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Analysis of Carbohydrate and Fatty Acid Marker Abundance in Ricin Toxin Preparations for Forensic Information

机译:蓖麻毒素毒素制品中碳水化合物和脂肪酸标记物含量的分析供法医学参考

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摘要

One challenge in the forensic analysis of ricin samples is determining the method and extent of sample preparation. Ricin purification from the source castor seeds is essentially a protein purification through removal of the nonprotein fractions of the seed. Two major, nonprotein constituents in the seed are the castor oil and carbohydrates. We used derivatization of carbohydrate and fatty acid markers followed by identification and quantification using gas chromatography/mass spectrometry (GC/MS) to assess compositional changes in ricin samples purified by different methods. The loss of ricinoleic acid indicated steps for oil removal had occurred, and a large decrease of ricinoleic acid was observed between unextracted mash and solvent extracted and protein precipitate preparations. Changes to the carbohydrate content of the sample were also observed following protein precipitation. The differential loss of arabinose relative to mannose was observed indicating the removal of the major carbohydrate fraction of the seed and enrichment of the protein content. When the data is combined and multivariate principle component analysis is applied, these changes in fatty acid and carbohydrate abundance are discriminating enough to be indicative of the preparation method used for each sample.
机译:蓖麻毒素样品的法医分析中的一项挑战是确定样品制备的方法和范围。从源蓖麻籽中纯化蓖麻毒素本质上是通过去除种子中非蛋白质部分的蛋白质纯化。种子中的两种主要的非蛋白质成分是蓖麻油和碳水化合物。我们使用了碳水化合物和脂肪酸标记物的衍生化,然后使用气相色谱/质谱(GC / MS)进行鉴定和定量,以评估通过不同方法纯化的蓖麻毒蛋白样品的组成变化。蓖麻油酸的损失表明已经进行了除油步骤,未萃取的液和溶剂提取物以及蛋白质沉淀物的制备之间观察到蓖麻油酸的大量减少。蛋白质沉淀后还观察到样品中碳水化合物含量的变化。观察到阿拉伯糖相对于甘露糖的差异损失,表明去除了种子的主要碳水化合物部分并增加了蛋白质含量。合并数据并应用多元主成分分析后,脂肪酸和碳水化合物丰度的这些变化足以区分出每个样品所用的制备方法。

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