Instrument and method for analysis of mannose 6-phosphate

摘要

Disclosed are an apparatus and method for separation analysis of mannose-6-phosphate (M6P) by post-column fluorescence detection method. The apparatus is based on chromatography, and includes a column with a solid phase having affinity for phosphate, a flow path for the eluate, a heater installed on the flow path for M6P and a basic amino acid to react by heating the eluate in the flow path, and a fluorescence detector installed downstream of the heater for continuously irradiating the eluate with excitation light and measuring the intensity of the emission, and may include in the flow path a supply channel for addition of a basic amino acid between the column and the heater. The method is characterized in that it uses the apparatus and a second mobile phase consisting of a second buffer containing phosphate of predetermined concentration and adjusted to a predetermined pH.

主权项

1. An apparatus for separation analysis of mannose-6-phosphate contained in a sample using column chromatography, comprising: a column with a solid phase having affinity for phosphate, a flow path for an eluate from the column, a heater for heating the eluate in the flow path to cause mannose-6-phosphate and a basic amino acid contained in the eluate to react, a fluorescence detector installed in the flow path downstream of the heater for continuously irradiating the eluate with excitation light and measuring the intensity of fluorescence emitted from the eluate; and a back-pressure generator installed in the flow path between the heater and the fluorescence detector.