Waters Carbamate Analysis Column User Manual

[ Care and Use ManUal ]

CarbaMate analysIs ColuMn

I. IntroduCtIon

The Waters Carbamate Analysis Column (3.9 x 150 mm) is packed

with a durable, high efficiency, 4μ spherical silica-based stationary

phase ideally suited for the reversed-phase separation of carbamate

pesticides and related compounds.

Waters exclusive sequential bonding and packing processes coupled

with stringent quality control procedures ensure precise surface

chemistry, reproducibility, and stability. When used as a component

of Waters Carbamate Analysis System with the Waters Carbamate

Analysis Method, this column is guaranteed to provide the resolution

and sensitivity needed for successful analysis of the analytes listed

in Figure 1.

Contents

I. IntroduCtIon

II. InstallatIon

a. Attaching the Column

b. Equilibration

III. MobIle phase and saMple guIdelIne

a. Solvent Preparation and Filtration

b. Sample Preparation and Filtration

IV. operatIon

a. Chromatography Guidelines

b. Efficiency Testing

V. Care and MaIntenanCe

a. Troubleshooting

b. Shutdown and Storage

VI. ColuMn and supplIes orderIng and InforMatIon

Figure 1: A Typical Separation of Carbamate Pesticides & Related
Compounds using Waters Carbamate Analysis System*

Please take a few moments to read this manual carefully. Use the

information it contains to ensure that you obtain quality results and

take full advantage of the features your Waters column offers.

Note: 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.

* For additional information on the complete method, please refer to Waters Carbamate

Analysis System Manual.

Carbamate Analysis Column 1

VII. Warranty/serVICe InforMatIon

[ Care and Use ManUal ]

Compression Screw or Nut

Ferrule

End must be straight
and smooth to achieve
maximum column efficiency

Tu be

Critical Distance to be determined by
the union or column fitting, etc.

Follow generally accepted procedures for quality control and methods

development when using this column.

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.

II. InstallatIon

a. Attaching the Column

1. If this column is to be used with an LC system previously used

for a purpose other than carbamate analysis, then before

installing the column in the flow path connect the column

inlet and outlet lines to each other with a union and flush the

lines, as well as the sample injector and injector loops, free of

previous solvents. Make certain that the final flushing solvent

is miscible with water. Remove the union.

2. Remove the end plugs from your column with a 5/16-inch

wrench.

5. Grasp the tubing on both sides of the scribe mark with cloth-

covered pliers (to prevent marring the tube surface) and gently

work the tub back and forth until it breaks cleanly. Check that

the end is straight and smooth with no burrs. Flush the tube

from the opposite end with mobile phase to remove any metal

particles that may have lodged in the interior of the tube.

6. Slide the compression screw head first followed by the ferrule

(large end of the taper first) over the tube. Insert the end of

the tube into the fitting seat to which it will be connected.

Tighten the compression screw in the fitting seat as in Step 3.

Assembly details are shown in Figure 2.

CAUTION: Properly bottom the tubing in the fitting seat

while tightening the compression screw to the column.

Tubing not completely seated will result in dead volume

which could cause excessive sample band spreading.

NOTE: Be sure to replace and tighten the end plugs when the

column is removed from the system for storage.

3. The column outlet is indicated by an arrow on the label

showing the direction solvent should flow. Thread the inlet and

outlet tubing fittings into the column until finger tight and then

tighten the fittings with a wrench turning each - turn.

CAUTION: Do not over-tighten. Over-tightening will damage

the connection. Follow the next three steps of this procedure

if you must remove a damaged compression screw or worn

ferrule.

4. Using a miniature tubing cutter, scribe deeply the

circumference of the tubing at the desired break point. Or,

alternatively, using a three-cornered file with a cutting edge,

cut 1/3 of the way through the tubing at the desired break

point.

Carbamate Analysis Column 2

Figure 2: Ferrule and Compression Screw Assembly

b. Equilibration

The Waters Carbamate Analysis Column is shipped in the mobile

phase used for column storage: 50/50 v/v methanol/acetonitrile.

For equilibration, flush the column with the initial mobile phase

used for the gradient analysis. This can be conveniently done while

the post-column reaction system, column oven, and detector are

warming up to stable operating conditions (about 30 minutes at 1.5

mL/min). Refer to the Waters Carbamate Analysis System Manual

for details.