Phenylboronic acid (>97.0% purity), 3-MCPD (>98% purity), 1,3-DCP (>98% purity), and d5 1,3-DCP (>98% purity) were purchased from Sigma-Aldrich (St. Louis, MO), and d5 3-MCPD (>98% purity) was purchased from Canadian isotopes (Point-Claire, Quebec, Canada). Sodium chloride crystals, acetonitrile, and LC-MS Optima water were purchased from Fisher Scientific (Fair Lawn, NJ, USA). For the QuEChERS method, salt extraction mylar pouches (AOAC 2007.1, 6 g of MgSO4 with 1.5 g of sodium acetate) and dSPE pouches (400 mg of PSA, 400 mg of C18) were purchased from United Chemical (Bristol, PA). Additionally, a FAPAS proficiency standard for 1,3-DCP and 3-MCPD in soy sauce was purchased from Sigma-Aldrich (St. Louis, MO). Stock solutions of 3-MCPD and d5 3-MCPD were initially prepared in 20% NaCl in water (PBA method) and 100% ethyl acetate (QuEChERS method) at concentrations of 2 mg/mL and further diluted for recovery and calibration solutions. Additionally, solutions of 1,3-DCP and d5 1,3-DCP were also prepared in ethyl acetate for the QuEChERS method. For the PBA and QuEChERS methods, seven-point calibration curves were prepared that corresponded to 3-MCPD and 1,3-DCP concentrations of 10–2600 μg/kg in soy sauce.
3 mcpd
Manufactured by Merck Group
Sourced in Macao, Germany
3-MCPD is a chemical compound used as a laboratory standard and analytical reagent in various applications. It serves as a reference material for the identification and quantification of similar compounds in research and testing environments. The core function of 3-MCPD is to provide a consistent and reliable point of comparison for analytical techniques.
Automatically generated - may contain errors
Lab products found in correlation
Microplate reader, by Thermo Fisher Scientific (2 mentions)
Phenylboronic acid, by Merck Group (1 mentions)
Acetonitrile, by Thermo Fisher Scientific (1 mentions)
Sodium chloride crystals, by Thermo Fisher Scientific (1 mentions)
Lc ms optima water, by Thermo Fisher Scientific (1 mentions)
Nec 1, by MedChemExpress (1 mentions)
Fer 1, by MedChemExpress (1 mentions)
Z vad fmk, by MedChemExpress (1 mentions)
Cck 8 reagent, by Solarbio (1 mentions)
C11 bodipy581 591, by Thermo Fisher Scientific (1 mentions)
Fluorescence microscope, by Zeiss (1 mentions)
7 protocols using 3 mcpd
1
Quantification of 3-MCPD and 1,3-DCP in Soy Sauce
2
Fabrication of Conductive ABS Filaments
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Commercially available ABS green filament and ABS conductive filament (graphene-based) of 1.75 mm were procured for FDM 3D printing. Iron(II) sulfate heptahydrate (FeSO4·7H2O), sodium borohydride (NaBH4), potassium chloride (KCl), potassium ferricyanide (K3[Fe(CN)6]), sodium hydroxide (NaOH), and 3-MCPD were purchased from Sigma-Aldrich (M) Sdn. Bhd. (Selangor, Malaysia). All analytical reagents used in this work were prepared without further refinement and used as obtained. Deionized (DI) water was used as solvent throughout this work.
3
Ionic Liquid Treatment of Olive Oil
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Example 5
The above general method was used as part of an ionic liquid treatment of a refined olive oil comprising less than 0.2 wt % FFA. The oil was doped with 3-MCPD obtained from Sigma-Aldrich at room temperature. Results of oil analysis before and after the ionic liquid treatment are provided in Table 1 below.
4
Chemical Preparation for Cell Viability Assays
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AB1 was obtained from Sigma-Aldrich (Steinheim, Germany; catalog no. A6636; purity ≥ 98%; lot 028M4088V), BaP from Supelco (Bellefonte, PA, USA; catalog no. 48564; analytical grade, lot LC15023V), Las from PhytoLab (Vestenbergsgreuth, Germany; catalog no. 80412; analytical grade; lot 12080), ME from Sigma-Aldrich (catalog no. 04607; analytical grade; lot BCBV6695), and NPYR also from Sigma-Aldrich (catalog no. 158240; purity ≥ 99%; lot MKCB3427).
