Waters Amino Acid Analysis Liquid Chromatography Column - Manual

[ Care and Use ManUal ]
I. Int roductIon
This manual provides the user with information necessary to obtain
the maximum efficiency and life from the Waters Amino Acid Analysis
Column. This column has been specifically designed for the separation
and analysis of free amino acids. The column is a precision device.
Waters Corporation uses very exacting manufacturing procedures
to achieve a small average particle size, narrow range of particle
size distribution and stable particle structure in the column. These
procedures create the high efficiency column-to-column reproducibility
and long separation life built into each amino acid column from Waters.
However, care must be taken to maintain packing density and protect
it from contamination by foreign material. The following information
will aid you in obtaining the highest performance and longest life from
your column.
II. Background InformatIon
Although there are some hydrophobic interactions between the column
and certain amino acids, the primary mode of separation is that
of ion-exchange. The cation-exchange resin utilized has a large
number of negatively charged (sulfonate) functional groups which
are covalently bound to it. These negatively charged groups, in turn,
readily associate with positively charged cations when they are
present in solution. Before the amino acid sample is injected onto the
column, the resin is equilibrated at a low pH with buffer containing
a high concentration of sodium (Na+) ions. This places the column
in the sodium form. As amino acids enter the column in this acidic
buffer, they are strongly positively charged and readily exchange with
the loosely bound sodium ions.
As the buffer gradually changes to a higher pH during the gradient run,
each amino acid becomes less positively charged reflecting its pK value
and differentially elutes accordingly.
contents
I. IntroductIon
II. Background InformatIon
a. Elution System 1
b. Elution System 2
c. General Preparation Guide for One Liter of Eluent
d. Use of Non-aqueous Solvents
e. Backpressure
f. Sample Considerations
g. SEP-PAK
® C
18
Cartridge Clean Up for Amino Acid Samples
h. Preparation Scheme
III. column care & use consIderatIons
a. Column Care Considerations
b. Column Storage
c. Column Regeneration
d. Troubleshooting
e. Regeneration of Protein and Lipid Contaminated Column
f. Regeneration of Heavy Metal Contaminated Column
g. Available Service and Applications Information
h. Chromatographic Troubleshooting
i. Typical Separation
IV. column Performance checks
V. column end fIttIngs
a. End Fitting Removal Procedure
b. Column Hookup
c. Storage
aPPendIx a. amIno acId analysIs column sPecIfIcatIons
amIno acId analysIs lIquId chromatography column