Pharmacokinetics and Excretion Study of Lycium barbarum Polysaccharides in Rats by FITC-Fluorescence Labeling
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals and Reagents
2.2. Animals
2.3. Instrumentation
2.4. Establishment of Quantitative Analysis Method for LBP-FITC
2.4.1. Fabrication of LBP-FITC
2.4.2. Preparation of LBP-FITC Standard Solution
2.5. Sample Collection
2.5.1. Urine and Fecal Samples
2.5.2. Plasma Sample
2.6. Sample Determination
2.6.1. Determination Conditions
2.6.2. Plasma Samples Determination
2.6.3. Urine Samples Determination
2.6.4. Fecal Samples Determination
2.7. Fabrication of Standard Curve
2.7.1. Plasma Samples
2.7.2. Urine Samples
2.7.3. Fecal Samples
2.8. Method Validation
2.8.1. Precision
2.8.2. Stability
2.8.3. Recovery Rate and Matrix Effect
2.9. Plasma Pharmacokinetics
2.9.1. Administration Method
2.9.2. Sampling Method
2.10. Excretion Study
2.10.1. Administration Method
2.10.2. Sample Collection and Processing
2.11. Sample Determination
2.12. Data Processing
2.12.1. Pharmacokinetic Data Processing
2.12.2. Data Processing of Excretion Experiment
3. Results and Discussion
3.1. Establishment and Validation of Quantitative Analysis Method
3.1.1. Standard Curve Establishment
3.1.2. Precision
3.1.3. Stability
3.1.4. Recovery and Matrix Effect
3.2. Pharmacokinetics Study
3.2.1. Plasma Concentration-Time Data
3.2.2. Mean Plasma Concentration–Time Curve
3.2.3. Pharmacokinetic Parameters
3.3. Excretion Study
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Conflicts of Interest
References
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Biological Sample | Theoretical Concentration (μg/mL) | Inter-Day, RSD (%) | Intra-Day, RSD (%) | ||||
---|---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | |||
Plasma | 0.2 | 7.28 | 6.9 | 5.37 | 4.88 | 4.92 | 7.75 |
2.0 | 3.45 | 4.01 | 4.92 | 2.67 | 5.39 | 4.83 | |
20.0 | 1.86 | 2.24 | 3.28 | 2.48 | 1.97 | 2.62 | |
Urine | 0.25 | 4.26 | 5.9 | 5.37 | 4.88 | 4.92 | 7.16 |
50.0 | 1.45 | 2.01 | 1.92 | 2.67 | 1.39 | 1.83 | |
500.0 | 0.86 | 1.04 | 0.58 | 0.68 | 0.47 | 0.78 | |
Feces | 5.0 | 3.68 | 3.92 | 6.21 | 4.86 | 3.9 | 6.12 |
50.0 | 1.66 | 1.91 | 2.04 | 1.69 | 1.12 | 2.86 | |
300.0 | 0.89 | 1.53 | 1.24 | 1.08 | 1.37 | 1.74 |
