franch by High-Speed Counter-Current Chromatography Using Singled Prepared Solvent System by GC

Lianhong Yin, Lina Xu, Xiaona Wang, Binan Lu, Yingnan Li, Mingming Hu, Yuetao Liu, Jinyong Peng

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College of Pharmacy, Dalian Medical University, 9 Western Lvshun South Road, Dalian 116044, China.

DOI: 10.4236/ajac.2011.24050                

Abstract

Coptis chinensis Franch, a widely used Traditional Chinese Medicine, shows various kinds of bioactivity. The major active components of the herb are considered to be alkaloids. Thus, preparative separation of these alkaloids is critical important for further pharmacology and mechanism studies. In the paper, five alkaloids from C. chinensis were purified by HSCCC using the solvent system composed of chloro-form-metha- nol-water (2:1:1, v/v/v) single prepared. The content of each solvent in solvent system were determined by gas chromatography (GC), then according the ratios of solvents in each phase to prepare the mobile and stationary phase respectively. And a comparative study was carried out between together preparation and single preparation of the solvent system. The purities and recoveries of all the products were over 98.5% and 92%. However, 134 mL chloroform, 336 mL methanol and 452 mL water were saved when the two phase were singled by GC. Our research showed an economical method for separating alkaloids from C. chinensis by HSCCC using the solvent system single prepared by GC.

Keywords

Alkaloid, Coptis Chinensis Franch, High Speed Counter-Current Chromaotgraphy, Gas Chromatography; Solvent System

