Direct determination of chloramphenicol acetyltransferase (CAT) activity in homogenates of transfected mammalian cells by high performance liquid chromatography.

H Siegert, B Wittig, S Wölfl
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Abstract

The conversion of chloramphenicol to its monoacetylated form by homogenates of pRSV CAT-transfected mammalian cells was assayed by HPLC. The method differed from conventional procedures, in that the extraction of chloramphenicol and its acetylated forms into an organic solvent was replaced by treatment of the cell homogenates with acetonitrile. This allows a rapid, direct analysis by HPLC with few experimental steps.

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高效液相色谱法直接测定转染动物细胞匀浆中氯霉素乙酰转移酶(CAT)活性。
用高效液相色谱法测定了经pRSV cat转染的哺乳动物细胞匀浆中氯霉素转化为单乙酰化形式的情况。该方法与传统方法的不同之处在于,将氯霉素及其乙酰化形式提取到有机溶剂中,而不是用乙腈处理细胞匀浆。这允许快速,直接的高效液相色谱分析与很少的实验步骤。
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