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3 protocols using propyl chloroformate

1

Metabolic Profiling of Biological Samples

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Tempol, short chain fatty acids, 1-propanol, propyl chloroformate, pyridine, sodium chloride, K2HPO4, and NaH2PO4 were purchased from Sigma-Aldrich Chemical Co. Ltd. (St. Louis, MO). Hexane and methanol were purchased from EMD Chemicals Inc (Gibbstown, NJ). The internal standard hexanoic acid-6,6,6-d3 was obtained from C/D/N Isotopes Inc (Pointe-Claire, Quebec, Canada). Sodium 3-trimethylsilyl [2,2,3,3-d4] propionate (TSP-d4) and D2O (99.9% in D) were purchased from Cambridge Isotope Laboratories (Miami, FL). Standard benzoic acid pellets were purchased from Parr Instrument Company (Moline, IL). All compounds were of the highest analytical grade available.
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2

HPLC-based Bile Acid and Metabolite Analysis

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Methanol (MeOH), acetonitrile and formic acid were of HPLC grade. Analytical grade of NaOH, propan-1-ol, pyridine, hexane and propylchloroformate (PCF) were purchased as well from Sigma-Aldrich (Saint Quentin Fallavier, France). Deionized water comes from a Milli-Q Elix system fitted with a LC-PaK and a MilliPak filter at 0.22 μm (Merck Millipore, Guyancourt, France). The following bile acid standards: cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), lithocholic acid (LCA), ursodeoxycholic acid (UDCA), hyodeoxycholic acid (HDCA), lycocholic acid (GCA), glycochenodeoxycholic acid (GCDCA), glycodeoxycholic acid (GDCA), glycolithocholic acid (GLCA), taurocholic acid (TCA), taurochenodeoxycholic acid (TCDCA), taurodeoxycholic acid (TDCA), tauroursodeoxycholic acid (TUDCA), and taurolithocholic acid (TLCA) and ammonium acetate were purchased from Sigma Chemical (St. Louis, MO, USA). All tryptophan reference, isotope labeled metabolites and SCFAs (acetate, propionate, butyrate, isobutyrate, valerate, isovalerate, acetate-D3, butyrate-13C2 and valerate-D9) were purchased from Sigma-Aldrich (Saint Quentin Fallavier, France). The stock solutions of bile acids, tryptophan metabolite and SCFAs were prepared separately in methanol at the concentration of 10 mmol/l.
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3

Synthesis of Acrylate Emulsion Polymers

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Model acrylate emulsion polymers were synthesized using radical emulsion polymerization of methyl methacrylate (MMA, Sigma Aldrich), n-butyl acrylate (BA, Sigma Aldrich), n-butyl methacrylate (BMA, Sigma Aldrich) and acrylic or methacrylic acid (AA or MAA, Sigma Aldrich), as outlined in Table 1. Sodium lauryl sulphate (32% in water, VWR, Amsterdam, The Netherlands) was used as emulsifier and ammonium persulphate (Evonik, Marl, Germany) as initiator. The samples are dispersed in water to approximately 30% by weight. Ammonia (25% in water, VWR) was used in a stoichiometric equivalent of 0.7 to partly neutralize the acid groups.
n-Methylpyrrolidone (NMP, peptide synthesis grade) and tetrahydrofuran (THF, uLCMS grade) were purchased from Biosolve (Valkenswaard, The Netherlands). Triethylamine (TEA, 99.5%), hydrochloric acid (HCl, 37%), N-methylmorpholine (NMM, >99%), chloro-dimethoxytriazine (CDMT, >98%), ethyl chloroformate (ECF, >99%), propyl chloroformate (PCF, >99%), 3-nitrobenzyl chloroformate (NBCF, >96%), p-bromophenacyl bromide (pBPB, 99.8%), p-nitrophenacyl bromide (pNPB, >98%) and 2-bromoacetophenone (PB, 99%) were purchased from Sigma-Aldrich (Zwijndrecht, The Netherlands).
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