柱前衍生化高效液相色谱法测定复方酮康唑乳膏中硫酸新霉素的含量

Determination of Neomycin Sulfate in Compound Ketoconazole Cream by Pre-column Derivatization HPLC

目的:建立柱前衍生化高效液相色谱(HPLC)法测定复方酮康唑乳膏中硫酸新霉素的含量。方法:采用安捷伦C18色谱柱(4.6 mm×250 mm,5μm),流动相为甲醇∶水(35∶65)(含0.2 mol·L^(-1)醋酸和0.013 mol·L^(-1)的庚烷磺酸钠),流速为1.0mL·min^(-1),柱温为30℃,检测波长为399 nm,进样量为20μL。结果:硫酸新霉素在0.004~0.06 mg·mL^(-1)(r=0.9997)范围内线性关系良好,6次重复性试验相对标准偏差(RSD)为0.88%,检测限为0.04μg,平均回收率为98.4%。结论:柱前衍生化HPLC法高效、便捷、准确,可作为抗生素微生物检定法的替代方法,用于测定复方酮康唑乳膏中硫酸新霉素的含量。

Objective： To establish a pre-column derivatization high performance liquid chromatography （HPLC） method for the determination of neomycin sulfate in compound ketoconazole cream. Methods： Agilent C18 column （4.6 mm ×250 ram, 5 μm） was used with a mobile phase methanol： water （35： 65） （containing 0.2 mol · L^-1 acetic acid and 0. 013 mol · L^-1 sodium heptane sulfonate） at a flow rate of 1.0 mL· min^-1. The column temperature was 30 ℃. The detection wavelength was set at 399 nm, and the injection volume was 20 μL. Results： The linear relationship between neomycin sulfate in the range of 0. 004 to 0.06 mg· mL^-1 （ r = 0. 9997） was good. The relative standard deviation （RSD） of the six replicates was 0. 88 %, the detection limit was 0.04 μg, and the average recovery was 98.4%. Conclusion： This pre-column derivatization HPLC method is an efficient, conven-ient and accurate method, and it can be used as an alternative method for antibiotic microbial assay to determine the content of neo-mycin sulfate in compound ketoconazole cream.