固相萃取-高效液相色谱法测定结合雌激素乳膏中主要成分含量

Determination of Contents of Main Components in Conjugated Estrogen Cream by Solid Phase Extraction-HPLC

采用固相萃取-高效液相色谱法(SPE-HPLC)测定结合雌激素乳膏中雌酮硫酸钠(Es)和马烯雌酮硫酸钠(Eqs)的含量。色谱柱为Agilent C18柱(4.6 mm×150 mm,5μm),流动相为乙腈-0.025 mol·L^-1 KH2PO4(1∶3,体积比),流速1.0 mL·min^-1,柱温35℃,检测波长205 nm,进样量20μL。结果表明,Es在6.89~220.5μg·mL^-1浓度范围内线性关系良好(R2=0.9996),Eqs在4.5~144.3μg·mL^-1浓度范围内线性关系良好(R2=0.9999);Es和Eqs峰面积RSD分别为0.15%和0.19%,结合雌激素含量RSD为1.25%(n=6),标示量80%、100%和120%的Es和Eqs的平均加标回收率分别为105.32%、103.10%、105.81%和95.15%、96.81%、96.37%。该方法准确度高,精密度和灵敏度均能满足测定要求,为结合雌激素乳膏质量控制提供了科学依据。

We determinated contents of sodium estrone sulphate(Es)and sodium equilin sulphate(Eqs)in conjugated estrogen cream by solid phase extraction-high performance liquid chromatography(SPE-HPLC).Agilent C18 column(4.6 mm×150 mm,5μm)is used with acetonitrile-0.025 mol·L^-1 KH2PO4(1∶3,volume ratio)as a mobile phase,and the flow rate is 1.0 mL·min^-1,the column temperature is 35℃,the detection wavelength is 205 nm,and the injection volume is 20μL.The results show that there exist a good linear relationship of Es in the concentration range of 6.89-220.5μg·mL^-1(R2=0.9996),and a good linear relationship of Eqs in the concentration range of 4.5-144.3μg·mL^-1(R2=0.9999),the RSDs of peak areas of Es and Eqs are 0.15%and 0.19%,respectively,the RSD of conjugated estrogen content is 1.25%(n=6),and the average adding standard recoveries of Es and Eqs are 105.32%,103.10%,105.81%and 95.15%,96.81%,96.37%,respectively.The method has the advantages of high accuracy,high precision,and high sensitivity,which can meet the requirements of the determination,and provides a scientific basis for the quality control of conjugated estrogen cream.