title: An efficient and inexpensive refrigerated LC system for H/D exchange mass spectrometry.
abstract: Loss of deuterium label during the LC step in amide hydrogen/deuterium exchange mass spectrometry (H/D-MS) is minimized by maintaining an acidic mobile phase pH and low temperature (pH 2.5, 0 °C). Here we detail the construction and performance of a low-cost, thermoelectrically refrigerated enclosure to house high-performance liquid chromatography (HPLC) components and cool mobile phases. Small volume heat exchangers rapidly decrease mobile phase temperature and keep the temperature stable to ±0.2 °C. Using a superficially porous reversed-phase column, we obtained excellent chromatographic performance in the separation of peptides with a median peak width of 4.4 s. Average deuterium recovery was 80.2% with an average relative precision of 0.91%.
authors.affiliation: Department of Chemistry, University of Kansas, Lawrence, KS 66045, USA.
authors.forename: Theodore R
authors.initials: TR
authors.lastname: Keppel
authors.forename: Martin E
authors.initials: ME
authors.lastname: Jacques
authors.forename: Robert W
authors.initials: RW
authors.lastname: Young
authors.forename: Kenneth L
authors.initials: KL
authors.lastname: Ratzlaff
authors.forename: David D
authors.initials: DD
authors.lastname: Weis
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase / Q000295:instrumentation / Q000379:methods*; D041961:Deuterium Exchange Measurement / Q000379:methods*; D004867:Equipment Design; D006863:Hydrogen-Ion Concentration; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D012034:Refrigeration / Q000295:instrumentation*; D013696:Temperature
chemical_list: D010455:Peptides