柱后衍生化 -HPLC 法测定硫酸卡那霉素注射液 及滴眼液的含量

目的 新建硫酸卡那霉素注射液及滴眼液含量的高效液相色谱 - 柱后衍生化 - 荧光检测方法。方法 色谱柱为 Zorbax Eclipse Plus C18 (4.6mm×100mm, 3.5μm)；流动相为 pH3.4 的缓冲液 ( 取庚烷磺酸钠一水合物 4.35g 和无水硫酸钠 16g，加水 溶解并稀释至成 1000mL，用冰醋酸调节 pH 值至 3.4±0.1)- 甲醇 (74:26)；柱温为 35℃；流速为 1mL/min。柱后衍生化试液为邻苯 二甲醛溶液；流速为 0.3mL/min；衍生化反应温度为 45℃；荧光激发波长为 340nm，发射波长为 455nm。结果 卡那霉素 A 在 0.0973~583.8μg/mL 浓度范围内线性关系良好 (r=0.9999)，硫酸卡那霉素注射液和滴眼液的平均回收率分别为 99.6% 和 99.3%， RSD 为 0.63% 和 0.66%(n=9)。结论 本方法检测灵敏度高，重复性好，简便快速，结果准确可靠，可作为硫酸卡那霉素注射液 及滴眼液的含量测定方法。

Determination of the content in kanamycin sulfate injection and eye drops by HPLC with post-column derivatization

Objective To establish a method of HPLC with post-column derivatization for the determination of the content in kanamycin sulfate injection and kanamycin sulfate eye drops. Methods A Zorbax Eclipse Plus C18 column(4.6mm×100mm, 3.5μm)was used, the mobile phase was buffer (containing 1-heptanesulfonate sodium salt and sodium sulfate)-methyl alcohol (74:26), the column temperature was 35℃ , and the flow rate was 1mL/min. The OPA solution was used for the post-column derivatization at a flow rate of 0.3mL/min. The temperature of the postcolumn reaction was 45℃ . The fluorescence was detected by an excitation and emission wavelength of 340 and 455nm respectively. Results The calibration curve was linear in the range of 0.0973~583.8μg/mL (r=0.9999). The average recovery rates of kanamycin sulfate injection and kanamycin sulfate eye drops were 99.6% and 99.3%, and RSD values were 0.63% and 0.66% (n=9). Conclusion The method is rapid and versatile, and ensures excellent results in terms of sensitivity and reproducibility. It enables the content determination of kanamycin sulfate injection and kanamycin sulfate eye