Extraction Optimization of Astragaloside IV by Response Surface Methodology and Evaluation of Its Stability during Sterilization and Storage
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Quantitation of Astragaloside I, Astragaloside II, and Astragaloside IV
2.3. Quantitative Validation Calibration Curves
2.3.1. Calibration Curves, Limits of Detection (LOD) and Quantification (LOQ)
2.3.2. Precision and Accuracy
2.4. Single-Factor Experiments of the Extraction Process
2.4.1. Ammonia Concentration
2.4.2. Solid–Liquid Ratio
2.4.3. Soaking Time
2.4.4. Extraction Time
2.4.5. Extraction Temperature
2.4.6. Extraction Times
2.5. Response Surface Method to Optimize Extraction Conditions
2.6. The Retention Rate of the Astragaloside IV Under Different Sterilization Conditions
2.7. The Retention Rate of Astragaloside IV in Different pH Value Solutions Stored at 4 or 25 °C
2.8. Statistical Analysis
3. Results and Discussion
3.1. Identification of Astragaloside I, Astragaloside II, and Astragaloside IV by UPLC–MS/MS
Quantitative Validation
3.2. Single-Factor Experiments
3.2.1. Ammonia Concentration
3.2.2. Solid–Liquid Ratio
3.2.3. Soaking Time
3.2.4. Extraction Time
3.2.5. Extraction Temperature
3.2.6. Extraction Times
3.3. Response Surface Methodology
3.3.1. Response Surface Experimental Design and Response
3.3.2. The ANOVA of Response Surface Methodology
3.3.3. The Impact of Different Factors on the Yield of Astragaloside IV
3.3.4. Optimization and Validation Procedures
3.4. The Changes in the Astragaloside IV Retention Rate Under Different Sterilization Conditions
3.5. Changes in the Astragaloside IV Retention Rate under Different Storage Conditions
4. Conclusions
Author Contributions
Funding
Conflicts of Interest
Sample Availability
Appendix A
Treatment Number | Ammonia Concentration (A) (%, v/v) | Solid–Liquid Ratio (B) (mL/g) | Soaking Time (C) (min) | Extraction Time (D) (min) | Content of Astragaloside IV (mg/g) | Astragaloside Ⅰ (mg/g) | Astragaloside Ⅱ (mg/g) |
---|---|---|---|---|---|---|---|
1 | 25 | 10 | 90 | 0 | 2.012 ± 0.073 | Not detected | Not detected |
2 | 25 | 20 | 120 | 30 | 2.301 ± 0.011 | Not detected | Not detected |
3 | 25 | 10 | 60 | 30 | 1.904 ± 0.023 | Not detected | Not detected |
4 | 25 | 15 | 120 | 0 | 2.235 ± 0.084 | Not detected | Not detected |
5 | 30 | 15 | 60 | 30 | 0.701 ± 0.036 | 0.107 ± 0.005 | 0.142 ± 0.010 |
6 | 25 | 20 | 90 | 0 | 2.049 ± 0.042 | Not detected | Not detected |
7 | 25 | 20 | 90 | 60 | 2.283 ± 0.086 | Not detected | Not detected |
8 | 25 | 15 | 90 | 30 | 2.215 ± 0.061 | Not detected | Not detected |
9 | 30 | 15 | 120 | 30 | 1.436 ± 0.021 | Not detected | 0.086 ± 0.002 |
10 | 30 | 20 | 90 | 30 | 0.938 ± 0.025 | 0.067 ± 0.004 | 0.103 ± 0.003 |
11 | 20 | 15 | 60 | 30 | 1.473 ± 0.077 | Not detected | 0.079 ± 0.002 |
12 | 25 | 15 | 90 | 30 | 2.157 ± 0.113 | Not detected | Not detected |
13 | 30 | 15 | 90 | 0 | 0.851 ± 0.032 | 0.076 ± 0.003 | 0.129 ± 0.004 |
14 | 30 | 10 | 90 | 30 | 0.940 ± 0.023 | 0.071 ± 0.002 | 0.116 ± 0.003 |
15 | 25 | 15 | 60 | 60 | 1.981 ± 0.049 | Not detected | Not detected |
16 | 20 | 20 | 90 | 30 | 1.702 ± 0.026 | Not detected | Not detected |
17 | 25 | 20 | 60 | 30 | 2.063 ± 0.036 | Not detected | Not detected |
18 | 25 | 10 | 120 | 30 | 2.602 ± 0.072 | Not detected | Not detected |
19 | 25 | 15 | 90 | 30 | 2.151 ± 0.034 | Not detected | Not detected |
20 | 25 | 15 | 90 | 30 | 2.220 ± 0.065 | Not detected | Not detected |
21 | 20 | 15 | 90 | 0 | 1.566 ± 0.073 | Not detected | 0.067 ± 0.002 |
22 | 20 | 15 | 90 | 60 | 1.757 ± 0.029 | Not detected | Not detected |
