Preparative Isolation of Seven Diterpenoid Alkaloids from Aconitum coreanum by pH-Zone-Refining Counter-Current Chromatography
Abstract
:1. Introduction
2. Results and Discussion
2.1. Selection of Two-Phase Solvent System
Solvent System (Pet–EtOAc–MeOH–H2O) (v/v/v/v) | K1 | K2 | K3 | K4 | K5 | K6 | K7 | |
---|---|---|---|---|---|---|---|---|
5:5:4:6 | K | 0.75 | 0.82 | 1.05 | 1.01 | 1.34 | 1.38 | 1.55 |
Kacid | 0.07 | 0.05 | 0.08 | 0.07 | 0.08 | 0.09 | 0.11 | |
5:5:2:8 | K | 1.52 | 1.52 | 1.75 | 1.78 | 2.12 | 2.23 | 2.25 |
Kacid | 0.12 | 0.12 | 0.14 | 0.12 | 0.15 | 0.16 | 0.19 | |
5:5:1:9 | K | 1.65 | 1.69 | 1.87 | 1.91 | 2.21 | 2.34 | 2.39 |
Kacid | 0.15 | 0.16 | 0.18 | 0.18 | 0.21 | 0.23 | 0.25 |
Results | Standard HSCCC | pH-Zone-Refining CCC | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
GFI | GFA | atisine | GFF | GFG | GFP | GFI | GFA | atisine | GFF | GFG | GFR | GFP | |
Purity (%) | 95.5 | 95.8 | 98.9 | 91.5 | 95.7 | 96.9 | 97.1 | 98.5 | 97.4 | 98.7 | 99.1 | 98.1 | 98.7 |
Yield (mg) | 25.7 | 11.9 | 8.9 | 9.5 | 9.2 | 10.4 | 356 | 578 | 74 | 94 | 423 | 67 | 154 |
Sample Size | 2 g | 3.5 g |
2.2. Identification of the Isolated Compounds
Compound | Formula | Selected Ion | Experimental m/z | Calcuated m/z | Error (mda) | Error (ppm) |
---|---|---|---|---|---|---|
GFI (1) | C23H9N5O2 | [M] | 387.0757 | 387.0756 | −0.07 | −0.19 |
GFA (2) | C20H11N7O5 | [M] | 429.0815 | 429.0822 | 0.66 | 1.54 |
atisine (3) | C17H13N9 | [M] | 343.1301 | 343.1294 | −0.75 | −2.17 |
GFF (4) | C33H15NO2 | [M] | 457.1088 | 457.1103 | 1.53 | 3.34 |
GFG (5) | C29H9N7O | [M] | 471.0861 | 471.0869 | 0.81 | 1.71 |
GFR (6) | C20H11N7O6 | [M] | 485.0995 | 4850985 | −0.98 | −2.02 |
GFP (7) | C37H13N3 | [M] | 499.1127 | 499.1109 | −1.8 | −3.6 |
3. Experimental Section
3.1. Apparatus
3.2. Reagents and Materials
3.3. Crude Alkaloids Extracted by Acid-Base Extraction Method
3.4. Determination of the Partition Coefficient (K)
3.5. pH-Zone-Refining CCC Separation Procedure
3.6. HPLC Analysis and Identification of CCC Fractions
3.7. TOF-MS and 1H-NMR for Identification
Parameter | Value |
---|---|
Capillary | 3500 V |
Nebulizer Pressure | 50 psig |
Drying Gas | 10 L/min |
Gas Temperature | 350 °C |
Fragmentor Voltage | 175 V |
Skimmer Voltage | 60 V |
Mass Range | 50–1000 |
Resolution | 9500 ± 500 (922.0098) |
Reference Masses | 121.0509, 922.0098 |
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
- Bessonova, I.A.; Yunusov, M.S.; Kondratèv, V.G.; Shreter, A.I. Alkaloids of Aconitum coreanum. I. Structure of acorine. Chem. Nat. Compd. 1987, 23, 573–575. [Google Scholar] [CrossRef]
- Editorial Committee of Chinese Bencao. Chinese Bencao; Shanghai Science and Technology Press: Shanghai, China, 1998; p. 499. [Google Scholar]
- Kai, J.; Yang, C.H.; Liu, J.H.; Tang, Q.F. Isolation and identification of Hetisine-type alkaloids from Aconitum coreanum by high speed countercurrent chromatography. Acta Pharm. Sin. 2006, 41, 128–131. [Google Scholar]
- Yang, C.H.; Zhang, H.H.; Liu, J.H. Alkaloid constituents from root of Aconitum coreanum. Chin. Tradit. Herb. Drugs 2004, 35, 1328–1330. [Google Scholar]
- Tang, Y.Q.; Yin, Y.M.; Huang, L.; Wang, M.H.; Yu, X.L.; Xu, J. Effects of Guanfu total base on experimental atrial fibrillation. Guide Chin. Med. 2011, 9, 249–251. [Google Scholar]
