Analysis of acidolysis products using HPLC Author/User Keywords: Equipment required: - Pump: Dionex P580 - Detector: Dionex UVD 340S - Autosampler: Dionex ASI - 100 - Oven: Dionex STH585 - Column Hamilton PRP-1 (4.1 x 250 mm, 7 μm) Analysis conditions: - sample dry weight (from 10 mg to 20 mg), - solvent: 93.34% acetonitrile, 6% THF, 0.66% 1% trifluoroacetic acid in MilliQ water, - The volume of the sample after dissolving: 1.5 mL, - Injection volume of 100 L, - Autosampler temperature: 40oC, - Mobile phase: acetonitrile (90%): 1% trifluoroacetic acid in MilliQ water (10%), - Flow rate of 0.8 ml / min, - Oven temperature: 25oC, - Wavelength - 210 nm Execution time: 30 min. Description: Samples for HPLC analysis ought to be taken directly from the liquid reaction products separated from the lipase preparation. To obtain about 10-20 mg dry weight of the lipids after evaporation of the solvents (in the thermoblock), place 120-140 μl samples of the liquid products in pre-weighed vials with the volume of 2 ml. Before HPLC analysis, dissolve each sample in 1.5 ml of acetonitrile (93.34%) supplemented with 6% of THF and 0.66% of 1% solution of trifluoroacetic acid in MilliQ water. Then place the dissolved samples in the autosampler (min. temp. 40oC) and conduct HPLC using the Hamilton PRP-1 column.