The extraction of the milk oligosaccharides was performed by solid phase extraction (SPE) using a graphitised carbon solid phase extraction cartridge (Supelclean ENVI-Carb, Supelco, Darmstadt, Germany, 3 mL). The SPE cartridges were first conditioned by passing 1.5 mL of 80% (v/v) acetonitrile containing 0.1% (v/v) trifluoroacetic acid (TFA), followed by washing with 1.5 mL of water. An aliquot of 100 µL milk serum was loaded onto the column, followed by 1.5 mL water for washing. Lactose and 3′-fucosyllactose (3′-FL) were eluted using 3 mL 3% (v/v) acetonitrile (Fraction A). The rest of the oligosaccharides were eluted in 1.5 mL 40% (v/v) acetonitrile containing 0.05% (v/v) TFA (Fraction B). Both fractions were dried under a stream of nitrogen and redissolved in 500 µL water.
Supelclean envi carb
Manufactured by Merck Group
Sourced in United States
Supelclean ENVI-Carb is a solid-phase extraction (SPE) sorbent used for the cleanup and concentration of organic compounds from environmental and biological samples. It is a porous carbon-based material with a high surface area that can adsorb a wide range of organic compounds.
Automatically generated - may contain errors
Lab products found in correlation
Hydrochloric acid (hcl), by Merck Group (2 mentions)
Ammonium formate, by Merck Group (2 mentions)
Hplc ms grade methanol, by Merck Group (1 mentions)
Ammonium hydroxide, by Merck Group (1 mentions)
Ammonium acetate, by Merck Group (1 mentions)
Milli q system, by Merck Group (1 mentions)
Methionine, by Merck Group (1 mentions)
Riboflavin, by Merck Group (1 mentions)
Trichloroacetic acid, by Thermo Fisher Scientific (1 mentions)
Hepes buffer, by Merck Group (1 mentions)
Perfluorooctanoic acid, by Thermo Fisher Scientific (1 mentions)
Na2co3, by Thermo Fisher Scientific (1 mentions)
Nitroblue tetrazolium nbt, by Merck Group (1 mentions)
Folin ciocalteu s phenol reagent, by Merck Group (1 mentions)
2 n morpholino ethanesulfonic acid, by Merck Group (1 mentions)
Ultrapure water, by Merck Group (1 mentions)
Deuterated zen zen d6, by Wellington Laboratories (1 mentions)
Lc ms grade methanol, by Merck Group (1 mentions)
Formic acid, by Merck Group (1 mentions)
Sodium acetate, by Merck Group (1 mentions)
10 protocols using supelclean envi carb
1
Milk Oligosaccharides Extraction by SPE
2
PFAS Quantification in Environmental Samples
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All the PFASs standards were obtained from National Measurement Institute (NMI), Australia. HPLC-MS grade methanol, water, ammonium hydroxide, ammonium acetate, perfluorooctanoic acid (PFOA), perfluorooctane sulfonate potassium salt (PFOS), potassium persulphate (K2S2O8), sodium hydroxide (NaOH) and hydrochloric acid (HCl) were procured from Sigma-Aldrich (Australia). Supelclean ENVI-Carb from Supelco was used for solid-phase extraction (SPE), with Supelclean™ ENVI-WAX. Glassware was intentionally avoided, and low-density polypropylene (PP) test tubes and pipette tips were used to conduct all experiments. Ultrapure water was used for dilution and cleaning purposes.
A list of analysed PFASs and internal standards in this study are presented inSupplementary Table S1 (Supporting Information).
A list of analysed PFASs and internal standards in this study are presented in
3
Comprehensive PFAA and Volatile PFAS Analysis
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The PFAAs and volatile PFASs
assessed and analyzed in this study are given inTables S1 (PFAAs) and S2 (volatile
PFASs) of theSupporting Information (SI) . PFAAs (50 μg/mL in methanol) were obtained from Greyhound
Chromatography (Merseyside, UK). Volatile PFASs (50 μg/mL in
methanol) were purchased from Chiron AS (Trondheim, Norway). Ultrapure
water originated from a Milli-Q system from Millipore (Watford, UK).
Ethyl acetate (HPLC, 054006) was supplied by Biosolve Chimie (Dieuze,
France). Acetonitrile (Chromasolve, 34851), Supelclean Envi-carb (Supelco,
957210-U), and ammonium formate (Bio ultra, 09735) were purchased
from Sigma-Aldrich (Zwijndrecht, The Netherlands). HPLC grade acetone
(J.T. Baker, 9254) and methanol (J.T. Baker, 8402) were obtained from
Boom (Meppel, The Netherlands).
assessed and analyzed in this study are given in
PFASs) of the
Chromatography (Merseyside, UK). Volatile PFASs (50 μg/mL in
methanol) were purchased from Chiron AS (Trondheim, Norway). Ultrapure
water originated from a Milli-Q system from Millipore (Watford, UK).
