Waters AccQ-Tag Amino Acid Analysis Column User Manual

[ CARE AND USE MANUAL ]

[ CARE AND USE MANUAL ]

ACCQ•TAG™ AMINO ACID ANALYSIS COLUMN

I. INTRODUCTION

This manual describes procedures for using the Waters AccQ•Tag™

Amino Acid Analysis column for the separation of hydrolysate amino

acids derivatized according to the AccQ•Tag

The Waters AccQ•Tag

™

Amino Acid Analysis column is manufactured

in Waters.

ISO 9002-certified facilities under strict guidelines to assure the high-

est quality products. Please take a few moments to read this manual

carefully. Follow its recommendations to ensure reproducible chroma-

tography and to maximize column lifetime.

a. Waters AccQ•Tag

The AccQ•Tag

™

Column Description

™

column separates the hydrolysate amino acid deriva-

tives produced in the reaction with Waters AccQ•Fluor

AccQ•Tag

™

column is a high-efficiency 4 μm Nova-Pak™ C18 column

specifically certified for use with the AccQ•Tag

When you use the AccQ•Tag

™

column as part of a Waters AccQ•Tag™

system, sub-picomole detection limits can be achieved.

™

Method.

™

Method.

™

reagent. The

CONTENTS

I. INTRODUCTION

a. Waters AccQ•Tag™ Column Description

b. Specifications

II. INSTALLING THE COLUMN

a. Preparing the System

b. Installing the Columns

III. USING THE COLUMN

a. Chromatography Guidelines

b. Efficiency Testing

™

c. Equilibrating the Column for the AccQ•Tag

Method

IV. CARE AND MAINTENANCE

a. Storing the Column

APPENDIX A. SPARE PARTS
APPENDIX B. WARRANTY/SERVICE INFORMATION

b. Specifications

Column AccQ•Tag™ Amino Acid Analysis Column

Dimensions 3.9 mm x 150 mm

Particle Size 4 μm

Packing Material Silica base bonded with C

18

Shipped in Acetonitrile/water (50%/50%)

[ CARE AND USE MANUAL ]

[ CARE AND USE MANUAL ]

II. INSTALLING THE COLUMN

a. Preparing the System

This procedure requires a zero dead-volume union. Before installing

the column in the flow path:

1. Directly connect the injector to the detector by replacing the old

column with a zero dead-volume union.

2. Flush the HPLC system with acetonitrile/water (60%/40%) at a

flow rate of 1.0 ml/min.

3. Remove the union.

4. Install the column in the column heater (see Section 2.2).

b. Installing the Columns

To install the column:

1. Remove the end plugs from the column and save them for use when

you store the column.

2. Screw the inlet and outlet fittings into the column until finger-

tight. An arrow on the label indicates the direction of flow.

To prepare a new stainless steel tubing-ferrule connection:

1. Use a file with a cutting edge or a tube cutter to scribe the

circumference of the steel tubing at the desired break point.

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

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

work the tube back and forth until it separates.

3. Ensure that the tubing end is straight, open, and free of burrs.

4. Slide the compression fitting, followed by the ferrule (large end of

the toper first) over the tube.

5. Properly bottom the tubing in the fitting seat. Dead volume results

in sample bandspreading.

If you are using a stainless steel system, use two 5/16 inch wrenches

to tighten the fittings 1/4 to 1/2 turn. Do not over tighten. Over tight-

ening damages the connection.

Preparing a new PEEK

Prepare a new tubing-ferrule connection (Figure 2-1) if the existing

connection is damaged or worn.

To prepare a new PEEK

1. Use a single-edge razor to make a straight, square cut in front of

the compression screw on the worn fitting.

2. Slide a compression screw and washer over the end of the

cut tubing.

Properly bottom the tubing in the fitting seat. Dead volume results in

sample bandspreading.

™

connection

™

tubing-ferrule connection:

Ferrule 0.009-inch I.D. Tubing (0.25 mm)

0.130-inch gap (3.3 mm)

Figure 1. Ferrule and Compression Screw Assembly

III. USING THE COLUMN

a. Chromatography Guidelines

Before running the first amino acid analysis on the new column,

perform the efficiency test in Section IV. b. Efficiency Testing.

General Considerations

The following operating guidelines will help you obtain the best

performance from your AccQ•Tag

•

Do not exceed an operating pressure of 26 MPa /260 atm or 4000 psi).

• Filter all eluents with a 0.22 pm membrane filter. Never use turbid

or cloudy eluents.

• Protect the column from vibration and mechanical shock.

™

column:

2