Identification and Determination of Aconitum Alkaloids in Aconitum Herbs and Xiaohuoluo Pill Using UPLC-ESI-MS
Abstract
:1. Introduction
2. Results and Discussion
2.1. LC-MS Fingerprints
2.1.1. UPLC-ESI-MS Fingerprint Analysis of Aconitum Herbs and Processed Products
Peak No. | tR (min) | m/z | Molecular Formula | Identification | VIP |
---|---|---|---|---|---|
1 | 2.07 | 408.4 | C23H37NO6 | cammaconine b | 3.91 |
2 | 3.08 | 424.4 | C23H37NO6 | senbusine A b | 5.55 |
3 | 3.14 | 378.4 | C22H35NO4 | aconosine b | 7.62 |
4 | 4.04 | 454.5 | C24H39NO7 | delcisine b | 6.43 |
5 | 4.37 | 408.4 | C23H37NO6 | isotalatizidine b | 7.37 |
6 | 4.45 | 422.5 | C24H39NO5 | talatizamine b | 9.46 |
7 | 5.00 | 590.7 | C31H43NO10 | benzoylmesaconitine a | 9.29 |
8 | 5.76 | 438.5 | C24H39NO6 | neoline b | 8.21 |
9 | 6.34 | 360.4 | C22H33NO3 | lepenine b | 5.68 |
10 | 6.38 | 574.7 | C31H43NO9 | benzoylhypaconitine a | 6.63 |
11 | 9.04 | 358.3 | C22H31NO3 | songorine b | 5.33 |
12 | 12.18 | 632.7 | C33H45NO11 | mesaconitine a | 9.12 |
13 | 12.91 | 616.8 | C33H45NO10 | hypaconitine a | 8.8 |
14 | 13.22 | 646.8 | C34H47NO11 | aconitine a | 5.59 |
15 | 13.61 | 630.8 | C34H47NO10 | deoxyaconitine b | 7.94 |
16 | 13.75 | 402.4 | C24H35NO4 | lucidusculine b | 3.59 |
2.1.2. Changes in Chemical Components of Aconitum Herbs and Processed Products
Peak No. | tR (min) | Identification | VIP |
---|---|---|---|
13 | 12.91 | hypaconitine | 13.70 |
12 | 12.18 | mesaconitine | 12.93 |
15 | 13.61 | deoxyaconitine | 12.43 |
7 | 5.00 | benzoylmesaconitine | 11.41 |
4 | 4.04 | delcisine | 9.75 |
5 | 4.37 | isotalatizidine | 9.75 |
14 | 13.22 | aconitine | 8.71 |
9 | 6.34 | lepenine | 7.74 |
2.1.3. Chemical Fingerprint of Xiaohuoluo Pill
2.2. Quantitative Analysis of Xiaohuoluo Pill
2.2.1. Optimization of UPLC-MS Conditions
2.2.2. Method Validation: Linearity, Limits of Detection, and Quantification
Analyte | Regressive equation a | r | Linear range (ng·mL−1) | LLOD a (ng·mL−1) | LLOQ b (ng·mL−1) |
---|---|---|---|---|---|
MA | y = 135.01x + 309.64 | 0.9993 | 1.41–501.00 | 0.45 | 1.41 |
AC | y = 54.337x – 21.315 | 0.9999 | 1.20–396.80 | 0.39 | 1.20 |
HA | y =148.34x – 130.21 | 0.9999 | 1.92–533.18 | 0.65 | 1.92 |
BMA | y = 6385.8x + 1390.3 | 0.9984 | 1004.00–20080.00 | 0.82 | 4.28 |
BAC | y = 15.025x + 314.98 | 0.9989 | 1.99–2490.00 | 0.54 | 1.99 |
BHA | y = 17.273x + 129.08 | 0.9993 | 2.02–1518.00 | 0.51 | 2.02 |
2.2.3. Validation: Precision, Repeatability, Stability, and Recovery
Compound | Precision RSD (%) (n = 6) | Reproducibility RSD (%) (n = 6) | Stability RSD (%) (n = 6) | Recovery (%) (n = 9)Mean ± RSD (%) |
---|---|---|---|---|
MA | 2.40 | 2.26 | 1.76 | 99.8 ± 2.49 |
AC | 2.36 | 3.78 | 1.42 | 100.9 ± 2.65 |
