Fox K E
Department of Pharmacology, School of Medicine, Oregon Health Sciences University, Portland 97201.
Antimicrob Agents Chemother. 1989 Apr;33(4):448-51. doi: 10.1128/AAC.33.4.448.
In most experiments when aminoglycoside antibiotic (AG) tissue levels are measured, the AG is extracted into buffered media. The data from this study reveal that buffer extraction results in only partial recovery of AG from tissues and that total recovery can be obtained after NaOH or trichloroacetic acid treatment. Tissues studied here included the whole bullfrog and guinea pig kidney and cochlear tissues after in vivo drug treatment. The AGs used were kanamycin, tobramycin, and gentamicin. Drug concentrations were determined by enzymatic assay and, in the case of tobramycin, also 3H-labeled radioactivity. Only a fraction of total concentration of AG in tissue was released into the supernatants of tissue homogenates. However, the remainder was recovered after NaOH solubilization of the residual pellet. Also, it was found that the G released from the pellet by NaOH was associated with protein. By contrast, trichloroacetic acid precipitation of tissue protein immediately released the drug into the supernatant.
在大多数测量氨基糖苷类抗生素(AG)组织水平的实验中,AG被提取到缓冲介质中。本研究的数据表明,缓冲液提取只能使组织中的AG部分回收,而在氢氧化钠或三氯乙酸处理后可实现完全回收。这里研究的组织包括经体内药物处理后的整个牛蛙、豚鼠肾脏和耳蜗组织。使用的AG有卡那霉素、妥布霉素和庆大霉素。药物浓度通过酶法测定,对于妥布霉素,还通过³H标记放射性测定。组织中AG总浓度只有一部分释放到组织匀浆的上清液中。然而,剩余部分在对残留沉淀进行氢氧化钠溶解后回收。此外,发现通过氢氧化钠从沉淀中释放出的G与蛋白质相关。相比之下,组织蛋白的三氯乙酸沉淀立即将药物释放到上清液中。