摘要
目的建立测定硫酸庆大霉素中4个C组分含量的高效液相色谱一串联质谱方法，并考察各组分对质谱检测器的响应因子。方法利用SynergiPolar-RPC18色谱柱（50mm×2．0mm，4gm，80A），以妥布霉素为内标，以流动相0．1％甲酸溶液-甲醇等度洗脱（10：90，V/V），流速0．2mL／min，柱温35℃，分析时间5min。在电喷雾离子化电离源上以选择反应监测方式进行正离子检测，用于定量分析的离子反应分别为m／z478．2→322．2（C1），464．2→322．2（C2＋C2a），450．2→322．2（C1a），和m／z468．2→163．1（内标）。结果庆大霉素浓度的线性范围为0．5～500μg／L，庆大霉素的定量下限为0．5gg／L。C1、C2＋C2a、Cla4个组分的批内和批间RSD分别为0．2％～8．1％和3．6％～6．4％，批内和批间准确度分别为93．1％～107．1％和97．6％～104．9％。庆大霉素4个C组分各自标准曲线的斜率值无显著性差异，4个C组分在质谱检测器上具有相同的响应因子。结论该方法特异性高、准确度好、精密度高，且4个C组分对质谱的响应因子一致，可以利用高效液相色谱－串联质谱对庆大霉素制剂中各组分进行准确定量。
Objective To establish a liquid chromatography-tandem mass spectrometry （LC-MS/MS） method for the quantification of gentamicin C components, and investigate the response factors of the four major gentamicin C components detected by mass detector. Methods Gentamicin C components and an internal standard tobromycin were separated on a Synergi Polar-RP C18 column （50mm×2.0mm, 4μm, 80A） using isocratic mobile phase of 0.1% formic acid and methanol at a flow rate of 0.2mL/min. At positive electrospray ionization mode, selected reaction monitoring of the precursor-product ion transitions m/z of 478.2→322.2（C1）, 464.2→322.2（C2＋C2a）, 450.2→322.2 （C 1a） for gentamicin and m/z of 468.2→ 163.1 for tobromycin was used for the quantification. Results The linear calibration curve of gentamicin was obtained in a concentration range of 0.5-500μg/L with a lower limit of quantification of 0.5pg/L. Intra- and inter-run precision of 4 components was less than 8.1% and 6.4%, respectively. Intra- and inter-run accuracy fell in the ranges of 93.1%-107.1% and 97.6%-104.9%, respectively. The slopes of 4 components calibration curves had no significant difference, and 4 different components showed identical responses in mass detector. Conclusion The method was specific, accurate, precise, and the response factors of 4 components detected by MS were consistent. In conclusion, the LC-MS/MS method was applied to the determination of gentamicin C components in formulation.
出处
《中国抗生素杂志》
CAS
CSCD
北大核心
2014年第2期122-127,共6页
Chinese Journal of Antibiotics
基金
国家"重大新药创制"科技重大专项"药物安全评价技术平台"(2012ZX09302001)
非人灵长类肾脏安全性生物标志物的国际联合研究
关键词
庆大霉素
妥布霉素
高效液相色谱-串联质谱法
响应因子
Gentamicin
Tobromycin
Liquid chromatography-tandem mass spectrometry （LC-MS/MS）
Response factor
