Using the Griess colorimetric nitrite assay for measuring aliphatic β-nitroalcohols.

摘要: Our recent studies suggest that aliphatic β-nitroalcohols (BNAs) may represent a useful class of compounds for topical therapeutic corneoscleral cross-linking agents. Thus, this study was undertaken in order to standardize simple method nitroalcohol quantitation based on denitration step followed by colorimetric Griess nitrite assay. Conditions necessary included pH 7-9 and heating 1 hour at 100°C. Standard curves two mono-nitroalcohols (2-nitroethanol 2-nitro-1-propanol), nitro-diol (2-methyl-2-nitro-1,3-propanediol), nitro-triol (2-hydroxymethyl-2-nitro-1,3-propanediol) showed excellent linearity the 100-500 M range with absorbance values 0.98. The lower limit detection ~20 M. Recovery from tissue homogenates (10mg/mL wet weight) rabbit cornea sclera as well solutions gelatin B (1mg/mL) ranged 89-103% scleral tissue, 68-106% corneal 90-99% B. assay can be successfully used quantitative determination BNAs is simpler use than conventional chromatographic techniques.