Valassis I N, Parissi-Poulou M, Macheras P
Department of Pharmacy, University of Athens, Greece.
J Chromatogr B Biomed Sci Appl. 1999 Jan 22;721(2):249-55. doi: 10.1016/s0378-4347(98)00468-x.
An isocratic reversed-phase HPLC method was developed to determine cefepime levels in plasma and vitreous fluid. Cefepime and the internal standard cefadroxil were separated on a Shandon Hypersil BDS C18 column by using a mobile phase of 25 mM sodium dihydrogen phosphate monohydrate (pH 3) and methanol (87:13, v/v). Ultraviolet detection was carried out at 270 nm. The retention times were 4.80 min for cefepime and 7.70 min for cefadroxil. This fast procedure which involves an efficient protein precipitation step (addition of HClO4), allows a quantification limit of 2.52 microg ml(-1) and a detection limit of 0.83 microg ml(-1). Recoveries and absolute recoveries of cefepime from plasma were 96.13-99.44% and 94-102.5% respectively. The intra-day and inter-day reproducibilities were less than 2% for cefepime at 10, 30, 50 microg ml(-1) (n=10). The method was proved to be suitable for determining cefepime levels in human plasma and was modified to measure vitreous fluid samples.
建立了一种等度反相高效液相色谱法来测定血浆和玻璃体液中的头孢吡肟含量。头孢吡肟和内标头孢羟氨苄在Shandon Hypersil BDS C18柱上分离，流动相为25 mM磷酸二氢钠一水合物（pH 3）和甲醇（87:13，v/v）。在270 nm处进行紫外检测。头孢吡肟的保留时间为4.80分钟，头孢羟氨苄的保留时间为7.70分钟。该快速方法包括一个高效的蛋白质沉淀步骤（加入高氯酸），定量限为2.52 μg ml⁻¹，检测限为0.83 μg ml⁻¹。头孢吡肟从血浆中的回收率和绝对回收率分别为96.13 - 99.44%和94 - 102.5%。在10、30、50 μg ml⁻¹（n = 10）浓度下，头孢吡肟的日内和日间重现性均小于2%。该方法被证明适用于测定人血浆中的头孢吡肟含量，并经过改进用于测量玻璃体液样本。
