HPLC法测定水合羟基化头孢克洛杂质研究

摘要：目的 建立HPLC法测定水合羟基化头孢克洛杂质方法。方法 采用C18(4.6 mm×250 mm，5 μm)色谱柱；流动相A为0.78%磷酸二氢钠溶液(取磷酸二氢钠7.8 g，加水溶解并稀释至1000 mL，用磷酸调pH至4.0)，流动相B为0.78%磷酸二氢钠溶液(pH 4.0)-乙腈(55:45)；流速1.2 mL/min；检测波长220 nm；柱温25℃。结果 水合羟基化头孢克洛杂质在9.108×10-4~3.4×10-2 mg/mL范围内线性关系良好(r=1.0000)，平均回收率为104.03%，RSD为0.82%(n=9)，定量限为17.721 ng。在回收率试验中加入的水合羟基化头孢克洛杂质，采用外标法和加校正因子的主成分自身对照法的结果一致，该杂质的相对保留时间约为1.11，校正因子为1.02。结论 该方法操作简便、准确度高、重现性好，可以采用不加校正因子的主成分自身对照法对水合羟基化头孢克洛杂质进行检测。

HPLC method for determination of hydroxylated cefaclor impurities

Abstract Objective To establish an HPLC method for the content determination of hydroxylated cefaclor impurity in cefaclor. Methods The determination was performed on HPLC, using a C18(4.6 mm×250 mm, 5 μm) column with the mobile phase A consisted of 0.78% sodium dihydrogen phosphate solution (taking sodium dihydrogen phosphate 7.8 g, dissolved in water and diluted to 1000mL, and pH adjusted to 4.0 with phosphoric acid) and the mobile phase B consisted of 0.78% sodium dihydrogen phosphate solution-acetonitrile (55:45). The flow rate was 1.2 mL/min, the detection wavelength was 254 nm, and the column temperature was 25℃. Results The method showed a good liner relationship within the range of 9.108×10-4~3.4×10-2 mg/mL of hydroxylated cefaclor impurity (r=1.0000), the average recovery was 104.03% with RSD=0.82% (n=9), and the limit of quantification was 17.721 ng. The contents of cefaclor hydroxylation degradation product impurities added in the recovery test using external standard method and the principal component self-control method with correction factor are consistent. The relative retention time of hydroxylated cefaclor impurity was 1.11. The relative correction factor values of hydroxylated cefaclor impurity was 1.02. Conclusion The method is simple and accurate. It has good reproducibility and could be used to determine the content of hydroxylated cefaclor impurity with the main component self-control method without correction factor in