Waters AccQ-Fluor Reagent Kit User Manual

[ Care and Use ManUal ]

ACCQ•Fluor reagent kIt

I. IntroduCtIon

The Waters AccQ•Fluor™ Reagent Kit contains five sets of the deriva-

tizing reagents used to perform the AccQ•Tag™ Amino Acid Analysis

Method. A set consists of one vial each of:

– AccQ•Fluor Borate Buffer - Vial 1. You add the buffer to the sam-

ples to generate the proper pH environment for the derivatizatlon.

– AccQ•Fluor Reagent Powder - Vial 2A. The reagent powder Is the

6-aminoquinolyl-N-hydroxysuccinimidyI carbamate (AQC) derivatiz-

ing reagent. It Is shipped dry for maximum stability.

– AccQ•Fluor Reagent Diluent - Vial 2B. Use the diluent(acetonitrile)

to reconstitute the reagent for derivatization.

The Waters AccQ•Fluor Reagent Kit (Figure 1) contains sufficient

reagent for up to 250 analyses.

Contents

I. IntroduCtIon

II. Waters accQ•tag Method overvIeW

III. reagent PurIty

Iv. storage

v. reconstItutIng accQ•Fluor reagent

vI. usIng Waters accQ•Fluor reagent

vII. derIvatIzIng the calIbratIon standard

vIII. reconstItutIng the hydrolyzed saMPle

IX. derIvatIzIng the saMPle

X. storIng derIvatIves

Figure 1: AccQ•Fluor Reagent Kit

AccQ•Fluor Reagent Kit 1

XI. sPecIFIcatIons

XII. technIcal lIterature

XIII. orderIng InForMatIon

[ Care and Use ManUal ]

II. Waters accQ•tag Method overvIeW

The AccQ•Tag Method is a precolumn derivatization and analysis

technique for peptide and protein hydrolysate amino acids. To

perform the AccQ-Tag Method:

1. Use Waters AccQ•Fluor Reagent (US Patent 5,296,599

and European Patent EP 0 533 200 B1) to derivatize the

amino acids.

2. Separate the derivatives by reversed-phase HPLC.

3. Quantitate the derivatives by fluorescence detection.

III. reagent PurIty

The Waters AccQ•Fluor Reagent Kit is manufactured in Waters’- ISO

9001-2006 certified facilities under strict guidelines to ensure the

lowest possible amino acid background. To retain this level of purity,

handle all reagents carefully.

6. Heat Vial 2A on top of heating block, vortex occasionally, until

the powder dissolves. Do not heat longer than 10 minutes.

Reconstituted AccQ•Fluor Reagent is approximately 10 mM

AQC in acetontrile.

Note: You can store the reconstituted AccQ•Tag reagent in a desiccator

at room temperature for up to one week. The AccQ•Tag reagent reacts

with atmospheric moisture so seal the container tightly when not in

use and DO NOT refrigerate.

Reconstituted AccQ•Fluor Reagent is approximately 10 mM AQC in

acetontrile.

IV. storage

If unopened, the AccQ•Fluor Reagent Kit may be stored at room

temperature for up to one year.

v. reconstItutIng accQ•Fluor reagent

To reconstitute the AccQ•Fluor Reagent (Figure 2):

1. Preheat a heating block to 55 °C.

2. Tap Vial 2A lightly before opening to ensure all AccQ•Fluor

Reagent Powder is at the bottom of the vial.

3. Rinse a clean micropipette by drawing and discarding 1 ml of

AccQ•Fluor Reagent Diluent from Vial 2B.

WARNING: Acetonitrile is flammable and toxic. Refer to the Material

Safety Data Sheet.

4. Transfer 1.0 ml of AccQ•Fluor Reagent Diluent From Vial 2B to

the AccQ•Fluor Reagent Powder In Vial 2A.

Figure 2: Reconstituting AccQ•fluor Reagent

storage

Store reconstituted AccQ•Fluor Reagent at room temperature in a

desiccator for up to one week.

Note: AccQ•Fluor reagent reacts with atmospheric moisture. Seal the

container tightly when not in use.

5. Cap Vial 2A lightly and vortex for 10 seconds.

AccQ•Fluor Reagent Kit 2