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2 protocols using n acetylputrescine

1

LC-MS Analysis of Polyamines and Related Compounds

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Acetonitrile (LC-MS grade) was purchased from Merck (Darmstadt, Germany), and methanol and formic acid (both LC-MS grade) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Chloroform, sodium bicarbonate, sodium carbonate, ammonium acetate, trifluoroacetic acid, perchloric acid, sodium hydroxide, acetone, and dansyl chloride were obtained from Sigma-Aldrich, whereas ethyl acetate was obtained from Merck. The water used throughout the study was purified in a Milli-Q system from Millipore (Burlington, MA, USA).
N1-acetylspermidine (N1-AcSPD), N8-acetylspermidine (N8-AcSPD), spermine-d20, spermine, spermidine-d6, spermidine, putrescine-d8, putrescine, N1,N12-diacetylspermine, N1,N8-diacetylspermidine (N1,N8-DiAcSPD), N8-acetylspermidine-d3, N1,N8-diacetylspermidine (N1,N8-DiAcSPD-d6), and agmatine were acquired from Toronto Research Chemicals (North York, ON, Canada). Cadaverine, N-acetylputrescine, N-acetylspermine (N1-AcSPM), lysine, ornithine, arginine, γ-aminobutyric acid, and 1,3-diaminopropane were purchased from Sigma-Aldrich. Lysine (13C6,15N2) and arginine (13C6,15N4) were supplied by Cambridge Isotope Laboratories (Tewksbury, MA, USA).
Oasis HLB cartridges (1 cc Vac Cartridge, 30 mg sorbent, and 30 µm particle size) were acquired from Waters (Milford, MA, USA).
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2

Analysis of Polyamines and Related Metabolites

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Ammonium acetate, sodium hydroxide, sodium chloride, methanol, dimethyl sulfoxide (DMSO) and isopropyl alcohol were from Sigma-Aldrich (St. Louis, MO, USA) and purchased with the highest purity available. MS grade formic acid and isopropyl alcohol were from Riedel-de Haën (Seelze, Germany) and acetonitrile was from Labscan (Gliwice, Poland). Deionized water was obtained by using a Milli-Q system from Millipore (Bedford, MA, USA). The following commercial standards were from Sigma-Aldrich: glutathione, glutathione oxidized, spermine, spermidine, putrescine, cadaverine, Nacetylspermine, ornithine, lysine, arginine, S-adenosylmethionine, adenine, N-acetylputrescine, Sadenosylhomocysteine, cysteine, cytidine, methionine, adenosine, 5'-deoxy-5'-(methylthio)adenosine, aminoisobutyric acid (GABA), tyramine and methionine sulfone.
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