Kohrman K A, MacGee J
J Assoc Off Anal Chem. 1977 Jan;60(1):5-8.
Diethylstilbestrol (DES) is readily extracted into toluene from biological fluids or tissue homogenates. Sodium carbonate is added to the initial mixture to eliminate potentially interfering substances. The toluene is extracted with a very small volume of aqueous trimethylphenylammonium hydroxide. This solution generates dimethyl DES in the vaporizer of a gas-liquid chromatograph. An internal standard, dienestrol (DI), is added at the beginning of the procedure and is partitioned and methylated in the same manner as DES. The DES and DI derivatives are well separated in less than 6 min on an ov-17 column. The entire analysis requires less than 15 min for a fluid specimen and less than 25 min for a solid tissue specimen. Seven samples can be analyzed each hour on a single column with a flame ionization detector. The relative standard deviations at levels from 2.5 to 100 ppm in bile are less than 5%. The lower limit of sensitivity is 8 ppb in a 1 ml bile sample.
己烯雌酚（DES）很容易从生物流体或组织匀浆中萃取到甲苯中。向初始混合物中加入碳酸钠以消除潜在的干扰物质。用极少量的氢氧化三甲基苯基铵水溶液萃取甲苯。该溶液在气液色谱仪的汽化器中生成二甲基己烯雌酚。在该程序开始时加入内标物己二烯雌酚（DI），并以与己烯雌酚相同的方式进行分配和甲基化。在ov - 17柱上，己烯雌酚和己二烯雌酚衍生物在不到6分钟的时间内就能很好地分离。对液体样本进行整个分析所需时间不到15分钟，对固体组织样本则不到25分钟。使用火焰离子化检测器时，每小时可以在一根柱子上分析7个样本。胆汁中浓度在2.5至100 ppm范围内的相对标准偏差小于5%。在1 ml胆汁样本中，灵敏度下限为8 ppb。
