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4 protocols using 4 aminobutyric acid

1

Metabolite Profiling of Biological Samples

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The following reagents were acquired from Sigma Aldrich (St. Louis, Missouri, USA) and Merck (Darmstadt, Germany): 3-phenylbutyric acid, methoxyamine hydrochloride (MOX-HCl), anhydrous sodium sulphate (Na2SO4), urease, glucose, 4-aminobutyric acid (GABA), L-alanine, L-leucine, L-phenylalanine, L-tryptophan, L-tyrosine, ascorbic acid, citric acid, succinic acid, palmitic acid, arabinose, ribose, N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) with 1% trimethylsilyl chloride (TMCS), and pyridine. Other chemicals used were acetonitrile, ethyl acetate, hydrogen chloride and diethyl ether from Burdick and Jackson brand (Honeywell International Inc., Muskegon, USA).
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2

Comprehensive Amino Acid Analysis Protocol

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The HPLC-grade acetonitrile, acetic acid, l-aspartic acid, l-glutamic acid, dl-serine, l-histidine, glysine, l-threonine, l-arginine, l-alanine, l-tyrosine, l-valine, l-methionine, l-isoleucine, l-leucine, l-phenylalanine, l-ornithine monohydrochloride, l-lysine monohydrochloride, 4-aminobutyric acid (as internal standard), and DEEMM were obtained from Merck (Darmstadt, Germany). High-purity water was supplied by the PURELAB Option-Q system from Elga Veolia (High Wycombe, UK); sodium acetate was purchased from Carlo Erba Reagents (Val-de-Reuil, France); and ltryptophan was purchased from Carl Roth (Karlsruhe, Germany).
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3

Fluorescent Erythromycin Derivative Synthesis

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The following reagents were used: chloramphenicol (Sigma, Steinheim, Germany), 1-hydroxysuccinimide (HOSu), N,N′-dicyclohexylcarbodiimide (DCC), 4-aminobutyric acid (GABA), triphenylphosphine, and 11-bromundecanoic acid (Sigma-Aldrich Chemie GmbH, Steinheim, Germany). The fluorescent erythromycin derivative, BODIPY-Ery, was obtained previously [69 (link)]. The alkyl-TPPs were obtained according to [17 (link)].
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4

Quantification of Biogenic Amines and Amino Acids

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L-norepinephrine hydrochloride, (±)−dopamine hydrochloride, serotonin hydrochloride, epinephrine hydrochloride, tryptamine, (±)−octopamine hydrochloride, (±)−synephrine, tyramine hydrochloride, β-phenethylamine, D,L-metanephrine hydrochloride, adenine riboside, 4-aminobutyric acid, L-tyrosine, L-tryptophan, L-tryptophan-d5 (TRP-D5) and 4-aminobutyric acid-2,2,3,3,4,4-d6 (GABA-D6) were purchased from Sigma-Aldrich Corporation (Milan, Italy). The deuterated internal standards, (±)−norepinephrine-2,5,6,α,β,β-d6 HCl (NE-D6), 2-(3,4-dihydroxyphenyl)ethyl-1,1,2,2-d4-amine (DA-D4), serotonin-α,α,β,β-d4 creatinine sulfate complex (5HT-D4), (±)−epinephrine-2,5,6,α,β,β-d6 (E-D6), tryptamine-α,α,β,β-d4 HCl (TRY-D4), (±)−p-octopamine-α,β,β-d3 HCl (OCT-D3), 2-(4-hydroxyphenyl)ethyl-1,1,2,2-d4-amine HCl (TYRA-D4), and 2-phenyl-d5-ethylamine (β-PEA-D5) were obtained from CDN Isotopes (Pointe-Claire, Quebec, Canada). The L-tyrosine-ring-d4 (TYRO-D4) was from Cambridge Isotope Laboratories, Inc. (Tewksbury, MA, USA). The purity of all analytes and labeled internal standards was ≥98%.
Water was purified with a Milli-Q Elix purification system (Millipore, Bedford, MA, USA). High-purity MS-grade solvents (formic acid, methanol, and acetonitrile) were obtained from Fluka (Milan, Italy) and used without further purification.
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