Development and Validation of a HPLC-MS/MS Method for Simultaneous Determination of Twelve Bioactive Compounds in Epimedium: Application to a Pharmacokinetic Study in Rats
Abstract
:1. Introduction
2. Results and Discussion
2.1. Method Development
2.2. Sample Preparation
2.3. Method Validation
2.3.1. Specificity
2.3.2. Calibration Curves
2.3.3. Precision and Accuracy
2.3.4. Extraction Recovery and Matrix Effect
2.3.5 Stability
2.4. Application
3. Experimental
3.1. Chemicals and Reagents
3.2. HPLC-MS/MS Analysis
3.3. Preparation of Extract from Epimedium
3.4. Working Solutions
3.5. Sample Preparation
3.6. Method Validation
3.6.1. Selectivity
3.6.2. Calibration Curve
3.6.3. Precision and Accuracy
3.6.4. Extraction Recovery and Matrix Effect
3.6.5. Stability
3.7. Pharmacokinetic Studies
4. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Sample Availability: The extracts of Epimedium are available from the authors. |
Compounds | Calibration Curve | r | Linear Range (ng/mL) | LLOQ(ng/mL) |
---|---|---|---|---|
baohuoside II | Y = 1.3944X + 4.6850 | 0.9930 | 1.0-200.0 | 1.0 |
baohuoside I | Y = 1.4452X + 0.0129 | 0.9943 | 2.0-400.0 | 2.0 |
sagittatoside A | Y = 0.5707X + 4.8545 | 0.9918 | 2.0-400.0 | 2.0 |
sagittatoside B | Y = 0.6968X + 7.8080 | 0.9908 | 2.0-400.0 | 2.0 |
epimedin A | Y = 0.1564X − 8.1949 | 0.9912 | 2.0-400.0 | 2.0 |
epimedin B | Y = 0.3559X + 5.3440 | 0.9939 | 2.0-400.0 | 2.0 |
epimedin C | Y = 0.1016X + 6.8756 | 0.9942 | 2.0-400.0 | 2.0 |
magnoflorine | Y = 0.0260X + 2.4814 | 0.9928 | 4.0-800.0 | 4.0 |
chlorogenic acid | Y = 0.1274X + 0.0016 | 0.9922 | 10.0-2000.0 | 10.0 |
neochlorogenic acid | Y = 1.7722 X + 0.0013 | 0.9956 | 4.0-800.0 | 4.0 |
cryptochlorogenic acid | Y = 0.0187X + 0.0010 | 0.9904 | 4.0-800.0 | 4.0 |
icariin | Y = 0.1178X + 0.0053 | 0.9966 | 4.0-800.0 | 4.0 |
Compounds | Spiked Concentration (ng/mL) | Intra-Day | Inter-Day | ||||
---|---|---|---|---|---|---|---|
Measured (ng/mL) | RE (%) | RSD (%) | Measured (ng/mL) | RE (%) | RSD (%) | ||
baohuoside II | 1 | 1.02 ± 0.08 | 2.0 | 7.8 | 1.01 ± 0.08 | 1.0 | 7.9 |
10 | 10.18 ± 0.88 | 1.8 | 8.6 | 10.09 ± 0.87 | 0.9 | 8.6 | |
200 | 197.54 ± 1.41 | −1.2 | 0.7 | 196.66 ± 3.62 | −1.7 | 1.8 | |
baohuoside I | 2 | 2.03 ± 0.07 | 1.5 | 3.4 | 2.03 ± 0.08 | 1.5 | 3.9 |
20 | 19.99 ± 0.98 | −0.1 | 4.9 | 19.54 ± 0.92 | −2.3 | 4.7 | |
