Introduction
Chinese name | Latin name |
---|---|
Danshen | Salvia miltiorrhiza Bge |
Danggui | Angelica sinensis (Oliv.) Diels |
Chishao | Paeonia lactiflora Pall |
Yanhusuo | Corydalis yanhusuo W.T.Wang |
Baijiangcao | Patrinia scabiosaefolia Fisch. ex Trev |
Xiangfu | Cyperus rotundus L |
Sanleng | Sparganium stoloniferum Buch.-Ham |
Wuyao | Lindera aggregate (Sims) Kos-term |
Gancao | Glycyrrhiza uralensis Fisch |
Tufuling | Smilax glabra Roxb |
Daxueteng | Sargentodoxa cuneata (Oliv.) Rehd. et Wils |
Materials and methods
Chemicals and reagents
Animal experimental
Plasma sample preparation
Plasma sample preparation of PRO, PE, AST, FE, and CH
Plasma sample preparation of SAB
LC–MS/MS method for pharmacokinetics study
Compound | Molecular weight | Parent ion | Fragment ion |
---|---|---|---|
AST | 450.408 | 449.3 | 151.1 |
FE | 194.19 | 193.0 | 177.6 |
PRO | 138.122 | 136.7 | 107.4 |
CH | 354.31 | 353.1 | 190.8 |
PE | 480.45 | 479.0 | 448.9 |
SAB | 718.62 | 717.4 | 519.4 |
IS | 580.53 | 579.1 | 271.0 |
Preparation of calibration standards and quality control (QC) samples
Bioanalytical method validation
PK-PD study in THP-1 Cells
Data analyses
Results
Specificity
Calibration curve and lower limit of quantification (LLOQ)
Compounds | Regression equations | Correlation coefficients | Linear ranges (ng/mL) | LLOQ (ng/mL) |
---|---|---|---|---|
AST | Y = 0.0006 X + 0.0099 | 0.9992 | 50—600 | 50 |
FE | Y = 0.0033 X + 0.1772 | 0.9992 | 25—500 | 25 |
PRO | Y = 0.0028 X + 0.0346 | 0.9993 | 10—500 | 10 |
CH | Y = 0.0112 X—0.0449 | 0.9990 | 50—600 | 50 |
PE | Y = 0.0001 X—0.0002 | 0.9994 | 50—600 | 50 |
SAB | Y = 0.0006 X—0.0030 | 0.9993 | 100—1000 | 100 |
Extraction recovery, matrix effect, precision, accuracy, and stability
Compounds | Concentration (ng/mL) | matrix effects (%) | Recovery (%) | Precision (RSD%) | Accuracy (%) | Room temperature 24 h | Three freeze- thaw cycles | In 56 ℃ for 1 h | ||
---|---|---|---|---|---|---|---|---|---|---|
Intra-day | Inter-day | Intra-day | Inter-day | measured value | measured value | measured value | ||||
100 | 96.55 ± 2.09 | 95.92 ± 2.07 | 5.85 ± 5.78 | 5.19 ± 5.18 | 98.73 ± 5.78 | 99.76 ± 5.18 | 99.68 ± 2.31 | 97.89 ± 4.06 | 98.46 ± 2.77 | |
AST | 200 | 102.99 ± 2.75 | 92.76 ± 2.47 | 2.85 ± 2.71 | 3.98 ± 3.87 | 95.05 ± 2.71 | 97.36 ± 3.87 | 199.01 ± 3.99 | 193.37 ± 7.61 | 194.85 ± 6.53 |
500 | 100.24 ± 2.28 | 94.96 ± 2.16 | 2.86 ± 2.82 | 2.43 ± 2.43 | 98.93 ± 2.82 | 99.96 ± 2.43 | 498.07 ± 3.95 | 497.15 ± 6.61 | 495.21 ± 8.43 | |
50 | 107.61 ± 2.45 | 89.83 ± 2.04 | 7.89 ± 8.08 | 6.33 ± 6.35 | 102.43 ± 8.08 | 100.4 ± 6.35 | 49.14 ± 2.35 | 49.41 ± 1.85 | 48.33 ± 2.76 | |
FE | 125 | 93.48 ± 4.51 | 90.13 ± 4.35 | 2.18 ± 2.19 | 2.61 ± 2.63 | 100.37 ± 2.19 | 100.68 ± 2.63 | 122.72 ± 2.65 | 124.36 ± 2.46 | 123.42 ± 1.76 |
400 | 88.56 ± 2.09 | 96.54 ± 2.74 | 1.45 ± 1.48 | 1.84 ± 1.88 | 102.23 ± 1.48 | 101.92 ± 1.88 | 403.59 ± 7.84 | 400.59 ± 6.95 | 399.13 ± 3.26 | |
20 | 92.25 ± 2.35 | 95.84 ± 2.44 | 8.48 ± 8.75 | 6.01 ± 6.25 | 103.23 ± 8.7 | 104 ± 6.25 | 20.81 ± 0.85 | 20.62 ± 1.06 | 19.88 ± 0.43 | |
