Department of Horticulture and Landscape Architecture, National Taiwan University, Taipei 10673, Taiwan.
Molecules. 2019 May 18;24(10):1920. doi: 10.3390/molecules24101920.
Stable-isotope dimethyl labeling is a highly reactive and cost-effective derivatization procedure that could be utilized in proteomics analysis. In this study, a liquid chromatography- tandem mass spectrometry in multiple reaction monitoring mode (LC-MS-MRM) platform for the quantification of kiwi allergens was first developed using this strategy. Three signature peptides for target allergens Act d 1, Act d 5, and Act d 11 were determined and were derivatized with normal and deuterated formaldehyde as external calibrants and internal standards, respectively. The results showed that sample preparation with the phenol method provided comprehensive protein populations. Recoveries at four different levels ranging from 72.5-109.3% were achieved for the H-labeled signature peptides of Act d 1 (SPA1-H) and Act d 5 (SPA5-H) with precision ranging from 1.86-9.92%. The limit of quantification (LOQ) was set at 8 pg mL for SPA1-H and at 8 ng mL for SPA5-H. The developed procedure was utilized to analyze seven kinds of hand-made kiwi foods containing 0.0175-0.0515 mg g of Act d 1 and 0.0252-0.0556 mg g of Act d 5. This study extended the applicability of stable-isotope dimethyl labeling to the economical and precise determination of food allergens and peptides.
稳定同位素双甲基标记是一种高反应性且具有成本效益的衍生化程序,可用于蛋白质组学分析。在这项研究中,首次利用该策略开发了用于定量猕猴桃过敏原的液相色谱-串联质谱多重反应监测模式(LC-MS-MRM)平台。确定了三个用于目标过敏原 Act d 1、Act d 5 和 Act d 11 的特征肽,并分别用正常和氘代甲醛将其衍生化作为外部校准物和内标。结果表明,采用苯酚法进行样品制备可提供全面的蛋白质群体。对于 H 标记的 Act d 1(SPA1-H)和 Act d 5(SPA5-H)的特征肽,回收率在 72.5-109.3%之间,精度在 1.86-9.92%之间。SPA1-H 的定量限(LOQ)设定为 8 pg mL,SPA5-H 的 LOQ 设定为 8 ng mL。该方法用于分析七种手工制作的猕猴桃食品,其中含有 0.0175-0.0515 mg g 的 Act d 1 和 0.0252-0.0556 mg g 的 Act d 5。这项研究扩展了稳定同位素双甲基标记在经济高效地测定食物过敏原和肽方面的适用性。