Simultaneous Quantification of Five Sesquiterpene Components after Ultrasound Extraction in Artemisia annua L. by an Accurate and Rapid UPLC–PDA Assay †
Abstract
:1. Introduction
2. Results
2.1. Single Factor Experiment for Extraction Optimization
2.2. Orthogonal Experiment for Extraction Optimization
2.3. Optimization of UPLC Separation
2.4. Regression Equation and Linearity
2.5. Precision, Accuracy, Stability, Repeatability and Recovery
2.6. Determination of Content
3. Discussion
4. Materials and Methods
4.1. Plant Materials
4.2. Chemicals and Reagents
4.3. Apparatus and Chromatographic Conditions
4.4. Preparation of Standard Solutions
4.5. Preparation of the Extracts and Sample Solutions
4.6. Calibration Curve and Lowest Limit of Quantification (LLOQ)
4.7. Precision, Accuracy, Stability, Repeatability and Recovery
Author Contributions
Funding
Conflicts of Interest
References
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Sample Availability: Samples of the compounds are available from the authors. |
Compound | Linear Range (μg/mL) | Regression Equation (y = ax + b) a) | R | LLOQ b) (μg/mL) | |
---|---|---|---|---|---|
Slope (a) | Intercept (b) | ||||
Art | 31.44–1572 | 0.8332 | 7.996 | 1.000 | 31.44 |
Art B | 25.48–1274 | 8.265 | 109.9 | 0.9996 | 25.48 |
Art C | 40.56–2028 | 0.4145 | 1.707 | 0.9998 | 40.56 |
DHAA | 31.44–1572 | 9.186 | 38.13 | 0.9998 | 31.44 |
AA | 26.88–1396 | 14.28 | 175.6 | 0.9990 | 26.88 |
Compound | Fortified Conc. (μg/mL) | Intra-Day (n = 5) | Inter-Day (n = 5) | ||||
---|---|---|---|---|---|---|---|
Observed Conc. (μg/mL) | Precision a) (%) | Accuracy b) (%) | Observed Conc. (μg/mL) | Precision (%) | Accuracy (%) | ||
Art | 786.0 | 817.6 | 0.21 | 104.0 | 807.9 | 1.27 | 102.8 |
314.4 | 330.1 | 0.15 | 105.0 | 324.3 | 2.42 | 103.1 | |
157.2 | 163.6 | 0.09 | 104.1 | 162.4 | 1.89 | 103.3 | |
Art B | 637.0 | 660.3 | 0.13 | 103.7 | 605.8 | 0.32 | 95.10 |
254.8 | 266.5 | 0.22 | 104.6 | 246.9 | 0.24 | 96.90 | |
127.4 | 129.8 | 0.10 | 101.9 | 123.0 | 1.12 | 96.60 | |
Art C | 1014 | 1068 | 0.63 | 105.3 | 1037 | 2.16 | 102.2 |
405.6 | 391.6 | 1.49 | 96.60 | 395.6 | 2.66 | 97.50 | |
202.8 | 212.9 | 0.18 | 105.0 | 197.4 | 2.88 | 97.30 | |
DHAA | 786.0 | 821.4 | 0.08 | 104.5 | 821.4 | 1.69 | 104.5 |
314.4 | 327.3 | 0.17 | 104.1 | 321.9 | 2.34 | 102.4 | |
157.2 | 161.7 | 0.15 | 102.9 | 158.3 | 2.85 | 100.7 | |
AA | 672.0 | 701.9 | 0.15 | 104.4 | 705.6 | 2.76 | 105.0 |
268.8 | 281.7 | 0.19 | 104.8 | 280.8 | 2.10 | 104.5 | |
134.4 | 136.0 | 0.24 | 101.2 | 130.3 | 2.37 | 97.00 |
Compound | Stability | Repeatability | RSD (%) |
---|---|---|---|
RSD (%) | Retention Time (min) | Peak Area (AU) | |
Art | 1.63 | 0.069 | 2.37 |
Art B | 1.65 | 0.060 | 2.42 |
Art C | 1.52 | 0.070 | 2.04 |
DHAA | 1.45 | 0.091 | 2.15 |
AA | 0.96 | 0.092 | 1.64 |
Compound | Original Conc. | Spiked Conc. | Found Conc. | Recovery a) ± SD | RSD (%) |
---|---|---|---|---|---|
(μg/mL) | (μg/mL) | (μg/mL) | (%) | ||
Art | 116.1 | 117.0 | 233.7 | 100.6 ± 1.28 | 1.27 |
Art B | 179.5 | 180.0 | 358.8 | 99.6 ± 0.83 | 0.84 |
Art C | 10.70 | 13.00 | 23.23 | 96.5 ± 2.72 | 2.82 |
