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.
Subject(s)
Amino Acids/isolation & purification , Chromatography, Reverse-Phase/methods , Mass Spectrometry/methods , Radiometric Dating/methods , Amino Acids/chemistry , Animals , Bone and Bones/chemistry , Carbon Isotopes/analysis , Carbon Isotopes/chemistry , Carbon Radioisotopes/analysis , Carbon Radioisotopes/chemistry , Chromatography, High Pressure Liquid/instrumentation , Chromatography, High Pressure Liquid/methods , Chromatography, Reverse-Phase/instrumentation , Collagen/chemistry , Collagen/isolation & purification , Hair/chemistry , Humans , Hydrolysis , Mass Spectrometry/instrumentation , Radiometric Dating/instrumentationABSTRACT
Single-compound analysis of stable or radio-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 compounds of interest.Here, we describe three complementary preparative HPLC procedures suitable for separating and isolating single amino acids from bone collagen or hair keratin with minimal isotopic contamination. Using preparative reversed-phase, ion-pair, or mixed-mode chromatography of underivatized amino acids in aqueous mobile phases, single amino acids can be isolated and further analyzed using mass spectrometric techniques.