氨苄西林、氨苄西林钠及注射用氨苄西林钠中二聚体及其他相关物质的HPLC检查法

Determination for dimer and related substances of ampicillin,ampicillin sodium,and ampicillin sodium injection by HPLC

目的：建立了HPLC测定氨苄西林、氨苄西林钠和注射用氨苄西林钠中氨苄西林二聚物及其他相关物质的方法。方法：用十八烷基硅烷键合硅胶为填充剂；取12％醋酸溶液0．5ml、0．2mol／L磷酸二氢钾溶液50ml和乙腈50ml置1000ml量瓶中，加水稀释至刻度，作为流动相A；取12％醋酸溶液0．5ml、0．2mol／L磷酸二氢钾溶液50ml和乙腈400ml置1000ml量瓶中，加水稀释至刻度，作为流动相B，进行梯度洗脱；流速：1．0ml／min；检测波长为254nm。结果：氨苄西林峰与二聚物峰和其他杂质峰分离良好，在2．2～177．4μg/ml浓度范围内，氨苄西林峰面积与样品浓度呈良好的线性关系，回归方程为Y=3．096×10^6X＋3．810×10^6，r=1．0000；在2．2～177．4μg／ml浓度范围内，氨苄西林二聚物峰面积与样品浓度线性关系良好，Y=3．266×10^6X＋3．751×10^63，r=0．9999。结论：本法可同时检测氨苄西林、氨苄西林钠和注射用氨苄西林钠中二聚物和其他相关物质，操作简便、快速，结果准确，重现性好。

OBJECTIVE To establish a simple, accurate and reproducible HPLC method for determination of ampicillin dimmer and related substances in ampicillin, ampicillin Sodium, ampicillin sodium for injection. METHODS A column packed with octadecylsilyl silica gel for chromatography R ; Mobile phase A ： A mixture of 0.5 ml of dilute acetic acid R, 50 ml of 0.2 mol/L potassium dihydrogen phosphate R and 50 ml of acetonitrile R, diluted to 1 000 ml with water R, Mobile phase B ： A mixture of 0.5 ml of dilute acetic acid R, 50 ml of 0.2 moL/L potassium dihydrogen phosphate R and 400 ml of acetonitrile R, diluted to 1 000 ml with water R, started the linear gradient at a flow rate of 1.0 ml/min; The UV detection wavelength was at 254 nm. RESUTS A sat- isfactory separation between ampicillin and related substances. The calibration curve was linear in the range of 10.6 - 159.0 μg/ml for ampicillin with good corelation coefficient, regression equation Y=3. 229 ×106 X ＋4. 054 × 103 ,r =0. 999 9. The calibration curve was linear in the range of 2.2 - 177.4 μg/ml for ampicillin dimer with good correlation coefficient, regression equation Y=3. 266 × 106X ＋3. 751 × 103 , r =0. 999 9. CONCLUSION This HPLC assay is simple and quick with high sensitivity, and will be used for the determination of ampicillin dimer and related substances in ampicillin, ampicillin Sodium, ampicillin sodium for iniection.