- Wang, M.N.; Zhu, J.N.; Yang, Y.; Li, J.D.; Huang, X.F.; Li, C.R.; Tian, Y.; Chen, X.S. Effect of acehytisine hydrochloride on vagotonic atrial fibrillation inmongrel dogs. Kokyu To Junkan 2008, 22, 148–152. [Google Scholar]
- Ito, Y. Golden rules and pitfalls in selecting optimum conditions for high-speed counter-current chromatography. J. Chromatogr. A 2005, 1065, 145–158. [Google Scholar] [CrossRef]
- Ito, Y. pH-zone-refining counter-current chromatography: Origin, mechanism, procedure and applications. J. Chromatogr. A 2013, 1271, 71–85. [Google Scholar] [CrossRef]
- Zheng, Z.J.; Wang, M.L.; Wang, D.J.; Duan, W.J.; Wang, X.; Zheng, C.C. Preparative separation of alkaloids from Nelumbo nucifera leaves by conventional and pH-zone-refining counter-current chromatography. J. Chromatogr. B 2010, 878, 1647–1651. [Google Scholar] [CrossRef]
- Tong, S.Q.; Yan, J.Z.; Jian, L.; Lou, J.Z. Separation of pyridine derivatives from synthetic mixtures by pH-zone-refining counter-current chromatography. J. Sep. Sci. 2007, 30, 1899–1904. [Google Scholar] [CrossRef]
- Fang, L.; Liu, Y.Q.; Yang, B.; Wang, X.; Huang, L.Q. Separation of alkaloids from herbs using high-speed counter-current chromatography. J. Sep. Sci. 2011, 34, 2545–2558. [Google Scholar] [CrossRef]
- Fang, L.; Zhou, J.; Lin, Y.L.; Wang, X.; Sun, Q.L.; Li, J.L.; Huang, L.Q. Large-scale separation of alkaloids from Gelsemium elegans by pH-zone-refining counter-current chromatography with a new solvent system screening method. J. Chromatogr. A 2013, 1307, 80–85. [Google Scholar] [CrossRef]
- Tang, Q.; Yang, C.; Ye, W.; Liu, J.; Zhao, S. Preparative isolation and purification of bioactive constituents from Aconitum coreanum by high-speed counter-current chromatography coupled with evaporative light scattering detection. J. Chromatogr. A 2007, 1144, 203–207. [Google Scholar] [CrossRef]
- Bessonova, I.A. Alkaloids of Aconitum coreanum IX. Tangutisine and 2,11,13-triacetyl-14-hydroxyhetisine. Chem. Nat. Comp. 1999, 35, 103–104. [Google Scholar] [CrossRef]
- Razakova, D.M.; Bessonova, I.A.; Yunosov, M.S. Atisine chloride and isoatisine from Aconitum coreanum and A. rotundifolium. Chem. Nat. Comp. 1988, 24, 266–267. [Google Scholar] [CrossRef]
- Liu, J.H.; Hao, Z.G.; Zhao, S.X.J. Identifica tion of the structure of Guanfu base F. China Pharm. Univ. 1988, 19, 238–239. [Google Scholar]
- Sample Availability: Samples of all the compounds are available from the authors.
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Wang, X.; Shu, X.; Wang, X.; Yu, J.; Jing, F. Preparative Isolation of Seven Diterpenoid Alkaloids from Aconitum coreanum by pH-Zone-Refining Counter-Current Chromatography. Molecules 2014, 19, 12619-12629. https://doi.org/10.3390/molecules190812619
Wang X, Shu X, Wang X, Yu J, Jing F. Preparative Isolation of Seven Diterpenoid Alkaloids from Aconitum coreanum by pH-Zone-Refining Counter-Current Chromatography. Molecules. 2014; 19(8):12619-12629. https://doi.org/10.3390/molecules190812619
Chicago/Turabian StyleWang, Xueyong, Xikai Shu, Xiao Wang, Jinqian Yu, and Feng Jing. 2014. "Preparative Isolation of Seven Diterpenoid Alkaloids from Aconitum coreanum by pH-Zone-Refining Counter-Current Chromatography" Molecules 19, no. 8: 12619-12629. https://doi.org/10.3390/molecules190812619