Masuda M, Minobe S, Fukui T, Nawata M, Watanabe T, Shibatani T, Ogawa Y
Research Laboratory of Applied Biochemistry, Tanabe Seiyaku Co., Ltd., Osaka, Japan.
Yakugaku Zasshi. 1995 Feb;115(2):136-44. doi: 10.1248/yakushi1947.115.2_136.
We improved the Limulus amebocyte lysate (LAL) test for endotoxins in parenteral drugs using immobilized histidine and a filtration plate. In order to deal with many samples at the same time and to apply to a routine assay, we used a filtration plate having 96 wells instead of a filter unit with a working volume of 2 ml. LAL test-affecting substances which are contained in a parenteral drug were separated from endotoxins by adsorbing endotoxins on immobilized histidine in the well of a filtration plate. Then the absorbed endotoxins were allowed to react with LAL reagent in the same well. We defined that this method had the higher precision than conventional methods and was not influenced by the concentrations of endotoxin and parenteral drugs. Hence we examined the recovery of endotoxin spiked to 23 kinds of parenteral drugs by this method, as a result, 100 +/- 25% of recovery was obtained from 17 kinds of them.
我们使用固定化组氨酸和过滤板改进了用于注射用药物中内毒素检测的鲎试剂检测法。为了同时处理多个样品并应用于常规检测，我们使用了具有96个孔的过滤板，而不是工作体积为2 ml的过滤单元。通过将内毒素吸附在过滤板孔中的固定化组氨酸上，将注射用药物中含有的影响鲎试剂检测的物质与内毒素分离。然后使吸附的内毒素在同一孔中与鲎试剂反应。我们确定该方法比传统方法具有更高的精密度，并且不受内毒素和注射用药物浓度的影响。因此，我们用该方法检测了添加到23种注射用药物中的内毒素的回收率，结果，其中17种药物的回收率为100±25%。
