A Strategy for Simultaneous Isolation of Less Polar Ginsenosides, Including a Pair of New 20-Methoxyl Isomers, from Flower Buds of Panax ginseng
Abstract
:1. Introduction
2. Results and Discussion
2.1. Purification of the Five Less Polar Ginsenosides from the Flower Buds of Panax Ginseng
2.1.1. Preparation of Crude Ginsenoside Extracts
2.1.2. Optimization of Solid Phase Extraction Conditions
2.1.3. Establishment of HPLC Isolation Conditions
2.2. Identification of the Isolated Ginsenosides
2.3. Method Validation of Quantification
2.4. Quantitative Analysis of the Five Less Polar Ginsenosides in FBPG
3. Materials and Methods
3.1. General
3.2. Plant
3.3. Sample Preparation
3.4. HPLC Analysis and Preparation
3.5. Structural Characterization of New Ginsenosides
3.6. Acid Hydrolysis
3.7. Validation
4. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
FBPG | Flower buds of Panax ginseng |
HMBC | Heteronuclear multiple bond correlation |
HSQC | Heteronuclear singular quantum correlation |
HR-ESIMS | High resolution electrospray ionization mass spectrometry |
LC-MS | Liquid chromatography coupled to mass spectrometry |
LOD | Limits of detection |
LOQ | Limits of quantification |
NMR | Nuclear magnetic resonance |
NOE | Nuclear overhauser effect |
ROESY | Rotating frame overhauser effect spectroscopy |
RSD | Relative standard deviation |
RP-HPLC | Reversed-phase high-performance liquid chromatography |
S/N | Signal-to-noise |
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Position | 1 | 2 | ||
---|---|---|---|---|
δH | δC | δH | δC | |
1 | 0.87 (1H, m) 1.58 (1H, m) | 39.2 | 0.78 (1H, m) 1.52 (1H, m) | 39.2 |
2 | 1.24 (1H, m) 1.86 (1H, m) | 26.3 | 1.29 (1H, m) 1.89 (1H, m) | 26.3 |
3 | 3.32 (1H, dd, 12.0, 4.5) | 89.0 | 3.32 (1H, dd, 11.5, 4.0) | 89.0 |
4 | - | 39.7 | - | 39.7 |
5 | 0.73 (1H, br d, 12.0) | 56.4 | 0.72 (1H, br d, 11.5) | 56.4 |
6 | 1.32 (1H, m) 1.47 (1H, dd, 12.0, 4.2) | 18.5 | 1.40 (1H, m) 1.52 (1H, dd, 12.0, 4.2) | 18.5 |
7 | 1.22 (1H, m) 1.51 (1H, m) | 35.2 | 1.24 (1H, m) 1.47 (1H, m) | 35.2 |
8 | - | 40.0 | - | 40.1 |
9 | 1.41 (1H, m) | 50.0 | 1.43 (1H, m) | 50.2 |
10 | - | 37.0 | - | 37.0 |
11 | 1.38 (1H, m) 2.03 (1H, m) | 31.2 | 1.45 (1H, m) 2.04 (1H, dd, 12.0, 7.0) | 31.4 |
12 | 3.76 (1H, dt, 9.5, 5.5) | 70.4 | 3.81 (1H, dt, 10.0, 3.0) | 70.6 |
13 | 1.86 (1H, t, 11.0) | 49.0 | 1.83 (1H, t, 10.5) | 49.7 |
