Determination of 28 food safety risk factors in novel foods by ultra-high-performance liquid chromatography-quadrupole/Orbitrap high-resolution mass spectrometry

An analytical method was established for the determination of 24 food safety risk factors (18 heterocyclic aromatic amines and six mycotoxins) in synthetic meat and synthetic protein samples, and four microcystins in microalgae samples using ultra-high-performance liquid chromatography-quadrupole/Orbitrap high-resolution mass spectrometry (UPLC-Q/Orbitrap HRMS). The novel synthetic food was extracted using 50% methanol solution and purified with 3 mL of n-hexane saturated with 50% (volume fraction) methanol solution. For the microalgae samples, extraction was performed with 80% (volume fraction) acetonitrile solution and purified using a PRiME HLB solid-phase extraction column. An ACQUITY HSS T3 column (100 mmx2.1 mm, 1.8 mu m) was used for analyte separation, with 5 mmol/L ammonium acetate containing 0.01% (volume fraction) formic acid and methanol selected as the mobile phase. Gradient elution was adopted. Data acquisition was conducted in both positive and negative modes in full-scan data-dependent secondary scan mode (full MS-ddMS2). Matrix-matched standards were employed for quantification in the external standard method. Additionally, mzVault and TraceFinder were used to establish a screening database for 28 food safety risk factors. The results showed that the relative deviation of the exact mass of 28 food safety risk factors was less than 5x10-6, with good linearity in the range of 1-100 mu g/kg, and a correlation coefficient (r2) of >= 0.9978. The detection limit ranged from 0.5 to 4 mu g/kg, and the quantification limit ranged from 1 to 10 mu g/kg. The method recovery ranged from 70.0% to 119.6% with relative standard deviations ranging from 0.4% to 14.6% (n=6). The method is simple, sensitive, accurate, and suitable for rapid screening of 28 food safety risk factors in novel foods.