Liquor fractions obtained from each pretreatment condition were neutralized and subjected to chromatography using a Luna® 5 μm C18(2) 100 Å LC Column 150 × 4.6 mm, together with a C18 4 × 2.0 mm ID guard column (both from Phenomenex, Cheshire, UK) to verify furfural and 5-hydroxymethyl-furfural content. Analyses were carried out using a Surveyor HPLC (Thermo Scientific, Hemel Hempstead, UK), with an elution system of acetonitrile by reversed-phase in an isocratic gradient (5% acetonitrile and 95% deionized water) at 1 mL/min. The eluted furfuraldehydes were detected by UV absorbance at 284 nm using a Finnigan Surveyor PDA Plus detector (Thermo Scientific; Hemel Hempstead, UK). The furfurals were quantified by interpolation of a calibration curve within the range of 0.005 μg/mL to 50 μg/mL in water.
Luna 5 μm c18 2 100 lc column 150 4.6 mm
Manufactured by Phenomenex
Sourced in United Kingdom, United States
The Luna® 5 μm C18(2) 100 Å LC Column 150 × 4.6 mm is a reversed-phase high-performance liquid chromatography (HPLC) column. It features a 5 μm particle size and a 100 Å pore size, with dimensions of 150 mm length and 4.6 mm internal diameter.
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2 protocols using luna 5 μm c18 2 100 lc column 150 4.6 mm
1
Quantification of Furfural Compounds
2
Quantifying Cellular Adenosine Nucleotides
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Cellular contents of adenosine nucleotides were determined as described previously (Ask et al.2013 (link)). Briefly, samples (two of 5 ml each) were quenched in 25 ml pure methanol at −40°C. Cellular ATP, ADP and AMP were extracted by incubation of cell pellets with 0.5 ml 0.52 M trichloroacetic acid containing 17 mM EDTA on ice for 15 min. Adenosine nucleotides were separated on a Luna® 5 μm C18(2) 100 Å LC column (150 × 4.6 mm) (Phenomenex Inc., Torrance, CA, USA) using acetonitrile and tetrabutylammonium buffer (0.005 M tetrabutylammonium hydrogensulfate, 0.01 M Na2HPO4, pH 7.0) as mobile phase at a flow rate of 1.0 ml min−1 at 20°C. The detection was performed with a photodiode array detector (PDA-3000; Dionex Corp.) at 260 nm. The energy charge was calculated according to the following equation: Energy charge = (ATP + ½ADP)/(ATP + ADP + AMP)
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