![]() ![]() Add 0.01 mL of acetaldehyde solution (0.3 in 100) to each cell, and mix. Stopper the cells, and allow to stand for 2 minutes at 25 ± 1. ![]() Add 2.5 mL of pH 9.0 buffer, 0.2 mL of -NAD solution, and 0.1 mL of pyrazole solution (0.68 in 100) to each cell, and mix. Pipet 0.1 mL of water into a second 1-cm spectrophotometric cell to provide the reagent blank. Pipet 0.1 mL of the Assay preparation into a 1-cm spectrophotometric cell. Dissolve an accurately weighed quantity of -nicotinamide adenine dinucleotide ( -NAD) in water to obtain a -NAD solution having a known concentration of about 20 mg per mL. Dissolve 3.3 g of potassium pyrophosphate, 15 mg of dithiothreitol, and 40 mg of edetate disodium in 70 mL of water, adjust with citric acid monohydrate solution (2.1 in 10) to a pH of 9.0 ± 0.1, dilute with water to 100 mL, and mix to obtain a pH 9.0 buffer. Use this solution as the Assay preparation. Transfer about 20 mg, accurately weighed, to a 200-mL volumetric flask, dissolve in 1 mL of water, dilute with an ice-cold solution of bovine serum albumin (1 in 100) to volume, and mix.
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