关键词: 快速滤过型净化, 超高效液相色谱-串联质谱, 全氟烷基化合物, 中华绒螯蟹

Abstract:

Perfluoroalkyl substances (PFASs) have become a new food-safety problem. Dietary intake is a major pathway of human exposure to PFASs. Chinese mitten crab (Eriocheir sinensis) is a high-end aquaculture product popular among consumers in China. Conventional extraction methods for PFASs are cumbersome and time consuming, and result in incomplete purification; thus, this technique does not meet the requirements for PFAS detection. Herein, an analytical strategy was established for the rapid detection of 14 PFASs in Chinese mitten crab based on multi-plug filtration cleanup (m-PFC) and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The carbon-chain length of the 14 PFASs analyzed in this study ranged from 4 to 14, and they are perfluorobutanoic acid (PFBA), perfluoro-n-pentanoic acid (PFPeA), perfluorohexanoic acid (PFHxA), perfluoroheptanoic acid (PFHpA), perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluoroundecanoic acid (PFUnDA), perfluorododecanoic acid (PFDoDA), perfluorotetradecanoic acid (PFTeDA), perfluoro-1-butane sulfonic acid (PFBS), perfluoro-1-hexane sulfonic acid (PFHxS), perfluoro-1-octane sulfonic acid (PFOS), and perfluoro-1-decanesulfonate (PFDS). The m-PFC column was prepared using carboxy-based multiwalled carbon nanotubes, and used to reduce the interference of sample impurities. The samples were extracted with 5 mL of 0.1% formic acid aqueous solution, 15 mL of acetonitrile and extraction salt (2 g Na₂SO₄ and 2 g NaCl). The supernatant (10 mL) was purified using the m-PFC column, concentrated to near dryness under nitrogen, and then redissolved in 1 mL of methanol. Finally, the sample solution was filtered through a 0.22 μm polypropylene syringe filter for UPLC-MS/MS analysis. The target analytes were separated using a Shimadzu Shim-pack G1ST-C18 chromatographic column (100 mm×2.1 mm, 2 μm) using methanol (A) and 5 mmol/L ammonium acetate aqueous solution (B) as the mobile phases via gradient elution. The linear gradient program were as follows: 0-0.5 min, 10%A-35%A; 0.5-3 min, 35%A-60%A; 3-5 min, 60%A-100%A; 5-6.5 min, 100%A; 6.5-7 min, 100%A-10%A. The target analytes were analyzed using negative electrospray ionization in multiple-reaction monitoring mode, and quantitative analysis was performed using the internal standard method.
In this study, we optimized the mobile-phase system as well as the extraction solvent, time, volume, and salt. The 14 PFASs exhibited good peak shapes and sensitivities when the 5 mmol/L ammonium acetate solution-methanol system was used as the mobile phase. Compared with acetonitrile or methanol alone, the extraction efficiencies of the 14 PFASs were significantly improved when 5 mL of 0.1% formic acid aqueous solution was added, followed by 15 mL of acetonitrile. The extraction efficiencies of the 14 PFASs did not differ significantly when the extraction time was within 3-15 min. The extraction salt (MgSO₄, Na₂SO₄, NaCl, (NH₄)₂SO₄, and Na₂SO₄+NaCl) significantly affected the extraction efficiencies of the 14 PFASs. The highest extraction efficiencies of the 14 PFASs, which ranged from 47.9% to 121.9%, were obtained when Na₂SO₄+NaCl was used as the extraction salt. Under the optimal experimental conditions, good linearities (R²=0.998-0.999) were obtained for seven PFASs (PFBS, PFHxA, PFHpA, PFHxS, PFDA, PFDoDA, PFTeDA) at 0.10-100 μg/L, and seven PFASs (PFBA, PFPeA, PFOA, PFOS, PFNA, PFUnDA, PFDS) at 0.50-100 μg/L. The average spiked recoveries for the 14 PFASs in Chinese mitten crabs at three levels ranged from 73.1% to 120%, with relative standard deviations (RSDs) in the range of 1.68%-19.5%(n=6). The limits of detection (LODs) and quantification (LOQs) of the 14 PFASs were in the range of 0.03-0.15 and 0.10-0.50 μg/kg, respectively. The developed method was applied to the analysis of crab samples collected from three farms in Shanghai, and PFASs with total concentrations of 3.52-37.77 μg/kg were detected in all samples. The detection frequencies for PFDA, PFUnDA, PFDoDA, PFTeDA, and PFOS were 100%. PFDA, PFUnDA, PFOS, and PFDoDA were the most abundant congeners, accounting for 31.2%, 30.6%, 15.0%, and 10.9%, respectively, of the 14 PFASs detected. The proposed method is simple, efficient, accurate, and suitable for the rapid analysis of 14 PFASs in Chinese mitten crabs.

Key words: multi-plug filtration cleanup (m-PFC), ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), perfluoroalkyl substances (PFASs), Chinese mitten crab