Analysis of acidolysis products using HPLC

Analysis of acidolysis products using HPLC
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.
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.