ANALYSIS OF MANNOSE-6-PHOSPHATE BY HPAEC

$1,305P41FY2010RRNIH

University Of Georgia, Athens GA

Investigators

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Abstract

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. As requested by the client, the standard was dissolved with 200 [unreadable]L H2O to obtain an initial concentration of 100 [unreadable]M, whereas each of the samples was dissolved with 50 [unreadable]L H2O. The standard was serially diluted from the initial concentration of 100 [unreadable]M to obtain concentrations of 50 [unreadable]M, 25 [unreadable]M and 12.5 [unreadable]M. A water-blank was injected in addition to the serially diluted standard and a calibration curve was established based on the area units generated from each concentration. The calibration standards and samples were analyzed for mannose-6-phosphate by HPAEC using a Dionex DX500 system equipped with a GP40 gradient pump, an ED40 electrochemical detector, Dionex CarboPac PA20 (3 x 150 mm) analytical column and a Thermo-Separation AS3500 autosampler containing a stainless steel needle. The autosampler was set to deliver an injection volume of 10 [unreadable]L and injection was made every 35 minutes. The eluents used in the gradient program were 100 mM NaOH and 1 M sodium acetate in 100 mM NaOH.

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