DHAA | 32.90 | 33.00 | 65.70 | 99.4 ± 2.67 | 1.69 |
AA | 282.5 | 285.0 | 561.6 | 97.9 ± 2.50 | 2.56 |
No. | Region | Art Content (mg/g) | Art B Content (mg/g) | Art C Content (mg/g) | DHAA Content (mg/g) | AA Content (mg/g) |
---|---|---|---|---|---|---|
1 | Xinjiang | 10.16 ± 0.011 | 5.347 ± 0.0023 | 2.700 ± 0.0025 | - | 3.449 ± 0.0015 |
2 | Hubei | 12.04 ± 0.017 | 8.108 ± 0.0015 | 2.228 ± 0.0021 | 0.8521 ± 0.00025 | 15.17 ± 0.0072 |
3 | Zhejiang1 | 5.457 ± 0.0031 | 4.581 ± 0.0020 | 1.153 ± 0.0016 | - | 6.459 ± 0.0016 |
4 | Shandong1 | 8.604 ± 0.0022 | 8.356 ± 0.0025 | 0.9970 ± 0.00036 | 0.6356 ± 0.00030 | 7.735 ± 0.0021 |
5 | Zhejiang2 | 27.59 ± 0.015 | 4.462 ± 0.0015 | 0.9026 ± 0.00021 | - | 3.022 ± 0.0019 |
6 | Hebei | 88.79 ± 0.010 | 1.289 ± 0.0020 | - | - | 1.406 ± 0.0021 |
7 | Shaanxi | 7.594 ± 0.0021 | 8.593 ± 0.0011 | 1.224 ± 0.0025 | - | 7.620 ± 0.0021 |
8 | Guizhou | 13.78 ± 0.010 | 3.332 ± 0.0015 | 1.980 ± 0.0019 | - | 4.499 ± 0.0020 |
9 | Hunan1 | 8.216 ± 0.0012 | 0.4597 ± 0.00070 | 0.9990 ± 0.00022 | 2.020 ± 0.0025 | 0.4449 ± 0.00024 |
10 | Beijing1 | 5.562 ± 0.0010 | 6.877 ± 0.0020 | 1.379 ± 0.0022 | 3.737 ± 0.0022 | 8.313 ± 0.0013 |
11 | Shandong2 | 4.457 ± 0.0020 | 7.391 ± 0.0014 | 1.494 ± 0.0016 | 0.4174 ± 0.00020 | 11.14 ± 0.0034 |
12 | Beijing2 | 2.303 ± 0.0019 | 3.562 ± 0.0021 | 0.2121 ± 0.00010 | 0.6527 ± 0.0019 | 5.606 ± 0.0024 |
13 | Hunan2 | 6.880 ± 0.0026 | 0.5579 ± 0.00041 | 0.8605 ± 0.00045 | 3.587 ± 0.0019 | 0.5311 ± 0.0078 |
14 | Liaoning | 3.649 ± 0.0016 | 6.323 ± 0.0022 | 1.338 ± 0.0017 | - | 6.477 ± 0.0029 |
15 | Hunan3 | 63.74 ± 0.010 | 4.028 ± 0.0015 | 2.967 ± 0.0015 | 13.58 ± 0.0057 | 3.231 ± 0.0021 |
Level | Factors | ||||
---|---|---|---|---|---|
A Method | B Solvent | C Ratio (g/mL) | D Time/h | E Cycles | |
1 | ultrasonic | methanol | 1:20 | 0.5 | 1 |
2 | Condensation reflux | petroleum ether | 1:30 | 1 | 2 |
3 | - | ethyl acetate | 1:40 | 2 | 3 |
Level | Factors | ||
---|---|---|---|
X ratio/(g/mL) | Y time/h | Z times | |
a | 1:10 | 0.25 | 1 |
b | 1:20 | 0.5 | 2 |
c | 1:30 | 1 | 3 |
No. | X | Y | Z |
---|---|---|---|
1 | a | a | a |
2 | a | b | b |
3 | a | c | c |
4 | b | a | b |
5 | b | b | c |
6 | b | c | a |
7 | c | a | c |
8 | c | b | a |
9 | c | c | b |
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Ruan, J.; Liu, Z.; Qiu, F.; Shi, H.; Wang, M. Simultaneous Quantification of Five Sesquiterpene Components after Ultrasound Extraction in Artemisia annua L. by an Accurate and Rapid UPLC–PDA Assay. Molecules 2019, 24, 1530. https://doi.org/10.3390/molecules24081530
Ruan J, Liu Z, Qiu F, Shi H, Wang M. Simultaneous Quantification of Five Sesquiterpene Components after Ultrasound Extraction in Artemisia annua L. by an Accurate and Rapid UPLC–PDA Assay. Molecules. 2019; 24(8):1530. https://doi.org/10.3390/molecules24081530
Chicago/Turabian StyleRuan, Jiaqi, Zhengyue Liu, Feng Qiu, Henan Shi, and Manyuan Wang. 2019. "Simultaneous Quantification of Five Sesquiterpene Components after Ultrasound Extraction in Artemisia annua L. by an Accurate and Rapid UPLC–PDA Assay" Molecules 24, no. 8: 1530. https://doi.org/10.3390/molecules24081530