HPLC法测定醋酸奥曲肽中醋酸和三氟乙酸的含量

Determination of Acetic Acid and Trifluoroacetic Acid in Octreotide Acetate by HPLC

目的建立高效液相色谱法同时测定醋酸奥曲肽原料药中醋酸和三氟乙酸的含量。方法色谱柱为YMCPack ODS-AQ(4.6 mm×250 mm,5μm),流动相A为磷酸溶液(取磷酸7 ml加水稀释至1000 ml,混匀,用氢氧化铵溶液调节pH至3.0,再加入甲醇10 ml,混匀),流动相B为甲醇,梯度洗脱,流速:1.0 ml·min^(-1),检测波长:210 nm,柱温:25℃。结果醋酸和三氟乙酸分别在10.092~1009.200μg·ml^(-1)(r=1.0000)、0.496~49.570μg·ml^(-1)(r=0.9999)范围内线性关系良好,定量限分别为30.9、16.6 ng,检测限分别为10.9、7.0 ng,平均回收率分别为99.78%、99.56%(n=9),RSD分别为0.36%、0.41%。结论该方法准确性高、专属性强、耐用性好,适用于测定醋酸奥曲肽中醋酸和三氟乙酸的含量。

Objective To establish an HPLC method for simultaneous determination of acetic acid and trifluoroacetic acid in octreotide acetate.Methods The HPLC gradient elution method was used on a YMC-Pack ODS-AQ column(4.6 mm×250 mm,5μm)with phosphate acid solution(7 ml of phosphate acid was diluted with water to 1000 ml,mixed,and pH was adjusted to 3.0 with ammonium hydroxide solution before 10 ml of methanol was added and mixed)as mobile phase A,and methanol as mobile phase B at a flow rate of 1.0 ml·min^(-1).The detection wavelength was 210 nm and the column temperature was 25℃.ResultsThe linear ranges of acetic acid and trifluoroacetic acid were 10.092-1009.200μg·ml^(-1)(r=1.0000)and 0.496-49.570μg·ml^(-1)(r=0.9999),respectively.The limits of quantitation were 30.9 ng and 16.6 ng respectively,while those of detection were 10.9 and 7.0 ng,respectively.The average recoveries(n=9)were 99.78%and 99.56%,and the RSDs were 0.36%and 0.41%,respectively.Conclusion This method is accurate,specific and durable,which can be used for the determination of acetic acid and trifluoroacetic acid in octreotide acetate.