400 | 390.27 ± 6.47 | −2.4 | 1.7 | 391.05 ± 7.15 | −2.2 | 1.8 | |
sagittatoside A | 2 | 2.06 ± 0.10 | 3.0 | 4.9 | 1.98 ± 0.12 | −1.0 | 6.0 |
20 | 20.77 ± 1.01 | 3.9 | 4.9 | 21.00 ± 0.90 | 5.0 | 4.3 | |
400 | 399.42 ± 5.65 | −0.1 | 1.4 | 396.13 ± 3.57 | −1.0 | 0.9 | |
sagittatoside B | 2 | 2.08 ± 0.06 | 4.0 | 2.9 | 2.14 ± 0.07 | 7.0 | 3.3 |
20 | 20.50 ± 1.36 | 2.5 | 6.6 | 21.07 ± 0.81 | 5.4 | 3.8 | |
400 | 397.48 ± 7.79 | −0.6 | 2.0 | 404.06 ± 7.29 | 1.0 | 1.8 | |
epimedin A | 2 | 2.13 ± 0.20 | 6.5 | 9.4 | 2.26 ± 0.15 | 13.0 | 6.6 |
20 | 19.62 ± 2.14 | −1.9 | 10.9 | 20.92 ± 1.98 | 4.6 | 9.5 | |
400 | 397.22 ± 7.08 | −0.7 | 1.8 | 405.26 ± 5.73 | 1.3 | 1.4 | |
epimedin B | 2 | 1.98 ± 0.08 | −1.0 | 4.0 | 2.00 ± 0.13 | 0.0 | 6.5 |
20 | 19.92 ± 1.03 | −0.4 | 5.2 | 20.87 ± 1.02 | 4.4 | 4.9 | |
400 | 399.43 ± 3.67 | −0.1 | 0.9 | 399.44 ± 5.76 | −0.1 | 1.4 | |
epimedin C | 2 | 2.02 ± 0.11 | 1.0 | 5.4 | 2.01 ± 0.07 | 0.5 | 3.5 |
20 | 19.90 ± 0.81 | −0.5 | 4.1 | 19.88 ± 0.89 | −0.6 | 4.5 | |
400 | 401.44 ± 3.03 | 0.4 | 0.8 | 400.94 ± 2.37 | 0.2 | 0.6 | |
magnoflorine | 4 | 4.05 ± 0.24 | 1.3 | 5.9 | 4.06 ± 0.18 | 1.5 | 4.4 |
40 | 41.67 ± 2.42 | 4.2 | 5.8 | 41.10 ± 1.87 | 2.8 | 4.5 | |
800 | 770.53 ± 16.22 | −3.7 | 2.1 | 797.84 ± 33.32 | −0.3 | 4.2 | |
chlorogenic acid | 10 | 9.86 ± 0.96 | −1.4 | 9.7 | 9.89 ± 0.62 | −1.1 | 6.3 |
100 | 94.73 ± 4.35 | −5.3 | 4.6 | 96.39 ± 6.66 | −3.6 | 6.9 | |
2000 | 1888.50 ± 52.19 | −5.6 | 2.8 | 1987.74 ± 96.63 | −0.6 | 4.9 | |
neochlorogenic acid | 4 | 4.16 ± 0.26 | 4.0 | 6.3 | 4.15 ± 0.19 | 3.8 | 4.6 |
40 | 39.69 ± 2.63 | −0.8 | 6.6 | 40.31 ± 1.47 | 0.8 | 3.6 | |
800 | 804.76 ± 7.42 | 0.6 | 0.9 | 807.99 ± 10.77 | 1.0 | 1.3 | |
cryptochlorogenic acid | 4 | 3.94 ± 0.12 | −1.5 | 3.0 | 3.92 ± 0.07 | −2.0 | 1.8 |
40 | 40.25 ± 1.39 | 0.6 | 3.5 | 40.63 ± 0.96 | 1.6 | 2.4 | |
800 | 780.52 ± 16.27 | −2.4 | 2.1 | 774.79 ± 13.82 | −3.2 | 1.8 | |
icariin | 4 | 4.02 ± 0.15 | 0.5 | 3.7 | 4.02 ± 0.11 | 0.5 | 2.7 |
40 | 39.30 ± 1.03 | −1.8 | 2.6 | 39.85 ± 1.13 | −0.4 | 2.8 | |
800 | 801.05 ± 3.35 | 0.1 | 0.4 | 802.21 ± 4.66 | 0.3 | 0.6 |
Compounds | Spiked Concentration (ng/mL) | Extraction Recovery (%) | RSD (%) | Matrix Effect (%) | RSD (%) |
---|---|---|---|---|---|
baohuoside II | 1 | 67.78 ± 1.75 | 2.6 | 119.03 ± 3.54 | 3.0 |
10 | 84.44 ± 3.81 | 4.5 | 115.54 ± 6.71 | 5.8 | |
