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3 protocols using debrisoquine sulfate

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

Lipid Standard Procurement and Characterization

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The lipid standard, phosphatidylserine PS (14:0/14:0), was purchased from Avanti® Polar lipids (Alabaster, AL). Debrisoquine sulfate, 4-nitrobenzoic acid (4-NBA) and UPLC-grade solvents such as acetonitrile, water and isopropanol were purchased from Fisher Scientific (Hanover Park, IL). Glyceric acid, glutaconic acid, pipecolic acid, homogentisic acid, sebacic acid, serotonin, hypoxanthine, α-ketobutyric acid, hippuric acid, taurine and hydroxyphenylpyruvic acid were purchased from Sigma-Aldrich® (Seelze, Germany). In addition, the MS/MS spectra provided by Scripps Center for Metabolomics (La Jolla, CA) were used as reference spectra for hydroquinone.
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3

Comprehensive Lipidomic and Metabolomic Analysis

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The following fatty acids and lipid standards
were purchased from Avanti Polar lipids (Alabaster, AL): sphingolipid
mix (SM) II, phosphatidylethanolamine PE (14:0/14:0), phosphatidylcholine
PC (14:0/14:0), phosphatidic acid PA (14:0/14:0), phosphatidylserine
PS (14:0/14:0), phosphatidylinositol PI (17:0/20:4), and lysophosphatidylcholine
LPC (17:1). Prostaglandin standard and leukotriene were purchased
from Cayman Chemical (Ann Arbor, MI), and fatty acid standard FA(17:1)
was purchased from Nu-Chek Prep (Elysian, MN). Debrisoquine sulfate,
4-nitrobenzoic acid (4-NBA), and UPLC-grade solvents such as acetonitrile,
water, and isopropanol were purchased from Fisher Scientific (Hanover
Park, IL). Glucose, riboflavin, arachidonic acid, linoleic acid, oleic
acid, palmitic acid, hippuric acid, nicotinic acid, lactic acid, uridine,
taurine, α-ketobutyric acid, uric acid, hydroxyphenylpyruvic
acid, acetylcarnitine, and carnitine were purchased from Sigma-Aldrich
(Seelze, Germany). The tandem MS spectrum of dityrosine provided by
Hanft et al. was used as a validation reference spectrum.8 (link) In addition, the MS/MS spectra provided by Scripps
Center for Metabolomics, METLIN, (La Jolla, CA) were used as reference
spectra for hydroquinone, inositol, gentisic acid, and dihydrolipoamide.
METLIN was also used to putatively identify the ion at m/z of 337.1048 as the sodium adduct of 7,8-dihydropteroic
acid.
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