Simultaneous Separation and Purification of Five Polymethoxylated Flavones from “Dahongpao” Tangerine (Citrus tangerina Tanaka) Using Macroporous Adsorptive Resins Combined with Prep-HPLC
Abstract
:1. Introduction
2. Results and Discussion
2.1. Static Adsorption and Desorption Capacities
2.2. Adsorption Kinetics
2.3. Adsorption Isotherms
2.4. Dynamic Adsorption and Desorption
2.4.1. Optimal Feeding Speed
2.4.2. Optimal Concentration of Total PMFs
2.4.3. Impurities Removal
2.4.4. Optimal Desorption Ethanol Concentration
2.4.5. Optimal Eluting Speed
2.5. Effect of Enrichment by HPD 300 Resin on PMFs Extraction
2.6. Separation and Purification of PMFs
2.6.1. Optimal Conditions of Prep-HPLC
2.6.2. Identification of Isolated PMF Monomers
3. Materials and Methods
3.1. Plant Materials and Reagents
3.2. Preparation of Crude Extracts from “Dahongpao” Tangerine
3.3. Determination of PMF Contents
3.4. Static Adsorption and Desorption Tests
3.4.1. Pretreatment of Macroporous Adsorptive Resins
3.4.2. Static Adsorption and Desorption Properties of the Resins
3.4.3. Adsorption Kinetics of Selected Resin
3.4.4. Adsorption Isotherms on the Selected Resin
3.5. Dynamic Adsorption and Desorption Tests
3.6. Comparison of PMFs before and after Enrichment
3.7. Separation of PMFs by MAR
3.7.1. Preparation of PMF Solutions
3.7.2. Optimization of Experimental Conditions for Prep-HPLC
3.7.3 Identification of PMFs
3.8. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Sample Availability: 5,6,7,4’- tetramethoxyflavone, nobiletin, tangeretin, sinensetin, and 5-hydroxy -6,7,8,3’,4’-pentamethoxyflavone are available from the authors. |
MAR Type | Polarity | Particle Diameter (mm) | Surface Area (m2/g) | Average Pore Dimeter (Å) |
---|---|---|---|---|
D 101 | Non-polar | 0.315–1.25 | 650–700 | 100–110 |
HPD 100 | Non-polar | 0.30–1.25 | 650–700 | 85–90 |
AB-8 | Weak polar | 0.30–1.25 | 480–520 | 130–140 |
HPD 300 | Weak polar | 0.30–1.25 | 800–870 | 50–55 |
HPD 400 | Middle polar | 0.30–1.25 | 500–550 | 75–80 |
DM 130 | Middle polar | 0.30–1.25 | 500–550 | 90–100 |
NKA-9 | Strong polar | 0.30–1.25 | 250–290 | 155–165 |
HPD 600 | Strong polar | 0.30–1.20 | 550–600 | 80 |
MAR Type | Adsorption Capacity (mg/g) | Desorption Capacity (mg/g) | Desorption Rate (%) |
---|---|---|---|
D 101 | 227.51 ± 5.87 c | 194.59 ± 19.49 c | 85.45 ± 6.36% ab |
HPD 100 | 271.52 ± 9.60 b | 248.17 ± 4.01 b | 91.43 ± 1.75% a |
AB-8 | 275.13 ± 3.67 b | 241.59 ± 1.51 b | 87.82 ± 1.73% ab |
HPD 300 | 301.75 ± 3.32 a | 274.01 ± 0.40 a | 90.81 ± 0.83% a |
HPD 400 | 265.57 ± 2.71 b | 239.17 ± 2.51 b | 90.06 ± 0.03% a |
DM 130 | 236.29 ± 1.82 c | 200.88 ± 2.66 c | 85.02 ± 1.78% ab |
NKA-9 | 209.03 ± 2.04 d | 173.06 ± 0.65 d | 82.80 ± 1.12% b |
HPD 600 | 238.03 ± 5.77 c | 206.98 ± 2.72 c | 87.00 ± 3.25% ab |
Models | Equations | Parameters | Values |
---|---|---|---|
Pseudo-first order model | ln(175.2235-Qt) = 5.1661 − 5.5651t | R2 | 0.7920 |
Qe (mg/g) | 175.2235 | ||
k1 (L/min) | 5.5651 | ||
Pseudo-second order model | t/Qt = 0.0054t + 0.0007 | R2 | 0.9999 |
Qe (mg/g) | 185.1852 | ||
k2 (g/(mg min)) | 0.0417 | ||
Intra-particle diffusion model | (first stage) | R2 | 0.9660 |