3-MCPD was purchased from Sigma-Aldrich (catalog no. 10721, purity 98%, lot MKCC7598), MPH was obtained from Supelco (catalog no. 44-2641; purity 99.9%, lot LC1217V), PFOA from Sigma-Aldrich (catalog no. 171468; purity 96%; lot 11419LE), and PFOS from abcr (Karlsruhe, Germany; catalog no. AB128838; purity 97%; lot 1307952).
For use in cell viability assays, 100× stock solutions of the test chemicals were prepared in dimethyl sulfoxide (DMSO; AppliChem, Darmstadt, Germany). DMSO concentrations in the experiments therefore never exceeded 1%. 3-MCPD was diluted in culture medium. For use in the RNA sequencing experiment, 1000× stock solutions were prepared in DMSO, while 3-MCPD was diluted in the medium and NPYR was used in its undiluted form.
3-MCPD was purchased from Sigma-Aldrich (catalog no. 10721, purity 98%, lot MKCC7598), MPH was obtained from Supelco (catalog no. 44-2641; purity 99.9%, lot LC1217V), PFOA from Sigma-Aldrich (catalog no. 171468; purity 96%; lot 11419LE), and PFOS from abcr (Karlsruhe, Germany; catalog no. AB128838; purity 97%; lot 1307952).
For use in cell viability assays, 100× stock solutions of the test chemicals were prepared in dimethyl sulfoxide (DMSO; AppliChem, Darmstadt, Germany). DMSO concentrations in the experiments therefore never exceeded 1%. 3-MCPD was diluted in culture medium. For use in the RNA sequencing experiment, 1000× stock solutions were prepared in DMSO, while 3-MCPD was diluted in the medium and NPYR was used in its undiluted form.
5
Ionic Liquid Treatment of Corn Oil
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Example 3
The above general method was used as part of an ionic liquid treatment of a refined corn oil comprising less than 0.2 wt % FFA. The oil was doped with 3-MCPD obtained from Sigma-Aldrich at room temperature. Results of oil analysis before and after the ionic liquid treatment are provided in Table 1 below.
6
Evaluating 3-MCPD Cytotoxicity in HUVECs
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To evaluate cell viability, HUVECs were seeded into 96-well plates (5×103 cells/well) and treated using 3-MCPD (MilliporeSigma) at concentrations of 0.1, 0.2, 0.4, 0.8, 1.6, 3.2 and 6.4 µg/ml for 24 h at 37°C. Then, 10 µl CCK-8 reagent (Beijing Solarbio Science & Technology Co., Ltd.) was added to each well for 4 h at 37°C. The absorbance was assessed at 450 nm using a microplate reader (Thermo Fisher Scientific, Inc.). Cell inhibition rate=(OD value of control group-OD value of experimental group)/OD value of control group ×100. The experiment was repeated three times and the half maximal inhibitory concentration (IC50) was calculated.
To assess which type of cell death was induced by 3-MCPD, HUVECs were pretreated using 1 µM ferrostatin-1 (Fer-1, ferroptosis inhibitor, MedChemExpress), 20 µM Z-VAD-FMK (pan caspase inhibitor, MedChemExpress), 10 µM Nec-1 (necrostasis inhibitor, MedChemExpress) and 10 µM 3-MA (autophagy inhibitor, MedChemExpress) at 37°C for 1 h. Cell viability was then assessed according to the aforementioned method.
To assess which type of cell death was induced by 3-MCPD, HUVECs were pretreated using 1 µM ferrostatin-1 (Fer-1, ferroptosis inhibitor, MedChemExpress), 20 µM Z-VAD-FMK (pan caspase inhibitor, MedChemExpress), 10 µM Nec-1 (necrostasis inhibitor, MedChemExpress) and 10 µM 3-MA (autophagy inhibitor, MedChemExpress) at 37°C for 1 h. Cell viability was then assessed according to the aforementioned method.
7
Lipid Peroxidation Assay in HUVECs
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HUVECs were seeded into 6-well plates (1×106 cells/well) and treated with 2.0 µg/ml 3-MCPD (MilliporeSigma) or untreated control cells for 24 h at 37°C. The cells were treated using 2 µM C11-BODIPY581/591 (lipid peroxidation) (Invitrogen; Thermo Fisher Scientific, Inc.) for 30 min at 37°C in the dark. The cells were washed with PBS to remove the unincorporated dye and evaluated using a fluorescence microscope (magnification, ×400; Carl Zeiss AG).
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