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本文介绍了应用荧先试剂二甲氨基-5-氯化磺酰萘(Dans-Cl)制备妇女尿中雌三醇(E_3)、雌二醇(E_2)与雌酮(E_1)的Dansyl衍生物,然后用以荧光检测的高效液相色谱 (HPLC)法测定其含量的分析方法。分析步骤:(1)取24小时妇女尿的1/1000;(2)酸水解尿中的雌激素缀合物;(3)用醚及碱液提取游离雌激素;(4)制备雌激素的Dansyl衍生物;(5)用HPLC法测定雌激素衍生物的峰面积,并以外标法计算尿中上述三种雌激素的含量。色谱条件:柱:青岛硅胶YWG-5μm,匀浆法装填150 mm×4.6 mm(i.d.);检测:荧光检测器,λex=362 nm,λem=511 nm;
This article describes the preparation of Dansyl derivatives of urinary estriol (E 3), estradiol (E 2) and estrone (E 1) in women using Dans-Cl , Followed by high performance liquid chromatography with fluorescence detection (HPLC) method for the determination of its content. Analysis steps: (1) taking a urine sample of 1 in 1,000 women for 24 hours; (2) hydrolyzing the estrogen conjugate in urine by acid hydrolysis; (3) extracting estrogenic free ether with ether and lye; (4) Dansyl derivatives; (5) HPLC method for the determination of estrogen derivatives peak area, and the external standard method to calculate urinary levels of the above three estrogen. Chromatographic conditions: Column: Qingdao Silica gel YWG-5μm, homogenization method filled 150 mm × 4.6 mm (i.d.); Detection: fluorescence detector, λex = 362 nm, λem = 511 nm;