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	H. H. Yu, K. J. Kim, J. D. Cha, H. K. Kim, Y. E. Lee, N. Y. Choi and Y. O. You, “Antimicrobial Activity of Ber-berine Alone and in Combination with Ampicillin or Oxacillin against Methicillin-Resistant Staphylococcus Aureus,” Journal of Medicial Food, Vol. 8, No. 4, 2005, pp. 454-461. doi:10.1089/jmf.2005.8.454
[2]	H. A. Jung., N. Y. Yoon, H. J. Bae, B. S. Min and J. S. Choi, “Ingibitory Activities of the Alkaloids from Coptidis Rhizoma against Aldose Reductase,” Archives of Pharmacal Research, Vol. 31, No. 11, 2008, pp. 1405- 1412. doi:10.1007/s12272-001-2124-z
[3]	J. Liu, C. He, K. Zhou, J. Wang and J. X. Kang, “Coptis extracts Enhance the Anticancer Effect of Estrogen Re-ceptor Antagonists on Human Breast Cancer Cells,” Bio-chemical and biophysical research communications, Vol. 378, No. 2, 2009, pp. 174-178. doi:10.1016/j.bbrc.2008.10.169
[4]	J. H. Chen, F. M. Wang, J. Liu, F. S. C. Lee, X. R. Wang and H. G. Yang, “Analysis of Alkaloids in Coptis Chi-nensis Franch by Accelerated Solvent Extraction Com-bined with Ultra Performance Liquid Chromatographic Analysis with Photodiode Array and Tandem Mass Spec-trometry Detections,” Analytia Chimuca Acta, Vol. 613, No. 2, 2008, pp. 184-195. doi:10.1016/j.aca.2008.02.060
[5]	H. L. Yang, S. S. Yu and Y. H. Pei, “Studies on Chemical Constituents from leaves of Lysidice Brevicalyx,” Zhongguo Zhong Yao Za Zhi, Vol. 33, No. 22, 2008, pp. 2633-2635.
[6]	C. L. Kuo, C. W. Chi and T. Y. Liu, “The Anti-Infam- matory Potential of Berberine in Vitro and in Vivo,” Cancer Letter, Vol. 203, No. 2, 2004, pp. 127-137. doi:10.1016/j.canlet.2003.09.002
[7]	L. Q. Tang, W. Wei, L. M. Chen and S. Liu, “Effects of Berberine on Diabetes Induced by Alloxan and a High-Fat/High-Cholesterol Diet in Rats,” Journal of Ethnopharmacology, Vol. 108, No. 1, 2006, pp. 109-115. doi:10.1016/j.jep.2006.04.019
[8]	L. P. Qu, G. R. Fan, J. Y. Peng and H. M. Mi, “Isolation of Six Isoflavones from Semen Sojae Praeparatum by Preparative HPLC,” Fitoterapia, Vol. 78, No 3, 2007, pp. 200-204. doi:10.1016/j.fitote.2006.11.002
[9]	Y. Q. Yin, Z. B. Shen and L. Y. Kong, “Studies on Chemical Constituents from Ipomoea Batatas,” Lishizhen Medicine and Materia Medica Research, Vol. 19, No. 11, 2008, pp.1501-1503.
[10]	Y. Ito and R. L. Bowman, “Counter-Current Chromato-graphy: Liquid-Liquid Partition Chromatography without Solid Support,” Science, Vol. 167, No. 3916, 1970, pp. 281-283. doi:10.1126/science.167.3916.281
[11]	Y. Ito, J. L. Sandlin and W.G. Bowers, “High-Speed Preparative Counter Current Chromatography with a Coil Planet Centrifuge,” Journal of Chromatography, Vol. 244, 1982, pp. 247-258. doi:10.1016/S0021-9673(00)85688-5
[12]	Y. Ito, “Counter Current Chromatography: Theory and Practice,” Marcel Dekker, New York, 1988, pp. 79-442.
[13]	Y. Ma, Y. Ito and A. Foucault, “Resolution of Gram Quantities of Racemates by High-Speed Counter-Current Chromatography,” Journal of Chromatography A, Vol. 704, No. 1, 1995, pp. 75-81. doi:10.1016/0021-9673(95)00148-G
[14]	L. J. Chen, Q. Zhang, G. L. Yang, L. Y. Fan, J. Tang, I. Garrard, S. N. Ignatova, D. Fisher and I. A. Sutherland. “Rapid Purification and Scale-Up of Honokiol and Mag-nolol Using High-Capacity High-Speed Counter-Current Chromatography,” Journal of Chromatography A, Vol. 1142, No. 2, 2007, pp. 115-122. doi:10.1016/j.chroma.2006.09.098
[15]	D. S. Abrams and J. M. Prausnitz, “Statistical Thermo-dynamics of Liquid Mixtures: A New Expression for the Excess Gibbs Free Energy of Partly or Completely Misc-ible Systems,” AIChE Journal, Vol. 21, 1975, pp. 116- 128. doi:10.1002/aic.690210115
