LGC, Queens Road, Teddington, Middlesex, TW11 0LY, UK.
Anal Biochem. 2011 May 1;412(1):40-6. doi: 10.1016/j.ab.2010.12.015. Epub 2010 Dec 23.
Over the past decade, a number of endogenous peptides and endogenous peptide analogs have been employed in therapeutics and as diagnostic markers. The use of peptides as standards for the absolute quantification of proteins has become commonly accepted. Consequently, the requirement for standard peptides traceable to the International System of Units with low associated measurement uncertainty, and for accurate methods of peptide quantification, has increased. Here we describe a method of peptide quantification involving microwave-assisted acid hydrolysis followed by gas chromatography-mass spectrometry that enables traceable quantification of a peptide by exact matching isotope dilution mass spectrometry where the total hydrolysis time required is only 3h. A solution of angiotensin I was quantified using this method, and the results were in agreement with those obtained previously using an oven hydrolysis liquid chromatography-tandem mass spectrometry method.
在过去的十年中，许多内源性肽和内源性肽类似物已被用于治疗和作为诊断标志物。肽作为蛋白质绝对定量的标准已被广泛接受。因此，需要与国际单位制可溯源、测量不确定度低的标准肽，以及准确的肽定量方法。在这里，我们描述了一种涉及微波辅助酸水解和气相色谱-质谱联用的肽定量方法，该方法通过精确匹配同位素稀释质谱法实现肽的可溯源定量，其中所需的总水解时间仅为 3 小时。使用该方法对血管紧张素 I 进行了定量，结果与以前使用炉水解液相色谱-串联质谱法获得的结果一致。
