Analysis of triacylglycerol composition
The TAG composition of the original MKF and the solid fraction of MKF
that crystallized at 5 min of isothermal time for the experiment both
without and with the application of HIU (70% amplitude) was analyzed
using high performance liquid chromatography (HPLC) (Shimadzu LC-20AD,
Shimadzu Corp, Japan) with system controller CBM-20A and diode array
detector SPD-M20A. The sample was vacuum filtered to obtain the solid
fraction that had crystallized during the 0 - 5 min. This was to
identify the TAG species that crystallized early in the sample without
and with HIU application. Two reverse C-18 columns (Inertsil ODS-3; 4.6
× 250 mm; 5 µm particle size; by GL Sciences Inc., Japan) were used in
series. Mobile phase was mixture of HPLC-grade acetone and acetonitrile
in a volume-to-volume ratio of 70:30 and the flow rate was fixed at 0.72
mL/min. The column temperature was set at 30 °C with a column heater
(Shimadzu CTO-10AS column oven). The injection volume was 20 µL.
Identification of TAGs was conducted based on the retention time of TAG
standards. All TAG contents were given in percentage area.