| Objective:Realgar is an arsenic-containing mineral medicine with a wide range of clinical applications.It is one of the traditional Chinese medicines commonly used in China.Long-term,excessive and irregular use of realgar or its preparations will cause the accumulation of arsenic in the body.Compatibility is an important method for the safe and rational application of toxic traditional Chinese medicine.It has been reported in the literature that platycodon grandiflorum has the effect of reducing realgar-induced hepatorenal toxicity in the prescription,but its specific mechanism is not clear.Based on these understandings,this article will explore the mechanism of realgar-platycodon grandiflorum compatibility from the following aspects:?1?established atomic fluorescence spectrophotometry?AFS?method to detect the concentration of soluble arsenic in different proportions of realgar andplatycodon grandiflorum and screened the best proportion according to the least soluble arsenic concentration;?2?a AFS method was established to investigate the effect of realgar-platycodon grandiflorum combinations on the fate of arsenic in vivo;?3?a high performance liquid chromatography-mass spectrometry?HPLC-MS?method was established to investigate the effect of realgar-platycodon grandiflorum combinations on the fate of Platycodin D?PD?in vivo.The study provide an experimental basis and new ideas for the mechanism research of the toxicity reducing and efficacy enhancing of realgar-platycodon grandiflorum combinations and its clinical application.Methods:1.A AFS method was established to determine the concentration of soluble arsenic in different proportions of realgar andplatycodon grandiflorum extract solution extracted by artificial gastric juice.The effects of ultrasonic extraction time and volume of artificial gastric juice on the dissolution of soluble arsenic were also investigated.2.A AFS method was established to determine the concentration of arsenic in blood and applied in the pharmacokinetics study of realgar and realgar-platycodon grandiflorum combinations.3.A HPLC-MS method was established to to measure the concentration of PD in plasma and applied in the pharmacokinetics study of platycodon grandiflorum and realgar-platycodon grandiflorum combinations.The data obtained from the experiment were expressed as average±standard.Differences were calcualted by SPSS19.0 software using One-sample t-test.The significance of differences between two groups was statistically significant.P<0.05 was considered statistically significant.Results:1.When the ultrasonic and extract time was 1.5 h,the volume of artificial gastric juice was 8 m L,realgar dissolved the most soluble arsenic,so it was the best experimental condition.With the increase of platycodon grandiflorum proportion,the concentration of soluble arsenic decreased gradually,and when the ratio of realgar toplatycodon grandiflorum was 1:6,the concentration of soluble arsenic was the lowest.2.The established AFS method can be used for the determination of arsenic in blood,and the method meet the requirements of biological sample analysis.There were significant differences in some pharmacokinetic parameters for arsenic in realgar group and realgar-platycodon grandiflorum group?P<0.05?.Both two abosorption peaks appeared in the blood concentration-time curves of realgar group and the realgar-platycodon grandiflorum group,and the Cmaxax value of the first peak was greater than the Cmaxax value of the second peak.Compared with the realgar group,the peak time of the second absorption peak in the compatible group was significantly reduced,and the Cmaxax of the second absorption peak increased from 7.72±1.62?g/m L to 11.3±3.74?g/mL?P<0.05?,suggesting that realgar and platycodon grandiflorum combinations accelerated the absorption of arsenic in rats'blood;the half-life of the first absorption peak(t1/2?1?)was reduced from 59.6±14.9 h to 18.1±6.46 h?P<0.05?,which indicated that the metabolic rate of arsenic in vivo was accelerated.3.Platycodin D?PD?,the main active component of platycodon grandiflorum,has a good linear relationship?R2>0.999?in the concentration range of 2.00-160 ng/mL,and the lower limit of quantification is 2.00ng/mL.The specificity,precision,accuracy,extraction recovery,matrix effect and stability of the established HPLC-MS method meet the requirements of biological sample analysis.The established HPLC-MS method for the analysis of PD in rat plasma was applied to the study of pharmacokinetics of platycodon grandiflorum group and realgar-platycodon grandiflorum group in rat plasma and the differences of pharmacokinetic behaviors of PD were compared.The results showed that the peak concentration Cmax?P<0.05?and the area under the drug concentration-time curve?AUC0-t?of PD in the realgar-platycodon grandiflorum group were significantly reduced as compared with the platycodon grandiflorum group?P<0.05?.The peak time Tmax,the average residence time MRT,and the half-life t1/2/2 increased?P<0.05?,suggesting that PD in the realgar-platycodon grandiflorum group showed slower absorption,distribution,and eliminate.Conclusion:1.The established AFS method is simple,fast,sensitive and reproducible,it can be used to determine the concentration of arsenic in medicine and blood.2.The established HPLC-MS method can be used to determine the concentration of PD in plasma.The method has the advantages of well seperation,good selectivity,high sensitivity,high accuracy and well repeatability.3.Platycodon grandiflorum can significantly reduce the dissolution of soluble arsenic in artificial gastric juice and accelerate the absorption and metabolism of arsenic in blood,therefore it can reduce the toxicity of realgar.PD can be absorbed and metabolized at a fast rate in vivo.Realgar can prolong the action time of PD in vivo. |
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