Comprehensive Quantitative Analysis of SQ Injection Using Multiple Chromatographic Technologies
Abstract
:1. Introduction
2. Results and Discussion
2.1. Qualitative Analysis of Constituents
2.2. Limitation of Qualitative Analysis Solely Using Mass Spectrometry
2.3. Method Validation
2.4. Quantification of Eighteen Analytes in Commercial Shenqi Fuzheng Injection (SQI) Samples
3. Materials and Methods
3.1. Chemicals and Materials
3.2. Sample Preparation
3.3. Ultra-Performance Liquid-Chromatography Tandem Mass Spectrometry (UPLC-MS) Conditions
3.4. HPLC-NH2P-ELSD Conditions
3.5. High Performance Gel Permeation Chromatography (HPGPC) Conditions
3.6. Acid Hydrolysis
3.7. Method Validation
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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No. | RT (Min) | Formula | [M + H]+ (Error, ppm) | [M − H]− (Error, ppm) | Fragment Ions in Positive Mode | Fragment Ions in Negative Mode | Identification |
---|---|---|---|---|---|---|---|
1 | 12.3 | C22H22O10 | 445.1150 (3.59) | 491.1215 [M + COOH]−, 481.0918 [M + Cl]−, 283.0623 [M − H − glc]−, 268.0375 [M − H − glc − H2O]−, 224.1435, 184.0517 | Calycosin-7-O-beta-d-glucoside | ||
2 | 14.9 | C9H16O4 | 187.0976 (3.21) | 169.0868, 125.0972 | Azelaic acid | ||
3 | 16.6 | C20H28O8 | 395.1728 (5.82) | 441.1773 [M + COOH]−, 431.1484 [M + Cl]−, 395.1701, 305.0586, 185.0970, 159.0812, 143.0708, 119.0351, 101.0243 | Lobetyolin | ||
4 | 19.2 | C23H28O10 | 463.1619 (3.24) | 509.1671 [M + COOH]−, 499.1383 [M + Cl]−, 345.9224, 301.1080, 254.0532, 135.0443 | Isomucronulatol-7-O-glucoside | ||
5 | 23.7 | C41H68O14 | 785.4671 (2.04) | 783.4554 (3.06) | (829.4593 [M + COOH]−, 819.4307 [M + Cl]−, 707.2920, 651.4105, 577.6834, 490.3592, 357.5564, 279.2332, 179.0567, 161.0450, 131.0343, 119.0352, 113.0245, 101.0246) | Astragaloside IV | |
6 | 23.8 | C41H68O14 | 783.4538 (1.02) | (829.4593 [M + COOH]−, 819.4297 [M + Cl]−, 621.4005, 489.3581, 394.8312, 279.2329, 161.0464, 113.0251, 101.0243) | Astragaloside III | ||
7 | 24.7 | C43H70O15 | 825.4628 (0.97) | (871.4710 [M + COOH]−, 861.4408 [M + Cl]−, 765.4429, 719.7998, 520.4337, 338.1722, 224.1057, 179.0572, 143.0355, 119.0348, 101.0252) | Astragaloside II | ||
8 | 25.2 | C48H78O18 | 943.5269 (0.32) | 941.5130 (2.23) | (879.5089, 733.4564, 615.3915, 457.3659, 362.4478, 247.0827, 163.0604) | Soyasaponin I | |
9 | 25.4 | C43H70O15 | 825.4659 (2.79) | (871.4702 [M + COOH]−, 861.4404 [M + Cl]−, 726.3011, 593.3695, 465.2882, 336.2658, 257.2371, 179.0583, 113.0243) | Isoastragaloside II | ||
10 | 26.0 | C43H70O15 | 825.4618 (2.18) | (871.4705 [M + COOH]−, 861.4405 [M + Cl]−, 765.4444, 603.3852, 335.0594, 223.8866, 179.0537, 161.0435, 143.0337, 113.0220) | Cyclocephaloside II | ||
11 | 1.8 | C10H13N5O4 | 268.1059 (5.22) | 211.9763, 178.0760, 136.0622, 119.0350 | Adenosine | ||
12 | 3.4 | C9H11NO2 | 166.0863 (3.01) | l-Phenylalanine | |||
13 | 5.1 | C14H21NO4 | 268.1556 (2.98) | 220.1272, 161.0537, 118.0810, 100.0709 | Codonopsine | ||
