To measure the concentration of short chain fatty acids (SCFs), 1 g of feces was extracted with 5 mL of methanol and centrifuged at 28,000 × g for 15 min. The extract was stored at −60°C until use. The stored sample was thawed, filtered through a 0.45-µm Millipore filter (Millipore, USA), and analyzed using a DB-FFAP 123-3253 (50 m × 0.32 mm × 0.50 µm), a flame ionization detector equipped with an Agilent 6890N gas chromatography system (Agilent Technologies, USA). Nitrogen gas was used as the carrier at a flow rate of 1.4 mL/min and a sample injection volume of 1 µL. Inlet and detector temperatures were 200°C and 240°C, respectively. Acetic acid, propionic acid, and butyric acid contents were evaluated by constructing a calibration curve using the respective standard reagents.
Db ffap 123 3253
Manufactured by Agilent Technologies
Sourced in United States
The DB-FFAP 123-3253 is a capillary column designed for gas chromatography. It is a polyethylene glycol-based stationary phase intended for the separation and analysis of volatile organic compounds.
Automatically generated - may contain errors
Lab products found in correlation
Millipore filter, by Merck Group (2 mentions)
Propionic acid, by Merck Group (2 mentions)
Butyric acid, by Merck Group (2 mentions)
Biomasher, by Takara Bio (2 mentions)
Acetic acid, by Merck Group (2 mentions)
Agilent 6890n gas chromatography system, by Agilent Technologies (1 mentions)
Flame ionization detector, by Hewlett-Packard (1 mentions)
0.45 μm syringe filter, by Advantec (1 mentions)
5 protocols using db ffap 123 3253
1
Quantification of Short Chain Fatty Acids in Feces
2
SCFA Analysis of Culture Medium
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The SCFA analysis was conducted according to a previous study report [10 (link)]. Samples (2 mL) were collected from the culture medium using a sterile syringe at 12 h intervals and stored at −80 °C until the SCFA analysis. Briefly, the samples were centrifuged (5000× g) at room temperature for 15 min, and the supernatant was used for analysis. Gas chromatography (Agilent Technologies, Santa Clara, CA, USA) was equipped with a GC column (DBFFAP 123-3253, 50 m × 0.32 mm × 0.50 μM), flame ionization detector, and an autosampler. The injector and detector port temperatures were 200 and 240 °C, respectively. The carrier gas was N2 at a flow rate of 1.4 mL/min. The SCFA concentration was expressed in mM.
3
Quantifying Short-Chain Fatty Acids in Mouse Cecum
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To evaluate the SCFA content of the mouse caecum, it was ground to a concentration of 1 g/mL in an 80% methanol solution using a BioMasher (TaKaRa, Shiga, Japan). Centrifugation was performed at 13,000 rpm for 10 min at 4 °C, and the supernatant was filtered using 0.45 μm syringe filters from ADVANTEC. The analysis was performed using a flame ionization detector (Hewlett Packard, Palo Alto, CA, USA) and a GC column (DB-FFAP 123-3253, 50 mm × 0.32 mm × 0.50 μm, Agilent Technologies, Inc., Santa Clara, CA, USA). SCFA concentrations were quantified using Acetic acid, propionic acid, and butyric acid as standards. The SCFA content in the cecum was determined using a calibration curve based on the corresponding standards. Acetic acid, propionic acid, and butyric acid were purchased from Sigma-Aldrich (St. Louis, MO, USA).
4
Quantification of Cecal SCFA Levels
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To evaluate the SCFA content in the mouse cecum, we ground the tissue using the BioMasher (TaKaRa, Shiga, Japan) in an 80% methanol solution. The ground solution was then centrifuged at 13,000 rpm for 10 min at 4 °C, and the supernatant was filtered using ADVANTEC’s 0.45 μm syringe filter. Subsequently, analysis was performed using a flame ionization detector (HP-5890/5971, Hewlett Packard, Palo Alto, CA, USA) and GC column (DB-FFAP 123-3253, Agilent Technologies, Inc., Santa Clara, CA, USA, 50 mm × 0.32 mm × 0.50 μm). The SCFA levels were quantified using Acetic acid, propionic acid, and butyric acid as standards. The content of SCFAs in the cecum was determined using a calibration curve based on the respective standards. Acetic acid, propionic acid, and butyric acid were purchased from Sigma-Aldrich (St. Louis, MO, USA).
5
Cecal Short-Chain Fatty Acid Analysis
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To determine the SCFA content, 1 g of cecal contents was collected 1 day before the end of the experiment, followed by SCFA extraction using 5 ml of methanol and filtering via a 0.45 μm Millipore filter (Millipore, USA). SCFAs were analyzed using a gas chromatography (GC) system (Agilent Technologies, Santa Clara, CA, USA) equipped with a GC column (DB-FFAP 123-3253, 50 m × 0.32 mm × 0.50 μM), flame ionization detector, and autosampler. Nitrogen was used as the carrier gas with a flow rate of 1.4 ml/min a split ratio of 10 : 1. The sample injection volume was 1 μl, and inlet and detector temperatures were 200°C and 240°C, respectively, based on the protocol by Demingne and Remesy [12 (link)]. Acetic acid, propionic acid, and butyric acid contents were used as standards.
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