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4.3
Apparatus
Oven with an accuracy better than 1- 2°C.
Vortex mixer.
Heating block (55 °C).
Millipore MillexR-HV syringe driven filter unit, Cat SLHV 013 NL, or equivalent.
UHPLC equipped with UV detector (260 nm).
4.4
Preparation of reagents
Volumes of glassware are purely indicative and may be modified as long as the proportion of
reagents is maintained.
4.4.1 The AccQ•Tag Ultra Reagent Kit
Prepare the reactives included in the kit following instructions of supplier.
4.4.2 Norvaline (Nva) as internal standard
To prepare a 10 mM internal standard stock solution of norvaline, add 117.16 mg of Nva to 100 mL
0.1 N HCl.
To prepare a 2.5 mM internal standard stock solution of norvaline, add 29.29 mg of Nva to 100 mL
0.1 N HCl.
Internal standard stock solution can be stored at -20 °C for up to 6 months.
4.4.3 Amino acid calibration standards with exception of cystine and taurine
Calibration standard stock solution is ready to use and contains 2.5 mM of each amino acid (cystine is
not used for quantification). Make aliquots of 150 μL, which can be stored at -20 °C for up to 6
months.
Prepare 0.5 mM of AA standard solution by diluting 5 times calibration standard stock solution with
0.1 M HCl.
Prepare 0.05 mM of AA standard solution by diluting 10 times AA standard solution at 0.5 mM with
0.1 M HCl.
AMINO-01
FOR ERP/WORKING GROUP USE ONLY
DO NOT DISTRIBUTE