Borner K, Borner E, Lode H
Institut für Klinische Chemie und Klinische Biochemie, Klinikum Steglitz, Freie Universität Berlin, Germany.
J Chromatogr. 1993 May 19;615(1):174-9. doi: 10.1016/0378-4347(93)80306-o.
A high-performance liquid chromatographic method for the determination of a new-broad spectrum cephalosporin (I, SCE-2787) has been developed. The analyte was extracted from serum by precipitation of serum proteins with acetonitrile. Acetonitrile was extracted from the protein-free supernatant by dichloromethane. Urine was simply diluted with mobile phase. Separation was performed by ion-pair chromatography on a reversed-phase column (Nucleosil 5C18; 125 mm x 4.0 mm I.D.; 5 microns average particle size). The guard column was Perisorb RP18 (30 mm x 4.0 mm I.D.; 30-40 microns particle size). The mobile phase was acetonitrile-buffer solution containing 15 mM heptanesulphonic acid (pH 3.2) (4.5:95.5, v/v). Detection was performed at 235 nm with a diode-array detector, which also served to record ultraviolet spectra. The assay was sensitive, precise, accurate and fast. Specificity was controlled by on-line recording the ultraviolet spectrum of I and also by enzymic degradation with beta-lactamase. No interferences were observed during the analysis of serum and urine of healthy volunteers in pharmacokinetic studies.
已开发出一种用于测定新型广谱头孢菌素（I，SCE - 2787）的高效液相色谱法。通过用乙腈沉淀血清蛋白从血清中提取分析物。用二氯甲烷从无蛋白上清液中提取乙腈。尿液只需用流动相稀释。在反相柱（Nucleosil 5C18；125 mm×4.0 mm内径；平均粒径5微米）上通过离子对色谱法进行分离。保护柱为Perisorb RP18（30 mm×4.0 mm内径；粒径30 - 40微米）。流动相为含有15 mM庚烷磺酸（pH 3.2）的乙腈 - 缓冲溶液（4.5:95.5，v/v）。用二极管阵列检测器在235 nm处进行检测，该检测器还用于记录紫外光谱。该测定方法灵敏、精密、准确且快速。通过在线记录I的紫外光谱以及用β - 内酰胺酶进行酶促降解来控制特异性。在药代动力学研究中对健康志愿者的血清和尿液分析过程中未观察到干扰。
