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目的:建立HPLC法测定小鼠不同组织和排泄物中痂囊腔菌素A浓度的方法。研究痂囊腔菌素A在小鼠体内组织的分布与粪、尿中的排泄动力学。方法:小鼠灌服30 mg痂囊腔菌素A后,采集样品,用HPLC定量方法测定痂囊腔菌素A在小鼠肝、肾、肺、脑及粪、尿中的浓度。结果:痂囊腔菌素A在10～1 200μg·L-1内线性关系良好(r=0.999 7～0.997 2),最低检出限量1μg·L-1,日间和日内RSD≤7.13%,提取回收率为75.5%～97.6%,方法回收率在97.9%～105.3%。小鼠灌服痂囊腔菌素A后,在组织中的分布浓度由高到低依次为肝、肺、肾、脑。尿和脑中未检测出痂囊腔菌素A。24 h痂囊腔菌素A在粪便中的累积排泄量为(83.7±3.9)%。结论:所建立的HPLC分析方法准确、灵敏、重复性好。小鼠灌服痂囊腔菌素A后,在肝、肺、肾中有分布,主要经肝、肠途径由粪便排出。
OBJECTIVE: To establish a HPLC method for the determination of cystatin A concentration in different tissues and excretions of mice. To investigate the distribution of cystatin A in mice and the excretion kinetics of excretion and urine. Methods: Mice were inoculated intraperitoneally with 30 mg of cystatin A. Samples were collected and the concentration of cystatin A in liver, kidney, lung, brain and feces and urine of mice was determined by HPLC quantitative method. Results: Cystatin A had a good linearity (r = 0.999 7 ~ 0.9972) in 10-1 200 μg · L-1 and the lowest limit of detection was 1 μg · L-1. The RSD was lower than or equal to 7.13% The recovery rate was 75.5% ~ 97.6%. The recovery rate was 97.9% ~ 105.3%. After mice were inoculated with cystatin A, the distribution in the tissues from high to low was liver, lung, kidney and brain. No cystatin A was detected in urine and brain. The cumulative excretion of stupidin A in feces was (83.7 ± 3.9)% in 24 h. Conclusion: The established HPLC method is accurate, sensitive and reproducible. After mice were inoculated with cystatin A, there was distribution in the liver, lung and kidney, and was mainly excreted in the feces through the liver and intestine.