Direct determination of chloramphenicol acetyltransferase (CAT) activity in homogenates of transfected mammalian cells by high performance liquid chromatography.
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.
Siegert
H
H
Institut für organische Chemie, Freie Universität Berlin.
Wittig
B
B
Wölfl
S
S
D000097:Acetonitriles; D000107:Acetylation; D000818:Animals; D002457:Cell Extracts; D002478:Cells, Cultured; D002701:Chloramphenicol / Q000378:metabolism; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D051379:Mice; D010957:Plasmids; D014162:Transfection
D000097:Acetonitriles; D002457:Cell Extracts; D002701:Chloramphenicol; D015500:Chloramphenicol O-Acetyltransferase; C032159:acetonitrile