新疆孕马尿中主要结合雌激素定性定量方法研究

目的: 建立孕马尿中主要结合雌激素的定性定量方法。 方法: 孕马尿样品通过HPD-400大孔树脂固相萃取处理后,采用银化硅胶薄层色谱法快速鉴别孕马尿中雌酮硫酸钠、马烯雌酮硫酸钠、17α-二氢马烯雌酮硫酸钠3种主要结合雌激素;采用merck整体化柱分离,测定样品中3种主要结合雌激素含量。 结果: 薄层色谱中,供试品色谱与对照品色谱在相应位置显相同颜色的条带,条带清晰,分离效果好,阴性无干扰;建立的含量快速测定方法,3种主要结合雌激素在10 min内可达到完全分离,浓度分别为0.8~6.4 μg · mL^-1 (r=0.9996),0.4~3.2 μg · mL^-1(r=0.9998),0.3~2.4 μg · mL^-1(r=0.9998)线性关系良好,平均回收率分别为96.1%,96.5%,98.2%。 结论: 该方法快速、准确、可靠,适合孕马尿中主要结合雌激素的定性、定量检测,对于孕马尿品质评价具有参考价值。

Objective: To establish a qualitative and quantitative method of main conjugated estrogens in pregnant mare urine. Methods: The macroporous adsorptive resins(HPD-400)was used to purify pregnant mare urine.The qualitative identification of sodium estrone sulfate,sodium equilin sulfate and sodium 17α-dihydroequilin sulfate were carried out by AgNO₃-TLC.The content of them were determined by HPLC with Merck Chromolith^ Performance HPLC column RP-18e (100 mm×4.6 mm). Results: The established TLC condition displayed a very well separation of three main conjugated estrogens and the bland solution had no interference;The three main conjugated estrogens could be well separated within 10 minutes with HPLC mothod.The content of three main conjugated estrogens showed good linearity in the ranges of 0.8-6.4 μg · mL^-1 (r=0.9996),0.4-3.2 μg · mL^-1(r=0.9998),0.3-2.4 μg · mL^-1(r=0.9998),respectively;The average recoveries were 96.1%,96.5%,98.2%,respectively. Conclusion: There qualitative and quantitative detection methods are rapid,accurate,reliable,and reproducible and can be effectively used for quality evaluation of main conjugated estrogens in pregnant mare urine