Quantitative Monosaccharide Analysis in Leaf Extracts

Free monosaccharides were extracted according to the method presented by Bell et al. (2017a) (link), with the exception that 0.2 g of lyophilized leaf powder was extracted. Extracts were analyzed on an Agilent 1100 series HPLC system equipped with a binary pump, degasser, and auto-sampler, with an external column heater (50°C). A Bio-Rad Aminex HPX-87H (300 × 7.8 mm, 9 μm particle size) column with a Micro-Guard Cation H guard column (Bio-Rad, Watford, United Kingdom) was used to achieve separation with an isocratic gradient of 5 mM sulfuric acid, and a flow rate of 0.6 mL per min. A Polymer Laboratories ERC-7515 refractive index detector (Church Stretton, United Kingdom) was used to detect monosaccharides. Compounds were quantified using authentic standards and analyzed with Agilent ChemStation software (Santa Clara, CA, United States).

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Bell L., Chadwick M., Puranik M., Tudor R., Methven L., Kennedy S, & Wagstaff C. (2020). The Eruca sativa Genome and Transcriptome: A Targeted Analysis of Sulfur Metabolism and Glucosinolate Biosynthesis Pre and Postharvest. Frontiers in Plant Science, 11, 525102.

Publication 2020

1100 series HPLC system Micro-Guard Cation H guard column

Hplc Leaf Micro guard Monosaccharides Polymer Powder Sulfuric acid

Corresponding Organization : University of Reading

Other organizations : Elsoms (United Kingdom)

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Variable analysis

independent variables

Amount of lyophilized leaf powder extracted (0.2 g)

dependent variables

Concentrations of free monosaccharides

control variables

Method of free monosaccharide extraction (Bell et al., 2017a)
HPLC system (Agilent 1100 series)
HPLC column (Bio-Rad Aminex HPX-87H, 300 × 7.8 mm, 9 μm particle size)
Guard column (Micro-Guard Cation H)
Isocratic gradient (5 mM sulfuric acid)
Flow rate (0.6 mL per min)
Column temperature (50°C)
Detector (Polymer Laboratories ERC-7515 refractive index detector)
Quantification method (using authentic standards)

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