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10
Table 9: Preparation of calibration standards of amino acids other than cystine.
Solution
25
pmol/µL
10
pmol/µL
5
pmol/µL
1
pmol/µL
0.5
pmol/µL
0
pmol/µL
Amino acid
solution
50*
100**
50**
100***
50***
0***
Taurine solution
50
a
100
b
50
b
100
c
50
c
0
c
2.5 mM Nva in
0.1 M HCl
20
20
20
20
20
20
0.1 M HCl
380
280
380
280
380
480
* 2.5 mM AA standard stock solution.
** 0.5 mM AA standard solution.
*** 0.05 mM AA standard solution.
a
2.5 mM Tau standard stock solution.
b
0.5 mM Tau standard solution.
c
0.05 mM Tau standard solution.
5.5
Derivatization
Derivatization converts the amino acids into highly stable derivatives. Standards and samples are
derivatized following instructions of supplier as described below:
1.
Preheat a heating block to 55 °C.
2.
Use a micropipettor to add 70 μL of AccQ•Tag Ultra Borate buffer (reagent 1) to a clean glass
vial with screw cap.
3.
Use a micropipettor to deliver 10 μL of calibration standard or neutralized sample solution to
the vial. Vortex mix briefly.
4.
Use a micropipettor to add 20 μL of reconstituted AccQ•Tag Ultra reagent to the sample vial.
Vortex mix immediately for several seconds and tap the vial.
5.
Let stand for 1 minute at room temperature.
6.
Heat the vial in a heating block or oven for 10 minutes at 55 °C.
AMINO-01
FOR ERP/WORKING GROUP USE ONLY
DO NOT DISTRIBUTE