HPLC法同时测定杆菌肽原料药中主成分的含量及有关物质

Simultaneous Determination of Main Components and Related Substance in Bacitracin by HPLC

目的：建立同时测定杆菌肽原料药中主成分的含量及有关物质的方法。方法：采用高效液相色谱法。色谱柱为XTerra MS？C18,流动相为水-0.2 mol/L磷酸氢二钾溶液（用磷酸调节p H至6.0）-甲醇-乙腈（28∶14∶52∶6,V/V/V/V）,主成分杆菌肽A检测波长为254 nm,杆菌肽F（杂质F）检测波长为300 nm,柱温为30℃,流速为1.0 ml/min,进样量为100μl。结果：杆菌肽A质量浓度在0.4~4 mg/ml范围内与峰面积呈良好的线性关系（r=0.999 9）;精密度、稳定性、重复性试验的RSD≤0.5%;平均加样回收率为100.1%,RSD为0.5%（n=9）。主成分杆菌肽A与其他成分分离度较好;杂质F含量在6.0%以下。结论：本方法操作简便、快捷,结果准确、可靠,可用于杆菌肽原料药的质量控制。

OBJECTIVE： To establish a method for simultaneous determination of main components and related substance in hacitracin. METHODS： HPLC method was adopted. The determination was performed on XTerra MS C18 column with mobile phase consisted of water-0.2 mol/L dibasic potassium phosphate solution （pH adjusted to 6.0）-methanol-acetonitrile （28 ： 14 ： 52 ： 6, V/V/V/V）. The detection wavelength for bacitracin A was set at 254 nm for main components and 300 nm for bacitracin F. The col- umn temperature was set at 30 ℃, the flow rate of 1.0 ml/min and the sample size was 100 μl. RESULTS：The linear range of baci- tracin A was 0.4-4 mg/ml（r=0.999 9） with an recovery of 100.1% （RSD=0.5% ,n=9）. RSDs of precision, stability and reproduc- ibility tests were lower than 0.5%. Main component bacitracin A was separated from other components well. The content of bacitra- cin F was less than 6.0%. CONCLUSIONS： The method is simple, rapid, accurate and reliable, and it is applicable for the quality control of bacitraein.