Dionex ase 350

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)].

Proximate composition of raw and extruded flours was determined in triplicate according to standard methods of the Association of Official Analytical Chemist AOAC [20 ]. Total nitrogen was determined by Leco FP628 (Leco Company, St. Joseph, MI, USA), and fat content determined on a Dionex ASE350 solvent extractor (Thermo Fisher Scientific, Inc., Waltham, MA, USA). Carbohydrate content was calculated by difference.

A detailed description of sampling and analytical methodologies of airborne PAHs can be found in the Supplemental Information. In brief, the airborne PAH concentrations of the 16 US EPA PAHs (Hussar et al. 2013 (link)) were determined in the PBZ and UAZ with a passive collection method. This passive method is based on cylindrical polyurethane foam samplers (PUF-cyl, length: 10 cm, diameter: 2.2 cm) (Strandberg et al. 2018 (link)).
The PUF-cyl passive sampler has previously been evaluated and calibrated with uptake factors for short sampling times (2–8 h) in other work environments (Bohlin et al. 2010 (link); Strandberg et al. 2018 (link)). Concurrent sampling with the passive PUF-cyl sampler and active sampling (NMAM 5515, (NIOSH 1994 )) in the UAZ were performed to determine the uptake-rates of PUF-cyl in the present work environment. The uptake factors were then used to determine the PAH levels from the passive sampling in the PBZ.
The PUF-cyl samplers were extracted using an Accelerated Solvent Extractor (Dionex ASE 350; Thermo Fisher Scientific), purified using an open column (ID 0.9 cm) with 1 g sodium sulfate, 4 g activated silica and 2 g activated alumina. PAHs were separated and detected by means of high-resolution gas chromatography/low resolution mass spectrometry (HRGC/LRMS) (Agilent Technologies, Inc.,CA, USA).

A precise 2.0 g powder sample was weighed, and 50 μL of the above five deuterated PAHs (25 ng each) were added as a standard surrogate mixture. The extraction was then performed with an accelerated solvent extraction (ASE) system (Dionex ASE 350, Thermo Fisher Scientific, Waltham, MA, USA). The conditions for ASE extraction, silica gel cleaning, and gel permeation chromatography were reported in our previous study [8 (link)]. Under mild N₂ purge, the cleaned-up extract was concentrated to about 50 μL. Finally, a syringe spike of 50 μL of Fluoranthene-d₁₀ (25 ng) and detection was performed using a high-resolution gas chromatography (GC) Orbitrap mass spectrometry (MS)-based method (Orbitrap GC/MS). A previous study described detailed Orbitrap GC/MS conditions for HPAHs and PAHs [8 (link)].

The carotenoids from each freeze-dried smoothie sample were extracted by using the accelerated solvent extractor Dionex ASE 350 (Thermo Fisher Scientific, Waltham, MA, USA) with acetone as the solvent. The extraction process was as follows: 1 g of each sample was weighed into extraction cells (34 mL), about 4 g of sand was added and mixed roughly. Pure acetone was used as the extraction solvent, with 3 cycles of 10 min for each sample. The pressure was 1500 psi and, after each cycle, the cells were washed with acetone. The process occurred at room temperature (23 °C). The obtained extract was evaporated and filled with acetone in a 10 mL volumetric flask.
The same HPLC system as for anthocyanin was used. The chromatographic conditions were as follows: injection volume 10 µL; flow rate: 0.5 mL/min; solvent A acetonitrile/0.05% triethylamine (v/v); solvent B methanol/ethyl acetate (11/9; v/v). The elution gradient was linear as follows: from 0 to 50 min solvent B increased from 5 to 40%, from 50 to 60 min solvent B increased from 40 to 80%, and from 60 to 75 min solvent B decreased from 80 to 5%. The detector was set for scanning in the range of 400 to 700 nm, while carotenoid quantification was performed at 454 nm. All measurements were performed in duplicate, and the results were expressed as µg of β-carotene/100 g of fresh weight of the sample.

Two topsoil samples (20 cm depth) were collected in 2018 in a 20 m² area in proximity to the largest industrial steel and iron ore sinter plant of Taranto (Apulia Region, South Italy) and to illegal dumping sites and the steel plant’s landfills nearby. Located in the south-east of Italy, the city of Taranto has been included among the National Priority Contaminated Sites and as an area of high risk of environmental crisis (defined within Italian National Laws D.Lgs. n. 426/98; D.M. 10 January 2000). A composite topsoil sample (5 kg) made up of 3 sub-samples taken in a 20 m² area was obtained using a stainless-steel spade. The resulting topsoil composite sample (60 g) was lyophilized, sieved (2 mm), and extracted (5 g) by accelerated solvent extraction (Dionex ASE 350, Thermo Fisher Scientific) using a standardized protocol previously described [31 (link)]. One topsoil extract was tested using the traditional luciferase measurement conducted by following the Dutch standard method RIKZ-SPECIE 07 according to the method reported for soil samples in [31 (link)]. The other topsoil extract, after the same procedure of extraction and purification, was used in proteomic analysis.

The bark was grounded into a fine powder (0.425 mm) by using a Wiley mill. Ten different solvents were used with three solvent‐dependent extraction techniques: water (1), methanol (2), ethanol (3), acetone (4), methylene chloride (5), ethyl acetate (6), chloroform (7), hexane (8), water–ethanol (9), and acid–base (10). The last solvent corresponds to a liquid–liquid extraction technique using several solvents described by Yubin, Miao, Bing, and Yao (Yubin, Miao, Bing, & Yao, 2014). This method allows mainly the extraction of alkaloids from the bark. The majority of the extracts (2, 3, 4, 5, 6, 7, 8) were obtained using the soxhlet extraction technique during 7 hr of reflux. The other two extracts (1, 9) were obtained using an accelerated solvent extractor (Dionex™ ASE™ 350; ThermoFisher). The solid bark was extracted with distilled water at 100°C and 1,500 psi over six cycles of 10 min each. For the water–ethanol extract (9), a liquid extraction with ethanol was carried out following a first extraction with water. The liquid extract was re‐extracted with ethanol at 120°C and 1,500 psi over six cycles of 5 min. Finally, all extracts were evaporated to dryness in a low temperature oven.