k3 (mg/(min1/2·g)) | 75.147 | ||
C(mg/g) | 73.414 | ||
(second stage) | R2 | 0.6008 | |
k3(mg/(min1/2·g)) | 2.2816 | ||
C(mg/g) | 174.45 |
Models | Equations | T (°C) | Parameters | ||||
---|---|---|---|---|---|---|---|
R2 | KL | Qm | KF | 1/n | |||
Langmuir | Ce/Qe = 0.00010Ce + 0.00425 | 25 | 0.9710 | 44.1981 | 235.1302 | - | - |
Ce/Qe = 0.00017Ce + 0.00448 | 35 | 0.9845 | 26.6570 | 223.2240 | - | - | |
Ce/Qe = 0.00021Ce + 0.00537 | 45 | 0.9889 | 26.1808 | 186.2802 | - | - | |
Freundlich | Qe = 314.5Ce0.2790 | 25 | 0.9637 | - | - | 314.5000 | 0.2790 |
Qe = 281.7663Ce0.3091 | 35 | 0.9473 | - | - | 281.7663 | 0.3091 | |
Qe = 224.44Ce0.2927 | 45 | 0.9251 | - | - | 224.4400 | 0.2927 |
No. | Contents | Before Enrichment (mg/g) | After Enrichment (mg/g) | Increased Times |
---|---|---|---|---|
1 | Isosinensetin | 1.239 ± 0.046 | 20.59 ± 1.65 | 16.62 |
2 | Sinensetin | 1.913 ± 0.066 | 37.56 ± 3.04 | 19.63 |
3 | 5,7,3′,4′-Tetrathoxyflavone | 0.056 ± 0.001 | 1.92 ± 0.60 | 34.29 |
4 | 5,6,7,4′-Tetrathoxyflavone | 1.207 ± 0.113 | 24.06 ± 3.79 | 19.93 |
5 | Nobiletin | 19.27 ± 0.743 | 355.09 ± 18.45 | 18.43 |
6 | 3,5,6,7,8,3′,4′-Hetamethoxyflavone | 0.554 ± 0.021 | 12.14 ± 1.56 | 21.91 |
7 | 5,7,4′-Trimethoxyflavone | 0.019 ± 0.005 | 1.67 ± 0.16 | 87.89 |
8 | 5-Hydroxy-6,7,8,3′,4′-pentamethoxyflavone | 1.555 ± 0.063 | 30.27 ± 2.57 | 19.47 |
9 | Tangeretin | 5.908 ± 0.229 | 111.27 ± 7.10 | 18.83 |
10 | Total PMFs | 31.720 ± 1.255 | 594.57 ± 28.05 | 18.74 |
Contents | CAS No. | Molecular Formula | Monoisotopic Mass (g/mol) | Purity |
---|---|---|---|---|
Isosinensetin | 17290-70-9 | C20H20O7 | 372.1209 | >98.0% |
Sinensetin | 2306-27-6 | C20H20O7 | 372.1209 | ≥98.0% |
5,7,3’,4’-Tetrathoxyflavone | 855-97-0 | C19H18O6 | 342.1103 | ≥97.0% |
5,6,7,4’-Tetrathoxyflavone | 1168-42-9 | C19H18O6 | 342.1103 | ≥98.0% |
Nobiletin | 478-01-3 | C21H22O8 | 402.1315 | ≥95.0% |
3,5,6,7,8,3’,4’-Hetamethoxyflavone | 1178-24-1 | C22H24O9 | 432.1420 | >98.0% |
5,7,4’-Trimethoxyflavone | 5631-70-9 | C18H16O5 | 312.0998 | ≥98.7% |
5-Hydroxy-6,7,8,3’,4’-pentamethoxyflavone | 2174-59-6 | C20H20O8 | 388.1158 | ≥98.0% |
Tangeretin | 481-53-8 | C20H20O7 | 372.1209 | ≥95.0% |
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Li, Z.; Zhao, Z.; Zhou, Z. Simultaneous Separation and Purification of Five Polymethoxylated Flavones from “Dahongpao” Tangerine (Citrus tangerina Tanaka) Using Macroporous Adsorptive Resins Combined with Prep-HPLC. Molecules 2018, 23, 2660. https://doi.org/10.3390/molecules23102660
Li Z, Zhao Z, Zhou Z. Simultaneous Separation and Purification of Five Polymethoxylated Flavones from “Dahongpao” Tangerine (Citrus tangerina Tanaka) Using Macroporous Adsorptive Resins Combined with Prep-HPLC. Molecules. 2018; 23(10):2660. https://doi.org/10.3390/molecules23102660
Chicago/Turabian StyleLi, Zhenqing, Ziyan Zhao, and Zhiqin Zhou. 2018. "Simultaneous Separation and Purification of Five Polymethoxylated Flavones from “Dahongpao” Tangerine (Citrus tangerina Tanaka) Using Macroporous Adsorptive Resins Combined with Prep-HPLC" Molecules 23, no. 10: 2660. https://doi.org/10.3390/molecules23102660