Lee-Benitez D, Leon C, Herrera J, Noriega-Guerra L, Bermudez J A
Arch Invest Med (Mex). 1975;6(1):55-64.
The aim of this report is to describe a quick method for measuring plasma estrogens by radio immunoassay in one day. Briefly, 1000 cpm of estradiol-3-H were added to 0.1 to 1.5 ml of plasma or serum to calculate losses during the procedure. After ether extraction and without any purification step, aliquots for tracer recoveries and the radioimmunoassay were taken. The antiserum working dilution was 1:100,000. The incubation of the antiserum at 37degreesC with the tritiated and cold estradiol lasts one hour. The free and bound fractions were separated with charcoal/dextran. With this method, 95% of estradiol, 16% of estrone and 6% of estriol are quantified. Other steroids normally present in plasma produce no interference. The minimal detectable level is 10 pg. Pasma and water blanks are indistinguishable of zero (C.V. 1%). Intra-analysis coefficient of variation is 6.1%. Mass recovery of estradiol (5 ng), estrone (5 ng) and estriol (5ng) are 4.75, 0.83 and 0.31 ng, respectively. Tracer recoveries are 92.6%. When plasma levels of estradiol are determined during menstrual cycle, normal and/or pathological pregnancy, by either the method described herein or measured selectively, the results show no significant differences. Finally, practical usefulness of this method is stressed, inasmuch results are obtained in six hours and one person can easily perform 200 samples in one week.
本报告的目的是描述一种通过放射免疫分析法在一天内快速测量血浆雌激素的方法。简而言之,将1000计数每分钟的雌二醇 - 3 - H添加到0.1至1.5毫升血浆或血清中,以计算该过程中的损失。经过乙醚萃取且无任何纯化步骤后,取出用于示踪剂回收率测定和放射免疫分析的等分试样。抗血清工作稀释度为1:100,000。抗血清在37℃下与氚化雌二醇和冷雌二醇孵育1小时。用活性炭/葡聚糖分离游离和结合部分。用这种方法,可定量95%的雌二醇、16%的雌酮和6%的雌三醇。血浆中通常存在的其他类固醇不会产生干扰。最低检测水平为10皮克。血浆和水空白与零无差异(变异系数1%)。分析内变异系数为6.1%。雌二醇(5纳克)、雌酮(5纳克)和雌三醇(5纳克)的质量回收率分别为4.75、0.83和0.31纳克。示踪剂回收率为92.6%。当通过本文所述方法或选择性测量来确定月经周期、正常和/或病理性妊娠期间的血浆雌二醇水平时,结果显示无显著差异。最后,强调了该方法的实际实用性,因为在6小时内即可获得结果,且一个人在一周内可轻松处理200个样本。