14 | 6.0 | C11H12N2O2 | [M + H − OH]+ 188.0943, (3.19) | Trytophan | |||
15 | 17.1 | C22H22O9 | 431.1343 (0.23) | 453.1158 [M + Na]+, 350.0444, 269.0813, 213.0919, 118.0416 | Ononin |
Analyte | EIC Ions | Linearity | LOD (µg/mL) | LOQ (µg/mL) | Repeatability RSD (%) (n = 6) | Stability | Spike Recovery (RSD %) (n = 3) | |||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
Regression Equation | R2 | Range (µg/mL) | Intra-Day | Inter-Day | RSD (%) (n = 6) | High | Middle | Low | ||||
1 | 283.07 | y = 85.094x − 47097 | 0.9997 | 3.04–12.16 | 0.15 | 0.49 | 1.61 | 3.74 | 3.12 | 101.00 (0.31) | 104.61 (2.25) | 95.32 (2.72) |
2 | 187.10 | y = 35.078x + 13967 | 0.9987 | 0.33–2.63 | 0.090 | 0.30 | 2.08 | 4.81 | 4.37 | 99.18 (3.01) | 99.56 (5.04) | 105.53 (1.83) |
3 | 441.18 | y = 31.521x − 15298 | 0.9981 | 1.22–3.80 | 0.14 | 0.47 | 2.03 | 2.27 | 3.18 | 97.11 (0.69) | 103.34 (2.45) | 96.33 (3.17) |
4 | 463.17 | y = 128.05x − 23035 | 0.9995 | 0.69–2.75 | 0.11 | 0.34 | 2.68 | 3.18 | 3.33 | 105.38 (0.71) | 104.63 (0.13) | 106.30 (0.53) |
5 | 829.47 | y = 595.32x + 39240 | 0.9992 | 0.50–4.00 | 0.0077 | 0.026 | 2.49 | 2.25 | 4.54 | 97.49 (0.80) | 105.62 (0.63) | 95.34 (2.88) |
6 | 829.47 | y = 453.52x − 9380.8 | 0.9991 | 0.19–0.93 | 0.0069 | 0.024 | 3.48 | 4.15 | 4.73 | 104.78 (1.25) | 101.74 (3.98) | 102.36 (3.40) |
7 | 871.48 | y = 410.39x − 220599 | 0.9990 | 0.57–2.85 | 0.11 | 0.36 | 1.32 | 1.33 | 1.48 | 100.62 (3.05) | 105.35 (1.31) | 95.35 (3.54) |
8 | 941.52 | y = 869.66x + 26058 | 0.9995 | 0.26–1.04 | 0.012 | 0.039 | 2.64 | 4.62 | 4.71 | 105.91 (5.70) | 106.59 (2.95) | 96.80 (3.58) |
9 | 871.48 | y = 393.52x + 9798.5 | 0.9992 | 0.52–2.60 | 0.14 | 0.49 | 1.69 | 2.90 | 3.88 | 94.97 (3.52) | 98.60 (2.60) | 105.16 (1.80) |
10 | 871.48 | y = 338.05x + 14751 | 0.9996 | 0.36–2.92 | 0.0048 | 0.016 | 1.59 | 2.39 | 2.75 | 102.57 (2.63) | 95.38 (3.06) | 96.58 (2.17) |
11 | 268.11 | y = 253.34x + 192461 | 0.9992 | 1.46–7.31 | 0.048 | 0.16 | 1.33 | 2.36 | 2.21 | 98.42 (1.79) | 99.91 (0.37) | 101.91 (1.91) |
12 | 120.08 | y = 159.12x + 104220 | 0.9991 | 1.48–7.38 | 0.102 | 0.34 | 1.38 | 2.45 | 2.16 | 98.82 (0.59) | 99.90 (3.16) | 98.75 (2.20) |
13 | 268.16 | y = 567.08x + 484891 | 0.9996 | 2.42–9.69 | 0.025 | 0.080 | 1.99 | 1.53 | 1.51 | 99.46 (0.68) | 99.75 (0.57) | 97.07 (6.77) |
14 | 188.09 | y = 1043.3x − 30639 | 0.9994 | 0.20–0.79 | 0.029 | 0.090 | 3.47 | 4.27 | 3.89 | 100.34 (0.94) | 101.60 (1.20) | 103.81 (4.25) |
15 | 431.14 | y = 125.32x − 18966 | 0.9996 | 0.87–2.24 | 0.17 | 0.56 | 3.74 | 4.33 | 4.31 | 103.98 (3.11) | 99.77 (3.71) | 105.11 (0.82) |
Analyte | SFI-1 | SFI-2 | SFI-3 | SFI-4 | SFI-5 | SFI-6 | SFI-7 | SFI-8 | SFI-9 | SFI-10 | SFI-11 | SFI-12 | SFI-13 |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | 7.53 | 7.60 | 7.52 | 7.60 | 7.69 | 7.90 | 7.25 | 7.22 | 7.46 | 7.45 | 5.67 | 5.83 | 5.63 |
2 | 0.36 | 0.42 | 0.43 | 0.38 | 0.41 | 0.44 | 0.38 | 0.35 | 0.38 | 0.32 | 1.04 | 1.28 | 1.27 |
3 | 2.48 | 2.71 | 2.79 | 2.76 | 2.66 | 2.63 | 2.58 | 2.16 | 2.50 | 2.48 | 1.94 | 2.81 | 2.46 |
