Preparative Separation of Alkaloids from Stem of Euchresta tubulosa Dunn. by High-Speed Counter-Current Chromatography Using Stepwise Elution
Abstract
:1. Introduction.
2. Results
2.1. Selection of Optimum Solvent Systems of HSCCC Separation
2.2. Sample Pretreatment of HSCCC Separation
2.3. HSCCC Separation
2.4. Results of HPLC-UV Analysis
2.5. Identification of Purified Alkaloids
2.6. Applicability Verification Test
2.6.1. Precision and Repeatability Test
2.6.2. Applicability Test
3. Materials and Methods
3.1. Reagents and Materials
3.2. Apparatus
3.3. Preparation of the Crude Sample
3.4. Selection of the Two-Phase Solvent Systems
- As: Peak area of a single alkaloid in the stationary phase;
- Am: Peak area of a single alkaloid in the mobile phase.
3.5. Preparation of HSCCC Two-Phase Solvent Systems and Sample Solution
3.6. HSCCC Separation Procedures
3.7. HPLC-UV Analysis
3.8. Applicability Verification Test
3.8.1. Precision and Repeatability Test
3.8.2. Applicability Test
- V1: Loss volume of the first phase separation;
- V2: Loss volume of the second phase separation;
- V3: Volume of upper phase retained in the instrument.
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Sample Availability: Samples of the compounds (1–4) are available from the authors. |
Solvent Systems | Volume Ratio (v/v) | K1 | K2 | K3 | K4 |
---|---|---|---|---|---|
CH2Cl2-MeOH-H2O | 2: 1: 1 | 0.23 | 0.53 | 0.51 | 0.68 |
5: 3: 2 | 0.25 | 0.56 | 0.65 | 0.82 | |
4: 3: 2 | 0.31 | 0.63 | 0.77 | 0.98 | |
CCl4-CH2Cl2-MeOH-H2O | 1: 4: 3: 2 | 0.38 | 0.81 | - | - |
2: 3: 3: 2 | 0.76 | 0.95 | - | - | |
3: 2: 3: 2 | 0.92 | 1.89 | - | - | |
4: 1: 3: 2 | 2.33 | 4.43 | - | - |
Rotation Speed (rpm) | Flow Velocity (mL/min) | Sf | Rotation Speed (rpm) | Flow Velocity (mL/min) | Sf |
---|---|---|---|---|---|
700 | 2 | 0.645 | 800 | 3 | 0.657 |
700 | 2.5 | 0.638 | 800 | 3.5 | 0.626 |
700 | 3 | 0.614 | 800 | 4 | 0.587 |
700 | 3.5 | 0.581 | 850 | 2 | 0.702 |
700 | 4 | 0.553 | 850 | 2.5 | 0.683 |
750 | 2 | 0.662 | 850 | 3 | 0.671 |
750 | 2.5 | 0.643 | 850 | 3.5 | 0.641 |
750 | 3 | 0.639 | 850 | 4 | 0.615 |
750 | 3.5 | 0.609 | 900 | 2 | 0.723 |
750 | 4 | 0.568 | 900 | 2.5 | 0.698 |
800 | 2 | 0.675 | 900 | 3 | 0.682 |
800 | 2.5 | 0.669 | 900 | 3.5 | 0.654 |
900 | 4 | 0.639 |
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Li, W.; Wang, H.; Dong, A. Preparative Separation of Alkaloids from Stem of Euchresta tubulosa Dunn. by High-Speed Counter-Current Chromatography Using Stepwise Elution. Molecules 2019, 24, 4602. https://doi.org/10.3390/molecules24244602
Li W, Wang H, Dong A. Preparative Separation of Alkaloids from Stem of Euchresta tubulosa Dunn. by High-Speed Counter-Current Chromatography Using Stepwise Elution. Molecules. 2019; 24(24):4602. https://doi.org/10.3390/molecules24244602
Chicago/Turabian StyleLi, Weixin, Huan Wang, and Aiwen Dong. 2019. "Preparative Separation of Alkaloids from Stem of Euchresta tubulosa Dunn. by High-Speed Counter-Current Chromatography Using Stepwise Elution" Molecules 24, no. 24: 4602. https://doi.org/10.3390/molecules24244602