Biological Sample | Theoretical Concentration (μg/mL) | 24 h at Room Temperature | Repeated Freeze-Thaw Test for 3 Times | Frozen at −20 °C for 15 Days | |||
---|---|---|---|---|---|---|---|
Measured Concentration (μg/mL) | RSD (%) | Measured Concentration (μg/mL) | RSD (%) | Measured Concentration (μg/mL) | RSD (%) | ||
Plasma | 0.2 | 0.196 ± 0.004 | 4.90 | 0.194 ± 0.005 | 4.98 | 0.191 ± 0.003 | 3.80 |
2.0 | 1.89 ± 0.02 | 2.07 | 1.93 ± 0.04 | 4.03 | 1.96 ± 0.03 | 3.24 | |
20.0 | 19.17 ± 0.34 | 3.74 | 19.03 ± 0.24 | 2.72 | 19.49 ± 0.17 | 1.87 | |
Urine | 0.25 | 0.246 ± 0.004 | 4.61 | 0.244 ± 0.005 | 4.98 | 0.247 ± 0.003 | 4.78 |
50 | 48.89 ± 0.82 | 1.07 | 49.93 ± 0.74 | 1.83 | 49.96 ± 0.83 | 1.34 | |
500 | 498.17 ± 3.34 | 0.74 | 496.03 ± 2.24 | 0.72 | 498.49 ± 3.17 | 0.89 | |
Feces | 5.0 | 4.92 ± 0.14 | 3.64 | 4.86 ± 0.12 | 4.02 | 4.96 ± 0.13 | 3.82 |
50.0 | 48.89 ± 0.22 | 2.11 | 46.93 ± 0.34 | 5.36 | 48.96 ± 0.23 | 4.64 | |
300.0 | 287.63 ± 0.34 | 3.28 | 279.08 ± 0.24 | 2.72 | 292.44 ± 0.31 | 2.81 |
Biological Sample | Theoretical Concentration (μg/mL) | Inter-Group | Intra-Group | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | |||||||||
Rate (%) (M ± SD) | RSD (%) | Rate (%) (M ± SD) | RSD (%) | Rate (%) (M ± SD) | RSD (%) | Rate (%) (M ± SD) | RSD (%) | Rate (%) (M ± SD) | RSD (%) | Rate (%) (M ± SD) | RSD (%) | ||
Plasma | 0.2 | 99.0 ± 7.9 | 7.28 | 96.0 ± 6.1 | 6.90 | 99.5 ± 7.8 | 5.37 | 104.0 ± 5.6 | 4.88 | 97.5 ± 7.2 | 4.92 | 100.2 ± 4.6 | 4.58 |
2.0 | 93.2 ± 3.8 | 3.45 | 87.1 ± 3.5 | 4.01 | 84.6 ± 2.9 | 4.92 | 95.2 ± 3.4 | 2.67 | 94.3 ± 5.2 | 5.39 | 91.6 ± 4.4 | 3.82 | |
20.0 | 98.4 ± 1.2 | 1.86 | 96.9 ± 1.8 | 2.24 | 98.9 ± 1.6 | 3.28 | 94.6 ± 0.9 | 2.48 | 97.2 ± 1.9 | 1.97 | 98.3 ± 2.8 | 2.04 | |
Urine | 0.25 | 99.2 ± 4.8 | 4.28 | 100.0 ± 5.1 | 5.90 | 98.4 ± 5.6 | 5.37 | 100.8 ± 4.8 | 4.88 | 98.0 ± 6.4 | 4.92 | 98.4 ± 1.4 | 1.42 |
50 | 97.7 ± 1.7 | 1.45 | 95.5 ± 1.6 | 2.01 | 97.4 ± 1.9 | 1.92 | 99.8 ± 1.7 | 2.67 | 95.8 ± 1.6 | 1.39 | 97.6 ± 1.5 | 1.64 | |
500 | 99.5 ± 0.5 | 0.86 | 99.1 ± 0.7 | 1.04 | 99.4 ± 0.4 | 0.58 | 98.8 ± 0.6 | 0.68 | 99.7 ± 0.7 | 0.47 | 99.3 ± 0.4 | 0.38 | |
Feces | 5.0 | 99.6 ± 4.78 | 3.68 | 97.2 ± 4.20 | 3.92 | 94.6 ± 2.72 | 6.21 | 99.2 ± 6.40 | 4.86 | 100.6 ± 3.18 | 3.90 | 98.2 ± 2.72 | 2.78 |
50.0 | 96.5 ± 1.64 | 1.66 | 96.3 ± 1.56 | 1.91 | 96.2 ± 1.86 | 2.04 | 95.6 ± 1.34 | 1.69 | 95.4 ± 1.06 | 1.12 | 98.3 ± 0.50 | 0.52 | |