4 | 1.80 | 1.89 | 1.86 | 1.86 | 1.91 | 1.91 | 1.89 | 1.84 | 1.85 | 1.80 | 1.64 | 1.86 | 1.84 |
5 | 2.61 | 2.95 | 2.81 | 2.90 | 2.83 | 2.88 | 2.83 | 2.83 | 2.85 | 2.83 | 2.12 | 2.32 | 2.29 |
6 | 0.44 | 0.49 | 0.45 | 0.47 | 0.46 | 0.45 | 0.46 | 0.45 | 0.45 | 0.45 | 0.34 | 0.31 | 0.31 |
7 | 2.26 | 2.61 | 2.43 | 2.50 | 2.50 | 2.51 | 2.48 | 2.48 | 2.49 | 2.52 | 1.98 | 1.91 | 1.90 |
8 | 1.31 | 1.37 | 1.40 | 1.38 | 1.39 | 1.28 | 1.35 | 1.38 | 1.31 | 1.33 | 0.93 | 1.06 | 1.04 |
9 | 0.97 | 1.21 | 1.10 | 1.12 | 1.10 | 1.12 | 1.14 | 1.08 | 1.13 | 1.14 | 0.77 | 0.78 | 0.72 |
10 | 1.12 | 1.44 | 1.30 | 1.31 | 1.30 | 1.30 | 1.26 | 1.32 | 1.38 | 1.37 | 0.94 | 0.95 | 0.87 |
11 | 7.39 | 7.82 | 7.92 | 7.64 | 7.42 | 7.23 | 7.38 | 7.31 | 8.04 | 8.18 | 7.06 | 8.48 | 8.60 |
12 | 5.52 | 5.82 | 5.77 | 5.70 | 5.74 | 5.47 | 5.36 | 5.46 | 5.15 | 5.55 | 5.49 | 5.32 | 5.25 |
13 | 5.10 | 4.99 | 5.02 | 4.78 | 4.79 | 4.84 | 5.16 | 4.89 | 4.93 | 4.92 | 6.10 | 4.93 | 4.87 |
14 | 0.33 | 0.30 | 0.27 | 0.36 | 0.31 | 0.29 | 0.30 | 0.37 | 0.26 | 0.26 | 0.19 | 0.31 | 0.43 |
15 | 1.92 | 1.82 | 1.73 | 1.77 | 1.86 | 1.71 | 1.32 | 1.63 | 1.85 | 1.76 | 1.17 | 1.18 | 1.15 |
Sub-total | 40.56 | 42.94 | 42.20 | 41.99 | 41.79 | 41.52 | 40.65 | 40.26 | 41.59 | 43.00 | 37.06 | 38.93 | 38.23 |
Fructose a | 7.13 | 7.10 | 7.14 | 7.14 | 7.14 | 7.17 | 7.15 | 7.17 | 7.18 | 7.63 | 7.49 | 7.41 | 7.13 |
Glucose a | 2.72 | 2.73 | 2.73 | 2.84 | 2.82 | 2.76 | 2.75 | 2.78 | 2.73 | 3.04 | 2.76 | 2.74 | 2.72 |
Sucrose a | 1.33 | 1.33 | 1.33 | 1.37 | 1.33 | 1.34 | 1.35 | 1.36 | 1.34 | 1.22 | 1.23 | 1.21 | 1.33 |
Salt a,b | 7.2 | 7.2 | 7.2 | 7.2 | 7.2 | 7.2 | 7.2 | 7.2 | 7.2 | 7.2 | 7.2 | 7.2 | 7.2 |
Dry weight a | 19.91 | 20.39 | 19.91 | 20.16 | 19.57 | 19.79 | 19.59 | 20.12 | 20.02 | 20.43 | 20.97 | 20.59 | 20.9 |
Content percentage | 92.52 | 90.25 | 92.63 | 92.22 | 94.69 | 93.54 | 94.39 | 92.20 | 92.37 | 93.65 | 89.26 | 90.33 | 88.13 |
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Chau, S.-L.; Huang, Z.-B.; Song, Y.-G.; Yue, R.-Q.; Ho, A.; Lin, C.-Z.; Huang, W.-H.; Han, Q.-B. Comprehensive Quantitative Analysis of SQ Injection Using Multiple Chromatographic Technologies. Molecules 2016, 21, 1092. https://doi.org/10.3390/molecules21081092
Chau S-L, Huang Z-B, Song Y-G, Yue R-Q, Ho A, Lin C-Z, Huang W-H, Han Q-B. Comprehensive Quantitative Analysis of SQ Injection Using Multiple Chromatographic Technologies. Molecules. 2016; 21(8):1092. https://doi.org/10.3390/molecules21081092
Chicago/Turabian StyleChau, Siu-Leung, Zhi-Bing Huang, Yan-Gang Song, Rui-Qi Yue, Alan Ho, Chao-Zhan Lin, Wen-Hua Huang, and Quan-Bin Han. 2016. "Comprehensive Quantitative Analysis of SQ Injection Using Multiple Chromatographic Technologies" Molecules 21, no. 8: 1092. https://doi.org/10.3390/molecules21081092
APA StyleChau, S.-L., Huang, Z.-B., Song, Y.-G., Yue, R.-Q., Ho, A., Lin, C.-Z., Huang, W.-H., & Han, Q.-B. (2016). Comprehensive Quantitative Analysis of SQ Injection Using Multiple Chromatographic Technologies. Molecules, 21(8), 1092. https://doi.org/10.3390/molecules21081092