PRO | 100 | 108.34 ± 1.53 | 90.61 ± 1.28 | 6.67 ± 6.62 | 4.78 ± 4.89 | 99.17 ± 6.62 | 102.15 ± 4.89 | 100.34 ± 1.48 | 97.77 ± 2.49 | 98.21 ± 2.33 |
400 | 105.24 ± 3.06 | 91.32 ± 2.65 | 4.3 ± 4.43 | 3.45 ± 3.48 | 103 ± 4.43 | 101.05 ± 3.48 | 398.27 ± 4.21 | 395.66 ± 7.14 | 394.26 ± 8.01 | |
100 | 103.81 ± 3.10 | 89.57 ± 2.67 | 4.51 ± 4.36 | 3.62 ± 3.56 | 96.56 ± 4.36 | 98.34 ± 3.56 | 99.91 ± 2.94 | 98.55 ± 2.93 | 97.22 ± 3.31 | |
CH | 200 | 102.10 ± 2.23 | 95.61 ± 3.02 | 4.96 ± 4.69 | 5.13 ± 4.96 | 94.62 ± 4.69 | 96.84 ± 4.96 | 200.03 ± 9.56 | 191.77 ± 8.91 | 195.56 ± 7.88 |
500 | 93.58 ± 1.13 | 97.97 ± 1.18 | 6.84 ± 6.75 | 4.57 ± 4.54 | 98.66 ± 6.75 | 99.58 ± 4.54 | 499.00 ± 6.02 | 485.90 ± 9.73 | 489.60 ± 8.91 | |
100 | 102.84 ± 2.17 | 90.62 ± 1.91 | 5.87 ± 5.68 | 4.09 ± 4.03 | 96.67 ± 5.68 | 98.7 ± 4.03 | 99.44 ± 2.43 | 99.68 ± 2.83 | 95.32 ± 1.06 | |
PE | 200 | 107.07 ± 0.71 | 98.69 ± 0.65 | 1.71 ± 1.75 | 2.53 ± 2.60 | 102.79 ± 1.75 | 102.68 ± 2.60 | 199.64 ± 2.59 | 197.55 ± 3.60 | 196.25 ± 4.08 |
500 | 100.02 ± 6.36 | 94.55 ± 6.01 | 1.83 ± 1.84 | 2.6 ± 2.62 | 100.82 ± 1.84 | 100.71 ± 2.62 | 499.23 ± 2.58 | 492.85 ± 7.60 | 495.87 ± 6.50 | |
200 | 91.94 ± 3.62 | 97.90 ± 3.85 | 3.83 ± 3.69 | 5.12 ± 5.10 | 96.23 ± 3.69 | 99.64 ± 5.10 | 208.88 ± 8.46 | 194.10 ± 8.56 | 196.22 ± 3.40 | |
SAB | 400 | 101.21 ± 1.16 | 92.44 ± 1.06 | 1.13 ± 1.03 | 3.2 ± 2.95 | 90.92 ± 1.03 | 92.09 ± 2.95 | 391.06 ± 6.16 | 388.99 ± 6.75 | 395.20 ± 10.70 |
800 | 102.70 ± 1.34 | 99.81 ± 1.30 | 3.57 ± 3.39 | 3.2 ± 2.97 | 94.98 ± 3.39 | 92.82 ± 2.97 | 792.27 ± 3.94 | 790.71 ± 3.52 | 791.11 ± 5.98 |
Pharmacokinetic and pharmacodynamic application
Parameter | Unit | AST | CH | FE | PE | PRO | SAB |
---|---|---|---|---|---|---|---|
AUC(0-t) | mg/L*h | 2.08 ± 0.20 | 0.92 ± 0.06 | 1.43 ± 0.23 | 2.21 ± 0.31 | 1.05 ± 0.06 | 3.40 ± 0.45 |
AUC(0-∞) | mg/L*h | 2.09 ± 0.20 | 0.94 ± 0.09 | 1.48 ± 0.24 | 2.22 ± 0.31 | 1.08 ± 0.07 | 3.70 ± 0.46 |
t1/2 | h | 3.42 ± 0.34 | 1.86 ± 0.21 | 0.90 ± 0.02 | 2.37 ± 0.20 | 0.93 ± 0.06 | 7.60 ± 3.34 |
tmax | h | 0.25 ± 0 | 0.25 ± 0 | 0.25 ± 0 | 0.25 ± 0 | 0.25 ± 0 | 0.29 ± 0.10 |
CLz/F | L/h/kg | 9.55 ± 0.81 | 21.33 ± 1.29 | 13.80 ± 2.37 | 9.15 ± 1.27 | 18.57 ± 1.07 | 5.48 ± 0.68 |
Vz/F | L/kg | 45.37 ± 6.31 | 57.19 ± 7.48 | 17.85 ± 3.28 | 31.26 ± 4.08 | 24.94 ± 1.96 | 59.83 ± 25.48 |
Cmax | mg/L | 2.27 ± 0.35 | 1.37 ± 0.06 | 1.20 ± 0.15 | 2.04 ± 0.59 | 1.43 ± 0.22 | 1.59 ± 0.33 |
Pharmacokinetic and pharmacodynamic model fitting
ingredients | fitting equation | r | ||
---|---|---|---|---|
effect fitting of blood concentration—IL-1β | effect fitting of blood concentration—TNF-α | effect fitting of blood concentration—IL-1β | effect fitting of blood concentration—TNF-α | |
AST | Y = 33.736—1.448 lnX | Y = 113.618—16.262 lnX | -0.746 | -0.900 |
CH | Y = 32.247—1.386 lnX | Y = 101.980—9.387 lnX | -0.776 | -0.891 |
FE | Y = 33.427—1.446 lnX | Y = 108.880—8.805 lnX | -0.764 | -0.867 |
PE | Y = 33.598—1.428 lnX | Y = 109.393—9.030 lnX | -0.782 | -0.858 |
PRO | Y = 33.031—1.454 lnX | Y = 107.704—8.778 lnX | -0.768 | -0.883 |
SAB | Y = 40.971—2.540 lnX | Y = 158.216—9.637 lnX | -0.781 | -0.893 |