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2 protocols using 4 guanidinobutyric acid

1

Comprehensive Metabolite Identification via UPLC-MS

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Debrisoquine sulfate, 4-nitrobenzoic acid (4 (link)–NBA) and UPLC-grade solvents such as acetonitrile, water and isopropanol were purchased from Fisher Scientific (Hanover Park, IL). 4-Guanidinobutyric acid, riboflavin, retinoic acid, (−)-epinephrine, hippuric acid, nicotinic acid, S-(5′-adenosyl)-L-homocysteine, taurine, uric acid, homovanillic acid, acetylcarnitine and carnitine were purchased from Sigma-Aldrich®, (St. Louis, MO). The METLIN (3 (link)) database (La Jolla, CA) was used for tandem MS (MS/MS) validations of 4-aminobutanoate, 3-hydroxybutanoate and quinolinic acid.
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2

Synthesis and Characterization of Galegine Hemisulfate

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Metformin hydrochloride (Cayman Chemical), buformin hydrochloride (Enamine), phenformin hydrochloride (Cayman Chemical), L-arginine hydrochloride (Acros), agmatine sulfate (Fluka), 4-guanidinobutyric acid (Fluka) and 3-guanidinopropionic acid (Sigma) were obtained with high purity (>97%).
Galegine hemisulfate was synthesized via the condensation reaction of a thiourea with the alkylamine using a procedure adapted from Williams et al.66 2-methyl-2-thiopseudourea hemisulfate, 7 mmol, (Sigma) was added to 8 mmol 3-methyl-2-buten-1-amine (A2B Chem) in 5 mL of water and stirred overnight at room temperature. The reaction produced precipitate which was dried on a porcelain plate and washed with methanol. The solids were then recrystallized in methanol to produce galegine hemisulfate in high purity (>95%) as determined by NMR. 1H-NMR (DMSO-d6, 400 MHz) δ (ppm) 1.62 (s, 3H), 1.68 (s, 3H), 3.64 (br, 2H), 5.16 (t, 1H), 7.62 (br, 4H), 8.46 (br, 1H). 1H-NMR experiments were conducted using the Varian Unity Inova 400 MHz NMR system and VnmrQ 2.2 software.
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