Quantitative Evaluation of Twelve Major Components of Sulfur-Fumigated Astragali Radix with Different Durations by UPLC-MS
Abstract
:1. Introduction
2. Experimental
2.1. Chemicals and Reagents
2.2. Sulfur-Fumigation of AR
2.3. Instruments
2.4. Standard Solutions Preparation
2.5. Sample Preparation
2.6. Method Validation
2.6.1. Calibration Curves, Limits of Detection and Quantification
2.6.2. Precision, Repeatability and Stability
2.6.3. Recovery Test
3. Results and Discussion
3.1. Optimization of Suitable LC-MS Conditions
3.2. Method Validation
3.2.1. Calibration Curves, Limits of Detection and Quantification
3.2.2. Precision, Repeatability and Stability
3.2.3. Accuracy
3.3. Quantification of the Major Components in AR with and without Sulfur-Fumigation
3.4. General Procedure for the Synthesis of Flavonoid Glycosides
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Sample Availability: Samples of the compounds 1–12 are available from the authors. |
No. | Analyte | Precursor Ion (m/z) | Product Ion (m/z) | DP a (V) | CE b (V) |
---|---|---|---|---|---|
1 | Calycosin | 285.2 | 213.1 | 70 | 45 |
2 | Calycosin-7-glucoside | 447.3 | 285.4 | 60 | 20 |
3 | Formononetin | 269.0 | 167.2 | 70 | 47 |
4 | Ononin | 431.1 | 269.1 | 65 | 20 |
5 | Methylnissolin | 301.2 | 167.2 | 54 | 20 |
6 | Astraisoflavan-7-O-β-d-glucoside | 463.3 | 167.4 | 60 | 40 |
7 | 7,2′-dihydroxy-3′,4′-dimethoxyisoflavane | 303.0 | 167.2 | 55 | 19 |
8 | 7,2′-dihydroxy-3′,4′-dimethoxy isoflavan-7-O-β-d-glucopyranoside | 463.3 | 301.0 | −74 | −24 |
9 | Astragaloside I | 867.9 | 59.1 | −250 | −83 |
10 | Astragaloside II | 825.7 | 59.1 | −110 | −70 |
11 | Astragaloside III | 783.8 | 160.9 | −150 | −47 |
12 | Astragaloside IV | 783.8 | 101.0 | −115 | −57 |
Analytes | Regression Equation (μg/mL) | Linear Range (μg/mL) | Correlation Coefficient (R2) | LOQ (ng/mL) | LOD (ng/mL) | Precision RSD a (%) | Repeatability RSD (%) | Stability RSD (%) | |
---|---|---|---|---|---|---|---|---|---|
Intra-Day | Inter-Day | ||||||||
Calycosin | y = 446765x + 322603 | 0.4752~11.88 | 0.9989 | 0.59 | 0.06 | 2.06% | 3.16% | 4.47% | 3.93% |
Calycosin-7-glucoside | y = 202558x + 365088 | 0.864~21.6 | 0.9992 | 1.08 | 0.11 | 1.31% | 2.09% | 3.43% | 2.95% |
Formononetin | y = 469388x + 141426 | 0.3482~17.41 | 0.9986 | 0.87 | 0.35 | 1.65% | 1.69% | 4.09% | 3.37% |
Ononin | y = 51545x + 4316.5 | 0.31~15.5 | 0.9994 | 3.10 | 1.55 | 1.42% | 3.86% | 4.63% | 4.05% |
Methylnissolin | y = 329224x + 84783 | 0.6976~17.44 | 0.9993 | 0.09 | 0.02 | 1.46% | 2.53% | 4.31% | 2.99% |
Astraisoflavan-7-O-β-d-glucoside | y = 130675x + 208291 | 1.5226~38.064 | 0.9991 | 0.08 | 0.04 | 2.03% | 2.78% | 4.49% | 3.73% |
7,2′-dihydroxy-3′,4′-dimethoxyisoflavane | y = 57437x + 11250 | 0.2571~6.427 | 0.9990 | 0.32 | 0.35 | 1.96% | 3.53% | 4.92% | 3.77% |