Ethyl acetate (HPLC, 054006) was supplied by Biosolve Chimie (Dieuze,
France). Acetonitrile (Chromasolve, 34851), Supelclean Envi-carb (Supelco,
957210-U), and ammonium formate (Bio ultra, 09735) were purchased
from Sigma-Aldrich (Zwijndrecht, The Netherlands). HPLC grade acetone
(J.T. Baker, 9254) and methanol (J.T. Baker, 8402) were obtained from
Boom (Meppel, The Netherlands).
4
Extraction and Quantification of Fluorinated Compounds
5
Analytical Determination of Mycotoxins
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Organic solvents of analytical grade, formic acid, nitric acid, ammonium formate, hydrochloric acid (reagent grade), iron (III) chloride hexahydrate, ethylene glycol, trisodium citrate, sodium acetate, poly(ethyleneglycol)-10k, and GCB (Supelclean ENVI-Carb, surface area: 100 m2/g, particle size: 120/400 mesh) were obtained from Sigma-Aldrich (St. Louis, MO, USA). LC-MS grade methanol and ultrapure water (resistivity 18.2 MΩ cm) were obtained from Sigma Aldrich and used for LC mobile phase.
Pure (purity ≥98%, unless differently specified) standards of the analytes AFB1, AFB2, AFG1, AFG2, OTA, and ZEN (purity ≥99%) were purchased from Sigma-Aldrich. The standards of aflatoxin M1 (AFM1) and deuterated OTA (OTA-d5) acquired from Sigma-Aldrich, and deuterated ZEN (ZEN-d6) acquired from Wellington Laboratories (Toronto, ON, Canada) were used as internal standards (ISs).
Individual stock standard solutions of the analytes were prepared at 200 ng µL−1 in methanol. A composite working standard solution of the six analytes was prepared in methanol at 10 pg µL−1 for AFB1, AFB2, AFG1, and AFG2, 30 pg µL−1 for OTA, and 750 pg µL−1 for ZEN. This mixture was renewed every two weeks. All the solutions were stored in the dark at −20 °C and brought to room temperature before use.
Pure (purity ≥98%, unless differently specified) standards of the analytes AFB1, AFB2, AFG1, AFG2, OTA, and ZEN (purity ≥99%) were purchased from Sigma-Aldrich. The standards of aflatoxin M1 (AFM1) and deuterated OTA (OTA-d5) acquired from Sigma-Aldrich, and deuterated ZEN (ZEN-d6) acquired from Wellington Laboratories (Toronto, ON, Canada) were used as internal standards (ISs).
Individual stock standard solutions of the analytes were prepared at 200 ng µL−1 in methanol. A composite working standard solution of the six analytes was prepared in methanol at 10 pg µL−1 for AFB1, AFB2, AFG1, and AFG2, 30 pg µL−1 for OTA, and 750 pg µL−1 for ZEN. This mixture was renewed every two weeks. All the solutions were stored in the dark at −20 °C and brought to room temperature before use.
6
Oligosaccharide Reduction and Analysis
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Reduction of oligosaccharides was conducted by mixing 10 µL of oligosaccharide sample from reaction 1 with 65 µL H2O (MilliQ) containing 700 µg NaBH4 or NaBD4. The reduction reaction was incubated over night at ambient temperature and then quenched by adding 20 µL 25 mM sodium acetate. Chromatographic analysis by HPAEC (see below) was done with the sample as is, whereas a further sample preparation for MALDI-ToF-MS was done as follows. Approximately 10 µL of a 1:1 (w/w) suspension of H2O:Supelclean ENVI Carb (Sigma-Aldrich) was packed in a pipette tip containing a C8-disc that was trapping the Supelclean material. The bed was conditioned with 50 µL H2O. The sample (5 µL) was applied, rinsed with 50 µL H2O and then eluted with 20 µL acetonitrile. The eluate was then analyzed by MALDI ToF-MS.
7
Glycomic Analysis of Insect Cell Lines
8
Milk Oligosaccharide Isolation and Lactose Quantification
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Solid phase extraction (SPE) was used to isolate the MOS by removing the majority of lactose. Dried milk carbohydrates were solubilized in nanopure water to a concentration of 50 g/L. SPE was performed using graphitized non-porous carbon cartridges (2 g bed weight, 12 mL volume, Supelclean Envi-carb, Sigma-Aldrich). Before samples were loaded, the SPE cartridges were washed with 1 tube volume (12 mL) of acetonitrile and conditioned with 2 tube volumes of nanopure water. Samples (100 mg of carbohydrates) were loaded onto the SPE cartridges, then salts were eluted with 2 tube volumes of water. Three tube volumes of 2% (v/v) acetonitrile/water solution were used to elute monomers and lactose from the cartridges. Then MOS were eluted in one tube volume of 40/60 (v/v) acetonitrile/water solution containing 0.05% (v/v) trifluoroacetic acid. The obtained oligosaccharides were dried to a constant weight in a centrifugal evaporator.
