Yamato S, Sugihara H, Shimada K
Department of Analytical Chemistry, Niigata College of Pharmacy, Japan.
Chem Pharm Bull (Tokyo). 1990 Aug;38(8):2290-2. doi: 10.1248/cpb.38.2290.
An alternative method for the assay of chloramphenicol using an enzymatic reaction coupled with a fluorescence detection system has been developed. Chloramphenicol was enzymatically acetylated by chloramphenicol acetyltransferase in the presence of acetyl-coenzyme A (acetyl-CoA) as the acetyl-donor, after which the liberated CoA-SH was derivatized with a fluorogenic reagent, 4-(aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole. The assay was linear over the range of 2.5-40 micrograms/ml. Analytical recoveries of chloramphenicol at concentrations of 7.5 and 22.5 micrograms/ml, added to human serum, plasma, or a slightly hemolyzed serum, were in the range of 93.3 to 106.2%. The enzymatic assay was not affected by the presence of ten other antibiotics tested.
已开发出一种使用酶促反应结合荧光检测系统来测定氯霉素的替代方法。在作为乙酰供体的乙酰辅酶A（acetyl-CoA）存在下，氯霉素被氯霉素乙酰转移酶进行酶促乙酰化，之后释放出的CoA-SH用荧光试剂4-（氨磺酰基）-7-氟-2,1,3-苯并恶二唑进行衍生化。该测定在2.5至40微克/毫升的范围内呈线性。添加到人体血清、血浆或轻度溶血血清中的浓度为7.5和22.5微克/毫升的氯霉素的分析回收率在93.3%至106.2%的范围内。该酶促测定不受所测试的其他十种抗生素存在的影响。
