Mini uniprep g2 syringeless filter

Aliquots (50 μg/L) of working standard mixture of HCAs containing internal standard (TriMeIQx = 0.05 μg/mL) were used as quality control for the extraction of samples with the accelerated solvent extractor (Dionex ASE 350, Thermo Scientific, MO, USA). Briefly, 25 uL of 5000 μg/L (5 μg/mL) working standard mixture was mixed with 2.5 mL of 0.5 M NaOH in MeOH/Water (70:30 v/v) and incubated for 2 h at room temperature with stirring in a 5 mL beaker. Diatomaceous earth (1:2 v/w) was added to the mixture and the paste loaded into 10 mL stainless steel ASE extraction cells for extraction with pure methanol. The ASE extraction program was set as follows: 2 cycles, 5 min heating time, 80 °C extraction temperature, 160 s purge time, and 1606 psi static pressure. The extracted analytes in 20 mL of methanol were evaporated to dryness with a rotary evaporator and nitrogen gas at room temperature. HCAs were re-suspended in 2.5 mL methanol and filtered (Mini-UniPrepTM G2 syringeless filter, 0.2 μm, Whatman, Buckinghamshire, UK) prior to analysis by UHPLC-HRMS/MS. The ASE extractions were performed in quadruplicates (n = 4) for each working standard mixture.

HCAs were extracted with an accelerated solvent extractor (Dionex ASE 350, Thermo Scientific, MO, USA). Grilled meat (1 g) was ground, spiked with internal standard (TriMeIQx = 0.05 μg/mL), mixed with 0.5 M NaOH in MeOH/Water (70:30 v/v) and incubated for 2 h at room temperature. Diatomaceous earth (1:2 w/w) was added to the mixture and HCAs were extracted with pure methanol. The ASE extraction program was as follows: 2 cycles, 5 min heating time, 80 °C extraction temperature, 160 s purge time, and 1606 psi static pressure. The extracted analytes were evaporated to dryness with a rotary evaporator and nitrogen gas at room temperature. HCAs were re-suspended in 2.5 mL methanol and filtered (Mini-UniPrepTM G2 syringeless filter, 0.2 μm, Whatman, Buckinghamshire, UK) prior to ultrahigh performance liquid chromatography coupled to high resolution accurate mass tandem mass spectrometry (UHPLC-HRMS/MS) analysis [14 (link)].