300.0 | 99.6 ± 1.05 | 0.89 | 99.3 ± 1.21 | 1.53 | 99.4 ± 1.50 | 1.24 | 98.9 ± 1.02 | 1.08 | 99.8 ± 1.24 | 1.37 | 99.4 ± 0.98 | 1.00 |
Concentration | 100 mg/kg | 50 mg/kg | 25 mg/kg | |||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Time(h) | 1 | 2 | 3 | 4 | 5 | 6 | 1 | 2 | 3 | 4 | 5 | 6 | 1 | 2 | 3 | 4 | 5 | 6 |
0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
0.5 | 5.176 | 5.722 | 6.454 | 6.425 | 6.69 | 7.426 | 4.933 | 5.082 | 5.358 | 5.527 | 4.333 | 5.632 | 3.514 | 4.050 | 4.180 | 4.132 | 3.121 | 2.645 |
1 | 5.305 | 7.451 | 7.084 | 7.679 | 7.463 | 7.975 | 4.724 | 6.331 | 6.174 | 6.564 | 5.666 | 5.963 | 4.391 | 4.641 | 4.578 | 4.427 | 3.519 | 3.054 |
2 | 5.160 | 7.543 | 7.624 | 7.706 | 6.878 | 8.228 | 4.623 | 6.494 | 6.634 | 7.214 | 6.078 | 6.747 | 4.741 | 5.976 | 5.403 | 5.194 | 5.404 | 4.898 |
3 | 6.085 | 6.343 | 5.407 | 5.023 | 5.219 | 6.677 | 5.902 | 6.056 | 5.401 | 4.98 | 5.146 | 7.021 | 4.496 | 5.184 | 5.315 | 4.413 | 5.038 | 4.635 |
4 | 5.677 | 5.158 | 5.321 | 5.186 | 4.93 | 6.001 | 4.897 | 5.235 | 5.111 | 4.666 | 4.789 | 5.336 | 4.193 | 4.237 | 4.872 | 3.938 | 4.524 | 3.816 |
6 | 5.555 | 4.09 | 5.231 | 4.844 | 4.394 | 6.578 | 4.206 | 4.004 | 4.859 | 4.758 | 4.143 | 5.282 | 3.988 | 3.320 | 4.855 | 4.672 | 4.273 | 2.974 |
8 | 4.045 | 4.713 | 4.88 | 4.784 | 4.875 | 5.714 | 3.981 | 3.651 | 4.546 | 4.256 | 4.254 | 4.967 | 3.728 | 3.019 | 4.026 | 3.947 | 3.949 | 2.874 |
10 | 3.582 | 3.11 | 3.726 | 4.987 | 4.027 | 4.832 | 3.515 | 2.897 | 3.669 | 4.034 | 3.812 | 4.185 | 3.413 | 2.478 | 3.640 | 3.773 | 3.213 | 2.376 |
12 | 2.903 | 3.923 | 2.76 | 4.557 | 3.777 | 3.658 | 2.808 | 2.682 | 3.528 | 3.879 | 3.221 | 3.241 | 3.051 | 2.566 | 3.449 | 3.296 | 2.642 | 1.511 |
24 | 2.592 | 2.631 | 2.756 | 2.612 | 2.551 | 3.146 | 2.349 | 2.476 | 2.743 | 2.581 | 2.756 | 2.248 | 3.026 | 2.126 | 2.996 | 2.551 | 2.638 | 1.239 |
36 | 1.884 | 1.858 | 2.507 | 1.748 | 2.161 | 2.668 | 1.964 | 1.869 | 2.721 | 2.511 | 2.108 | 2.046 | 3.018 | 1.992 | 2.671 | 2.498 | 2.088 | 1.034 |
48 | 1.592 | 1.161 | 2.065 | 1.499 | 1.118 | 2.689 | 1.362 | 1.558 | 2.014 | 2.123 | 1.964 | 1.337 | 2.094 | 1.423 | 1.941 | 2.039 | 1.901 | 0.892 |
Parameter | Unit | 100 mg/kg | 50 mg/kg | 25 mg/kg | |||
---|---|---|---|---|---|---|---|
