用亲水性的C_(16)硅胶反相色谱柱分离测定血清样品中的雌激素  

Determination of Several Estrogens in Serum by High-Performance Liquid Chromatography with Ultraviolet Detection

作　　者：蔡亚岐[1] 王玲[1] 梁立娜[1] 刘京生[1] 史亚利[2] 牟世芬[1] 江桂斌[1] 

机构地区：[1]中国科学院生态环境研究中心环境化学与生态毒理学国家重点实验室,北京100085 [2]北京科技大学化学系,北京100083

出　　处：《分析化学》2005年第7期913-916,共4页Chinese Journal of Analytical Chemistry

基　　金：国家重点基础研究计划(973项目)(No.2003CB415001);国家自然科学基金(No.20475060)资助课题

摘　　要：将亲水性较强的C16硅胶反相色谱柱应用于血清样品中β雌二醇、雌三醇、雌酮和17α乙炔基雌二醇的分离。实验对分离条件进行了优化,得到的最佳色谱条件是:柱温40℃,流速1mL/min,以40%乙腈水溶液作等度洗脱。在此条件下4种雌激素可在大约26min内实现基线分离,得到的4个色谱峰峰型对称。分离后的4种雌激素用紫外检测器在200nm处进行测定,方法对β雌二醇、雌三醇、雌酮和17α乙炔基雌二醇的检出限分别为0.024、0.015、0.012和0.016mg/L;校正曲线的线性范围为2～3个数量级,相关系数为0.998以上。方法应用于血清样品的测定,β雌二醇、雌三醇、雌酮和17α乙炔基雌二醇的标准加入回收率分别为96.3%、103.7%、100.1%和95.2%。A simple and direct method for determination of four estrogens beta-estradiol, estriol, estrone and 17 alpha-ethynylestradiol in serum sample by high-performance liquid chromatography coupled with ultraviolet detection was developed. The four target analytes were easily and completely separated on a polar-enhanced C-16 silica stationary phase packed reversed-phase column at a flow-rate of 1 mL/min by isocratic elution with mobile phase of acetonitrile/water (40:60, V/V) in about 26 min. The column temperature was kept at 40 degrees C. The separated four analytes were detected directly by UV detector at 200 nm wavelength. Under the optimized conditions, when the injection volume was 20 mu L, the detection limits (signal-to-noise ratio equal to 3) for beta-estradiol, estriol, estrone and 17 alpha-ethynylestradiol were 0.024, 0.015, 0.012 and 0.016 mg/L, respectively. The calibration graphs for the four analytes were linear over two to three orders of magnitude with correlation coefficients greater than 0.998. The analytical method had been applied to the determination of beta-estradiol, estriol, estrone and 17 alpha-ethynylestradiol in real serum samples. The accuracy of the proposed method was tested by recovery measurements on spiked samples and good recovery results (95.2% similar to 103.7%) were obtained.

关 键 词：血清样品 雌激素 色谱柱 亲水性 17Α-乙炔基雌二醇 分离测定 反相 硅胶 紫外检测器 雌三醇 分离条件 色谱条件 基线分离 线性范围 校正曲线 相关系数 方法应用 标准加入 雌酮 min 水溶液 色谱峰 检出限 数量级 回收率 

分 类 号：R446.1[医药卫生—诊断学]