UHPLC-MS/MS Quantification Combined with Chemometrics for Comparative Analysis of Different Batches of Raw, Wine-Processed, and Salt-Processed Radix Achyranthis Bidentatae
Abstract
1. Introduction
2. Results
2.1. Optimization of Extraction Conditions
2.2. Optimization of Chromatographic and Mass Spectrometric Conditions
2.3. Method Validation
2.4. Quantitative Analysis and Boxplot Analysis of Raw, Wine-Processed, and Salt-Processed Products
2.5. Chemometric Analysis
2.5.1. Hierarchical Clustering Analysis
2.5.2. Principal Component Analysis
3. Materials and Methods
3.1. Chemicals and Reagents
3.2. Preparation of Sample Solutions
3.3. UHPLC–MS/MS Analysis Conditions
3.4. Validation of the Method
3.5. Qualitative and Quantitative Analysis
3.6. Applications for Different Batches of Raw, Wine-Processed, and Salt-Processed Products
3.7. Data Processing
4. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Sample Availability: Samples are available from the authors. |
Analytes | tR (min) | [M − H]− | SRM Transitions (Precursor→Product) | Collision Energy (eV) |
---|---|---|---|---|
β-ecdysterone | 2.85 | 479.519 | 479.519→319.0 | 28.54 |
25-R inokosterone | 3.24 | 479.519 | 479.519→319.0 | 28.54 |
25-S inokosterone | 3.52 | 479.519 | 479.519→319.0 | 28.54 |
ginsenoside R0 | 8.65 | 955.745 | 955.745→793.5 | 55.00 |
chikusetsusaponin IVa | 8.75 | 793.678 | 793.678→631.4 | 47.04 |
glycyrrhizin (I.S.) | 8.52 | 255.207 | 255.207→119.0 | 25.43 |
Analytes | Calibration Curves | r2 | Linearity Ranges (mg/mL) | LOD (mg/mL) | LOQ (mg/mL) |
---|---|---|---|---|---|
β-ecdysterone | y = 0.0545x − 0.0558 | 0.9993 | 1.90–494.0 | 0.365 | 1.085 |
25-S inokosterone | y = 0.3174x − 0.3118 | 0.9995 | 1.35–270.0 | 0.332 | 0.995 |
25-R inokosterone | y = 0.3814x − 0.5633 | 0.9993 | 1.75–315.0 | 0.499 | 1.485 |
ginsenoside R0 | y = 0.4588x − 0.4343 | 0.9994 | 2.10–210.0 | 0.325 | 0.955 |
Chikusetsusaponin IVa | y = 0.6858x − 0.7556 | 0.9994 | 1.85–185.0 | 0.369 | 1.105 |
Analytes | Precision | Repeatability | Stability | Recovery (%, mean/RSD, n = 3) | |||
---|---|---|---|---|---|---|---|
Intra-Day | Inter-Day | Low | Medium | High | |||
β-ecdysterone | 1.57 | 2.25 | 2.95 | 2.68 | 98.91 (0.01) | 99.31 (0.07) | 99.29 (0.07) |
25-S inokosterone | 2.46 | 1.51 | 2.25 | 2.54 | 98.98 (0.47) | 101.17 (0.85) | 98.58 (1.86) |
25-R inokosterone | 1.83 | 1.72 | 2.58 | 2.97 | 101.48 (0.14) | 98.68 (0.42) | 99.81 (0.90) |
ginsenoside R0 | 2.34 | 1.68 | 2.16 | 2.58 | 101.09 (0.28) | 100.57 (0.72) | 100.89 (0.15) |
Chikusetsusaponin IVa | 1.78 | 3.00 | 3.00 | 2.63 | 101.41 (0.23) | 99.75 (0.84) | 99.97 (0.78) |
