庆大霉素C1a生产过程中不同样品的HPLC法检测  

Determination of Different Samples in the Preparation of Gentamicin C1a by HPLC

作　　者：丁碧粉 张韬[1,3] 李继安[1] 陈代杰 林惠敏[1] 

机构地区：[1]中国医药工业研究总院上海医药工业研究院,创新药物与制药工艺国家重点实验室,上海201203 [2]上海师范大学生命与环境科学学院,上海200234 [3]中国药科大学,江苏南京211198 [4]中国医药工业研究总院,上海201203

出　　处：《中国医药工业杂志》2016年第3期310-315,共6页Chinese Journal of Pharmaceuticals

基　　金：国家“重大新药创制”科技重大专项(2012ZX09201101-008);国家自然科学基金(81273413);上海市自然科学基金(14ZR1440200)

摘　　要：参照美国药典USP37-NF32版,采用柱前衍生化-HPLC法测定庆大霉素C1a(1)对照品及生产阶段的不同样品。在检测经离子交换树脂HZ-732富集后的洗脱液样品B时出现1个新的杂质峰,且检测所得1的浓度明显偏低。经LCMS分析,推测该杂质为邻苯二甲醛(OPA)不足的情况下1与巯基乙酸在碱性条件下的结合产物。经方法优化,通过稀释样品、增加衍生化试剂量及增加衍生化试剂中OPA的量,均可准确检测不同样品中1的浓度和纯度。向衍生化试剂中增加OPA,更适用于1发酵生产过程中不同阶段样品的检测。According to the USP37-NF32, the gentamicin Cla （1） reference substance and different samples during the fermentation process were detected by HPLC with pre-column derivatization respectively. There was a new peak detected in the sample B, which was collected after the enrichment by ion exchange resin HZ-732, and the concentration of I was significantly lower than the actual value. Through analyzing by LC-MS, the result indicated that the new peak was a reaction product of 1 with thioglycolic acid in the alkaline condition when o-phthalaldehyde （OPA） was insufficient. After optimization, the concentration and purity of 1 in different samples could be detected accurately by means of sample dilution, increasing the amount of derivatization agent, or increasing the amount of OPA in the original derivatization agent. The last method was more suitable for the detection of different samples during the fermentation of I.

关 键 词：庆大霉素C1a 发酵 柱前衍生化 高效液相色谱 测定 

分 类 号：O657.72[理学—分析化学] R978[理学—化学]