| Sanhuang dispersible tablet is a kind of dosge forms from Traditional Chinese medicine(TCM) preparation by modern extractive technique and preparation means, researched by institute of TCM laboratory pharmaceutical ,including rhubarb total anthraquinones extractive,baicalin and berberine as main medicine compositions. It possesses pharmacologic action of purging phathogenic fire, dejections, heat-clearing and detoxicating. Compared with the traditional San-huang sugar-coated tablets, it possess better qualities of quality-contolled and stable curative effect. But the research is less for San-huang dispersible tablets on pharmacokinetic. Then pharmacokinetical research on TCM preparations has also evoked more and more attention. So it is essential to study this aspect deeply for San-huang dispersable tablets.Objective: To establish research methods on pharmacokinetics in vivo of normal, febrile and inflame rat model for San-huang dispersible tablet;and evaluate the correlation between plasma concentration of effective composition in vivo and pharmacodymatics.Methods: (1) to determine content of bacalin, aloe-emodin, and rhein in rats plasma by release phase high performance liquid chromatograph(HPLC)mehod. (2) to choose wistar male rats as experimental animals, divided into thirteen groups, randomly, and five rats each group. To establish rats voix pedis engorgement model with 1% crn as making phlegmonosis agent, and inspect the effect on rats voix pedis engorgement of San-huang dispersible tablets. (3) to establish rats febrility model with 20% dride yeast suspend solution as fever agent, and inspect the effect on rats body temperature of San-huang dispersible tablets after administrated intragastrically. (4) to choose wistar male rats as experimental animals, divided into thirteen groups randomly, and five rats each group, and administrated San-huang dispersible tablets suspending solution as the dosage of 1650mg/kg intragastrically, then recipe blood from eyeball at 5min, 15min, 30min, 1h, 2h, 3h, 5h, 7h, 9h, 11h,14h,18h,24h after intragastric administration to determine the bacalin, aloe-emodin, and rhein concentration in plasma by Rp-HPLC method. (5) to handle the data of medicine concentration in plasma using 3p97 and Topfit pharmacokinetic software, and calculate pharmacokinetic parameter for baicalin, aloe-emodin and rhein in San-huang dispersible tablets in normal, febrile and inflame rat model, respectively, then compared the parameters among different model to find whether it exit difference or not for baicalin, aloe-emodin and rhein, respectively. (6) to establish quantity-effect curve to find whether the concentration of baicalin, aloe-emodin and rhein in San-huang dispersible tablets in plasma at different time is correlated with the change value of rat body temperature and voix pedis engorgement degree or not.Result: (1) In the blank plasma samples of rats,the linear range of baicalin was from 0.32μg/mL to 6.4μg/mL. The lowest detection limitation was 0.1μg/mL at s/N≥3. The precision of low, middle, and high concentration plasma sample of bacalin within and between days was good, and RSD7.3%,4.2%,5.3%;6.9%,4.7%,3.4%,respectively. The accuracy is -2.3%,9.9%,11.7% for low, middle,and high, respectively;the extraction recovery rate of low, middle, high concentration plasma sample of baicalin is 73.50%,74.17%,78.27%,respectively, and the RSD is 3.11%,4.08%,1.49%;the low, middle, high concentration plasma sample of baicalin is stable for 24h at room temperature, and the residue of three kinds of baicalin plasma sample is stable for 48h at 4℃, but the solution of low concentration sample is not stable at 4℃, while the middle and high plasma sample is stable.