Stool samples were collected from the cecum of burn-injured and sham mice. Samples were processed and analyzed as previously described (23 (link)). Briefly, weighed cecal specimens were diluted in 25% meta-phosphoric acid at a 1:5 ratio. Samples were then centrifuged at 16,000 × g for 15 min at 4°C, filtered through 0.45-um syringe tip filter (Thermo Fischer Scientific; Waltham, MA), and stored at −80°C until analysis. Butyrate concentrations performed by high-performance liquid chromatography using a 0.01 N sulfuric acid mobile phase (apparent pH 2.0) pumped at a flow rate of 0.7 mL/min through a Rezex ROA-organic acid H+ (8%) 300 × 7.8 mm analytical column (Phenomenex, Torrance, CA) maintained at 65°C. The effluent following a 20-μL injection was monitored with a UV detector set at 210 nm. All equipment was made by Shimadzu (Kyoto, Japan). Peak area of butyric acid was compared to standards ranging for 1 to 100 mM. Butyrate concentrations were corrected for dilution and fecal weight and expressed as μmol per gram of wet weight feces (23 (link), 24 (link)).
Rezex roa organic acid h 8 300 7.8 mm analytical column
Manufactured by Phenomenex
Sourced in United States
The Rezex ROA-organic acid H+ (8%) 300 × 7.8 mm analytical column is a high-performance liquid chromatography (HPLC) column designed for the analysis of organic acids. The column features a stationary phase composed of a sulfonated polystyrene-divinylbenzene copolymer with a particle size of 8%, and dimensions of 300 mm length and 7.8 mm internal diameter.
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4 protocols using rezex roa organic acid h 8 300 7.8 mm analytical column
1
Butyrate Profiling in Burn Mouse Model
2
Butyric Acid Levels in Scald Injury Mice
3
UHPLC-based Glucose Quantification
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Quantification of glucose resulting from reactions with the Celluclast® enzyme cocktail was achieved using a Dionex Ultimate 3000 UHPLC system (Dionex, Sunnyvale, CA, USA) equipped with a Rezex ROA-Organic Acid H + (8%), 300 × 7.8 mm analytical column and a SecureGuard Carbo-H + 4 × 3.0 mm guard column (Phenomenex, Torrance, CA, USA) operated at 65 °C. Sample components were eluted isocratically over 22 min using 5 mM sulfuric acid as mobile phase with a flow rate of 0.6 mL/min. Products were detected using a refractive index (RI) detector 101 (Shodex, Tokyo, Japan) and data collection and analysis were carried out with the Chromeleon 7.0 software.
4
Glucose Quantification in Cellulosic Hydrolysis
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Glucose released during saccharification of cellulosic substrates was analyzed by high-performance liquid chromatography (HPLC) utilizing a Dionex Ultimate 3000 (Dionex, Sunnyvale, USA) coupled to a refractive index (RI) detector 101 (Shodex, Japan). Hydrolysis products generated from Avicel were separated at 85 °C, with 5 mM H2SO4 as the mobile phase at 1 mL min−1 flow rate, using a Rezex RFQ-Fast Acid H+ (8%) 100 × 7.8 mm analytical column (Phenomenex, Torrance, USA). Hydrolysis products released from Norway spruce and SEB were separated using a Rezex ROA-organic acid H+ (8%), 300 × 7.8 mm analytical column (Phenomenex), operated at 65 °C and 0.6 mL min−1 of 5 mM H2SO4. Glc4gemGlc was quantified by high-performance anion exchange chromatography (HPAEC) using a Dionex ICS 3000 coupled to a pulsed amperometric detector (PAD, Dionex), as described by Müller and colleagues and discussed in this study [16 (link)].
AscA was measured spectrophotometrically at 265 nm (Agilent Cary 60 spectrophotometer) using a standard curve for quantification that was prepared using AscA concentrations ranging from 5 to 150 µM. A buffer-enzyme mixture was used as a blank.
AscA was measured spectrophotometrically at 265 nm (Agilent Cary 60 spectrophotometer) using a standard curve for quantification that was prepared using AscA concentrations ranging from 5 to 150 µM. A buffer-enzyme mixture was used as a blank.
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