200 | 93.58 ± 4.14 | 4.4 | 103.39 ± 7.55 | 7.3 | |
baohuoside I | 2 | 99.45 ± 6.74 | 6.8 | 93.08 ± 5.89 | 6.3 |
20 | 97.78 ± 1.36 | 1.4 | 103.85 ± 6.55 | 6.3 | |
400 | 98.50 ± 1.73 | 1.8 | 98.21 ± 3.20 | 3.3 | |
sagittatoside A | 2 | 74.52 ± 7.02 | 9.4 | 112.96 ± 8.54 | 7.6 |
20 | 79.30 ± 3.64 | 4.6 | 118.72 ± 9.92 | 8.4 | |
400 | 90.06 ± 3.35 | 3.7 | 107.33 ± 10.01 | 9.3 | |
sagittatoside B | 2 | 99.77 ± 4.19 | 4.2 | 98.77 ± 3.97 | 4.0 |
20 | 82.34 ± 4.82 | 5.9 | 112.35 ± 7.63 | 6.8 | |
400 | 86.73 ± 2.69 | 3.1 | 111.49 ± 6.28 | 5.6 | |
epimedin A | 2 | 60.66 ± 6.63 | 10.9 | 113.02 ± 10.87 | 9.6 |
20 | 68.52 ± 0.66 | 1.0 | 119.11 ± 13.22 | 11.1 | |
400 | 84.85 ± 3.63 | 4.3 | 112.83 ± 9.14 | 8.1 | |
epimedin B | 2 | 70.66 ± 4.43 | 6.3 | 119.84 ± 6.79 | 5.7 |
20 | 94.72 ± 0.75 | 0.8 | 107.49 ± 8.41 | 7.8 | |
400 | 75.30 ± 1.12 | 1.5 | 119.30 ± 3.05 | 2.6 | |
epimedin C | 2 | 96.35 ± 1.00 | 1.0 | 102.59 ± 3.00 | 2.9 |
20 | 70.91 ± 2.30 | 3.2 | 112.31 ± 11.51 | 10.2 | |
400 | 81.57 ± 1.92 | 2.4 | 118.74 ± 6.52 | 5.5 | |
magnoflorine | 4 | 61.95 ± 6.86 | 11.1 | 115.27 ± 15.25 | 13.2 |
40 | 70.29 ± 4.68 | 6.7 | 114.36 ± 5.95 | 5.2 | |
800 | 81.33 ± 4.59 | 5.6 | 110.84 ± 12.78 | 11.5 | |
chlorogenic acid | 10 | 97.79 ± 4.36 | 4.5 | 98.55 ± 7.49 | 7.6 |
100 | 89.78 ± 8.65 | 9.6 | 98.36 ± 5.32 | 5.4 | |
2000 | 78.81 ± 5.22 | 6.6 | 114.50 ± 13.84 | 12.1 | |
neochlorogenic acid | 4 | 82.92 ± 2.58 | 3.1 | 113.90 ± 5.98 | 5.3 |
40 | 78.53 ± 2.88 | 3.7 | 114.36 ± 5.95 | 5.2 | |
800 | 84.04 ± 3.36 | 4.0 | 115.44 ± 8.99 | 7.8 | |
cryptochlorogenic acid | 4 | 91.38 ± 10.31 | 11.3 | 96.33 ± 6.89 | 7.2 |
40 | 76.05 ± 6.75 | 8.9 | 118.92 ± 11.70 | 9.8 | |
800 | 76.95 ± 3.26 | 4.2 | 116.51 ± 12.06 | 10.4 | |
icariin | 4 | 96.32 ± 4.65 | 4.8 | 95.68 ± 5.32 | 5.6 |
40 | 88.84 ± 9.11 | 10.3 | 111.87 ± 10.96 | 9.8 | |
800 | 91.78 ± 4.42 | 4.8 | 105.94 ± 8.98 | 8.5 |
Compounds | Spiked Concentration (ng/mL) | Room Temperature for 2 h | Three Freeze-Thaw Cycles | Auto-Sampler for 12 h | −70 °C for 7 Days | ||||
---|---|---|---|---|---|---|---|---|---|
Measured (ng/mL) | RSD (%) | Measured (ng/mL) | RSD (%) | Measured (ng/mL) | RSD (%) | Measured (ng/mL) | RSD (%) | ||
baohuoside II | 1 | 0.96 ± 0.09 | 9.4 | 1.09 ± 0.02 | 1.8 | 1.10 ± 0.11 | 10.0 | 1.04 ± 0.12 | 11.5 |
10 | 9.99 ± 0.31 | 3.1 | 8.94 ± 0.63 | 7.0 | 10.38 ± 0.60 | 5.8 | 8.84 ± 0.31 | 3.5 | |
