Testing extraction and storage parameters for a fecal hormone method
Testing extraction and storage parameters for a fecal hormone method
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测试粪便激素方法的提取和储存参数
DOI：
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发表时间：
2010
影响因子：
2.4
通讯作者：
J. Beehner
中科院分区：
文献类型：
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作者：
David J. Pappano;E. K. Roberts;J. Beehner
进行了四项实验，以检测“田间友好”粪便激素提取方法的不同方面，该方法利用田间甲醇提取，然后储存在C18固相萃取柱上。粪便样本收集自布朗克斯动物园饲养的狒狒（狒狒），实验在实验室环境中进行，以确保最大限度的控制。这些实验的目的是模拟粪便样本在野外工作中所处的条件，或者改进现有的方案。实验检测了粪便激素代谢物保存/回收率与以下因素之间的关系：（1）样品在环境温度下储存的时间;（2）样品经历的冻融循环次数;（3）不同提取溶液的有效性;以及（4）不同检测盒清洗的有效性。对于每项实验，通过放射免疫测定法测定粪便糖皮质激素（GC）和睾酮（T）代谢产物的样本。每个实验的结果如下。首先，在环境温度下储存直到储存4周才影响激素水平，GC和T代谢物在4周时均显着增加。第二，GC在两次冻融循环后，T在六次冻融循环后，样品中的激素水平显著降低。第三，对于GC和T，使用各种甲醇溶液的激素提取显著高于使用100%乙醇。最后，使用20%甲醇溶液清洗柱显著增加GC水平，但对T水平无影响。这些结果表明，当利用C18盒粪便类固醇储存，研究人员应该考虑几种方法选择，以优化激素的保存和回收粪便样品。Am. J. Primatol. 72：934-941，2010.© 2010 Wiley利斯公司
Four experiments were conducted to test different aspects of a “field‐friendly” fecal hormone extraction method that utilizes methanol extraction in the field followed by storage on C18 solid‐phase extraction cartridges. Fecal samples were collected from geladas (Theropithecus gelada) housed at the Bronx Zoo, and the experiments were conducted in a laboratory setting to ensure maximum control. The experiments were designed to either simulate the conditions to which fecal samples are subjected during fieldwork or improve on an existing protocol. The experiments tested the relationship between fecal hormone metabolite preservation/recovery and: (1) the amount of time a sample is stored at ambient temperature; (2) the number of freeze/thaw cycles a sample undergoes; (3) the effectiveness of different extraction solutions; and (4) the effectiveness of different cartridge washes. For each experiment, samples were assayed by radioimmunoassay for fecal glucocorticoid (GC) and testosterone (T) metabolites. Results for each of the experiments were as follows. First, storage at ambient temperature did not affect hormone levels until 4 weeks of storage, with significant increases for both GC and T metabolites at 4 weeks. Second, hormone levels significantly decreased in samples after two freeze/thaw cycles for GCs and six freeze/thaws cycles for T. Third, for both GCs and T, hormone extraction using various methanol solutions was significantly higher than using 100% ethanol. Finally, using a 20% methanol solution to wash cartridges significantly increased GC levels but had no effect on T levels. These results suggest that, when utilizing C18 cartridges for fecal steroid storage, researchers should consider several methodological options to optimize hormone preservation and recovery from fecal samples. Am. J. Primatol. 72:934–941, 2010. © 2010 Wiley‐Liss, Inc.
影响因子：
2.7
作者：
Wasser, SK;Hunt, KE;Monfort, SL
通讯作者：
Monfort, SL