HPLC-UV-CAD同时测定卡前列素氨丁三醇注射液中主要成分含量

Simultaneous determination of the main components in carboprost tromethamine injection by HPLC-UV-CAD

目的：建立高效液相色谱联用紫外和电喷雾检测器同时检测卡前列素氨丁三醇注射液中卡前列素、氨丁三醇和苯甲醇含量的方法。方法：采用资生堂CAPCELL PAK CR 1∶4混合模式色谱柱（150 mm×4.6 mm,5μm）,以超纯水为流动相A,0.1 mol·L-1醋酸铵缓冲液（p H 6.2）为流动相B,甲醇为流动相C,梯度洗脱;柱温40℃;进样量10μL;流速1.0 m L·min-1;利用电喷雾检测器（charged aerosol detection,CAD）检测卡前列素和氨丁三醇,雾化室温度35℃,采样频率30 Hz;利用UV检测器检测苯甲醇,检测波长258 nm,采样频率5 Hz。结果：卡前列素的线性范围为9.38~56.2μg·m L-1（r=0.997 1,n=6）,最低检测限为0.3μg·m L-1,最低定量限为0.6μg·m L-1,方法平均回收率为104.7%（n=9）;氨丁三醇的线性范围为5.19~23.3μg·m L-1（r=0.999 2,n=6）,最低检测限为1.0μg·m L-1,最低定量限为2.5μg·m L-1,方法平均回收率为95.5%（n=9）;苯甲醇的线性范围为0.297~1.78 mg·m L-1（r=1,n=6）,最低检测限为5.0μg·m L-1,最低定量限为10.0μg·m L-1,方法平均回收率为100.1%（n=9）。实际样品测定结果与采用美国药典标准方法结果一致。结论：该方法不需衍生操作,经济、灵敏、简便、准确性和重现性良好,能够同时准确检测卡前列素氨丁三醇注射液中卡前列素、氨丁三醇和苯甲醇的含量,适用于卡前列素氨丁三醇注射液的质量控制。

Objective： To establish an HPLC-UV-CAD method for the simultaneous determination of carboprost, tromethamine and benzyl alcohol in carboprost tromethamine injection. Methods： Carboprost, tromethamine and benzyl alcohol were separated on a CAPCELL PAK CR 1：4 column （150 mm× 4.6 mm,5 μm）. The mobile phase was composed of ultrapure water, 0.1 mol· L-1 ammonium acetate buffer （ pH 6.2） and methanol with gradient elution mode at a flow about 1.0 mL·min-1 ; the injection volume was 10 μL and column temperature was 40℃. CAD was used to detect earboprost and tromethamine, in which nebulization temperature was 35 ℃ and collection frequency was 30 Hz. UV was used to detect benzyl alcohol, in which detection wavelength was set at 258 nm and collection frequency was 5 Hz. Results： The linear range of carboprost was 9.38 - 56.2 μg·mL-1 （ r = 0. 997 1, n = 6） ; the lowest limit of detection was 0.3 μg·mL-1 ; LOQ was 0. 6μg·mL-1 ; average recovery was 104.7%. The linear range of tromethamine was 5. 19 - 23.3 μg·mL-1（r = 0. 999 2, n = 6） ; the lowest limit of detection was 1μg·mL-1 ; LOQ was 2.5μg·mL-1; average recovery was 95.5 %. The linear range of benzyl alcohol was 0.297 - 1.78 mg·mL-1 （r = 1, n =6） , the lowest limit of detection was 5.0μg·mL-1 ; LOQ was 10.0μg·mL-1 ; average recovery was 100.1%. The actual measurement results of the sample were consistent with the standard method currently used. Conclusion： The HPLC method established does not require derivative operations, and is cheap, sensitive, simple, accurate, reproducible and accurate. This method can simultaneously detect carboprost, tromethamine and benzyl alcohol in carboprost tromethamine injection.