7,2′-dihydroxy-3′,4′-dimethoxy isoflavan-7-O-β-d-glucopyranoside | y = 127060x + 236161 | 2.496~62.4 | 0.9988 | 0.01 | 0.001 | 1.25% | 2.37% | 3.68% | 1.50% |
Astragaloside I | y = 1009.3x − 10607 | 8.56~214 | 0.9991 | 0.09 | 0.04 | 2.36% | 3.18% | 4.84% | 2.52% |
Astragaloside II | y = 10750x + 25435 | 1.028~25.2 | 0.9993 | 0.03 | 0.002 | 1.65% | 3.78% | 2.66% | 2.03% |
Astragaloside III | y = 12640x + 30515 | 1.128~28.2 | 0.9993 | 0.03 | 0.01 | 1.41% | 2.19% | 2.65% | 1.87% |
Astragaloside IV | y = 6748.6x + 164720 | 4.584~114.6 | 0.9989 | 0.06 | 0.01 | 2.18% | 1.98% | 4.67% | 2.06% |
Analyte | Initial Amount (μg) | Added Amount (μg) | Detected Amount (μg) | Recovery (%) | RSD (%) |
---|---|---|---|---|---|
0.61 | 1.90 | 103.50% | 3.2 | ||
Calycosin | 1.22 | 1.22 | 2.48 | 101.50% | 1.5 |
1.83 | 3.01 | 98.50% | 2.3 | ||
3.735 | 4.79 | 96.70% | 2.7 | ||
Calycosin-7-glucoside | 7.47 | 7.47 | 14.51 | 97.10% | 0.9 |
11.205 | 19.28 | 103.20% | 1.7 | ||
1.21 | 3.47 | 95.40% | 1.6 | ||
Formononetin | 2.42 | 2.42 | 5.10 | 105.20% | 3.1 |
3.63 | 6.01 | 99.20% | 2.7 | ||
0.87 | 2.61 | 99.60% | 0.4 | ||
Ononin | 1.74 | 1.74 | 3.53 | 101.40% | 0.9 |
2.61 | 4.56 | 104.70% | 1.5 | ||
2.29 | 7.12 | 103.90% | 3.6 | ||
Methylnissolin | 4.57 | 4.57 | 9.36 | 102.30% | 3.7 |
6.86 | 12.06 | 105.50% | 3.0 | ||
8.54 | 26.67 | 104.10% | 2.6 | ||
Astraisoflavan-7-O-β-d-glucoside | 17.08 | 17.08 | 34.02 | 99.60% | 3.4 |
25.62 | 42.23 | 98.90% | 2.6 | ||
1.99 | 6.24 | 104.50% | 1.5 | ||
7,2′-dihydroxy-3′,4′-dimethoxyisoflavane | 3.98 | 3.98 | 8.15 | 102.30% | 1.6 |
5.97 | 10.11 | 101.60% | 3.2 | ||
1.26 | 3.87 | 102.70% | 3.1 | ||
7,2′-dihydroxy-3′,4′-dimethoxy | 2.51 | 2.51 | 4.96 | 98.80% | 2.3 |
3.77 | 6.39 | 101.80% | 5.7 | ||
isoflavan-7-O-β-d-glucopyranoside | 2.01 | 6.12 | 101.70% | 0.6 | |
Isoflavan-7-O-β-d-glucopyranoside | 4.01 | 4.01 | 7.90 | 98.40% | 2.3 |
6.02 | 9.58 | 95.50% | 2.6 | ||
1.23 | 3.79 | 103.10% | 3.1 | ||
Astragaloside I | 2.45 | 2.45 | 4.88 | 99.60% | 4.1 |
3.68 | 6.06 | 98.90% | 2.8 | ||
1.07 | 3.16 | 98.60% | 1.5 | ||
Astragaloside III | 2.14 | 2.14 | 4.40 | 102.90% | 2.7 |
3.21 | 5.30 | 99.10% | 3.1 | ||
4.33 | 13.50 | 104.10% | 1.8 | ||
Astragaloside IV | 8.65 | 8.65 | 16.83 | 97.30% | 3.1 |
12.98 | 22.44 | 103.80% | 2.2 |
Compounds | AR with and without Sulfur-Fumigation. | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 h | 2 h | 4 h | 6 h | 8 h | 12 h | 16 h | 24 h | 36 h | 48 h | 60 h | 72 h | Non-Fumigated | |
1 | 7.49 ± 0.03 | 5.04 ± 0.04 | 6.40 ± 0.02 | 7.36 ± 0.07 | 4.91 ± 0.00 | 5.97 ± 0.01 | 7.44 ± 0.10 | 5.37 ± 0.02 | 5.42 ± 0.05 | 4.86 ± 0.01 | 5.28 ± 0.06 | 6.03 ± 0.03 | 13.55 ± 0.68 |
