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Placental tissue metabolome analysis by GC-MS: Oven-drying is a viable sample preparation method

机译:GC-MS胎盘组织代谢物分析:烘箱干燥是一种可行的样品制备方法

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摘要

Analysis of the human placenta metabolome has great potential to advance the understanding of complicated pregnancies and deleterious fetal outcomes in remote populations, but samples preparation can present unique challenges. Herein, we introduce oven-drying as a simple and widely available method of sample preparation that will facilitate investigations of the placental metabolome from remote and under-studied populations. Placentae from complicated and uncomplicated pregnancies were prepared in three ways (oven-dried at 60 degrees C, fresh, lyophilized) for metabolome analysis via gas chromatography-mass spectrometry (GC-MS). Multiple computer models (e.g. PLS-DA, ANN) were employed to classify and determine if there was a difference in placentae metabolome and a group of metabolites with high variable importance in projection scores across the three preparations and by complicated vs. control groups. The analyses used herein were shown to be thorough and sensitive. Indeed, significant differences were detected in metabolomes of complicated vs. uncomplicated pregnancies; however, there were no statistical differences in the metabolome of placentae prepared by oven-drying vs. lyophilization vs. fresh placentae. Oven-drying is a viable sample preparation method for placentae intended for use in metabolite analysis via GC-MS. These results open many possibilities for researching metabolome patterns associated with fetal outcomes in remote and resource-poor communities worldwide.
机译:对人胎盘代谢物的分析有可能推进对遥远人群的复杂妊娠和有害胎儿结果的理解,但样品准备可以呈现独特的挑战。在此,我们将烘箱干燥作为一种简单且广泛可选择的样品制剂方法,其将促进胎盘代谢物从遥远和研究的群体的研究。通过三种方式(在60℃,新鲜,冻干)以三种方式(新鲜,冻干)制备胎盘,通过气相色谱 - 质谱(GC-MS)进行代谢分析。使用多种计算机模型(例如PLS-DA,ANN)来分类和确定胎盘代谢物和一组具有高可变性的代谢物,在三种制剂中的投影分数和对照组复杂的对照组。这里使用的分析显示为彻底和敏感。实际上,在复杂的对比的复杂性妊娠的代谢中检测到显着差异;然而,通过烘箱干燥与新鲜胎盘的封闭剂对冻干组制备的胎盘代谢物中没有统计学差异。烘箱干燥是胎盘的可行样品制备方法,用于通过GC-MS用于代谢物分析。这些结果开辟了研究与全世界偏远和资源贫困社区中的胎儿结果相关的代谢模式的许多可能性。

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