200 | 193.54 ± 3.74 | 1.9 | 195.26 ± 1.32 | 0.7 | 190.62 ± 5.99 | 3.1 | 188.79 ± 7.35 | 3.9 | |
baohuoside I | 2 | 1.93 ± 0.03 | 1.6 | 1.97 ± 0.08 | 4.1 | 2.01 ± 0.07 | 3.5 | 2.03 ± 0.09 | 4.4 |
20 | 19.56 ± 0.23 | 1.2 | 19.71 ± 0.49 | 2.5 | 19.24 ± 0.96 | 5.0 | 19.48 ± 0.52 | 2.7 | |
400 | 389.39 ± 7.29 | 1.9 | 393.71 ± 5.02 | 1.3 | 387.09 ± 9.42 | 2.4 | 391.78 ± 3.49 | 0.9 | |
sagittatoside A | 2 | 2.08 ± 0.12 | 5.8 | 2.11 ± 0.07 | 3.3 | 1.93 ± 0.09 | 4.7 | 1.98 ± 0.12 | 6.1 |
20 | 20.97 ± 0.09 | 0.4 | 19.84 ± 0.81 | 4.1 | 21.29 ± 0.18 | 0.8 | 19.43 ± 0.40 | 2.1 | |
400 | 397.27 ± 8.71 | 2.2 | 393.69 ± 3.71 | 0.9 | 397.42 ± 6.71 | 1.7 | 395.24 ± 11.34 | 2.9 | |
sagittatoside B | 2 | 2.04 ± 0.16 | 7.8 | 2.09 ± 0.05 | 2.4 | 2.07 ± 0.08 | 3.9 | 2.03 ± 0.14 | 6.9 |
20 | 21.32 ± 0.59 | 2.8 | 20.07 ± 0.95 | 4.7 | 21.16 ± 0.51 | 2.4 | 18.84 ± 0.52 | 2.8 | |
400 | 398.03 ± 9.83 | 2.5 | 394.07 ± 3.55 | 0.9 | 407.64 ± 17.60 | 4.3 | 381.68 ± 5.87 | 1.5 | |
epimedin A | 2 | 2.10 ± 0.09 | 4.3 | 2.07 ± 0.13 | 6.3 | 2.06 ± 0.04 | 1.9 | 2.03 ± 0.09 | 4.4 |
20 | 20.29 ± 0.43 | 2.1 | 19.58 ± 0.94 | 4.8 | 20.35 ± 0.78 | 3.8 | 19.70 ± 1.54 | 7.8 | |
400 | 400.51 ± 0.52 | 0.1 | 392.00 ± 2.85 | 0.7 | 398.73 ± 3.31 | 0.8 | 400.95 ± 4.79 | 1.2 | |
epimedin B | 2 | 2.01 ± 0.04 | 2.0 | 2.07 ± 0.09 | 4.3 | 2.18 ± 0.05 | 2.3 | 2.07 ± 0.09 | 4.3 |
20 | 21.51 ± 0.97 | 4.5 | 20.27 ± 1.82 | 9.0 | 21.01 ± 0.50 | 2.4 | 21.00 ± 0.78 | 3.7 | |
400 | 400.89 ± 3.17 | 0.8 | 400.51 ± 2.96 | 0.7 | 397.07 ± 2.81 | 0.7 | 403.31 ± 2.82 | 0.7 | |
epimedin C | 2 | 2.10 ± 0.05 | 2.4 | 2.03 ± 0.10 | 4.9 | 2.02 ± 0.14 | 6.9 | 2.14 ± 0.05 | 2.3 |
20 | 20.24 ± 0.83 | 4.1 | 20.43 ± 0.90 | 4.4 | 19.73 ± 0.37 | 1.9 | 21.37 ± 0.10 | 0.5 | |
400 | 399.97 ± 1.97 | 0.5 | 397.64 ± 8.12 | 2.0 | 396.56 ± 4.69 | 1.2 | 396.46 ± 8.82 | 2.2 | |
magnoflorine | 4 | 3.99 ± 0.08 | 2.0 | 4.01 ± 0.13 | 3.2 | 4.37 ± 0.19 | 4.3 | 4.02 ± 0.10 | 2.5 |
40 | 39.29 ± 2.92 | 7.4 | 41.99 ± 2.30 | 5.5 | 41.70 ± 1.06 | 2.5 | 40.27 ± 1.70 | 4.2 | |
800 | 776.84 ± 1.85 | 0.2 | 780.80 ± 15.38 | 2.0 | 809.27 ± 12.34 | 1.5 | 775.71 ± 3.82 | 0.5 | |
chlorogenic acid | 10 | 9.93 ± 0.03 | 0.3 | 9.52 ± 0.84 | 8.8 | 9.26 ± 0.68 | 7.3 | 8.88 ± 0.39 | 4.4 |
100 | 101.51 ± 11.83 | 11.7 | 91.99 ± 6.26 | 6.8 | 98.92 ± 4.58 | 4.6 | 85.73 ± 4.01 | 4.7 | |
