Optimized Extraction of Total Triterpenoids from Jujube (Ziziphus jujuba Mill.) and Comprehensive Analysis of Triterpenic Acids in Different Cultivars
Abstract
:1. Introduction
2. Results and Discussion
2.1. UAE Process Optimization
2.1.1. Model Fitting
2.1.2. Model Validation
2.2. Triterpenic Acid Contents in the 99 Jujube Samples
2.3. HCA and PCA
3. Materials and Methods
3.1. Plant Materials
3.2. Chemicals
3.3. Determination of Total Triterpenoid Content (TTC)
3.4. Analysis of Triterpenic Acids by UPLC–MS
3.5. Analysis of Antioxidant Activities
3.6. UAE Procedures
3.7. RSM Experimental Design
3.8. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Run | X1: Temperature (°C) | X2: Ethanol Concentration (%) | X3: Time (min) | X4: Liquid-to-Solid Ratio (mL/g) | Y: Total Triterpenoid Yield (mg/g) |
---|---|---|---|---|---|
1 | 40 | 90 | 35 | 35 | 16.60 |
2 | 50 | 80 | 40 | 35 | 18.68 |
3 | 40 | 85 | 35 | 25 | 14.13 |
4 | 60 | 85 | 40 | 35 | 17.72 |
5 | 50 | 85 | 30 | 25 | 16.82 |
6 | 40 | 85 | 35 | 45 | 16.40 |
7 | 50 | 85 | 35 | 35 | 19.25 |
8 | 40 | 85 | 30 | 35 | 16.13 |
9 | 50 | 80 | 35 | 25 | 17.18 |
10 | 60 | 80 | 35 | 35 | 17.96 |
11 | 50 | 85 | 35 | 35 | 19.18 |
12 | 50 | 90 | 40 | 35 | 18.58 |
13 | 60 | 90 | 35 | 35 | 18.08 |
14 | 50 | 90 | 35 | 45 | 18.93 |
15 | 50 | 85 | 35 | 35 | 19.12 |
16 | 50 | 85 | 40 | 25 | 16.78 |
17 | 50 | 80 | 35 | 45 | 18.81 |
18 | 50 | 90 | 30 | 35 | 18.84 |
19 | 40 | 80 | 35 | 35 | 16.15 |
20 | 60 | 85 | 35 | 45 | 18.01 |
21 | 50 | 80 | 30 | 35 | 18.64 |
22 | 50 | 90 | 35 | 25 | 17.04 |
23 | 50 | 85 | 30 | 45 | 18.73 |
24 | 50 | 85 | 35 | 35 | 19.05 |
25 | 60 | 85 | 30 | 35 | 17.95 |
26 | 40 | 85 | 40 | 35 | 16.42 |
27 | 50 | 85 | 35 | 35 | 19.08 |
28 | 50 | 85 | 40 | 45 | 18.69 |
29 | 60 | 85 | 35 | 25 | 16.38 |
Source | Sum of Squares | df | Mean Square | F Value | p-value |
---|---|---|---|---|---|
Model | 45.29 | 14 | 3.24 | 229.64 | <0.0001 |
X1: Temperature | 8.79 | 1 | 8.79 | 623.9 | <0.0001 |
X2: Ethanol concentration | 0.035 | 1 | 0.035 | 2.5 | 0.1362 |
X3: Time | 0.0048 | 1 | 0.0048 | 0.34 | 0.5687 |
X4: Liquid-to-solid ratio | 10.53 | 1 | 10.53 | 747.33 | <0.0001 |
X1X2 | 0.027 | 1 | 0.027 | 1.93 | 0.1862 |
X1X3 | 0.068 | 1 | 0.068 | 4.8 | 0.0459 |
X1X4 | 0.1 | 1 | 0.1 | 7.27 | 0.0174 |
X2X3 | 0.023 | 1 | 0.023 | 1.6 | 0.2269 |
X2X4 | 0.017 | 1 | 0.017 | 1.2 | 0.2919 |
X3X4 | 0 | 1 | 0 | 0 | 1 |
X12 | 21.3 | 1 | 21.3 | 1512.04 | <0.0001 |
X22 | 0.089 | 1 | 0.089 | 6.32 | 0.0248 |
X32 | 0.61 | 1 | 0.61 | 43.09 | <0.0001 |
X42 | 7.37 | 1 | 7.37 | 523.14 | <0.0001 |
Residual | 0.2 | 14 | 0.014 | ||
Lack of Fit | 0.17 | 10 | 0.017 | 2.67 | 0.1785 |
Pure Error | 0.026 | 4 | 0.00643 | ||
Cor Total | 45.49 | 28 | |||
Adeq Precision | 56.589 | ||||
R2 = 0.9957; Adj R2 = 0.9913; Pred R2 = 0.9774 |
Peak No. | Retention Time (min) | Compound | [M + H]− (m/z) | Regression Equation | R2 |
---|---|---|---|---|---|
1 | 1.72 | Maslinic acid isomer-1 | 471.34 | — | — |
2 | 2.16 | Maslinic acid isomer-2 | 471.34 | — | — |
3 | 2.56 | Maslinic acid isomer-3 | 471.34 | — | — |
4 | 2.76 | Maslinic acid isomer-4 | 471.34 | — | — |
5 | 3.29 | Alphitolic acid | 471.34 | y = 138799x + 1595.7 | 0.9994 |
6 | 3.99 | Maslinic acid | 471.34 | y = 140553x + 1065.1 | 0.9996 |
7 | 4.34 | 2α-hydroxy ursolic acid | 471.34 | — | — |
8 | 5.04 | Maslinic acid isomer-5 | 471.34 | — | — |
9 | 5.41 | Oleanolic acid isomer-1 | 455.35 | — | — |
10 | 5.72 | Maslinic acid isomer-6 | 471.34 | — | — |
11 | 6.3 | Maslinic acid isomer-7 | 471.34 | — | — |
12 | 7.64 | Betulinic acid | 455.35 | y = 125572x + 1121.5 | 0.9991 |
