Granich G G, Krogstad D J
Antimicrob Agents Chemother. 1987 Mar;31(3):385-8. doi: 10.1128/AAC.31.3.385.
We developed a high-performance liquid chromatographic assay to measure ceftriaxone in serum, urine, and cerebrospinal fluid. Ion pairing was used because ceftriaxone is a relatively polar compound which is poorly retained on C18 columns in standard reverse-phase high-performance liquid chromatography and which produces trailing peaks in the absence of ion-pairing agents. The mobile phase was a combination of acetonitrile and water (46:54), adjusted to pH 9.0 with 10 mM K2HPO4, which contained 10 mM hexadecyltrimethylammonium bromide as the ion-pairing agent. Moxalactam (200 micrograms/ml) was used as the internal standard. A silica-packed precolumn (3 cm long) was used to prevent rapid deterioration of the analytical column (30 by 0.4 cm) by the alkaline pH of the mobile phase, and it significantly extended the life of the analytical column. The assay was linear with ceftriaxone concentrations of 1 to 250 micrograms/ml (r = 0.999) and correlated well with an agar diffusion bioassay (r = 0.990). Reproducibility was good, with intrarun coefficients of variation from 2.3 to 6.4% and interrun coefficients of variation from 3.2 to 21.4%. The absolute recoveries of ceftriaxone and moxalactam were 91 to 97 and 96 to 98%, respectively. No interferences were observed with more than 40 commonly prescribed drugs, including 10 cephalosporins (cefotaxime, cefoperazone, ceftazidime, ceftizoxime, cefoxitin, cefamandole, cephalothin, cefazolin, cephapirin, and cephalexin), or with sera from patients with renal or hepatic disease.
我们开发了一种高效液相色谱法来测定血清、尿液和脑脊液中的头孢曲松。采用离子对法是因为头孢曲松是一种相对极性的化合物，在标准反相高效液相色谱中，它在C18柱上保留性较差，并且在没有离子对试剂的情况下会产生拖尾峰。流动相是乙腈和水（46:54）的混合物，用10 mM磷酸氢二钾调节至pH 9.0，其中含有10 mM十六烷基三甲基溴化铵作为离子对试剂。莫西沙星（200微克/毫升）用作内标。使用硅胶填充的预柱（3厘米长）以防止分析柱（30×0.4厘米）因流动相的碱性pH值而快速劣化，并且它显著延长了分析柱的使用寿命。该测定法在头孢曲松浓度为1至250微克/毫升时呈线性（r = 0.999），并且与琼脂扩散生物测定法相关性良好（r = 0.990）。重现性良好，批内变异系数为2.3%至6.4%，批间变异系数为3.2%至21.4%。头孢曲松和莫西沙星的绝对回收率分别为91%至97%和96%至98%。未观察到超过40种常用处方药的干扰，包括10种头孢菌素（头孢噻肟、头孢哌酮、头孢他啶、头孢唑肟、头孢西丁、头孢孟多、头孢噻吩、头孢唑林、头孢匹林和头孢氨苄），也未观察到来自患有肾脏或肝脏疾病患者血清的干扰。
