To analyze saccharides and xylitol, the culture samples were centrifuged at the indicated time points, and the supernatant was used for the quantification by HPLC, using a Jasco HPLC 2000 plus series (Biolab, Vienna, Austria) with an Aminex hpx 87 h column at 65 °C. H2SO4 (c = 5 mmol l−1) was used as eluent at an isocratic flow rate of 0.8 ml min−1. Data acquisition was performed with a refractive index detection. Data were analyzed with ChromPass (Version 1.8.6.1, Jasco Europe, Italy).
Chrompass
Manufactured by Jasco
Sourced in Italy
ChromPass is a lab equipment designed for high-performance liquid chromatography (HPLC) applications. It is a compact and reliable system that provides efficient separation and analysis of complex mixtures. ChromPass ensures accurate and consistent results, making it a valuable tool for various scientific and industrial sectors.
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Lab products found in correlation
Luna phenyl hexyl column, by Phenomenex (1 mentions)
Luna pfp, by Phenomenex (1 mentions)
Luna pentafluorophenyl column, by Phenomenex (1 mentions)
Hpx 87h column, by Aminex (1 mentions)
Xylitol, by Merck Group (1 mentions)
Furfural, by Merck Group (1 mentions)
Formic acid, by Carl Roth (1 mentions)
Arabinose, by Carl Roth (1 mentions)
Xylose, by Carl Roth (1 mentions)
Acetic acid, by Carl Roth (1 mentions)
Pu 980 plus pump, by Jasco (1 mentions)
Lg 980 02 gradient unit, by Jasco (1 mentions)
Kinetex pfp column, by Phenomenex (1 mentions)
4 protocols using chrompass
1
Saccharides and Xylitol Quantification
2
HPLC Analysis of Complex Samples
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The HPLC devices (Jasco, Gross-Umstadt, Germany) consisted of the following equipment: binary high-pressure HPLC pump system (PU-2080 Plus), autosampler (AS-2055), degasser (DG-2080-53; 3-line), DAD (diode array detection) detector (MD-2010 Plus) or UV/VIS detector (UV-2075), and injection valve (Rh 7725i type Rheodyne; Bensheim, Germany). Chromatographic gradient development and peak separation was performed on an analytical 250 × 4.6 mm, 5 µm Luna® phenyl-hexyl column (Phenomenex Ltd., Aschaffenburg, Deutschland) or Luna® pentafluorophenyl column (Phenomenex Ltd., Aschaffenburg, Deutschland). Furthermore, (sub)fractionation was carried out through chromatographic separation on a preparative 250 × 21.2 mm Luna® phenyl-hexyl column with a particle size of 5 µm (Phenomenex Ltd., Aschaffenburg, Deutschland) and a semipreparative 250 × 10 mm, 5 µm Luna® PFP (Phenomenex Ltd., Aschaffenburg, Deutschland) or a semipreparative 250 × 10 mm, 5 µm Luna® phenyl-hexyl column (Phenomenex Ltd., Aschaffenburg, Germany). Each column was attached to a suitable precolumn. The analyzed data was processed by ChromPass (version 1.9; Jasco Groß-Umstadt, Germany) or Galaxie software (version 1.10; Agilent Technologies, Oberhaching, Germany).
3
Quantification of Saccharides and Inhibitors in Lignocellulosic Hydrolysates
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To analyze saccharides and inhibitory compounds, the sterilefiltered hydrolysate samples were centrifuged and the supernatant was used for the quantification by HPLC, using a Jasco HPLC 2000 plus series (Biolab, Vienna, Austria) with an Aminex HPX 87H column at 65 °C. H 2 SO 4 (c = 5 mmol l -1 ) was used as eluent at an isocratic flow rate of 0.8 ml min -1 . Sugars were detected with a refractive index and organic acids and inhibitors by a UV detector. Data were analyzed with ChromPass (Version 1.8.6.1, Jasco Europe, Italy). For the preparation of standards, chemicals were obtained from the following suppliers: xylitol from Sigma-Aldrich (Steinheim, Germany), glucose, xylose, arabinose, acetic acid and formic acid from Carl Roth (Karlsruhe, Germany), cellulose form Machery-Nagel (Düren, Germany), furfural from Merck (Hohenbrunn, Germany), and hydroxymethylfurfural (HMF) from Alfa Aesar (Kandel, Germany). Resulting saccharide yields (g) per 100 g wheat straw were calculated as following: g ½ saccharide per 100 g wheat straw ¼ g ½ saccharide per L as measured by HPLC*100 % ½ dry mass of wheat straw* g ½ wheat straw loading per L
4
Quantification of Tocopherols in Biological Samples
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Tocopherols were quantified in plasma (collected at slaughter), liver, and endometrium by a validated method described earlier [62] . Briefly, tissue samples were saponified for 30 min at 70 • C in a shaking water bath. To both tissue and plasma samples butylated hydroxytoluene (BHT; 25 μL of a 1 mg/ml ethanolic solution) was added. Then all samples were extracted twice with n-Hexane. The supernatants were pooled and evaporated (Christ SpeedDry; Christ, Osterode Germany). The dried residues were resuspended and injected into a Jasco HPLC system (AS-950 Plus autosampler, PU-980 Plus pump, FP-950 Plus fluorescence detector, LG-980-02 gradient unit, and a 3-line degasser; Jasco, Groß-Umstadt, Germany). The tocopherols were separated on a Kinetex PFP column (2.6 μm, 150 × 4.6 mm; Phenomenex, Aschaffenburg, Germany) using a methanol: water (85:15, vol/vol) mobile phase. The fluorescence detector was set to an excitation wavelength of 296 nm and emission wavelength of 325 nm. Peaks were recorded and integrated using ChromPass version 1.8.6.1 (Jasco). The concentrations of tocopherols were quantified against external standard curves with authentic compounds (Sigma Aldrich, St. Louis, MO).
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