The amount of residual lactose in the MOS samples following SPE was quantified to adjust total MOS. The dried MOS were solubilized in nanopure water to a concentration of 4 g/L then diluted 1:500 with water. HPAEC-PAD was used to quantify the lactose content in post-SPE milk samples as described above.
The amount of residual lactose in the MOS samples following SPE was quantified to adjust total MOS. The dried MOS were solubilized in nanopure water to a concentration of 4 g/L then diluted 1:500 with water. HPAEC-PAD was used to quantify the lactose content in post-SPE milk samples as described above.
9
Quantification of Oxylipin Standards
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Analytical standards of OLE, DIGO, DIGI, CON, and OUB octahydrate were purchased from Sigma-Aldrich (Buchs, Switzerland). Individual stock solutions were prepared by dissolving the crystalline standards in MeOH at a concentration of 1 mg/mL. A methanolic solution of DIGO-D (1 mg/mL) was obtained from Cayman Chemicals (Ann Arbor, MI, USA). Intermediate solutions were prepared by diluting the stock solutions in MeOH. The stock and the intermediate solutions were stored at −20 °C.
The MeOH absolute ULC-MS, ACN ULC-MS, and HCOOH 99% ULC-MS were purchased from Biosolve (Valkenswaard, the Netherlands). The ammonia solution 28–30% was obtained from Merck (Darmstadt, Germany), while NH4HCO3 LC-MS and HCOONH4 LC-MS were supplied by Sigma-Aldrich (Steinheim, Germany). H2O was purified by a Milli-Q purification system (Millipore Corp., Bedford, MA, USA).
Oasis® MAX (3 cc, 60 mg) LP and Oasis® HLB (3 cc, 60 mg) extraction cartridges were provided by Waters (Wexford, Ireland). Discovery® DSC-18 (6 mL, 500 mg) and Supelclean™ ENVI-Carb™ (6 mL, 500 mg) SPE cartridges were obtained from Sigma-Aldrich. VWR (Randor, PA, USA) was the supplier of 15 and 50 mL centrifuge PP tubes and centrifugal filters (modified nylon, 0.2 µm, 500 µL).
The MeOH absolute ULC-MS, ACN ULC-MS, and HCOOH 99% ULC-MS were purchased from Biosolve (Valkenswaard, the Netherlands). The ammonia solution 28–30% was obtained from Merck (Darmstadt, Germany), while NH4HCO3 LC-MS and HCOONH4 LC-MS were supplied by Sigma-Aldrich (Steinheim, Germany). H2O was purified by a Milli-Q purification system (Millipore Corp., Bedford, MA, USA).
Oasis® MAX (3 cc, 60 mg) LP and Oasis® HLB (3 cc, 60 mg) extraction cartridges were provided by Waters (Wexford, Ireland). Discovery® DSC-18 (6 mL, 500 mg) and Supelclean™ ENVI-Carb™ (6 mL, 500 mg) SPE cartridges were obtained from Sigma-Aldrich. VWR (Randor, PA, USA) was the supplier of 15 and 50 mL centrifuge PP tubes and centrifugal filters (modified nylon, 0.2 µm, 500 µL).
10
Synthesis of Magnetic Graphitized Carbon-Titania Composite
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The mGCB@TiO2 composite was prepared based on a previous report for carbon nanotubes with some modification. 29 GCB (Supelclean ENVI-Carb, Sigma-Aldrich, St. Louis, MO) was first activated by treating 400 mg with 50 mL concentrated nitric acid, under stirring at room temperature for 7 h. Afterwards, the material was extensively washed with water until neutral pH and dried overnight at 50 °C. The just described oxidation with nitric acid was skipped for non-oxidized GCB material. GCB, either oxidized or non-oxidized (150 mg), was then magnetized treating with FeCl3•6H2O (810 mg) trisodium citrate (150 mg), sodium acetate (3.6 g), and poly(ethylene glycol)-10k (1.0 g) in 40 mL of ethylene glycol solution. The mixture was sonicated for 3 h and then sealed in a 125 mL autoclave for 10 h at 200 °C. When cold, the autoclave was opened and the product washed with water and ethanol. Finally, 30 mg of the magnetic graphitized carbon black (mGCB) was sonicated in 50 mL isopropyl alcohol for 30 min and added with 20 µL diethylamine. After stirring for 5 min, the mixture was added with 1.5 mL of titanium isopropoxide, transferred into the autoclave and kept at 200 °C for 24 h. The final product was washed with water and ethanol, dried and finally calcined at 400 °C for 2 h.
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