高效液相色谱法测定生物样本中头孢噻肟和舒巴坦的浓度
【关键词】 头孢噻肟；舒巴克坦；色谱法,高压液相；血浆；尿
【Abstract】 AIM: To establish a determination method of concentrations in human biologic sample following intravenous administration of cefotaxime and sulbactam. METHODS: A RPHPLC method was developed and validated to determine cefotaxime and sulbactam in plasma and urine. The mobile phase of cefotaxime and sulbactam consisted of 1 g/L triethylamine (regulated with phosphate acid to pH=6.2) and methanol in a 68∶32 and 84∶16 (v/v) respectively and pumped at a flow rate of 1 mL/min. The plasma samples were deproteinated by addition of methanol, the urine samples were diluted with super pure water, then 20 μL of supernatant was injected onto a C18 column and monitored with UV detector at 254 nm and 220 nm respectively. RESULTS: The assay of cefotaxime in human plasma and urine were linear over the range of 5.3-530.0 mg/L and 5.45-545.00 mg/L respectively, with the detection limit of 1 mg/L and 0.25 mg/L respectively. The assay of sulbactam in human plasma and urine were linear over the range of 1.04-208.00 mg/L, with the detection limit of 0.50 mg/L and 0.25 mg/L respectively. The average recovery of the method was (89-110)%, and the RSD within and between days were all less than 5%. The cefotaxime and sulbactam sample of plasma and urine stored in a -20℃ refrigerator became stable within 3 d. CONCLUSION: The assay method is rapid, accurate and usable for the determination of cefotaxime and sulbactam concentrations in plasma and urine.
【Keywords】 cefotaxime; sulbactam; chromatography, high pressure liquid; plasma; urine
【摘要】 目的： 为静滴注射用头孢噻肟钠舒巴坦钠后在血浆和尿液中的药物浓度分析提供检测方法. 方法： 采用HPLC测定头孢噻肟钠血浆和尿液中浓度. 在血样中加入甲醇沉淀蛋白后，尿样用超纯水稀释后取上清液进样，色谱分析条件: 以C18反相柱作分析柱；柱温：室温；流速： 1 mL/min；头孢噻肟和舒巴坦的流动相分别为：1g/L的三乙胺水溶液（用磷酸调pH值6.2）∶甲醇＝68∶32和84∶16；头孢噻肟和舒巴坦的`紫外检测波长分别为254 nm和220 nm. 结果： 头孢噻肟血样和尿样的线性范围分别为5.3～530.0 mg/L和5.45～545.00 mg/L，最低检测浓度分别为1.00 mg/L和0.25 mg/L；舒巴坦血样和尿样的线性范围为1.04～208.00 mg/L，最低检出浓度分别为0.5 mg/L和0.25 mg/L；方法平均提取回收率在 (89～110)%之间；日内、日间RSD均小于5%. 头孢噻肟和舒巴坦血浆和尿液样品置于-20℃冰箱3 d内稳定. 结论： 本方法具有快速、简便、灵敏、准确等特点,适合于头孢噻肟和舒巴坦血浆和尿液浓度测定.
【关键词】 头孢噻肟；舒巴克坦；色谱法，高压液相；血浆；尿
0引言
由湘北威尔曼制药有限公司开发研制的一类复方新药注射用头孢噻肟钠舒巴坦钠已经国家食品药品监督管理局批准进行Ⅰ期临床试验（临床研究批件号：2005L00727）. 头孢噻肟钠属第三代半合成头孢菌素类，其抗菌谱广，对革兰阴性菌、革兰阳性菌、需氧菌和某些厌氧菌均有较好的抗菌活性，特别对革兰阴性菌的杀灭作用更强. 随着头孢噻肟在临床的广泛使用，许多临床病原菌通过产生β内酰胺酶对头孢噻肟产生耐药性而导致临床治疗方案的失败［1-2］. 舒巴坦为半合成β内酰胺酶抑制剂,与头孢噻肟合用时出现协同作用，使对头孢噻肟耐药的细菌的最低抑菌浓度下降到敏感范围之内. 为了评价静滴注射用头孢噻肟钠舒巴坦钠后该药品在健康志愿受试者的耐受性以及药代动力学特点，我们采用高效液相色谱法测定了生物样本中头孢噻肟和舒巴坦的浓度.
