| Objective:Realgar is a traditional Chinese medicine(TCM)containing arsenic and is widely used in clinical practice.Although the oral utilization of realgar is relatively low,drug-induced arsenic poisoning incidents caused by unscientific(long-term,excessive and irregular)rational use of single realgar or its compound preparations have been reported from time to time.However;the theory of compatibility,detoxification and synergism of TCM plays an important guiding role in the rational application of toxic TCM.Literature reports that Scutellaria has the effect of reducing realgar-induced liver and kidney toxicity in the prescription,but the pharmacokinetic research after Realgar-Scutellaria compatibility is rarely reported.Therefore,this study studied the mechanism of attenuation and synergism of Realgar-Scutellaria compatibility from the following aspects:(1)Establish Atomic Fluorescence Spectrometry(AFS)to determine the effect of different compatibility ratios of realgar and Scutellaria on soluble arsenic content in realgar,and screen out the best detoxification ratio of Realgar-Scutellaria.(2)Establish an AFS method for the determination of arsenic in blood and study the effect of Realgar-scutellaria compatibility on the process of arsenic in vivo.(3)Establish a high performance liquid chromatography-mass spectrometry(HPLC-MS)method for the determination of flavonoids in plasma to study the effect of Realgar-Scutellaria compatibility on the in vivo process of flavonoids.In order to clarify the scientific connotation of the mechanism of toxicity reduction and synergism of Scutellaria and realgar compatibility,experimental basis is provided,and reference is provided for reasonable and safe medication of compound traditional Chinese medicine containing realgar.Methods:1.AFS analysis method was established to determine soluble arsenic in different proportions of Realgar-Scutellaria drug pairs.At the same time,the effects of artificial gastric juice volume and ultrasonic extraction time on soluble arsenic dissolution in realgar were investigated.2.Establish AFS analysis method to determine arsenic concentration in blood of realgar group and Realgar-Scutellaria group,draw blood concentration-time graph of arsenic,and compare the difference of pharmacokinetic parameters.3.Establish a high performance liquid chromatography-tandem mass spectrometry(HPLC-MS)method for the determination of flavonoids in rat plasma to determine flavonoids in plasma of Scutellaria group and Realgar-Scutellaria group,and to draw drug time curves and compare the differences of pharmacokinetic parameters.The experimental data were expressed by mean ± standard deviation((?)),and the data were analyzed and processed by SPSS 19.0 statistical software.The significance test of the difference between groups was carried out by one-sample t-test.The difference was statistically significant(P<0.05).Results:1.Under the conditions of 8 mL artificial gastric juice volume and 1.5 h ultrasonic extraction time,the dissolution amount of soluble arsenic in single realgar was the highest,and the soluble arsenic content gradually decreased with the increase of the proportion of Scutellaria in different compatibility ratios of realgar to Scutellaria.When the ratio of realgar to Scutellaria was 1:4,the soluble arsenic content reached the lowest value.2.The established AFS analysis method can be used for the determination of arsenic content in medicinal materials and blood with good precision,accuracy and recovery rate,which meets the analysis requirements of biological samples.From the blood arsenic curve and parameters of realgar and Realgar-Scutellaria group,the blood arsenic drug time curves of realgar group and Realgar-Scutellaria group in rats showed "double peaks",and the first absorption peak value was higher than the second peak value.Compared with realgar group,the second peak concentration(Cmax)of realgar-Scutellaria group increased(P<0.05),and the second peak time(Tmax)was significantly earlier(P<0.05),indicating that Scutellaria accelerated the absorption of arsenic in rats.The second half-life(T1/2)of arsenic in Realgar-Scutellaria group was significantly lower than that in Realgar group(P<0.05),and the clearance rate(CLz)was significantly increased(P<0.05),indicating that Scutellaria accelerated the metabolic rate of arsenic in vivo.3.To establish HPLC-MS analysis method for the determination of baicalin,baicalein and baicalein in Scutellaria.According to the results of the methodological investigation,the linear relationship between baicalin and baicalein was good(R2?0.99)in the range of baicalin concentration 0.230-9.20 ?g/mL and baicalein concentration 0.137-5.50 ?g/mL.The specificity,precision,accuracy,extraction recovery rate,matrix effect and stability all meet the requirements of biological sample analysis,and can be used for pharmacokinetic study of flavonoids of Scutellaria in rats,From the baicalin and baicalein drug curve and parameters of the Scutellaria group and Realgar-Scutellaria group,Baicalin and baicalein in Scutellaria group and Realgar-Scutellaria group have "double peaks" in rats.Compared with Scutellaria group,Realgar-Scutellaria group reduced the first peak concentration Cmax,advanced T(1/2)1,delayed the peak time of the second peak,decreased AUC0-t and CLz.In the Realgar-Scutellaria group,the peak concentration of baicalein at the first peak decreased,the half-life of the second peak T(1/2)2 prolonged,MRT,Vz and CLz all increased to different degrees,which indicates that Realgar can slow down the absorption,distribution and elimination time of baicalin and baicalein,the active components of Scutellaria,and prolong their action time in vivo.Conclusion:The established AFS analysis method can be used for the determination of arsenic content in medicinal materials and blood.The method has the advantages of high sensitivity and simple operation.2.The established HPLC-MS analysis method can be used for the determination of flavonoids in plasma.The method has the advantages of good separation effect,good selectivity,high sensitivity,high accuracy and good reproducibility.3.Scutellaria has significant inhibitory effect on the dissolution of soluble arsenic in realgar,which can accelerate the absorption and metabolism of arsenic in realgar in rats and reduce the toxicity of realgar.After the compatibility of realgar and Scutellaria,the action time of baicalin and baicalein in Scutellaria in rats is prolonged and the metabolic time is slow,suggesting that realgar can prolong the action time of baicalin and baicalein in vivo. |
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