Mean | SD | Mean | SD | Mean | SD | ||
Cmax a | mg/L | 7.44 | 0.72 | 6.56 | 0.51 | 5.27 | 0.44 |
Tmax b | h | 2 | 0.63 | 2.33 | 0.52 | 2 | 0 |
AUC(0–t) c | mg/L.h | 151.09 | 15.10 | 141.25 | 12.02 | 128.21 | 27.64 |
AUC(0–∞) d | mg/L.h | 230.49 | 73.26 | 236.18 | 35.08 | 242.57 | 64.09 |
MRT(0–t) e | h | 18.38 | 1.01 | 19.15 | 0.84 | 20.07 | 1.49 |
t1/2 f | h | 31.39 | 14.64 | 38.09 | 9.43 | 45.76 | 4.57 |
CLz/F g | L/h/kg | 0.47 | 0.12 | 0.22 | 0.03 | 0.11 | 0.04 |
Vz/F h | L/kg | 18.96 | 3.66 | 11.57 | 2.16 | 7.29 | 2.38 |
Time | Urine | Feces | ||
---|---|---|---|---|
Amount (μg) | % of Dose | Amount (μg) | % of Dose | |
0–4 | 3.87 ± 0.04 | 0.038 ± 0.008 | 0.05 ± 0.02 | 0.052 ± 0.018 |
0–10 | 6.44 ± 0.06 | 0.064 ± 0.012 | 4.81 ± 1.21 | 48.28 ± 9.349 |
0–24 | 7.49 ± 0.09 | 0.075 ± 0.015 | 6.94 ± 1.18 | 69.53 ± 6.483 |
0–48 | 8.36 ± 0.13 | 0.084 ± 0.022 | 8.88 ± 1.12 | 89.24 ± 5.487 |
0–72 | 9.38 ± 0.23 | 0.094 ± 0.036 | 9.23 ± 0.50 | 92.18 ± 3.609 |
Time (h) | Δt (h) | Urinary | Feces | ||
---|---|---|---|---|---|
Excretion Amount (mg) | Excretion Rate (mg/h) | Excretion Amount (mg) | Excretion Rate (mg/h) | ||
0 | 0 | ||||
4 | 4 | 3.87 ± 0.06 | 0.968 ± 0.211 | 0.05 ± 0.02 | 0.013 ± 0.005 |
10 | 6 | 2.57 ± 1.24 | 0.428 ± 0.107 | 4.76 ± 1.12 | 0.793 ± 0.187 |
24 | 14 | 1.05 ± 0.89 | 0.075 ± 0.064 | 2.13 ± 1.08 | 0.152 ± 0.077 |
48 | 24 | 0.87 ± 1.03 | 0.036 ± 0.043 | 1.94 ± 0.51 | 0.081 ± 0.021 |
72 | 24 | 1.02 ± 0.92 | 0.043 ± 0.038 | 0.35 ± 0.13 | 0.015 ± 0.005 |
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Xia, H.; Yang, C.; Zhou, B.; Tang, H.; Yang, L.; Liao, W.; Sun, G. Pharmacokinetics and Excretion Study of Lycium barbarum Polysaccharides in Rats by FITC-Fluorescence Labeling. Foods 2021, 10, 2851. https://doi.org/10.3390/foods10112851
Xia H, Yang C, Zhou B, Tang H, Yang L, Liao W, Sun G. Pharmacokinetics and Excretion Study of Lycium barbarum Polysaccharides in Rats by FITC-Fluorescence Labeling. Foods. 2021; 10(11):2851. https://doi.org/10.3390/foods10112851
Chicago/Turabian StyleXia, Hui, Chao Yang, Beijia Zhou, Huali Tang, Ligang Yang, Wang Liao, and Guiju Sun. 2021. "Pharmacokinetics and Excretion Study of Lycium barbarum Polysaccharides in Rats by FITC-Fluorescence Labeling" Foods 10, no. 11: 2851. https://doi.org/10.3390/foods10112851