高效液相-氢化物发生-原子荧光光谱法分析六神丸在大鼠肝脏中代谢的砷形态  

Analysis of Arsenic Metabolites of Liushen Pill in Rat Liver by HPLC-HG-AFS

作　　者：陈吴越 崔欣语 周昊言 王佳佳[1] 周婧[1] 吕翔[1] 马宏跃[1] CHEN Wuyue;CUI Xinyu;ZHOU Haoyan;WANG Jiajia;ZHOU Jing;LYU Xiang;MA Hongyue(Nanjing University of Chinese Medicine,Jiangsu Key Laboratory for High Technology Research of TCM Formula,Collaborative Innovation Center of Chinese Medicinal Resources Industrialization of Jiangsu Province,Nanjing 210023,China)

机构地区：[1]南京中医药大学,江苏省方剂高技术研究重点实验室,江苏省中药资源产业化过程协同创新中心,江苏南京210023

出　　处：《中医药信息》2021年第3期26-29,共4页Information on Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81673563,81274199);江苏省方剂高技术研究重点实验室/江苏省中药资源产业化过程协同创新中心资助项目(FJGJS-2015-15,ZDXM-14);六神丸质量标准化项目(ZDXM-1-14,FJGJS-2015-15);江苏省青蓝工程学术带头人项目。

摘　　要：目的:建立大鼠肝脏组织中四种砷形态的高效液相色谱-氢化物发生-原子荧光光谱(HPLC-HG-AFS)测定法,并用于大鼠灌胃给予六神丸后肝脏组织中砷形态研究。方法:乙酸-水(100∶1)作为提取溶剂,采用超声或细胞破碎进行前处理,离心取上清,重复提取3次,合并上清液。采用HamiltonPRP-X100色谱柱,以17.5 mmol/L(NH_(4))_(2)HPO_(4)(pH=5.9)为流动相,进行HPLC-HG-AFS分析。结果:四种不同砷形态在12 min内得到了良好的分离。在5~50 ng/mL范围内,线性关系良好,相关系数R 2均大于0.99。大鼠灌胃给不同剂量的六神丸,测得肝脏中砷形态为二甲基砷DMA,说明无机砷在大鼠体内发生了形态转变。同时发现砷在大鼠肝组织中存在蓄积现象,肝脏中二甲基砷浓度随给药量增加而增加。结论:该方法简单易行,灵敏度高,可用于准确检测大鼠肝脏中砷形态化合物。Objective:To establish the method using high performance liquid chromatography-hydride generation-atomic fluorescence spectrometry(HPLC-HG-AFS)to determine arsenic speciation in rat liver after the administration of Liushen Pill.Methods:Ultrasonic ice bath or ice cell disrupter was used to pretreat the samples with acetic acid-water(100∶1)as extraction agent.The supernatant after centrifugation was injected for HPLC-HG-AFS analysis.The separation of arsenicals were performed on a Hamilton PRP-X100 anion-exchange column with isocratic elution by 17.5 mmol/L(NH_(4))_(2)HPO_(4)(pH=5.9).Results:The separation of the four arsenic species were achieved within 12 minutes.Good linearities of arsenic speciations were in the ranges from 5 ng/mL to 50 ng/mL with the correlation coefficients of more than 0.99.Dimethyarsinic acid(DMA)was found to be the main species in liver after different doses of Liushen Pill intervention.The results showed that inorganic arsenic transformed to organic arsenic.Accumulation of arsenic was founded in rat liver.The concentration of DMA in liver increased with the increased dosing of Liushen Pill.Conclusion:The method is simple and sensitive,which can be used for the accurate analysis of arsenic speciations in rat liver.

关 键 词：六神丸 砷形态 肝组织 高效液相-氢化物发生-原子荧光光谱法 

分 类 号：R285.5[医药卫生—中药学]