| Objective: The purpose of the present work is to develop a simple, sensitive and specific HPLC assay for the quantitation of active components of traditional Chinese medicine in rat plasma and its application in pharmacokinetics study, and to investigate whether the pharmacokinetics of ferulic acid and ferulic acid in Chuanxiong extraction of different solutions were similary or not.Methods: The separation was performed on a Cosmosil-C18 column. The mobile phase consisted of methanol-3% acetic acid solution in a volume 32:68 (v:v). The mobile phase was delivered at a flow rate of 0.8 mL/min. Detection was performed at a wavelength of 323 nm at room temperature. The sample injection volume was 20μL. The HPLC was used to determine the plasma concentration of ferulic acid after oral administration or bolus injecting ferulic acid solution in rat plasma. Moreover, their pharmacokinetics parameters were estimated by DAS 1.0 software.Results: (1) The retention time for internal standard and ferulic acid were approximately 7.7 min and 12.5 min, respectively The linear regression analysis of the standard calibration plot for rat plasma were y = 0.7139 x-0.3738 for the high concentration range (5.0-50.0μg/mL) and y = 0.6517 x + 0.0066 for the low concentration (0.1-5.0μg/mL) with r values of 0.9999 and 0.9994, respectively. The RSD of ferulic acid was lower than 7.76% for intra-day and inter-day assays. Assay accuracy was higher than 91.95%. The average recovery of ferulic acid was more than 87.93%. The limit of quantitation was deemed to be 0.03μg/mL.(2) The mean maximum concentration of ferulic acid and ferulic acid in Chuanxiong decoction in plasma was 32.67 and 2.29μg/mL at 9.0 and 2.4 min after oral administration with a dose of 30 mg/kg or equivalent to 30 mg/kg ferulic acid of Chuanxiong extraction, respectively. The t1/2a, t1/2β, Vd/F, CL/F, AUC0-t, Tmax and Cmax have significantly differences following oral administration of Chuanxiong extraction of different solutions. The AUC0-t were 1.27±0.55 mg/L*h, 1.43±0.44 mg/L*h and 2.89±0.44 mg/L*h after oral administration of aqueous solution, ethyl acetate and alcoholic extraction. respectively. The t1/2a and t1/2β were 1.62±1.21 h, 0 .58±1.19 h, 0.42±0.97 h and 6.44±0.50 h, 7.02±0.36 h, 3.21±0.17 h after oral administration of aqueous solution, alcoholic and ethyl acetate extraction, respectively.Conclusions: (1) The maximum concentration and time of maximum concentration of oral administration ferulic acid and ferulic acid in Chuanxiong decoction had significant differences. The maximum concentration of ferulic acid in plasma was more 14 times than that in Chuanxiong decoction, but the peak time of maximum concentration of ferulic acid was less 5 times than that in Chuanxiong decoction. The results suggested that the absorption of ferulic acid after oral administration of ferulic acid in Chuanxiong decoction was much lower than ferulic acid in rat. Therefore, the pharmacokinetics parameters of monomer following oral administration cannot simply presume those of active component of traditional Chinese medicine.(2) With statistics analysis, the ferulic acid of pharmacokinetics parameter t1/2a, t1/2β, AUC0-t had significant differences in Chuanxiong extraction with different solutions in rat vivo. The AUC0-t and bioavailability of alcoholic extraction was larger than those of aqueous solution extraction or ethyl acetate extraction. The t1/2a aqueous solution extraction and t1/2β of alcoholic extraction were slow. The choice of different solutions should be noticed in extraction process of traditional Chinese medicine, and avoided influencing the physiological disposition of active components.New ideas: (1) Pharmacokinetics of ferulic acid of Chuanxiong extracted with different solutions was investigated for the first time. (2) The absorption of ferulic acid was estimated in rat vivo by deconvolution methods for the first time. |
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