[16]	S. Y. Shi, K. L. Huang, Y. P. Zhang and S. Liu, “Prepara-tive Isolation and Purification Of Two Flavonoid Glyco-sides from Taraxacum Mongolicum by High-Speed Counter-Current Chromatography,” Separation and puri-fication technology, Vol. 60, No. 1, 2008, pp. 81-85. doi:10.1016/j.seppur.2007.07.047
[17]	J. Y. Peng, G. R. Fan, L. P. Qu, Z. Xin and Y.T. Wu, “Application of Preparative High-Speed Counter-Current Chromatography for Isolation and Separation of Schizan-drin and Gomisin A from Schisandra Chinensis,” Journal of Chromatography A, Vol. 1082, No. 2, 2005, pp. 203- 207. doi:10.1016/j.chroma.2005.05.091
[18]	H. B. Li, F. Chen, T. Y. Zhang, F. Q. Yang and G. Q. Xu, “Preparative Isolation and Purification of Lutein from the Microalga Chlorella Vulgaris by High-Speed Counter- Current Chromatography,” Journal of Chromatography A, Vol. 905, No. 1-2, 2001, pp. 151-155. doi:10.1016/S0021-9673(00)00987-0
[19]	C. Seger, K. Eberhart, S. Sturm, H. Strasser and H. Stuppner, “Apolar Chromatography on Sephadex LH-20 Combined with High-Speed Counter-Current Chromato-graphy: High Yield Strategy for Structurally Closely Re-lated an Analyses-Destruxin Derivatives from Metarhi- zium Anisopliae as a Case Study,” Journal of Chromato-graphy A, Vol. 1117, No. 1, 2006, pp. 67-73. doi:10.1016/j.chroma.2006.03.055
[20]	W. B. Zhi, Q. Y. Deng, J. N. Song, M. Gu and F. Ouyang, “One-Step Purification of Alphamylase from the Cultiva-tion Supernatant of Recombinant Bacillus Subtilis by High-Speed Counter-Current Chromatography with Aqueous Polymer Two-Phase Systems,” Journal of Chromatography A, Vol. 1070, No. 1-2, 2005, pp. 215- 219. doi:10.1016/j.chroma.2005.02.065
[21]	E. Pérez, M. J. Santos and C. Minguillón, “Application of Cellulose and Amylose Arylcarbamates as Chiral Selectors in Counter-Current Chromatography,” Journal of Chromatography A Vol. 1107, No. 1-2, 2006, pp. 165- 174. doi:10.1016/j.chroma.2005.12.061
[22]	F. Q. Yang, T. Y. Zhang, R. Zhang and Y. Ito, “Applica-tion of Analytical and Preparative High-Speed Counter- Current Chromatography for Separation of Alkaloids from Coptis Chinensis Franch,” Journal of Chromatography A, Vol. 829, No.1-2, 1998, pp. 137-141. doi:10.1016/S0021-9673(98)00776-6
[23]	J. Y. Peng, X. Han, Y. W. Xu, Y. Qi, L. N. Xu and Q. W. Xu, “New Approach of Application High Speed Coun-tercurrent Chromatography Coupled with Direct Injection the Powders of a Raw Materials without Any Preparation, for Isolation and Separation of Four Alkaloids with High Recoveries from Coptis Chinensis Franch,” Journal of liquid chromatography & related technologies, Vol. 30, No. 17-20, 2007, pp. 2929-2940. doi:10.1080/10826070701588984
[24]	P. Jacobs, W. Dewé, A. Flament, M. Gibella and A. Cec-cato, “A New Validation Approach Applied to the GC Determination of Impurities in Organic Solvents,” Journal of Pharmaceutical and Biomedical Analysis, Vol. 40,No. 2, 2006, pp. 294-304. doi:10.1016/j.jpba.2005.06.036
[25]	F. J. Arrebola, M. J. González-Rodríguez, A. Garrido Frenich, A. Marín-Juan and J. L. Martínez Vidal. “De-termination of Halogenated Solvents Content in Olive Oil by Two Completely Automated Headspace Techniques Coupled to Gas Chromatography-Mass Spectrometry,” Analytica chimica acta, Vol. 552, No. 1-2, 2005, pp. 60- 66. doi:10.1016/j.aca.2005.07.054
[26]	I. J. Garrard, L. Janaway and D. Fisher, “Minimising Solvent Usage in High Speed, High Loading, and High Resolution Isocratic Dynamic Extraction,” Journal of liquid chromatography & related technologies, Vol. 30, No. 1-4, 2007, pp. 151-163. doi:10.1080/10826070601077252
