Citation: LAN Hao-Hui, WANG Xin-Cen, ZHANG Xiao-Mei, ZHANG Hong-Wei, CONG Pei-Xu, XU Jie, XUE Chang-Hu. Lipid Profile Analysis Method Based on Ultra High Performance Liquid Chromatography-Triple Time-of-Flight Tandem Mass Spectrometry and Its Application in Gadus Macrocephalus[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(7): 903-911. doi: 10.19756/j.issn.0253-3820.191595 shu

Lipid Profile Analysis Method Based on Ultra High Performance Liquid Chromatography-Triple Time-of-Flight Tandem Mass Spectrometry and Its Application in Gadus Macrocephalus

1.
College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China;
2.
Technology Center of Qingdao Customs District, Qingdao 266000, China

Received Date: 11 October 2019
Revised Date: 27 April 2020

Fund Project: This work was supported by the National Key R&D Program of China (No. 2018YFD0901101).

A method for analysis of lipid profile by ultra high performance liquid chromatography-triple time-of-flight tandem mass spectrometry (UPLC-Triple TOF-MS/MS) was established. The chromatographic column was selected, and the concentration of ammonium formate in mobile phase and the collision energy of mass spectrometry were optimized. The ACQUITY UPLC CSH C₁₈ chromatographic column (150 mm×2.1 mm, 1.7 μm) was used. The mobile phase A was acetonitrile/water (65:35, V/V), the mobile phase B was isopropanol/acetonitrile (85:15, V/V), and both of which contained 5 mmol/L ammonium formate and 0.1% formic acid. IDA automatic secondary scanning mode was adopted for mass spectrometry. Through methodological verification, lipid standards maintained good linearity in certain concentration ranges. The correlation coefficients (R²) were all above 0.995, the limits of detection (LOD) were 0.001-0.25 μmol/L, the limits of quantification (LOQ) were 0.0025-0.2 μmol/L, and the relative standard deviations were less than 10%. Most of the recovery rates were more than 80%. The lipid of Pacific cod (Gadus macrocephalus) was extracted by chloroform-methanol (2:1, V/V). A total of 498 lipid molecular species were identified in Pacific cod, including 255 phospholipid molecular species, 52 sphingolipid molecular species and 191 glyceride molecular species, of which the main subclasses were phosphatidylethanolamine, sphingomyelin and diglyceride. The results showed that the method had high sensitivity, high resolution and high throughput, and could be applied to the analysis of lipid profile of Pacific cod, which laid a foundation for the application of lipidomics in the field of aquatic products.
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沈阳化工大学材料科学与工程学院沈阳 110142