No. | Region or Pharmacy (Specimen No.) | Content of Investigated Components (n = 3, mg/g ± SD) | ||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
β-Ecdysterone (1) | 25-R Inokosterone (2) | 25-S Inokosterone (3) | Ginsenoside R0 (4) | Chikusetsusaponin IVa (5) | ||||||||||||
RRAB | WRAB | SRAB | RRAB | WRAB | SRAB | RRAB | WRAB | SRAB | RRAB | WRAB | SRAB | RRAB | WRAB | SRAB | ||
S1 | Wenxian, Henan (NWH2016-01) | 12.778 ± 0.318 | 31.214 ± 0.198 | 88.051 ± 0.410 | 0.636 ± 0.006 | 1.453 ± 2.545 | 4.667 ± 0.791 | 0.880 ± 0.009 | 1.935 ± 0.025 | 6.153 ± 0.084 | 0.740 ± 0.024 | 4.396 ± 0.020 | 11.221 ± 0.009 | 0.956 ± 0.010 | 2.943 ± 0.001 | 13.864 ± 0.226 |
S2 | Zhengzhou, Henan (NZH2016-02) | 26.493 ± 0.167 | 32.577 ± 0.002 | 81.066 ± 0.387 | 1.153 ± 0.034 | 1.477 ± 0.001 | 4.117 ± 0.097 | 1.458 ± 0.019 | 1.897 ± 0.029 | 41.570 ± 0.716 | 0.400 ± 0.002 | 3.880 ± 0.024 | 12.268 ± 0.068 | 0.444 ± 0.002 | 1.905 ± 0.007 | 13.608 ± 0.448 |
S3 | Zhoukou, Henan (NZH2016-03) | 38.670 ± 0.194 | 38.554 ± 0.002 | 81.746 ± 0.335 | 1.680 ± 0.002 | 1.798 ± 0.073 | 4.417 ± 0.030 | 2.216 ± 0.016 | 2.308 ± 0.003 | 5.852 ± 0.011 | 2.232 ± 0.002 | 3.033 ± 0.076 | 9.5488 ± 0.006 | 1.319 ± 0.002 | 1.784 ± 0.003 | 12.519 ± 0.019 |
S4 | Jiefang, Henan (NJH2016-04) | 23.406 ± 0.123 | 30.819 ± 0.017 | 50.605 ± 0.404 | 1.083 ± 0.002 | 1.423 ± 0.003 | 2.982 ± 0.560 | 1.437 ± 0.036 | 1.844 ± 0.003 | 3.979 ± 0.096 | 7.520 ± 0.005 | 5.866 ± 0.001 | 12.925 ± 0.108 | 3.250 ± 0.197 | 4.525 ± 0.014 | 16.179 ± 0.006 |
S5 | Wushe, Henan (NWH2016-05) | 26.280 ± 0.262 | 35.231 ± 0.021 | 92.555 ± 0.613 | 1.141 ± 0.067 | 1.516 ± 0.003 | 4.677 ± 0.061 | 1.554 ± 0.002 | 2.045 ± 0.011 | 6.177 ± 0.085 | 6.108 ± 0.030 | 5.323 ± 0.023 | 11.986 ± 0.049 | 1.715 ± 0.001 | 3.131 ± 0.004 | 13.634 ± 0.017 |
S6 | Heshan, Henan (NHH2016-06) | 14.076 ± 0.229 | 43.287 ± 0.025 | 77.559 ± 0.796 | 0.727 ± 0.004 | 1.948 ± 0.001 | 4.149 ± 0.014 | 0.986 ± 0.010 | 2.495 ± 0.001 | 5.545 ± 0/243 | 0.406 ± 0.002 | 0.580 ± 0.001 | 12.358 ± 0.383 | 0.483 ± 0.01 | 2.438 ± 0.002 | 13.716 ± 0.09 |
S7 | Jinshui, Henan (NJH2016-07) | 11.332 ± 0.015 | 39.833 ± 0.020 | 70.805 ± 0.042 | 0.599 ± 0.013 | 1.754 ± 0.001 | 3.794 ± 0.018 | 0.812 ± 0.004 | 2.271 ± 0.003 | 4.892 ± 0.196 | 0.385 ± 0.005 | 3.604 ± 0.008 | 6.401 ± 0.054 | 0.504 ± 0.006 | 2.078 ± 0.017 | 6.0731 ± 0.022 |
S8 | Luoyang, Henan (NLH2016-08) | 24.191 ± 0.036 | 36.948 ± 0.003 | 68.983 ± 0.428 | 1.115 ± 0.002 | 1.578 ± 0.004 | 3.789 ± 1.459 | 1.472 ± 0.003 | 2.035 ± 0.003 | 4.952 ± 0.144 | 6.654 ± 0.014 | 4.394 ± 0.001 | 13.239 ± 3.978 | 1.322 ± 0.008 | 2.544 ± 0.002 | 13.989 ± 0.004 |