[27]	W. Y. Liu, “Pharmaceutical Analysis,” People’s Medical Publishing House, 2006, pp. 75-80.
[28]	Y. M. Liu and S. J. Shen, “Determination of Coptisine, Berberine and Palmatine in Traditional Chinese Medicinal Preparations by Capillary Electrophoresis,” Journal of Chromatography A, Vol. 639, No. 2, 1993, pp. 323-328. doi:10.1016/0021-9673(93)80269-E
[29]	S. W. Sun and H. M. Tseng, “Improved Detection of Coptidis Alkaloids by Field-Amplified Sample Stacking in Capillary Electrophoresis,” Journal of Pharmaceutical and Biomedical Analysis, Vol. 36, No. 1, 2004, pp. 43-48. doi:10.1016/j.jpba.2004.04.017
[30]	S. Yu, X.Y. Pang, Y. X. Deng, L. Liu, Y. Liang, X. D. Liu, L. Xie, G. J. Wang and X. T. Wang, “A Sensitive and Specific Liquid Chromatography Mass Spectrometry Method for Simultaneous Determination of Berberine, Palmatine, Coptisine, Epiberberine and Jatrorrhizine from Coptidis Rhizoma in Rat Plasma,” International Journal of Mass Spectrometry, Vol. 268, No. 1, 2007, pp. 30-37. doi:10.1016/j.ijms.2007.08.003
[31]	S. W. Sun and H. M. Tseng, “Sensitivity Improvement on Detection of Coptidis Alkaloids by Sweeping in Capillary Electrophoresis,” Journal of Pharmaceutical and Bio-medical Analysis, Vol. 37, No. 1, 2005, pp. 39-45. doi:10.1016/j.jpba.2004.09.042
[32]	W. C. Chuang, D. S. Young, L. K. Liu and S. J. Sheu, “Liquid Chromatographic-Electrospray Mass Spectrome-tric Analysis of Coptidis Rhizoma,” Journal of Chroma-tography A, Vol. 755, No. 1, 1996, pp. 19-26. doi:10.1016/S0021-9673(96)00591-2
[33]	Y. T. Deng, Q. F. Liao, S. H. Li, K. H. Bi, B. Y. Pan and Z. Y. Xie, Simultaneous Determination of Berberine, Palmatine and Jatrorrhizine by Liquid Chromatogra-phy-Tandem Mass Spectrometry in Rat Plasma and Its Application in a Pharmacokinetic Study after Oral Ad-ministration of Coptis-Evodia Herb Couple,” Journal of Chromatography B, Vol. 863, No. 2, 2008, pp. 195-205. doi:10.1016/j.jchromb.2007.12.028
[34]	H. L. Li, W. D. Zhang, R. H. Liu, C. Zhang, T. Han, X. W. Wang, X. L. Wang, J. B. Zhu and C. L. Chen, “Si-multaneous Determination of Four Active Alkaloids from a Traditional Chinese Medicine Corydalis Saxicola Bunting. (Yanhuanglian) in Plasma and Urine Samples by LC-MS-MS,” Journal of Chromatography B, Vol. 831, No. 1-2, 2006, pp. 140-146. doi:10.1016/j.jchromb.2005.11.049
[35]	H. S. Lee, Y. E. Eom and D. O. Eom, “Narrowbore High Performance Liquid Chromatography of Berberine and Palmatine in Crude Drugs and Pharmaceuticals with Ion-Pair Extraction Using Cobalt Thiocyanate Reagent,” Journal of Pharmaceutical and Biomedical Analysis, Vol. 21, No. 1, 1999, pp. 59-63. doi:10.1016/S0731-7085(99)00113-2
[36]	Y. X. Luo, D. X. Wen, S. J. Jiang, X. L. Zeng, J. N. Jiang, Y. C. Xu and P. D. Xie, “Studies on HPLC Fingerprint of Herba Corydalis Saxicolae,” Chinese Journal of Phar-macutical Analysis, Vol. 27, No. 11, 2007, pp. 1749- 1751.
[37]	J. Y. Ling, G. Y. Zhang, Z. J. Cui and C. K. Zhang, “Su-percritical Fluid Extraction of Quinolizidine Alkaloids from Sophora Flavescens Ait. and Purification by High-Speed Counter-Current Chromatography,” Journal of Chromatography A, Vol. 1145, No. 1-2, 2007, pp. 123-127. doi:10.1016/j.chroma.2007.01.080
[38]	F. Y. Yang and Y. Ito, “Preparative Separation of Lap-paconitine, Ranaconitine, N-deacetyllappaconitine and N-deacetylranaconitine from Crude Alkaloids of Sample Aconitum Sinomontanum Nakai by High-Speed Counter- Current Chromatography,” Journal of Chromatography A, Vol. 943, No. 2, 2002, pp. 219-225. doi:10.1016/S0021-9673(01)01464-9