HSCCC Separation of the Two Iridoid Glycosides and Three Phenolic Compounds from Veronica ciliata and Their in Vitro Antioxidant and Anti-Hepatocarcinoma Activities
Abstract
:1. Introduction
2. Results
2.1. Optimization of UPLC Analysis
2.2. Selection of HSCCC Experimental Conditions
2.3. HSCCC Separation of Sample
2.4. Structure Identification
2.5. Antioxidant Activity
2.5.1. DPPH• Scavenging Capacity
2.5.2. ABTS Radical-Scavenging Capacity
2.5.3. Reducing Power Assay
2.6. The Anti-Hepatocarcinoma Activity of Five Compounds
3. Discussion
4. Materials and Methods
4.1. HSCCC Apparatus
4.2. Reagents and Materials
4.3. Preparation of the Crude Extract
4.4. Separation and Purification by HSCCC
4.4.1. Selection of Two-Phase Solvent System
4.4.2. Preparation of Two-Phase Solvent System and Sample Solution
4.4.3. HSCCC Separation
4.5. Purification of Compounds 1–3 by Preparative HPLC
4.6. Peak Identification
4.6.1. UPLC-PDA Analysis
4.6.2. MS Analysis
4.6.3. NMR Analysis
4.7. In Vitro Antioxidant and Anti-Hepatocarcinoma Activity
4.7.1. DPPH• Scavenging Capacity [33]
4.7.2. ABTS Radical-Scavenging Capacity
4.7.3. Reducing Power Assay [35]
4.7.4. In Vitro Anti-Hepatocarcinoma Activity [2,12]
Cell Culture
Cell Proliferation Inhibition Assay
4.8. Statistical Analysis
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Solvent System (n-Hexane/n-Butanol/Water) | k-Values | ||||
---|---|---|---|---|---|
k1 | k2 | k3 | k4 | k5 | |
1.5:5:5 | 0.009 | 0.13 | 0.19 | 0.52 | 1 |
1.5:4:5 | 0.019 | 0.17 | 0.55 | 0.56 | 1.7 |
1.5:3:5 | 0.02 | 0.19 | 0.59 | 0.67 | 2.1 |
1.5:2:5 | 0.1 | 0.2 | 0.6 | 1.4 | 3.8 |
3:2:5 | 0.21 | 0.32 | 0.95 | 2.7 | 5.8 |
Carbon | Compound | ||||
---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | |
1 | 121.95 | 123.8 | 92.9 | 95.6 | |
2 | 164.2 | 131.98 | 116.6 | ||
3 | 103.6 | 115.56 | 146.8 | 141.2 | 142.8 |
4 | 182.4 | 162.02 | 152.0 | 101.6 | 103.6 |
5 | 162.7 | 115.56 | 118.6 | 35.3 | 36.8 |
6 | 99 | 131.98 | 124.8 | 80.1 | 82.1 |
7 | 164.5 | 170.9 | 58.3 | 59.9 | |
8 | 94 | 65.7 | 67.2 | ||
9 | 158.1 | 41.7 | 43.7 | ||
10 | 104.7 | 58.5 | 61.8 | ||
1′ | 123.1 | 97.8 | 99.6 | ||
2′ | 113.5 | 73.3 | 75.6 | ||
3′ | 145.8 | 77.5 | 78.5 | ||
4′ | 149.4 | 70.4 | 72.4 | ||
5′ | 116 | 76.5 | 77.5 | ||
6′ | 119.5 | 61.5 | 63.9 | ||
1″ | 119.4 | 122.4 | |||
2″ | 131.6 | 116.6 | |||
3″ | 115.4 | 146.4 | |||
4″ | 162.4 | 152.4 | |||
5″ | 115.4 | 117.4 | |||
6″ | 131.6 | 124.6 | |||
7″ | 165.5 | 168.8 |
Samples | IC50 (μg/mL) | Reducing Power | |
---|---|---|---|
DPPH | ABTS | ||
luteolin | 4.168 | 5.587 | 0.0028 |
4-hydroxy benzoic acid | 6.838 | 58.78 | 0.0012 |
3,4-dihydroxy benzoic acid | 0.703 | 5.186 | 0.013 |
catalposide | 20.16 | 43.83 | 0.0004 |
verproside | 6.502 | 16.9 | 0.0018 |
Vc | 0.57 | 4.645 | |
BHT | 0.0016 |
Compounds | HepG2 |
---|---|
luteolin | 102.356 ± 2.01 |
4-hydroxy benzoic acid | 444.759 ± 2.65 |
3,4-dihydroxy benzoic acid | 186.033 ± 2.27 |
catalposide | 184.592 ± 2.27 |
verproside | 177.147 ± 2.25 |
5-FU | 35.420 ± 1.56 |
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Lu, Q.; Sun, Y.; Shu, Y.; Tan, S.; Yin, L.; Guo, Y.; Tang, L. HSCCC Separation of the Two Iridoid Glycosides and Three Phenolic Compounds from Veronica ciliata and Their in Vitro Antioxidant and Anti-Hepatocarcinoma Activities. Molecules 2016, 21, 1234. https://doi.org/10.3390/molecules21091234
Lu Q, Sun Y, Shu Y, Tan S, Yin L, Guo Y, Tang L. HSCCC Separation of the Two Iridoid Glycosides and Three Phenolic Compounds from Veronica ciliata and Their in Vitro Antioxidant and Anti-Hepatocarcinoma Activities. Molecules. 2016; 21(9):1234. https://doi.org/10.3390/molecules21091234
Chicago/Turabian StyleLu, Qiuxia, Yiran Sun, Yueyue Shu, Shancai Tan, Li Yin, Yiran Guo, and Lin Tang. 2016. "HSCCC Separation of the Two Iridoid Glycosides and Three Phenolic Compounds from Veronica ciliata and Their in Vitro Antioxidant and Anti-Hepatocarcinoma Activities" Molecules 21, no. 9: 1234. https://doi.org/10.3390/molecules21091234