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[ CARE AND USE MANUAL ]
3. Example chromatography of glyphosate
solvent standard achieved on the Anionic
Polar Pesticide Column
4.32
100
%
0
-0.00
2.00
4.00
III. COLUMN USE
To ensure the continued high performance of the Anionic Polar
Pesticide Columns, follow these guidelines:
a. Sample Preparation
It is highly recommended to use the methods provided with this
column (www.waters.com/polarpesticides) however if you are
developing a method of your own, it is important to remember
the following for proper performance and column longevity.
1. Consider preparing the sample in organic solvents (Methanol
or Acetonitrile) for the best peak shape and sensitivity. It is
not recommended to use 100% water as a diluent since the
separation is operated under HILC condition. Using organic
sample diluents may avoid peak distortion due to "strong
solvent effects". In particular, stronger solvents can impact
peak shapes of low retaining analytes.
2. If the sample is not dissolved in the mobile-phase modifier,
ensure that the sample, solvent and mobile phases are
miscible in order to avoid sample precipitation. All samples
must be filtered before analysis to ensure removal of
particulates.
3. Please use polypropylene vials (p/n: 186005222) for this
application as glyphosate fixates to glass.
6.00
8.00
10.00 min.
b. Solvents
To maintain maximum column performance, use high
quality LC-MS grade solvents. Solvents containing
suspended particulate materials will generally clog the
outside surface of the inlet distribution frit of the column.
This will result in higher operating pressure, reduced
column lifetime, and compromised performance.
c. Additives
The Anionic Polar Pesticide Column can safely be used
with commonly used high purity acidic and basic additives
used in LC-MS such as formic acid, ammonium formate
and ammonium hydroxide. Consider the volatility, solubility,
and detector compatibility when choosing an appropriate
additive. Additives tend to improve peak shape and control
the retention characteristics of analytes but can also impart
different selectivities. It is recommended to flush and
remove additives and salts from the column before placing
the column into storage.
d. Pressure
The Anionic Polar Pesticide Columns is packed to pressures
designed for use on UPLC/UHPLC/HPLC instrumentation.
*Maximum
Column
operating column
dimensions
pressure
2.1 x 50 mm
1,800 psi
2.1 x 100 mm
3,600 psi
2.1 x 150 mm
4,800 psi
* Pressure drop across the column, does not include added system pressure
e. Temperature
The general guideline for maximum temperature for the
Anionic Polar Pesticide Column is 60° C with most common
LC mobile phases and additives. However, some data
shows good stability beyond 60° C. This would need to be
verified experimentally.
f. pH
The Anionic Polar Pesticide Columns can be used routinely
under HILIC conditions between pH 2 to 7. As with any LC
column, operating at the extremes of pH, pressures and
temperatures will result in decreased column lifetime.
g. Flow rate
The Anionic Polar Pesticide Columns can be used with flow
rates between 0.1 and 0.7 mL/min. Any flow rates should be
acceptable as long as they fall within the pressure limits.
Anionic Polar Pesticide Column
Maximum
operating column
pressure
124 bar
248 bar
331 bar
4
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