Insoluble dietary fiber (IDF), soluble dietary fiber (SDF), and low molecular weight soluble dietary fiber (LMSDF) were determined using the enzymatic gravimetric/chromatographic method according to McCleary et al. [30 (link)]. LMSDF was estimated by using high performance liquid chromatography in combination with a refractive index detector (HPLC-RI, Hitachi, Düsseldorf, Germany). Parameters for HPLC-RI analysis were as follows: column, TSKgel G2500PWxl (3000 mm × 7.8 mm, Tosoh, Grießheim, Germany), 2× in series; eluent, double distilled water with a constant flow rate of 0.4 mL/min; oven temperature, 80 °C. Total dietary fiber (TDF) was calculated from IDF, SDF, and LMSDF. To structurally characterize IDF and SDF, a preparative isolation procedure of dietary fiber was performed (resulting in preparative IDF/SDF) using, among others, a thermostable α-amylase instead of a pancreatic α-amylase as detailed in [31 (link)]. Because analyses of IDF, SDF, and LMSDF were carried out in duplicate, range/2 was reported as a suitable measure to indicate statistical dispersion.
Hplc ri
Manufactured by Hitachi
Sourced in Germany
The HPLC-RI is a high-performance liquid chromatography system with a refractive index detector. It is a laboratory instrument used for the separation, identification, and quantification of various compounds in a liquid mixture.
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Lab products found in correlation
4 protocols using hplc ri
1
Comprehensive Dietary Fiber Analysis
2
Separation of Oligo-/Polysaccharides by HPLC-RI
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SDF was dissolved in 50 mM sodium nitrate (2 g•L -1 ) and treated for 24 h at 40 °C. The supernatant was used for the separation of oligo-/polysaccharides based on their hydrodynamic volumes using HPLC-RI (Hitachi, Merck, Darmstadt, Germany) equipped with a guard column (TosohTSK-gel PWxl 40 mm × 6.0 mm, particle size 12 μm, Tosoh, Tokyo, Japan) and two size-exclusion columns in series (TosohTSKgel G6000PWxl 300 mm × 7.8 mm, particle size 13 μm; TSKgel G4000PWxl 300 mm × 7.8 mm, particle size 10 μm, Tosoh, Tokyo, Japan). Sodium nitrate was used as eluent at a flow rate of 0.5 mL•min -1 at 50 ℃. Dextrans were used for calibration [13] (link).
3
Molecular Weight Distribution of SDF Polymers
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The molecular weight distribution of SDF polymers was determined by dissolving SDF in 50 mM sodium nitrate for 24 h at 40 °C, followed by centrifugation and separation of the supernatant according to hydrodynamic volume using two size exclusion columns in series (TosohTSK-gel G6000PWxl 300 mm × 7.8 mm, particle size 13 µM; TSKgel G4000PWxl 300 mm × 7.8 mm, particle size 10 µM) and a guard column (TosohTSK-gel PWxl 40 mm × 6.0 mm, particle size 12 µM). Sodium nitrate was used as eluent (flow rate: 0.5 mL min -1 , 50 °C), detection was carried out by HPLC-RI (Hitachi, Merck, Darmstadt, Germany) [17] (link).
4
Molecular Weight Distribution of SDF Polymers
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The molecular weight distribution of SDF polymers was determined by dissolving SDF in 50 mM sodium nitrate for 24 h at 40 °C, followed by centrifugation and separation of the supernatant according to hydrodynamic volume using two size exclusion columns in series (TosohTSK-gel G6000PWxl 300 mm × 7.8 mm, particle size 13 µM; TSKgel G4000PWxl 300 mm × 7.8 mm, particle size 10 µM) and a guard column (TosohTSK-gel PWxl 40 mm × 6.0 mm, particle size 12 µM). Sodium nitrate was used as eluent (flow rate: 0.5 mL min -1 , 50 °C), detection was carried out by HPLC-RI (Hitachi, Merck, Darmstadt, Germany) [17] (link).
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