2 | 4.24 ± 0.05 | 5.45 ± 0.02 | 5.10 ± 0.00 | 4.83 ± 0.01 | 4.61 ± 0.04 | 4.27 ± 0.02 | 4.07 ± 0.05 | 4.39 ± 0.03 | 4.33 ± 0.04 | 4.48 ± 0.01 | 3.92 ± 0.02 | 3.87 ± 0.02 | 3.21 ± 0.01 |
3 | 0.59 ± 0.01 | 0.59 ± 0.03 | 0.75 ± 0.02 | 0.78 ± 0.00 | 0.50 ± 0.02 | 0.64 ± 0.01 | 0.50 ± 0.03 | 0.62 ± 0.00 | 0.60 ± 0.02 | 0.56 ± 0.01 | 0.61 ± 0.02 | 0.63 ± 0.01 | 1.37 ± 0.04 |
4 | 0.81 ± 0.02 | 1.19 ± 0.09 | 1.04 ± 0.01 | 1.05 ± 0.04 | 0.88 ± 0.05 | 0.80 ± 0.07 | 1.06 ± 0.02 | 0.89 ± 0.00 | 0.91 ± 0.02 | 0.95 ± 0.01 | 0.76 ± 0.02 | 0.75 ± 0.01 | 0.40 ± 0.02 |
5 | 0.30 ± 0.01 | 0.25 ± 0.00 | 0.27 ± 0.02 | 0.27 ± 0.00 | 0.21 ± 0.00 | 0.22 ± 0.01 | 0.35 ± 0.02 | 0.22 ± 0.05 | 0.23 ± 0.01 | 0.22 ± 0.02 | 0.23 ± 0.00 | 0.22 ± 0.01 | 0.24 ± 0.01 |
6 | 0.55 ± 0.00 | 0.90 ± 0.01 | 0.69 ± 0.01 | 0.64 ± 0.02 | 0.59 ± 0.00 | 0.58 ± 0.05 | 0.53 ± 0.07 | 0.58 ± 0.00 | 0.52 ± 0.02 | 0.67 ± 0.00 | 0.47 ± 0.00 | 0.44 ± 0.01 | 0.32 ± 0.02 |
7 | 4.18 ± 0.06 | 2.61 ± 0.01 | 2.99 ± 0.05 | 3.44 ± 0.00 | 2.93 ± 0.02 | 3.29 ± 0.04 | 3.06 ± 0.01 | 2.51 ± 0.00 | 3.17 ± 0.02 | 2.60 ± 0.00 | 2.54 ± 0.00 | 3.24 ± 0.01 | 4.47 ± 0.10 |
8 | 0.86 ± 0.00 | 1.59 ± 0.00 | 1.38 ± 0.05 | 1.20 ± 0.07 | 1.13 ± 0.06 | 1.16 ± 0.02 | 0.95 ± 0.01 | 1.27 ± 0.00 | 1.08 ± 0.04 | 1.42 ± 0.06 | 0.97 ± 0.00 | 0.75 ± 0.00 | 0.68 ± 0.00 |
9 | 0.71 ± 0.01 | 0.74 ± 0.02 | 0.70 ± 0.00 | 0.71 ± 0.05 | 0.70 ± 0.00 | 0.70 ± 0.02 | 0.74 ± 0.04 | 0.69 ± 0.07 | 0.70 ± 0.00 | 0.68 ± 0.01 | 0.67 ± 0.02 | 0.70 ± 0.04 | 0.73 ± 0.04 |
10 | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND |
11 | 0.78 ± 0.06 | 0.84 ± 0.02 | 0.79 ± 0.01 | 0.70 ± 0.04 | 0.71 ± 0.00 | 0.73 ± 0.01 | 0.75 ± 0.00 | 0.59 ± 0.02 | 0.60 ± 0.00 | 0.64 ± 0.01 | 0.57 ± 0.02 | 0.58 ± 0.01 | 0.95 ± 0.03 |
12 | 3.97 ± 0.00 | 4.17 ± 0.02 | 3.73 ± 0.01 | 3.38 ± 0.01 | 3.59 ± 0.00 | 3.53 ± 0.02 | 3.61 ± 0.01 | 2.68 ± 0.05 | 2.81 ± 0.02 | 3.03 ± 0.04 | 2.62 ± 0.00 | 2.58 ± 0.01 | 4.94 ± 0.02 |
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Xing, X.; Sun, Z.; Yang, M.; Zhu, N.; Yang, J.; Ma, G.; Xu, X. Quantitative Evaluation of Twelve Major Components of Sulfur-Fumigated Astragali Radix with Different Durations by UPLC-MS. Molecules 2018, 23, 2609. https://doi.org/10.3390/molecules23102609
Xing X, Sun Z, Yang M, Zhu N, Yang J, Ma G, Xu X. Quantitative Evaluation of Twelve Major Components of Sulfur-Fumigated Astragali Radix with Different Durations by UPLC-MS. Molecules. 2018; 23(10):2609. https://doi.org/10.3390/molecules23102609
Chicago/Turabian StyleXing, Xiaoyan, Zhonghao Sun, Meihua Yang, Nailiang Zhu, Junshan Yang, Guoxu Ma, and Xudong Xu. 2018. "Quantitative Evaluation of Twelve Major Components of Sulfur-Fumigated Astragali Radix with Different Durations by UPLC-MS" Molecules 23, no. 10: 2609. https://doi.org/10.3390/molecules23102609