Quantification Method for Triglyceride Molecular Species in Fish Oil with High Performance Liquid Chromatography Ultraviolet Detector - Evaporative Light Scattering Detector

摘要

We reported the quantification method for the triglyceride (TG) molecular species in fish oil using high performance liquid chromatography (HPLC)-ultraviolet detector (UV) system with isocratic elution. However, the system spent much time for one run and TG molecular species in fish oil consisting of saturated and/or monounsaturated fatty acids, such as triolein, could not be quantified because those hardly absorb UV. Consequently, the improved method using tandem jointed HPLC-UV-evaporative light scattering detector (ELSD) system with gradient elution was developed. The quantification was carried out with internal standard method and the respective calibration curves for TG molecular species were calculated with chromatogram area ratio between internal standard and TG molecular species versus amount ratio between them. As the results, the second order calibration curves were acquired and all the coefficient of correlations for the calibration curves were higher than 0.99. Several kinds of TG molecular species in bigeye tuna body oil were quantified. The results by UV and ELSD were almost the same for all TG molecular species. In contrast, the results of the chromatogram area percent by UV and ELSD, historically used method for the indication of absolute content of TG molecular species in oil, were completely different. This method would play an important role for the clarification of the nutritional function of fish oil from the TG molecular structure perspectives.