23 | 20 | 15 | 120 | 30 | 1.861 ± 0.016 | Not detected | Not detected |
24 | 25 | 15 | 90 | 30 | 2.205 ± 0.035 | Not detected | Not detected |
25 | 25 | 10 | 90 | 60 | 2.102 ± 0.039 | Not detected | Not detected |
26 | 30 | 15 | 90 | 60 | 0.882 ± 0.026 | 0.069 ± 0.003 | 0.125 ± 0.002 |
27 | 25 | 15 | 120 | 60 | 2.590 ± 0.108 | Not detected | Not detected |
28 | 20 | 10 | 90 | 30 | 1.660 ± 0.025 | Not detected | 0.062 ± 0.002 |
29 | 25 | 15 | 60 | 0 | 1.801 ± 0.049 | Not detected | Not detected |
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The Factors | Level | ||
---|---|---|---|
−1 | 0 | 1 | |
Ammonia concentration (A) (%, v/v) | 20 | 25 | 30 |
Solid–liquid ratio (B) (mL/g) | 10:1 | 15:1 | 20:1 |
Soaking time (C) (min) | 60 | 90 | 120 |
Extraction time (D) (min) | 0 | 30 | 60 |
Qualitative Ion Pair | Quantitative Ion Pair | Cone Hole Voltage/V | Collision Energy/eV | |
---|---|---|---|---|
Astragaloside I | 891.5 > 891.5 | 891.5 > 891.5 | 13 | 10 |
Astragaloside II | 849.4 > 849.4 | 849.4 > 849.4 | 14 | 9 |
827.5 > 827.5 | 14 | 10 | ||
827.5 > 143 | 14 | 11 | ||
Astragaloside IV | 785.4 > 473.39 | 785.4 > 143.11 | 14 | 12 |
785.4 > 143.11 | 14 | 10 |
Parameters | Substances | ||
---|---|---|---|
Astragaloside I | Astragaloside II | Astragaloside IV | |
Calibration curve | y = 400.12x + 1309.5 | y = 655.11x + 1353.9 | y = 1047.7x + 197.46 |
Test range (μg/mL) | 0.083–2.67 | 0.042–1.33 | 0.026–0.83 |
r2 | 0.9991 | 0.9935 | 0.9997 |
LOD (μg) | 0.006 | 0.003 | 0.002 |
LOQ (μg) | 0.024 | 0.015 | 0.009 |
Intra-day RSD (%) (n = 5) | 1.5 | 3.7 | 1.4 |
Inter-day RSD (%) (n = 5) | 1.8 | 1.2 | 1.1 |
Recoveries (%) | 94.1 ± 4.3 | 98.4 ± 5.8 | 96.5 ± 1.9 |
Content of Astragaloside IV | Sum of Squares | Degrees of Freedom | Mean Square | F-Value | p-value |
---|---|---|---|---|---|
Model | 7.71 | 14 | 0.55 | 135.37 | <0.0001 |
A | 1.52 | 1 | 1.52 | 373.33 | <0.0001 |
B | 8.00 × 10−4 | 1 | 8.00 × 10−4 | 0.20 | 0.6642 |
C | 0.80 | 1 | 0.80 | 196.77 | <0.0001 |
D | 0.099 | 1 | 0.099 | 24.42 | 0.0002 |
AB | 6.25 × 10−4 | 1 | 6.25 × 10−4 | 0.15 | 0.7010 |
AC | 0.029 | 1 | 0.029 | 7.10 | 0.0185 |
AD | 6.40 × 10−3 | 1 | 6.40 × 10−3 | 1.57 | 0.2304 |
BC | 0.053 | 1 | 0.053 | 13.00 | 0.0029 |
BD | 5.77 × 10−3 | 1 | 5.77 × 10−3 | 1.42 | 0.2533 |
CD | 8.1 × 10−3 | 1 | 8.1 × 10−3 | 1.99 | 0.1801 |
A2 | 4.81 | 1 | 4.81 | 1181.04 | <0.0001 |
B2 | 9.87 × 10−4 | 1 | 9.87 × 10−4 | 0.24 | 0.6301 |
C2 | 9.33 × 10−3 | 1 | 9.33 × 10−3 | 2.29 | 0.1523 |
D2 | 0.031 | 1 | 0.031 | 7.50 | 0.0160 |
Residual | 0.057 | 14 | 4.07 × 10−3 | ||
Lack of Fit | 0.052 | 10 | 5.25 × 10−3 | 4.64 | 0.0761 |
Pure Error | 4.52 × 10−3 | 4 | 1.13 × 10−3 | ||
Cor Total | 7.77 | 28 | |||
R-Squared | 0.9927 | ||||
Adj R-Squared | 0.9853 | ||||
C.V. % | 3.52 | ||||
Adequate Precision | 41.691 |
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Xu, L.; Wei, K.; Jiang, J.; Zhang, L. Extraction Optimization of Astragaloside IV by Response Surface Methodology and Evaluation of Its Stability during Sterilization and Storage. Molecules 2021, 26, 2400. https://doi.org/10.3390/molecules26082400
Xu L, Wei K, Jiang J, Zhang L. Extraction Optimization of Astragaloside IV by Response Surface Methodology and Evaluation of Its Stability during Sterilization and Storage. Molecules. 2021; 26(8):2400. https://doi.org/10.3390/molecules26082400
Chicago/Turabian StyleXu, Lin, Kongjiong Wei, Jiaolong Jiang, and Lianfu Zhang. 2021. "Extraction Optimization of Astragaloside IV by Response Surface Methodology and Evaluation of Its Stability during Sterilization and Storage" Molecules 26, no. 8: 2400. https://doi.org/10.3390/molecules26082400