(q)
Water –Acetonitrile25:75 solution: Add50mL
1mLofwater to150mL
1mLof ACN in
aglassbottleandmix.
(r)
Eluent B: Ammonium formate (50mmol/L pH4.4): Add2.3g
0.1g (1.89mL) of formic acid
(100%) in a beaker containing 800mL of water. Adjust the pH to 4.40
0.05 with
ammoniumhydroxide solution (25%). Transfer quantitatively to a 1000mL volumetric flask,
anddilute to thevolumewithwater.
(s)
2AB labeling reagent: 2AB [0.35mol/L] - NaBH3CN [1.0mol/L] in DMSO - acetic acid [30%]
solution. Pipette the volume of dimethylsulfoxide (DMSO) and acetic acid in a 10mL glass
tube according to the number of tests to perform (see table below for quantities). Mix the
solutionusinga vortexmixer.Weigh theamount of anthranilic acidamide (2AB) and sodium
cyanoborohydride (NaBH3CN) (see table below for quantities) in another glass tube of
10mL, then add the corresponding volume of 30% acetic acid in DMSO. Mix on a Vortex
anduseanultrasonicbath for completedissolution (about 10min).
Example of quantities for 2AB reagent preparation
Max. number
of tests
30%Aceticacid inDMSO
2AB (0.35M),NaBH
3
CN(1M)
in30%Acetic acid/DMSO
DMSO (mL)
Acetic acid
100% (mL)
30%AceticAcid in
DMSO (mL)
2AB
(mg)
NaBH
3
CN
(mg)
11
2.10
0.90
2.50
118
5
157
5
22
4.20
1.80
5.00
236
10
314
10
33
6.30
2.70
7.50
354
10
471
10
(t)
AmmoniumAcetate buffer: 0.1mol/L pH 5.5 In a 100-mL beaker, dissolve 0.771g
0.005g
of anhydrous ammonium acetate (CH3COONH4) with 80mL
5mL of water and adjust pH
to 5.50
0.05 with acetic acid. Transfer quantitatively in a 100-mL volumetric flask and
complete to themarkwithwater.
(u)
Amyloglucosidase enzyme solution: 60U/mL in ammonium acetate solution (0.1mol/L pH
5.5): Weigh the amount of AMG enzyme corresponding to 180 Units
20 U and dissolve
with3.0mLof ammoniumacetatebuffer 0.1mol/LpH5.5 (6.8).
Notes regardingAmyloglucosidase (AMG)
GOS-01
FORWORKINGGROUP/ERPUSEONLY
DONOTDISTRIBUTE