2000 | 1866.65 ± 25.87 | 1.4 | 1883.55 ± 25.34 | 1.3 | 1924.54 ± 82.95 | 4.3 | 1849.43 ± 30.88 | 1.7 | |
neochlorogenic acid | 4 | 4.00 ± 0.14 | 3.5 | 3.97 ± 0.05 | 1.3 | 3.85 ± 0.14 | 3.6 | 4.13 ± 0.06 | 1.5 |
40 | 39.79 ± 2.13 | 5.4 | 38.73 ± 1.14 | 2.9 | 41.77 ± 1.37 | 3.3 | 39.14 ± 1.82 | 4.6 | |
800 | 802.82 ± 7.13 | 0.9 | 791.74 ± 0.87 | 0.1 | 803.36 ± 2.53 | 0.3 | 804.80 ± 4.45 | 0.6 | |
cryptochlorogenic acid | 4 | 4.00 ± 0.14 | 3.5 | 4.03 ± 0.13 | 3.2 | 4.01 ± 0.22 | 5.5 | 4.01 ± 0.06 | 1.5 |
40 | 40.59 ± 1.78 | 4.4 | 39.42 ± 1.80 | 4.6 | 41.58 ± 0.51 | 1.2 | 39.06 ± 1.65 | 4.2 | |
800 | 773.95 ± 5.78 | 0.7 | 781.42 ± 2.45 | 0.3 | 768.52 ± 23.65 | 3.1 | 759.75 ± 7.73 | 1.0 | |
icariin | 4 | 4.07 ± 0.10 | 2.5 | 4.09 ± 0.09 | 2.2 | 4.11 ± 0.10 | 2.4 | 4.00 ± 0.06 | 1.5 |
40 | 40.07 ± 0.42 | 1.0 | 40.45 ± 1.98 | 4.9 | 40.34 ± 1.48 | 3.7 | 40.22 ± 0.40 | 1.0 | |
800 | 799.61 ± 7.41 | 0.9 | 796.41 ± 5.25 | 0.7 | 808.06 ± 2.49 | 0.3 | 801.34 ± 5.26 | 0.7 |
Compounds | Tmax1 (h) | Tmax2 (h) | Cmax1 (ng/mL) | Cmax2 (ng/mL) | t1/2 (h) | AUC(0-tn) (h·ng/mL) | AUC(0-∞) (h·ng/mL) | MRT(0-tn) (h) | MRT(0-∞) (h) |
---|---|---|---|---|---|---|---|---|---|
baohuoside II | 0.18 ± 0.03 | 10.67 ± 1.03 | 6.87 ± 4.58 | 29.17 ± 7.70 | 6.93 ± 2.94 | 245.95 ± 133.39 | 354.89 ± 143.76 | 11.47 ± 1.99 | 16.50 ± 3.06 |
baohuoside I | 0.38 ± 0.14 | 10.00 ± 1.79 | 65.20 ± 33.56 | 170.73 ± 66.97 | 13.25 ± 4.80 | 1448.99 ± 615.56 | 1675.75 ± 598.59 | 10.81 ± 1.21 | 15.68 ± 4.10 |
sagittatoside A | 0.18 ± 0.03 | 11.67 ± 0.82 | 20.35 ± 10.26 | 203.50 ± 97.03 | 13.81 ± 7.09 | 1614.13 ± 348.72 | 1782.28 ± 351.94 | 12.02 ± 0.71 | 14.26 ± 2.00 |
sagittatoside B | 0.20 ± 0.04 | 11.00 ± 1.67 | 19.38 ± 9.79 | 112.97 ± 44.50 | 9.38 ± 2.11 | 853.80 ± 171.13 | 1036.71 ± 201.70 | 11.67 ± 1.17 | 14.89 ± 2.09 |
epimedin A | 0.22 ± 0.04 | 9.33 ± 0.94 | 82.79 ± 7.61 | 59.34 ± 26.24 | 12.50 ± 6.85 | 550.27 ± 199.35 | 600.14 ± 218.86 | 10.82 ± 2.33 | 14.06 ± 3.98 |
epimedin B | 0.19 ± 0.07 | 7.67 ± 0.75 | 316.73 ± 59.06 | 129.90 ± 77.04 | 3.64 ± 3.68 | 945.89 ± 250.22 | 1066.50 ± 212.44 | 6.98 ± 1.14 | 8.37 ± 2.39 |
epimedin C | 0.22 ± 0.04 | 7.67 ± 0.75 | 135.22 ± 31.26 | 103.20 ± 33.10 | 17.85 ± 8.23 | 850.40 ± 268.63 | 919.35 ± 214.91 | 9.33 ± 2.34 | 12.54 ± 2.59 |