HA | 2.82 | 1.79 | 1.11 | 101.7 ± 1.94 |
BMA | 3.11 | 2.56 | 1.44 | 100.3 ± 1.06 |
BAC | 3.07 | 3.07 | 1.05 | 99.7 ± 1.32 |
BHA | 2.89 | 2.50 | 3.09 | 98.1 ± 1.08 |
2.2.4. Sample Analysis
Analytes/Contents (μg·g−1) | ||||||
---|---|---|---|---|---|---|
Samples | MA | AC | HA | BMA | BAC | BHA |
S-01 | 0.31 ± 0.01 | 7.55 ± 0.16 | 8.05 ± 0.10 | 295.20 ± 6.20 | 88.37 ± 2.16 | 45.45 ± 0.55 |
S-02 | N.D. a | 0.18 ± 0.003 | 0.67 ± 0.01 | 327.18 ± 7.71 | 62.10 ± 0.53 | 54.75 ± 2.03 |
S-03 | 0.60 ± 0.02 | 0.78 ± 0.01 | 5.22 ± 0.12 | 343.38 ± 1.03 | 27.46 ± 0.21 | 52.02 ± 0.23 |
S-04 | 0.02 ± 0.002 | 0.07 ± 0.01 | 0.13 ± 0.01 | 28.52 ± 2.32 | 3.57 ± 0.29 | 3.11 ± 0.24 |
S-05 | 0.24 ± 0.004 | 2.15 ± 0.02 | 2.16 ± 0.02 | 55.71 ± 0.82 | 11.12 ± 0.13 | 9.49 ± 0.01 |
S-06 | 0.11 ± 0.003 | 0.13 ± 0.01 | 0.35 ± 0.01 | 14.71 ± 0.80 | 1.28 ± 0.05 | 1.34 ± 0.08 |
S-07 | 0.18 ± 0.003 | 0.80 ± 0.004 | 0.96 ± 0.01 | 117.80 ± 8.21 | 13.31 ± 1.09 | 11.61 ± 0.60 |
S-08 | 0.19 ± 0.002 | 0.84 ± 0.01 | 0.70 ± 0.01 | 137.56 ± 4.35 | 16.47 ± 0.40 | 14.34 ± 0.44 |
S-09 | 0.19 ± 0.002 | 0.83 ± 0.01 | 0.99 ± 0.01 | 129.54 ± 1.43 | 15.33 ± 0.01 | 13.36 ± 0.01 |
3. Experimental
3.1. Chemicals and Reagents
3.2. Materials
Name | No. | Source | Batch No. |
---|---|---|---|
Concentrated pill | S-01 | Foci, Lanzhou | 10B5 |
S-02 | Taibao, Lanzhou | 62101204 | |
S-03 | Dalu, Shandong | 110101 | |
Honey pill | S-04 | Wohua, Shandong | 110302 |
S-05 | Shiyitang, Harbin | 1101147 | |
S-06 | Tongrentang, Beijing | 0013070 | |
S-07 | Hongjitang, Jinan | 1011001 | |
S-08 | Hongjitang, Jinan | 1104001 | |
S-09 | Hongjitang, Jinan | 1104002 |
3.3. UPLC-ESI-MS Conditions
3.3.1. UPLC-MS Qualitative Conditions
3.3.2. UPLC-MS Quantitative Conditions
3.4. Preparation of Standard and Quality Control Samples
3.5. Preparation of Sample Solution
3.5.1. Xiaohuoluo Pill
3.5.2. Aconitum Herbs and Processed Products
4. Conclusions
Acknowledgments
References
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Cui, P.; Han, H.; Wang, R.; Yang, L. Identification and Determination of Aconitum Alkaloids in Aconitum Herbs and Xiaohuoluo Pill Using UPLC-ESI-MS. Molecules 2012, 17, 10242-10257. https://doi.org/10.3390/molecules170910242
Cui P, Han H, Wang R, Yang L. Identification and Determination of Aconitum Alkaloids in Aconitum Herbs and Xiaohuoluo Pill Using UPLC-ESI-MS. Molecules. 2012; 17(9):10242-10257. https://doi.org/10.3390/molecules170910242
Chicago/Turabian StyleCui, Ping, Han Han, Rui Wang, and Li Yang. 2012. "Identification and Determination of Aconitum Alkaloids in Aconitum Herbs and Xiaohuoluo Pill Using UPLC-ESI-MS" Molecules 17, no. 9: 10242-10257. https://doi.org/10.3390/molecules170910242