S9 | Xiayi, Henan (NXH2016-09) | 25.973 ± 0.215 | 35.182 ± 0.055 | 67.382 ± 0.603 | 1.230 ± 0.067 | 1.594 ± 0.002 | 29.085 ± 0.013 | 1.601 ± 0.006 | 1.984 ± 0.006 | 4.955 ± 0.049 | 12.930 ± 0.011 | 4.867 ± 0.023 | 22.730 ± 0.091 | 1.611 ± 0.007 | 3.099 ± 0.034 | 24.632 ± 0.006 |
S10 | Qinyang, Henan (NQH2016-10) | 16.420 ± 0.106 | 4.202 ± 0.024 | 79.573 ± 0.177 | 0.806 ± 0.034 | 1.779 ± 0.001 | 4.242 ± 0.472 | 1.080 ± 0.002 | 2.354 ± 0.005 | 5.639 ± 0,.181 | 6.467 ± 0.108 | 9.142 ± 0.007 | 28.838 ± 0.164 | 2.272 ± 0.008 | 6.523 ± 0.034 | 30.468 ± 0.503 |
S11 | Kaifeng, Henan (NKH2016-11) | 30.524 ± 0.013 | 21.687 ± 0.204 | 92.820 ± 0.568 | 1.397 ± 0.004 | 1.051 ± 0.001 | 4.966 ± 0.033 | 1.829 ± 0.004 | 1.359 ± 0.079 | 6.458 ± 0.035 | 4.089 ± 0.030 | 3.540 ± 0.042 | 4.935 ± 0.004 | 1.142 ± 0.002 | 1.616 ± 0.001 | 7.0181 ± 0.007 |
S12 | Lankao, Henan (NLH2016-12) | 12.690 ± 0.101 | 37.268 ± 0.031 | 56.059 ± 0.068 | 0.646 ± 0.002 | 1.712 ± 0.057 | 3.267 ± 0.004 | 0.882 ± 0.001 | 2.208 ± 0.025 | 4.335 ± 0.019 | 1.310 ± 0.001 | 3.470 ± 0.004 | 16.011 ± 0.037 | 0.728 ± 0.001 | 1.656 ± 0.021 | 10.215 ± 0.016 |
S13 | Shanyang, Henan (NSH2016-13) | 28.123 ± 0.053 | 29.411 ± 0.175 | 44.903 ± 0.086 | 1.255 ± 0.043 | 1.315 ± 0.001 | 2.648 ± 0.047 | 1.635 ± 0.001 | 1.715 ± 0.005 | 3.567 ± 0.109 | 10.360 ± 0.001 | 6.113 ± 0.001 | 6.812 ± 0.153 | 2.427 ± 0.010 | 4.202 ± 0.147 | 4.517 ± 0.07 |
S14 | Nanyang, Henan (NNH2016-14) | 7.6519 ± 0.064 | 26.536 ± 0.157 | 68.005 ± 0.099 | 0.446 ± 0.006 | 1.173 ± 0.001 | 3.612 ± 0.581 | 0.621 ± 0.002 | 1.562 ± 0.008 | 4.790 ± 0.014 | 1.100 ± 0.001 | 4.006 ± 0.003 | 17.141 ± 0.070 | 0.700 ± 0.001 | 2.133 ± 0.024 | 8.525 ± 0.002 |
S15 | Macun, Henan (NMH2016-15) | 16.737 ± 0.003 | 24.189 ± 0.034 | 78.139 ± 0.497 | 0.768 ± 0.014 | 1.057 ± 0.057 | 3.823 ± 0.289 | 1.078 ± 0.001 | 1.468 ± 0.010 | 5.214 ± 0.015 | 10.151 ± 0.08 | 7.249 ± 0.004 | 17.032 ± 0.052 | 2.862 ± 0.008 | 4.830 ± 0.007 | 16.423 ± 0.008 |
S16 | Longting, Henan (NLH2016-16) | 23.450 ± 0.014 | 39.351 ± 0.050 | 59.081 ± 0.377 | 1.051 ± 0.015 | 1.726 ± 0.001 | 3.246 ± 0.003 | 1.414 ± 0.001 | 2.272 ± 0.014 | 4.353 ± 0.109 | 2.905 ± 0.135 | 4.513 ± 0.003 | 9.875 ± 0.227 | 1.212 ± 0.010 | 1.750 ± 0.006 | 3.0279 ± 0.001 |
S17 | Gulou, Henan (NGH2016-17) | 15.539 ± 0.174 | 30.572 ± 0.003 | 67.816 ± 0.331 | 0.716 ± 0.003 | 1.279 ± 0.003 | 3.343 ± 0.001 | 0.995 ± 0.004 | 1.722 ± 0.09 | 4.628 ± 0.0119 | 2.034 ± 0.206 | 3.646 ± 0.006 | 5.127 ± 0.042 | 1.005 ± 0.001 | 1.253 ± 0.001 | 6.234 ± 0.029 |