(2)Also,the linear range of aloe-emodin and rhein was 0.1077~2.6928μg/mL ,0.2016~5.04μg/mL, respectively. The lowest detection limitation was 0.08μg/mL and 0.14μg/mL at s/N≥3, respectively. The precision of low, middle, and high concentration plasma sample of aloe-emodin and rhein within and between days was good, and RSD10.1%,5.7%,5.1%(within days) 10.8%,0.88%,7.5% (between days);1.7%,5.6%,9.6%(within days), 10.5%,4.0%,5.6%(between days), respectively. the accuracy of aloe-emodin and rhein is 17.6%,-13.5%,7.9% and 7.3%,-15.7%,14.9% for low, middle,and high, respectively;the extraction recovery rate of low, middle, high concentration plasma sample of aloe-emodin and rhein is 92.66%,65.48%,67.84%,58.37%,59.64%,63.45%, respectively, RSD为5.86%,2.86%,2.97%, 2.37% , 1.22% , 4.03% respectively. The low, middle, high concentration plasma sample of aloe-emodin and rhein is stable for 24h at room temperature, and the residue of three kinds of aloe-emodin and rhein plasma sample is also stable for 48h at 4℃. (3) San-huang dispersible tablets can inhibit rat feet swelling degree by crn. (4) San-huang disperbal tablets can also inhibit rat body temperature elevated degree induced by dried yeast. (5) the absorption of baicalin in San-huang dispersible tablets consistent with two-compartment model in vivo of normal, febrile and inflame rat. Among the total, the pharmacokinetic parameters of baicalin in inflame model are T1/2(α)214.9min,T1/2(β) 435.9min,T1/2(ka)168.0min,AUC 1879μg·min/mL,V 275mL,CL(s)0.88,respectively;the febrile model is T1/2(α)33.8min,T1/2(β)705min,T1/2(ka)13.9min,AUC 971μg·min/mL,V 1098mL,CL(s)1.70;and the normal is T1/2(α)272min,T1/2(β) 353min,T1/2(ka)102.1min,AUC 1431.9μg·min/mL,V 584mL,CL(s)1.15. Calculate by Topfit soft, the pharmacokinetic parameter of rhein in inflame model is,T1/2262min,AUC(0-t)642.15μg·min/mL,V 944mL,CL(s)2.50L/min,MRT(0-t)482min,MRT(0-∞)520min;the febrile is T1/2665min,AUC(0-t) 455.53μg·min/mL,V 2690mL,CL(s)2.80L/min,MRT(0-t)510min,MRT(0-∞)940min;and the normal is T1/2183min,AUC(0-t) 563.23μg·min/mL,V 772mL,CL(s)2.93L/min,MRT(0-t)358min,MRT(0-∞)358min;there is no absorbed peak in plasma of normal rats for aloe-emodin,while the pharmacokinetic parameter of aloe-emodin in inflame model is T1/2969min,AUC(0-t) 528.7μg·min/mL,V 3010mL,CL(s)2.15L/min,MRT(0-t)627min,MRT(0-∞)1313min;the febrile model is T1/21310min,AUC(0-t)691.08μg·min/mL,V 2300mL,CL(s)1.22L/min, MRT(0-t)768min,MRT(0-∞)2020min.Conclusion:(1) Established Rp-HPLC method to determine baicalin, rhein and aloe-emodin in plasma samples, this method possesses high sensitivity, strong specificity, operated easily, and the result is accuracy and reliable.(2) San-huang dispersible tablets possess anti-inflammatory action. (3) San-huang dispersible tablets possess antifebric action. (4) the pharmacokinetics of baicalin in San-huang dispersible tablets all consistent with two-compartment model in vivo of normal, inflame, and febrile model rat. And there is less difference on rate of elimination, peak time, peak concentration and AUC of baicalin between normal rat and inflame rat , but there is some difference on major pharmacokinetic parameter, such as rate of elimination, peak time, peak concentration and AUC, between normal and febrile rat, the same as the inflame and febrile model rat. But the rate of elimination is slow in vivo of febrile model rat for rhein in San-huang dispersible tablets, different from the normal and inflame model rat, and the other major pharmacokinetic parameters are also different among the three kinds of model, but having no statistic sense. Aloe-emodin can not be determined in the plasma sample of normal rat. Furthermore the pharmacokinetics exist some difference between inflame and febrile model rat, Compared with the febrile rat, the rate of elimination is faster, the peak time is shorter, and the distribution is wider in vivo of the inflame rat. (5) The plasma concentration of Baicalin possesses negative correlation with feet swelling degree of rats with lag time. Also there is negative correlation between the plasma concentration of aloe-emodin and body temperature chang values of rats, while it is positive correlation for rhein.But the reason that the concentration of rhein is positive correlation with body temperature is not clear , we will research it.
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