13 | 9.17 | Oleanolic acid | 455.35 | y = 90033x - 1164.7 | 0.9992 |
14 | 9.49 | Ursolic acid | 455.35 | y = 113372x + 1035.8 | 0.9997 |
15 | 11.09 | Betulonic acid | 455.35 | — | — |
16 | 12.59 | Oleanonic acid + Ursonic acid | 455.35 | — | — |
Ma1 | Ma 2 | Ma3 | Ma4 | Aa | Ma | 2αHa | Ma5 | Oa1 | Ma6 | Ma7 | Ba | Oa | Ua | Ba’ | Oa’ + Ua’ | Total | ABTS | DPPH | |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
ABTS | 0.1274 | 0.1943 | –0.0398 | 0.2576 a | 0.9285 b | 0.5205 b | 0.0146 | 0.1462 | –0.0792 | –0.0141 | –0.051 | 0.4838 b | 0.1736 | 0.3378 b | 0.464 b | –0.009 | 0.694 b | - | 0.5205 b |
FRAP | 0.1174 | 0.0713 | 0.2629 | 0.1938 | 0.5549 b | 0.9475 b | 0.1955 | –0.1852 | 0.0911 | –0.013 | –0.0387 | 0.3508 b | 0.6123 b | 0.2048 a | 0.0111 | 0.1888 | 0.583 b | 0.4993 b | - |
Variables | Cluster 1 | Cluster 2 | Cluster 3 | Cluster 4 | Cluster 5 |
---|---|---|---|---|---|
Maslinic acid isomer-1 | 0.046 b | 0.022 | 0.040 | 0.011 a | 0.020 |
Maslinic acid isomer-2 | 0.355 | 1.706 | 7.917 b | 0.332 | 0.266 a |
Maslinic acid isomer-3 | 25.352 | 17.883 | 161.294 b | 6.081 a | 65.097 |
Maslinic acid isomer-4 | 11.305 b | 2.839 | 2.802 | 0.655 | 0.316 a |
Alphitolic acid | 1535.713 b | 1245.100 | 1189.970 | 555.637 a | 826.015 |
Maslinic acid | 449.873 b | 203.267 | 328.307 | 136.641 a | 287.112 |
2α-hydroxy ursolic acid | 40.014 | 29.682 | 216.845 b | 29.278 a | 127.938 |
Maslinic acid isomer-5 | 82.301 | 149.580 | 216.391 b | 79.385 | 49.260 a |
Oleanolic acid isomer-1 | 12.814 b | 8.929 | 7.839 | 7.487 a | 8.074 |
Maslinic acid isomer-6 | 49.880 | 74.380 | 160.215 b | 51.110 | 38.100 a |
Maslinic acid isomer-7 | 2.833 a | 9.938 | 70.101 b | 3.801 | 16.411 |
Betulinic acid | 1827.349 | 1909.479 | 3158.536 b | 1043.670 a | 1411.514 |
Oleanolic acid | 317.869 | 217.850 | 568.121 b | 159.657 a | 308.816 |
Ursolic acid | 92.502 a | 92.296 a | 530.525 b | 112.549 | 216.542 |
Betulonic acid | 78.186 | 109.488 b | 107.236 | 48.038 | 42.063 a |
Oleanonic acid + Ursonic acid | 40.246 | 33.499 | 86.389 b | 26.385 a | 50.759 |
Total | 4572.323 | 4104.121 | 6814.528 b | 2258.410 a | 3448.962 |
ABTS | 3.346 | 2.938 | 3.066 b | 1.488 a | 2.132 |
FRAP | 3.931 b | 2.645 | 3.172 | 2.336 a | 3.208 |
Component | Eigenvalue | Percentage of Variance (%) | Cumulative (%) |
---|---|---|---|
PC1 | 6.04 | 31.79 | 31.79 |
PC2 | 3.89 | 20.46 | 52.25 |
PC3 | 2.70 | 14.23 | 66.49 |
PC4 | 1.34 | 7.04 | 73.53 |
Code | X1: Temperature (°C) | X2: Ethanol Concentration (%) | X3: Time (min) | X4: Liquid-to-solid Ratio (mL/g) |
---|---|---|---|---|
−1 | 40 | 80 | 30 | 25:1 |
0 | 50 | 85 | 35 | 35:1 |
+1 | 60 | 90 | 40 | 45:1 |
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Song, L.; Zhang, L.; Xu, L.; Ma, Y.; Lian, W.; Liu, Y.; Wang, Y. Optimized Extraction of Total Triterpenoids from Jujube (Ziziphus jujuba Mill.) and Comprehensive Analysis of Triterpenic Acids in Different Cultivars. Plants 2020, 9, 412. https://doi.org/10.3390/plants9040412
Song L, Zhang L, Xu L, Ma Y, Lian W, Liu Y, Wang Y. Optimized Extraction of Total Triterpenoids from Jujube (Ziziphus jujuba Mill.) and Comprehensive Analysis of Triterpenic Acids in Different Cultivars. Plants. 2020; 9(4):412. https://doi.org/10.3390/plants9040412
Chicago/Turabian StyleSong, Lijun, Li Zhang, Long Xu, Yunjian Ma, Weishuai Lian, Yongguo Liu, and Yonghua Wang. 2020. "Optimized Extraction of Total Triterpenoids from Jujube (Ziziphus jujuba Mill.) and Comprehensive Analysis of Triterpenic Acids in Different Cultivars" Plants 9, no. 4: 412. https://doi.org/10.3390/plants9040412