S18 | Xiangfu, Henan (NXH2016-18) | 30.854 ± 1.209 | 33.943 ± 0.003 | 96.379 ± 0.332 | 1.304 ± 0.003 | 1.513 ± 0.046 | 4.800 ± 0.020 | 1.768 ± 0.001 | 1.991 ± 0.006 | 6.297 ± 0.198 | 5.200 ± 0.147 | 3.600 ± 00.004 | 11.494 ± 0.029 | 1.797 ± 0.001 | 1.635 ± 0.004 | 5.432 ± 0.001 |
S19 | Mengxian, Henan (NMH2016-19) | 22.038 ± 0.009 | 26.264 ± 0.013 | 83.783 ± 0.210 | 1.022 ± 0.015 | 1.159 ± 0.001 | 4.127 ± 0.272 | 1.343 ± 0.005 | 1.536 ± 0.004 | 5.558 ± 0.073 | 5.119 ± 0.040 | 3.545 ± 1.156 | 13.331 ± 0.006 | 2.164 ± 0.001 | 2.846 ± 0.005 | 10.735 ± 0.001 |
S20 | Boai, Henan (NBH2016-20) | 10.088 ± 0.009 | 38.828 ± 0.003 | 86.465 ± 1.138 | 0.536 ± 0.002 | 1.695 ± 0.001 | 4.360 ± 0.023 | 0.743 ± 0.001 | 2.202 ± 0.086 | 5.632 ± 0.074 | 2.009 ± 0.011 | 5.517 ± 0.023 | 21.533 ± 0.012 | 1.157 ± 0.001 | 4.126 ± 0.015 | 15.114 ± 0.001 |
S21 | Hongqi, Henan (NHH2016-21) | 18.976 ± 1.070 | 27.053 ± 0.020 | 35.614 ± 2.982 | 0.833 ± 0.020 | 1.141 ± 0.001 | 2.210 ± 0.083 | 1.152 ± 0.02 | 1.569 ± 0.012 | 3.121 ± 0.068 | 11.546 ± 0.037 | 6.552 ± 0.016 | 13.514 ± 0.001 | 3.277 ± 0.002 | 4.511 ± 0.179 | 8.721 ± 0.017 |
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Yang, L.; Jiang, H.; Yan, M.; Xing, X.; Guo, X.; Yang, B.; Wang, Q.; Kuang, H. UHPLC-MS/MS Quantification Combined with Chemometrics for Comparative Analysis of Different Batches of Raw, Wine-Processed, and Salt-Processed Radix Achyranthis Bidentatae. Molecules 2018, 23, 758. https://doi.org/10.3390/molecules23040758
Yang L, Jiang H, Yan M, Xing X, Guo X, Yang B, Wang Q, Kuang H. UHPLC-MS/MS Quantification Combined with Chemometrics for Comparative Analysis of Different Batches of Raw, Wine-Processed, and Salt-Processed Radix Achyranthis Bidentatae. Molecules. 2018; 23(4):758. https://doi.org/10.3390/molecules23040758
Chicago/Turabian StyleYang, Liu, Hai Jiang, Meiling Yan, Xudong Xing, Xinyue Guo, Bingyou Yang, Qiuhong Wang, and Haixue Kuang. 2018. "UHPLC-MS/MS Quantification Combined with Chemometrics for Comparative Analysis of Different Batches of Raw, Wine-Processed, and Salt-Processed Radix Achyranthis Bidentatae" Molecules 23, no. 4: 758. https://doi.org/10.3390/molecules23040758
APA StyleYang, L., Jiang, H., Yan, M., Xing, X., Guo, X., Yang, B., Wang, Q., & Kuang, H. (2018). UHPLC-MS/MS Quantification Combined with Chemometrics for Comparative Analysis of Different Batches of Raw, Wine-Processed, and Salt-Processed Radix Achyranthis Bidentatae. Molecules, 23(4), 758. https://doi.org/10.3390/molecules23040758