用于同位素分析的未衍生化氨基酸的制备型高效液相色谱分离

Preparative HPLC Separation of Underivatized Amino Acids for Isotopic Analysis.

作者信息

Tripp Jennifer A, Devièse Thibaut, McCullagh James S O

机构信息

Department of Chemistry, University of San Francisco, San Francisco, CA, USA.

Oxford Radiocarbon Accelerator Unit, Research Laboratory for Archaeology and the History of Art, University of Oxford, Oxford, UK.

出版信息

Methods Mol Biol. 2019;2030:69-83. doi: 10.1007/978-1-4939-9639-1_7.

DOI:10.1007/978-1-4939-9639-1_7

PMID:31347111

Abstract

Single-compound analysis of stable or radioactive isotopes has found application in a number of fields ranging from archaeology to forensics. Often, the most difficult part of these analyses is the development of a method for isolating the compound(s) of interest, which can derive from a wide range of sample types including the hair, nails, and bone.Here we describe three complementary preparative HPLC techniques suitable for separating and isolating amino acids from bone collagen and hair keratin. Using preparative reversed-phase, ion-pair, or mixed-mode chromatography in aqueous carbon-free mobile phases, or those from which carbon can easily be removed, underivatized single amino acids can be isolated and further analyzed using mass spectrometric techniques.

摘要

稳定或放射性同位素的单化合物分析已在从考古学到法医学等多个领域得到应用。通常，这些分析中最困难的部分是开发一种分离目标化合物的方法，目标化合物可能来自多种样品类型，包括头发、指甲和骨骼。在此，我们描述了三种互补的制备型高效液相色谱技术，适用于从骨胶原蛋白和毛发角蛋白中分离和提纯氨基酸。使用制备型反相、离子对或混合模式色谱，在无碳水性流动相或易于去除碳的流动相中，可以分离出未衍生化的单一氨基酸，并使用质谱技术进行进一步分析。