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3 protocols using lc ms grade ammonium hydroxide

1

Quantification of Heterocyclic Amines

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Harman, Norharman, AαC, MeAαC, Trp-P-1, Trp-P-2, Glu-P-1, Glu-P-2, IQ, PhIP and their corresponding isotope-labeled analogs—Harman-13C2, 15N; Norharman-D7; AαC-15N3; MeAαC-D3; Trp-P-1-13C2,15N; Trp-P-2-13C2,15N; Glu-P-1-13C3; Glu-P-2-13C2,15N; IQ-D3; and PhIP-D3—were purchased from Toronto Research Chemicals Inc. (North York, ON, Canada). LC/MS grade acetonitrile (ACN) from Fisher Scientific (Pittsburgh, PA, USA) and LC/MS grade water from J.T. Baker (Center Valley, PA, USA) were used for mobile phase preparation. LC/MS grade ammonium hydroxide from Sigma-Aldrich (St. Louis, MO, USA) was used for pH adjustment. All other reagents were at least high performance liquid chromatography (HPLC) grade. Ninety-six-well plates of ISOLUTE SLE+ (supported liquid extraction) and EVOLUTE® CX (mix mode of cation exchange resin, CX) for sample preparation were purchased from Biotage Inc (Charlotte, NC, USA).
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2

Liquid Chromatography-Mass Spectrometry Preparation

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LC-MS grade ammonium hydroxide was purchased from Sigma-Aldrich (Overijse, Belgium), and ULC-MS grade ammonium acetate, ULC-MS grade acetonitrile (ACN), and HPLC-S grade ACN were from Biosolve (Valkenswaard, The Netherlands). The water was produced by a Milli-Q system (Millipore, Overijse, Belgium). Two-mL syringes, 50 mL PP (polypropylene) tubes, AR-grade ammonium carbonate, AR-grade magnesium sulfate (MgSO4), and AR-grade sodium chloride (NaCl) were purchased from VWR (Haasrode, Belgium). Two-mL amber glass vials were from Waters (Waters Corp., Milford, MA. USA). Lastly, 0.2 μm polytetrafluoroethylene (PTFE) syringe filters were from Phenomenex (Utrecht, The Netherlands).
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3

Organic Compound Synthesis with Reagents

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TEMPO 98%, sodium borohydride 98 atom%, hexamethyldisilazane, chlorotrimethylsilane, methanol 99.9%, Dowex 50 W × 8 resin, sodium acetate 99%, acetic anhydride 99% and LC-MS-grade ammonium hydroxide 25% were obtained from Sigma-Aldrich (Sigma-Aldrich Oy, Helsinki, Finland). Sodium bromine 99%, ammonia 25%, acetic acid 99.7%, sodium hydroxide 99%, sodium nitrate 99% and LC-MS-grade acetonitrile were purchased from Merck (Merck Oy, Espoo, Finland). Pyridine, sodium hypochlorite 18%, and potassium dihydrogenphosphate 99.9% were provided by VWR (VWR International Oy, Helsinki, Finland). Hydrochloric acid was supplied by J.T. Baker (Avantor Performance Materials, Center Valley, PA, USA), while technical grade ethanol was purchased from Altia Oyj (Rajamäki, Finland) .
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