14 | - | 51.5 | - | 51.8 |
15 | 1.05 (1H, t, 11.0) 1.57 (1H, m) | 30.9 | 1.05 (1H, t, 10.0) 1.54 (1H, m) | 31.3 |
16 | 1.73 (1H, m) 2.23 (1H, m) | 26.8 | 1.85 (1H, m) 2.22 (1H, m) | 26.8 |
17 | 2.42 (1H, td, 9.0) | 50.0 | 2.35 (1H, td, 7.5) | 47.2 |
18 | 0.95 (3H, s) | 15.9 | 1.00 (3H, s) | 15.9 |
19 | 0.83 (3H, s) | 16.3 | 0.86 (3H, s) | 16.4 |
20 | - | 79.9 | - | 80.2 |
21 | 1.19 (3H, s) | 21.2 | 1.17 (3H, s) | 18.5 |
22 | 1.67 (1H, m) 2.04 (1H, m) | 35.1 | 1.44 (1H, m) 1.66 (1H, m) | 35.7 |
23 | 2.05 (1H, m) 2.30 (1H, m) | 22.8 | 2.13 (1H, m) n.d. | 21.7 |
24 | 5.25 (1H, t, 6.5) | 125.5 | 5.23 (1H, t, 7.5) | 125.0 |
25 | - | 131.3 | - | 131.4 |
26 | 1.67 (3H, s) | 25.8 | 1.75 (3H, s) | 25.8 |
27 | 1.64 (3H, s) | 17.7 | 1.67 (3H, s) | 17.7 |
28 | 1.32 (3H, s) | 28.2 | 1.33 (3H, s) | 28.2 |
29 | 1.13 (3H, s) | 16.7 | 1.14 (3H, s) | 16.6 |
30 | 0.96 (3H, s) | 17.1 | 0.98 (3H, s) | 17.4 |
20-OCH3 | 3.27 (3H, s) | 48.8 | 3.20 (3H, s) | 48.4 |
1′ | 4.95 (1H, d, 7.5) | 105.1 | 4.96 (1H, d, 7.5) | 105.2 |
2′ | 4.26 (1H, m) | 83.5 | 4.25 (1H, m) | 83.5 |
3′ | 4.28 (1H, m) | 78.0 | 4.27 (1H, m) | 78.0 |
4′ | 4.18 (1H, t, 10.0) | 71.7 | 4.17 (1H, t, 9.0) | 71.7 |
5′ | 3.96 (1H, m) | 78.3 | 3.96 (1H, m) | 78.3 |
6′ | 4.50 (1H, m) 4.58 (1H, br d, 11.0) | 62.9 | 4.49 (1H, m) 4.58 (1H, br d, 11.5) | 62.9 |
1″ | 5.40 (1H, d, 7.5) | 106.1 | 5.41 (1H, d, 7.5) | 106.1 |
2″ | 4.16 (1H, t, 10.0) | 77.2 | 4.16 (1H, t, 9.0) | 77.2 |
3″ | 4.33 (1H, m) | 78.4 | 4.34 (1H, m) | 78.4 |
4″ | 4.36 (1H, m) | 71.8 | 4.36 (1H, m) | 71.8 |
5″ | 3.94 (1H, m) | 78.2 | 3.94 (1H, m) | 78.1 |
6″ | 4.32 (1H, m) 4.50 (1H, m) | 62.8 | 4.38 (1H, br d, 11.5) 4.50 (1H, m) | 62.8 |
Ginsenoside | C-17 | C-20 | C-21 | C-22 |
---|---|---|---|---|
20(R)-methoxyl-ginsenoside Rg3 | 47.2 | 80.2 | 18.5 | 35.7 |
20(S)-methoxyl-ginsenoside Rg3 | 50.0 | 79.9 | 21.2 | 35.1 |
Δ20(R)-20(S)δ | −2.8 | 0.3 | −2.7 | 0.6 |
20(R)-ginsenoside Rg3 [27] | 50.7 | 73.0 | 22.8 | 43.3 |
20(S)-ginsenoside Rg3 | 54.8 | 73.0 | 27.1 | 35.9 |
Δ20(R)-20(S)δ | −4.1 | 0 | −4.3 | 7.4 |
20(R)-ginsenoside Rf2 [27] | 51.8 | 73.4 | 22.9 | 43.6 |
20(S)-ginsenoside Rf2 | 54.7 | 72.5 | 27.2 | 36.4 |
Δ20(R)-20(S)δ | −2.9 | 0.9 | −4.3 | 7.2 |
20(R)-ginsenoside Rh2 [27] | 52.2 | 73.4 | 23.0 | 43.7 |
20(S)-ginsenoside Rh2 | 54.8 | 72.9 | 26.9 | 35.2 |
Δ20(R)-20(S)δ | −2.6 | 0.5 | −3.9 | 8.5 |
