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The remaining pellet was incubated overnight in 1.2 mL of acetate buffer (40 mM sodium acetate, 60 mM acetic acid) and 0.2 mL of enzymes solution (3 units of amyloglucosidase and 0.25 units of α-amylase); 0.5 mL of the supernatant was mixed with 0.5 mL of water and 1 mL of phenol (5/95; v/v).
Maximum and constant absorbance value was achieved at pH (4.5-5.5 4.5-5.52.5 ml of acetate busing.
The stock was prepared by mixing of 0.05 ml of 1% Acridine orange with 5 ml of acetate buffer with 0.2 M (pH 4.0).
Glycine were mixed with 2 mL of acetate buffer (0.6 M, pH = 4.5) and 2 mL of 10 mg/mL ninhydrin ethanol solution.
A working solution was freshly prepared by mixing 25 mL of acetate buffer, 2.5 mL of TPTZ solution, and 2.5 mL of FeCl3·6H2O solution.
Therefore 2.5 ± 0.5 ml of acetate buffer of pH 5 was chosen as the optimum volume of the buffer throughout this approach.
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A further 20 mL of sodium acetate-acetic acid buffer solution were added to achieve pH 5.3, before dilution to 100 mL with water.
To a solution of N-Boc- R -α-alkyl-β- sec-amino)alaN-Boc- R -α-alkyl-β- sec-aminof ethyl acetate, 1 ml of 4 N-Boc- R -α-alkyl-β- sec-aminoed.
The residue was partitioned between 50 mL of ethyl acetate and 50 mL of saturated sodium bicarbonate solution and the aqueous layer extracted with 30 mL of ethyl acetate.
The solid was briefly warmed to 65 °C with 25 ml of ethyl acetate and allowed to slowly cool.
For quantifying linalool and geraniol in various commercial beers, 2 mL of ethyl acetate was added to 8 mL of the beer in glass tubes (Kimble Chase, Rockwood, TN, USA).
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