Waters Carbamate Analysis Column - Manual (Page 3)

[ Care and Use ManUal ]
Carbamate Analysis Column 3
III. MobIle phase and saMple guIdelIne
a. Solvent Preparation and Filtration
1. Use only HPLC Grade or better solvents suitable for high sensitivity
fluorescence analysis, filtered to remove micro-particulate matter
about 0.45 µm.
NOTE: All glassware used for solvent and sample preparation must
be scrupulously clean. Detergent, fingerprint, cigarette smoke, breath
residues, etc., may contain amines which can cause interference with the
analysis.
Acetonitrile and methanol, even HPLC grade, may contain traces of
amines or ammonia, which will react with OPA/mercaptoethanol to
form highly fluorescent impurities. These derivatives may cause base-
line shifts or increased baseline noise. If this becomes a problem, clean
the reservoirs and use fresh solvent. If necessary, switch to a different
lot of solvent or to a different solvent vendor until a suitable grade is
found.
Distill or HPLC grade water. De-ionized water is not acceptable because
it contains organic compounds which may alter column selectivity.
2. Use vacuum filtration, sonication, and/or helium sparging to remove
dissolved gases, which could affect your solvent delivery system.
Waters Solvent Clarification Kit is designed to assist in the degassing
and preparation of mobile phases. Waters Carbamate Analysis System
has provision for continuous sparging of each mobile phase component.
3. Use a Waters In-Line Pre column Filter to capture system particulates
and extend column life. Note: a Waters In-Line Pre column Filter is
supplied with the Waters Carbamate Analysis System.
b. Sample Preparation and Filtration
1. Use a Waters Sample Clarification Kit or 0.45 μ Membrane Filter
Units to filter samples and prevent excessive pressure buildup.
2. Do not inject a sample that is dissolved in a solvent, which is not
miscible with the mobile phase.
3. If samples contain contaminants which become irreversibly bound
to the column packing under normal operating conditions, it may be
desirable to use Waters Sep-Pak
® Cartridges or Waters Guard-Pak™ Pre
column module and Guard-Pak Cartridges to remove the contaminants
off-line or on-line, respectively.
IV. operatIon
a. Chromatography Guidelines
Liquid chromatography columns have a finite life which, is directly
related to the care and use they receive. Column life is affected
by contamination from; samples and solvents, frequent solvent
changeovers, and improper handling and storage.
If you observe a change in peak shape, retention of a particular
compound, or resolution between two compounds, take immediate
steps to determine the reason for the changes. Until the cause of the
change is determined, do not rely on the results of the analyses.
Follow generally accepted procedures for quality control and
methods development when using this column
NOTE: Before running the first analysis on your new column, perform
the test sample separation given in Section IV. b., Efficiency Testing
PRECAUTIONS
a. Pressure
Maximum pressure should not exceed: 28 Mpa (4000 psi or 275
bars). Typical operating pressure in the Waters Carbamate Analysis
System: 10-20 Mpa (1500-3000 psi or 100-200 bars).
b. Temperature
Recommended column operating temperature range: 20 °C - 40
°C. Typical operating pressure in the Waters Carbamate Analysis
System: 30 °C.
c. Flow Rate
There are not flow rate restrictions as long as the recommended
pressure limits are not exceeded. Typical operating flow rate is
1.5mL/min. Flow rate should be increased gradually (in 0.5 mL/min
increments) to reach operating flow rate and decreased gradually to
zero upon system shutdown.
d. pH Range
Maintain pH of mobile phase and samples between 3 and 8. Avoid
using concentrated acids or bases.