The quantification of alcohols and sugars was carried out by HPLC (Thermo Fisher Scientific) using a refraction index detector equipped with a HyperREZ™ XP Carbohydrate H+ 8 column (Thermo Fisher Scientific) coupled with a HyperRETZ™ XP Carbohydrate Guard (Thermo Fisher Scientific). The chromatography conditions were the same as described by Minebois et al. (2020b (link)). Briefly, 1 ml of samples was diluted with Milli‐Q water depending on the remaining sugar amount and filtered through a 0.22‐μm nylon filter. The run conditions were 1.5 mM of H2SO4 at 0.6 ml/min flux with 35 bars of pressure, and the oven temperature was maintained at 50°C. The concentration of the different compounds was determined through their corresponding standard calibration curve.
Hyperrez xp carbohydrate h 8 column
Manufactured by Thermo Fisher Scientific
The HyperREZ™ XP Carbohydrate H+ 8 column is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of carbohydrates. It features an 8-column configuration and is optimized for the separation of monosaccharides, disaccharides, and oligosaccharides.
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2 protocols using hyperrez xp carbohydrate h 8 column
1
HPLC Quantification of Alcohols and Sugars
2
Yeast Fermentation Analysis Techniques
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Fermentations were carried out in an incubator from Selecta (Barcelona, Spain) using Saccaromyces cerevisiae cells from Lallemand Inc (Montreal, Quebec, Canada).
For density measurements, a Densito 30 PX densitometer from Mettler-Toledo GmbH (Greifensee, Switzerland) was employed. Musts and wine composition was analysed with a Surveyor Plus HPLC chromatography system from Thermo Fisher Scientific (Waltham, MA, USA) equipped with refraction index and UV-vis detectors. A Hyper REZ XP carbohydrate H+8 column, also from Thermo Fisher Scientific, was employed as stationary phase at 50 °C. Samples were filtered with 0.2 µm nylon filter devices.
A series of buffers and solutions were employed. Coating buffer: 50 mM carbonate-bicarbonate buffer, pH 9.6; PBS: 10 mM phosphate buffer, pH 7.4, with 140 mM NaCl; PBST: PBS containing 0.05% (v/v) Tween 20; PB: 100 mM sodium phosphate buffer, pH 7.4; washing solution: 150 mM NaCl containing 0.05% (v/v) Tween 20; enzyme substrate buffer: 25 mM citrate and 62 mM sodium phosphate buffer, pH 5.4.
For density measurements, a Densito 30 PX densitometer from Mettler-Toledo GmbH (Greifensee, Switzerland) was employed. Musts and wine composition was analysed with a Surveyor Plus HPLC chromatography system from Thermo Fisher Scientific (Waltham, MA, USA) equipped with refraction index and UV-vis detectors. A Hyper REZ XP carbohydrate H+8 column, also from Thermo Fisher Scientific, was employed as stationary phase at 50 °C. Samples were filtered with 0.2 µm nylon filter devices.
A series of buffers and solutions were employed. Coating buffer: 50 mM carbonate-bicarbonate buffer, pH 9.6; PBS: 10 mM phosphate buffer, pH 7.4, with 140 mM NaCl; PBST: PBS containing 0.05% (v/v) Tween 20; PB: 100 mM sodium phosphate buffer, pH 7.4; washing solution: 150 mM NaCl containing 0.05% (v/v) Tween 20; enzyme substrate buffer: 25 mM citrate and 62 mM sodium phosphate buffer, pH 5.4.
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