AOAC SPIFAN Review Team (February 2017)

Single Laboratory Validation Report for Total Amino Acids by UHPLC-UV in Infant Formulas and Adult Nutritionals

Jaudzems, Lahrichi, and Fuerer

–

January 2017

page 3/20

- Sodium hydroxide solution, 1 M (e.g. Merck 109137)

- Optional: sodium hydroxide solution, 6 M (e.g. Merck 137032)

- Hydrochloric acid fuming 37 % (12 M) GR for analysis (e.g. Merck 100317)

- Hydrochloric acid, 1 M (e.g. Merck 109057)

- Hydrochloric acid, 0.1 M (e.g. Merck 109060)

- Laboratory Water Grade Type 1

3.2

Reagents

3.2.1 Solutions to prepare

- Sodium hydroxide (NaOH) solutions at 6 M, 0.2 M, and 0.05 M

- Hydrochloric acid (HCl) solution at 0.2 M

- 1% DDP in 0.2 M NaOH

- 0.1% phenol in 12 M HCl

3.2.2 AccQ•Tag™ Ultra Derivatization kit

Pre

pare the reagents included in the kit following the manufacturer’s

instructions.

3.2.2.1 AccQ•Tag™ Ultra Borate buffer (reagent 1)

Ready-to-use solution.

Alternative reagent: 5 %

(W/V)

sodium tetraborate in water.

3.2.2.2 AccQ•Tag™ Ultra reagent (vial 2A and 2B)

Reconstitute AccQ•Tag™ Ultra reagent (vial 2A) according to the manufacturer’s instructions:

a. Preheat a heating block to 55 °C.

Tap vial 2A lightly before opening to ensure all AccQ•Tag™ Ultra reagent powder

is at the bottom of the vial.

c. Rinse a clean micropipette by drawing and discarding 1

mL of AccQ•Tag™ Ultra

reagent diluent from vial 2B (ready-to-use solution). Repeat 2 times.

d. Draw 1.0

mL from vial 2B and transfer it to the AccQ•Tag™ Ultra reagent powder

in vial 2A. Cap the vial tightly.

e. Vortex mix for approximately 10 seconds.

f. Heat vial 2A on top of the preheated heating bl

ock until the AccQ•Tag™ Ultra

reagent powder is dissolved. Do not heat the reagent for longer than 10 minutes.

Once reconstituted, the AccQ•Tag™ Ultra reagent is approximately 10

mM. Store

reconstituted AccQ•Tag™ Ultra reagent in a desiccator at room temp

erature for up to 1 week.

Caution:

AccQ•Tag™ Ultra reagent reacts with atmospheric moisture. Seal the container

tightly when not in use. Do not refrigerate. Do not use discolored reagent, especially if it is

yellow or green.

Alternative reagent: Into a 4 mL vial, weigh out approximately 3.0 to 4.0 mg of 6-

Aminoquinolyl-N-hydroxysuccinimidyl carbamate. Continue with step c in section

3.2.2.2

above using LC-grade acetonitrile instead of the

AccQ•Tag™ Ultra reagent diluent.

AMINO-01 (FEBRUARY 2017)

SLV REPORT

FOR ERP USE ONLY

DO NOT DISTRIBUTE