Application Notes
Amino Acids L-Histidine Standard by HPLC/ELSD, UV
The amino acid L-histidine was separated by reversed phase HPLC and detected by evaporative light scattering and UV at 220nm. The ELSD was able to resolve the L-histidine from impurities while the UV was not.
Amino Acids L-Histidine Standard, Alltech Application Note 0011E, June 23, 2000.
A 99% pure L-histidine amino acid standard was separated from its impurities isocratically in around 5 minutes by reversed phase HPLC on an Absorbosphere C18, 5µm, 250 x 2.1mm column (Part No. 9945) using methanol/water and was detected by UV at 254nm and by evaporative light scattering using an Alltech 500 ELSD (Evaporative Light Scattering Detector.)
The UV detection did not resolve the L-histidine from the impurities, while the ELSD produced distinct peaks for L-histidine and the impurities.
In ELSD, the mobile phase is first evaporated. Solid particles remaining from the sample are then carried in the form of a mist into a cell where they are detected by a laser.
Key Analyte(s)
L-histidine
Equipment
Alltech Adsorbosphere ColumnsAlltech Adsorbosphere® Columns are designed using high temperature bonding for exceptional surface coverage and stability for common applications. They are available in pore sizes from 60-90Å and come in four varieties.
Technique
HPLCHigh Performance Liquid Chromatography (HPLC)is a technique for separating components in a sample on the basis of interactions between the component, a liquid mobile phase and a solid stationary phase.
Source
Alltech Associates, Inc.Alltech Associates, Inc., one of the leading suppliers of HPLC columns, manufactures and distributes HPLC, GC and IC columns, accessories and consumables and maintains a large searchable database of application notes and chromatograms on its website.
