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4 protocols using methanol meoh of hplc grade

1

Quantification of 8-OHdG and 8-OHG

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Methanol (MeOH) of HPLC grade was obtained from Merck KGaA (Darmstadt, Germany). Formic acid (HCOOH), acetic acid (CH3COOH), and 8-OHdG were bought from Sigma-Aldrich (St Louis, MO, USA). [15N5]8-Hydroxy-2′-deoxyguanosine ([15N5]8-OHdG) was purchased from Cambridge Isotope Laboratories Inc. (Andover, MA, USA). 8-OHG and [13C15N2]8-hydroxyguanosine ([13C15N2]8-OHG) were bought from Toronto Research Chemical (Toronto, Canada). Water was gained from a Milli-Q water purification apparatus (Millipore, Milford, MA, USA).
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2

HPLC Analysis of Aspartate Transaminase

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An Agilent 1200 series Gradient HPLC with diode-array detection (DAD), Agilent Chemstation software (Agilent, Us) was used for the separation, qualitative and quantitative determination of AST. Methanol (MeOH) of HPLC grade and other chemicals at analytical grade were obtained from Merck, Germany.
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3

Comprehensive Metabolite Analysis Protocol

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Methanol (MeOH) of HPLC grade, 2-picolylamine
(2-PA), dipyridyl disulfide (DPDS), triphenylphosphine (TPP), acetic
acid, propionic acid, butyric acid, valeric acid, caproic acid, lactic
acid, succinic acid, isobutyric acid, isovaleric acid, 2-methylbutyric
acid, 3,3-dimethylbutyric acid, 2-methylvaleric acid, 3-methylvaleric
acid, 4-methylbutyric acid, indole-3-acetic acid, indole-3-butyric
acid, indole-3-lactic acid, 2-ethylbutyric acid (2-EtB), d4-acetic acid, glucose, galactose, fructose, arabinose, fucose, rhamnose,
glucuronic acid, galacturonic acid, Nacetylglucosamine, N-acetylgalactosamine, mannose, allose, ribose, 3-methyl-1-phenyl-2-
pyrazoline-5-one (PMP), trifluoroacetic acid (TFA), and ammonium acetate
were purchased from Sigma-Aldrich (St. Louis, MO). d2-indole-3-propionic
acid was purchased from Toronto Research Chemicals (Toronto, Canada).
Algal starch (U–13C, 98%+), 13C6 glucose, and unlabeled algal starch were purchased from Cambridge
Isotope Laboratories (Tewksbury, MA). Isopropanol of LC/MS grade was
purchased from Fisher Scientific (Waltham, MA). Acetonitrile (HPLC-grade)
was purchased from Honeywell (Muskegon, MI).
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4

Metabolite Sourcing and Preparation

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Labeled and non-labeled metabolites came from Sigma-Aldrich (St-Quentin-Fallavier, France) or Eurisotop (Saint-Aubin, France). Methanol (MeOH) of HPLC-grade was obtained from Sigma-Aldrich and formic acid (FA) from Carlo Erba reagents (Val de Reuil, France). Water filtered with an Elga Purelab (Flex system, High Wycombe, UK) was used in all experiments.
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