Compounds * | Calibration Curve | Correlation Coefficient (R2) | Test Range (μg/mL) | LOD (μg) | LOQ (μg) |
---|---|---|---|---|---|
1 | y = 2359.3x + 65.67 | 0.9998 | 8–80 | 0.086 | 0.262 |
2 | y = 2714.9x + 46.11 | 0.9998 | 100–1000 | 0.074 | 0.275 |
3 | y = 6665.5x + 270.39 | 0.9998 | 400–1500 | 0.092 | 0.258 |
4 | y = 3332.6x + 139.30 | 0.9999 | 1000–5000 | 0.069 | 0.232 |
5 | y = 2640.6x + 140.72 | 0.9994 | 100–1000 | 0.088 | 0.263 |
Compounds | Precision | |||
---|---|---|---|---|
Intra-Day (n = 3) | Inter-Day (n = 5) | |||
Content (μg/mL) | RSD (%) | Content (μg/mL) | RSD (%) | |
1 | 80.34 ± 1.02 | 1.51 | 80.58 ± 1.24 | 1.54 |
2 | 200.26 ± 1.52 | 0.76 | 200.47 ± 1.82 | 0.91 |
3 | 501.72 ± 2.55 | 0.51 | 500.29 ± 2.78 | 0.56 |
4 | 1401.89 ± 4.93 | 0.35 | 1401.25 ± 5.78 | 0.41 |
5 | 203.57 ± 1.58 | 0.78 | 202.15 ± 2.51 | 1.24 |
Compounds | Original (μg) | Spiked (μg) | Found (μg) | Recovery (%) | RSD (%) |
---|---|---|---|---|---|
1 | 5.70 | 35.00 | 40.58 ± 0.41 | 97.89 | 1.00 |
2 | 106.92 | 150.00 | 256.26 ± 2.08 | 99.38 | 0.81 |
3 | 512.03 | 500.00 | 1010.06 ± 5.23 | 99.61 | 0.52 |
4 | 3123.73 | 400.00 | 3521.27 ± 5.65 | 99.92 | 0.16 |
5 | 240.24 | 200.00 | 439.16 ± 5.36 | 99.54 | 1.22 |
Compounds | Content (μg/g FBPG) (n = 9) (mean ± SD) |
---|---|
1 | 1.77 ± 0.93 |
2 | 33.26 ± 7.46 |
3 | 159.31 ± 17.12 |
4 | 971.28 ± 28.05 |
5 | 74.65 ± 8.42 |
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Share and Cite
Li, S.-S.; Li, K.-K.; Xu, F.; Tao, L.; Yang, L.; Chen, S.-X.; Gong, X.-J. A Strategy for Simultaneous Isolation of Less Polar Ginsenosides, Including a Pair of New 20-Methoxyl Isomers, from Flower Buds of Panax ginseng. Molecules 2017, 22, 442. https://doi.org/10.3390/molecules22030442
Li S-S, Li K-K, Xu F, Tao L, Yang L, Chen S-X, Gong X-J. A Strategy for Simultaneous Isolation of Less Polar Ginsenosides, Including a Pair of New 20-Methoxyl Isomers, from Flower Buds of Panax ginseng. Molecules. 2017; 22(3):442. https://doi.org/10.3390/molecules22030442
Chicago/Turabian StyleLi, Sha-Sha, Ke-Ke Li, Fei Xu, Li Tao, Li Yang, Shu-Xiao Chen, and Xiao-Jie Gong. 2017. "A Strategy for Simultaneous Isolation of Less Polar Ginsenosides, Including a Pair of New 20-Methoxyl Isomers, from Flower Buds of Panax ginseng" Molecules 22, no. 3: 442. https://doi.org/10.3390/molecules22030442