magnoflorine | 0.18 ± 0.03 | 10.67 ± 1.63 | 411.40 ± 110.77 | 275.67 ± 89.36 | 17.77 ± 8.25 | 3676.19 ± 874.51 | 4003.32 ± 969.59 | 9.86 ± 0.75 | 12.52 ± 1.72 |
chlorogenic acid | 0.19 ± 0.05 | 540.28 ± 225.18 | 0.45 ± 0.30 | 879.68 ± 365.08 | 886.48 ± 376.98 | 3.85 ± 1.28 | 3.91 ± 1.32 | ||
neochlorogenic acid | 0.19 ± 0.05 | 205.96 ± 75.12 | 0.64 ± 0.27 | 263.82 ± 68.24 | 268.58 ± 70.86 | 3.18 ± 1.57 | 3.25 ± 1.97 | ||
cryptochlorogenic acid | 0.19 ± 0.05 | 269.77 ± 114.07 | 0.53 ± 0.24 | 397.96 ± 170.58 | 404.11 ± 168.92 | 3.75 ± 1.91 | 3.82 ± 2.48 | ||
icariin | 0.16 ± 0.05 | 350.28 ± 195.96 | 0.41 ± 0.19 | 322.75 ± 204.68 | 324.78 ± 209.27 | 2.71 ± 0.79 | 2.75 ± 0.97 |
Compounds | Precursor Ion (m/z) | Product Ion (m/z) | Frag. (V) | C.E. (V) |
---|---|---|---|---|
baohuoside II | 499.2 | 353.0 | 140 | 20 |
baohuoside I | 513.2 | 366.0 | 140 | 20 |
sagittatoside A | 675.2 | 367.0 | 145 | 32 |
sagittatoside B | 645.2 | 366.2 | 145 | 30 |
epimedin A | 675.1 | 365.8 | 130 | 32 |
epimedin B | 645.1 | 365.6 | 145 | 30 |
epimedin C | 659.2 | 365.7 | 145 | 30 |
magnoflorine | 340.1 | 310.1 | 145 | 22 |
chlorogenic acid | 353.0 | 191.0 | 90 | 10 |
neochlorogenic acid | 352.9 | 191.0 | 115 | 10 |
cryptochlorogenic acid | 353.1 | 172.9 | 100 | 10 |
icariin | 721.0 | 513.2 | 145 | 10 |
liquiritin (IS) | 417.1 | 255.0 | 145 | 13 |
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Sun, M.; Yin, Y.; Wei, J.; Chen, X.; Ouyang, H.; Chang, Y.; Gao, X.; He, J. Development and Validation of a HPLC-MS/MS Method for Simultaneous Determination of Twelve Bioactive Compounds in Epimedium: Application to a Pharmacokinetic Study in Rats. Molecules 2018, 23, 1322. https://doi.org/10.3390/molecules23061322
Sun M, Yin Y, Wei J, Chen X, Ouyang H, Chang Y, Gao X, He J. Development and Validation of a HPLC-MS/MS Method for Simultaneous Determination of Twelve Bioactive Compounds in Epimedium: Application to a Pharmacokinetic Study in Rats. Molecules. 2018; 23(6):1322. https://doi.org/10.3390/molecules23061322
Chicago/Turabian StyleSun, Mengjie, Yanwei Yin, Juan Wei, Xiaopeng Chen, Huizi Ouyang, Yanxu Chang, Xiumei Gao, and Jun He. 2018. "Development and Validation of a HPLC-MS/MS Method for Simultaneous Determination of Twelve Bioactive Compounds in Epimedium: Application to a Pharmacokinetic Study in Rats" Molecules 23, no. 6: 1322. https://doi.org/10.3390/molecules23061322