The SCFAs from colon microbiota were determined by gas chromatography (Trace 1300, Thermo Fischer), equipped with a flame ionization detector. Standards of acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid, valeric acid, and 2-methyl valeric acid in 5% formic acid (as internal standard) were purchased from Sigma–Aldrich (Sigma–Aldrich, St. Louis, MO, United States) and were of HPLC grade with >99% purity. Before the SCFAs analysis, 500 mg of colon content were mixed with 500 μl of cold internal standard solution and sonicated for 5 min in cold water. Then, centrifugate at 4°C with 15,000 g 15 min, the supernatant was transferred to a spin column (45 kDa) centrifugate and the resulting supernatant was injected into a capillary column (30 m ×250 μm × 0.25 μm, Restek Corporation, Bellefonte, PA, USA). The column starting temperature was 90°C (2 min) followed by 10°C/min until 150°C, then 50°C/min until 250°C (1 min).
2 methylvaleric acid
Manufactured by Merck Group
Sourced in Sao Tome and Principe, United States, United Kingdom
2-methylvaleric acid is a chemical compound with the formula CH3CH2CH(CH3)COOH. It is a carboxylic acid that can be used as a reagent or intermediate in various chemical processes.
Automatically generated - may contain errors
Lab products found in correlation
Propionic acid, by Merck Group (2 mentions)
Butyric acid, by Merck Group (2 mentions)
Isovaleric acid, by Merck Group (2 mentions)
Valeric acid, by Merck Group (2 mentions)
Isobutyric acid, by Merck Group (2 mentions)
Acetic acid, by Merck Group (2 mentions)
Trace 1300, by Thermo Fisher Scientific (1 mentions)
Ribose, by Merck Group (1 mentions)
Rhamnose, by Merck Group (1 mentions)
Mannose, by Merck Group (1 mentions)
Glucuronic acid, by Merck Group (1 mentions)
Arabinose, by Merck Group (1 mentions)
Indole 3 acetic acid, by Merck Group (1 mentions)
Galacturonic acid, by Merck Group (1 mentions)
Galactose, by Merck Group (1 mentions)
Indole 3 butyric acid, by Merck Group (1 mentions)
3 methylvalericacid, by Merck Group (1 mentions)
Fucose, by Merck Group (1 mentions)
N acetylglucosamine, by Merck Group (1 mentions)
N acetylgalactosamine, by Merck Group (1 mentions)
5 protocols using 2 methylvaleric acid
1
Quantification of Colon SCFAs by GC
2
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).
(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).
3
Quantitative Analysis of SCFAs
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The analysis of SCFAs was performed as previously described (Menezes et al., 2010 (link)) with some modifications. Frozen fecal samples (500 mg) were extracted with 1.5 mL of acetonitrile that included 0.05% 2-methyl-valeric acid (internal standard) (Sigma-Aldrich, WI, United States) and 12% HClO4, centrifuged at 11,000 × g for 20 min at 4°C, and filtered with the 0.2 μm PVDF filters (Millipore, MA, United States). The supernatants were injected (3 μL; split 1:10) into a Plus HP 6890 CG system (Hewlett-Packard, DE, United States) coupled to a flame ionization detector (FID) and a capillary-fused silica column (CP7747, Varian, CA, United States). The injector and FID temperatures were maintained at 270°C and 300°C, respectively. The analysis was performed using a temperature ramp from 115°C to 250°C (13 min) under constant pressure. Identification and quantification of the SCFAs was performed by comparing with a mixture of external standards (Volatile free acid mix, Supelco, PA, United States).
4
Quantification of Volatile Fatty Acids in Milk
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Short chain FA standards (acetic, propionic, iso-butyric, butyric, iso-valeric, valeric and caproic acids) as well as 2-methylvaleric acid (Sigma-Aldrich Company Ltd., Dorset, UK) were weighed (250 mg) and placed in 50 mL tubes and dissolved with approximately 50 mL of distilled water. The volatile FA (acetic, propionic, iso-butyric, butyric, iso-valeric, valeric and caproic acids) were mixed and 0.5 ml of this mixture was added to 0.5 mL of 2-methylvaleric acid solution to be used as the internal standard (Yang and Choong, 2001) (link). This mixture (0.1µl) was injected into a Hewlett Packard HP6890 GC (Agilent Technologies Inc. Germany) equipped with a flame ionization detector and utilising a capillary column (30.0 m x 250 µm x 0.25 µm) supplied by Greyhound Chromatography and Allied Chemicals (Merseyside, UK).
The milk samples were thawed at room temperature and 1 mL of each sample was transferred to a GC vial and 50 µl of the 2-methylvaleric acid (0.5%) aqueous solution was added, then mixed thoroughly. There was 0.1 µl of the mixture subsequently injected into the GC. The concentration of each VFA (mg/mL) was determined using the equation of Yang and Choong (2001) (link).
The milk samples were thawed at room temperature and 1 mL of each sample was transferred to a GC vial and 50 µl of the 2-methylvaleric acid (0.5%) aqueous solution was added, then mixed thoroughly. There was 0.1 µl of the mixture subsequently injected into the GC. The concentration of each VFA (mg/mL) was determined using the equation of Yang and Choong (2001) (link).
5
Authentic Standards for Quantification
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Authentic standards used for quantification in this study included E-2-decenal, 2-heptanone, nonanal, 2-nonanone, 2-methylisoborneol, 2-methyl-3-heptanone, α-terpineol, E,E-2,4-decadienal, 2-methylvaleric acid, and endoborneol (Sigma-Aldrich, St. Louis, MO). Diethyl ether was obtained from Fisher Scientific (Somerville, NJ).
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