Simultaneous Measurement of Four Hydroxyoctadecadienoic Acid Isomers in Meat Products by Normal-phase High Performance Liquid Chromatography
LIU Ting, XIONG Qiang, GENG Zhiming, XU Weimin
2021, 42(4):
271-277.
doi:10.7506/spkx1002-6630-20190703-039
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In this study, a high performance liquid chromatography (HPLC) method was established for simultaneous determination of 13-hydroxy-9Z,11E-octadecadienoic acid (13-Z,E-HODE), 13-hydroxy-9E,11E-octadecadienoic acid (13-E,E-HODE), 9-hydroxy-10Z,12E-octadecadienoic acid (9-Z,E-HODE) and 9-hydroxy-10E,12E-octadecadienoic acid (9-E,E-HODE) in meat products. The analytes were extracted from samples with methanol, purified by Sep-Pak C18 column, and then separated using isocratic elution on an Absolute SiO2 column (250 mm × 4.6 mm, 5 μm) with a mobile phase consisting of n-hexane, isopropanol and acetic acid (98.3:1.6:0.1, V/V) before being detected at 234 nm using a photo-diode array (PDA). The results indicated that the method displayed a good linearity within the concentration ranges of 0.5–20.0 μg/mL (R2 = 0.999 4) for 13-Z,E-HODE, and 0.25–10.0 μg/mL (R2 = 0.999 2) for 13-E,E-HODE, 0.75–12.5 μg/mL (R2 = 0.999 2) for 9-Z,E-HODE and 0.5–7.5 μg/mL (R2 = 0.999 6) for 9-E,E-HODE, respectively. The detection limits (LODs) for 13-Z,E-HODE, 13-E,E-HODE, 9-Z,E-HODE and 9-E,E-HODE were 0.075, 0.035, 0.090 and 0.060 μg/g, respectively, and the limits of quantification (LOQs) were 0.25, 0.12, 0.32 and 0.20 μg/g, respectively. The average recoveries from spiked samples were 89.03%, 89.03%, 89.33% and 87.93%, respectively. When this method was used to analyze 18 meat products, all the samples contained 13-Z,E-HODE, 13-E,E-HODE, 9-Z,E-HODE and 9-E,E-HODE at levels of 1.73–9.10, 0.56–5.79, 2.37–11.02 and 0.78–5.82 μg/g, respectively. In conclusion, this method was proved to be fast, accurate and reproducible, and could be used for the simultaneous quantitative analysis of 13-Z,E-HODE, 13-E,E-HODE, 9-Z,E-HODE and 9-E,E-HODE in meat products.