Commission Regulation (EC) No 900/2008Show full title

3. REAGENTS

(Use double-distilled water)

3.1. 0,5 N sodium hydroxide solution (0,5 mol/l).

3.2. 96 % (minimum) glacial acetic acid.

3.3. Solution of amyloglucosidase:

Immediately before use, dissolve approximately 10 mg of amyloglucosidase (EC 3.2.1.3) (60 U per mg) in one ml of water(1).

3.4. Triethanolamine buffer solution:

dissolve 14,0 g of triethanolamine hydrochloride [tris(2-hydroxyethyl)ammonium chloride] and 0,25 g of magnesium sulphate MgSO₄7H₂O) in 80 ml of water, add approximately 5 ml of 5 N sodium hydroxyde solution (5 mol/l) and adjust the pH to 7,6, using a 1 N sodium hydroxide solution (1 mol/l).

Make up to 100 ml with water. This buffer solution can be kept for at least four weeks at 4 °C.

3.5. NADP (nicotinamide adenine dinucleotide phosphate, disodium salt) solution:

dissolve 60 mg of NADP in 6 ml of water. This solution can be kept for at least four weeks at 4 °C.

3.6. ATP (adenosine-5′-triphosphate, disodium salt) solution:

dissolve 300 mg of ATP. 3H₂O and 300 mg of sodium hydrogen carbonate (NaHCO₃) in 6 ml of water. This solution can be kept for at least four weeks at 4 °C.

3.7. Suspension HK/G6P-DH [Hexokinase (EC 2.7.1.1) and glucose-6-phosphate-dehydrogenase (EC 1.1.1.49)]:

suspend 280 U HK and 140 U of G6P-DH in 1 ml of 3,2 M ammonium sulphate solution. This suspension can be kept for at least one year at 4 °C.

(1)

U is the international unit of enzyme activity.