Soluble sugars were extracted from flour samples (400 mg DW), following the method of [43 (link)]. Samples were homogenized in 20 mL of cold H2O, stirred for 30 min on ice, sonicated for 5 min and centrifuged (15,000 g, 20 min, 4 °C). The supernatant was collected, and extraction procedure was repeated with the pellet. Both supernatants were joined and cleared with nylon syringe filters (0.45 µm) before injection. Sugars separation was performed with an HPLC system coupled to a refractive index detector (Model 2414, Waters, MA, USA), using a SugarPak 1 column (300 mm length × 6.5 mm diameter, Waters) at 90 °C, with H2O as eluent (containing 50 mg EDTA-Ca L−1 H2O) and a flow rate of 0.5 mL min−1. Standard curves were used for the quantification of each sugar.
Model 2414
Manufactured by Waters Corporation
Sourced in United States
The Model 2414 is a refractive index (RI) detector designed for high-performance liquid chromatography (HPLC) applications. It provides a precise and stable measurement of the refractive index of the analytes eluting from the HPLC column. This detector can be used to detect a wide range of compounds, making it a versatile tool in various analytical workflows.
Automatically generated - may contain errors
Lab products found in correlation
Sugar pak 1 column, by Waters Corporation (5 mentions)
Hpx 87h column, by Bio-Rad (3 mentions)
Dionexcarbopac pa1, by Thermo Fisher Scientific (2 mentions)
Alliance 2795 system, by Waters Corporation (2 mentions)
X brigetm amide column, by Waters Corporation (2 mentions)
Astra 5 software, by Wyatt Technology (1 mentions)
Model e2695, by Waters Corporation (1 mentions)
Acrodisc syringe filter, by Pall Corporation (1 mentions)
Nylaflo nylon membrane filter, by Pall Corporation (1 mentions)
Model 2996, by Waters Corporation (1 mentions)
Waters alliance hplc system, by Waters Corporation (1 mentions)
Sugar pak 1, by Waters Corporation (1 mentions)
Tsk g 3000 pwxl column, by Tosoh (1 mentions)
Tsk g6000 pwxl column, by Tosoh (1 mentions)
Breeze 2, by Waters Corporation (1 mentions)
Styragel column, by Waters Corporation (1 mentions)
Model 515, by Waters Corporation (1 mentions)
G3000 pwxl, by Waters Corporation (1 mentions)
E2695, by Waters Corporation (1 mentions)
Empower tm 3 chromatography data software, by Waters Corporation (1 mentions)
22 protocols using model 2414
1
Soluble Sugars Extraction and Quantification
2
Quantification of Soluble Sugars in Leaf Samples
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Soluble sugars (sucrose, fructose, glucose, and sorbitol) were determined in approximately 400 mg of powdered frozen leaf material, based on the method previously described [63 (link)] with alterations as in [64 (link)]. Briefly, the samples were homogenized in 4 mL of cold H2O with 50 mg of polyvinylpolypyrrolidone, left to extract for 20 min on ice at 100 rpm and centrifuged (12,000× g, 5 min, 4 °C). The supernatant was boiled to denature the proteins (3 min), placed on ice (6 min) and centrifuged again. The obtained clear solution was then filtered (0.45 µm, nylon) before the injection of a 50 μL aliquot into an HPLC system equipped with a refractive index detector (Model 2414, Waters, Milford, MA, USA). The separation of sugars was performed using a SugarPak 1 column (300 × 6.5 mm, Waters) at 90 °C, with H2O as the eluent (containing 50 mg EDTA-Ca L−1 H2O) and a flow rate of 0.5 mL min−1 for 22 min. Standard curves were used for the quantification of each sugar.
3
Enzymatic Production of Xylooligosaccharides from Rice Husk
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Enzymatic hydrolysis of rice husk xylan was done by adding 1 mL of commercial xylanase solution in 9 mL of alkaline extracted xylan solution in 50 mM citric acid-Na2HPO4 buffer (pH 6.0), and incubated at 50 °C for various time interval based on BBD model. After incubation reaction was arrested by placing the reaction tube in boiling water bath for 5 min. Then, the XOS mixtures were centrifuged at 6000 rpm for 10 min and filtered through 0.2 μm syringe filter to remove the residues. The samples were quantified by HPLC equipped with refractive index detector (Model 2414, Waters Corporation) using Shodex SUGAR SH1011 column. The samples were eluted with 0.05 M sulfuric acid with flow rate of 0.8 mL/min. The concentration of sugars was determined using peak area ratio of the mixture xylooligosaccharide standards including xylobiose (X2), xylotriose (X3), xylotetraose (X4), and xylopentaose (X5). Glycerol was used as the internal standard. The xylooligosaccharide concentration of samples was expressed as mg XOS/mL xylan.
4
SEC-MALLS Analysis of Hyaluronic Acid Polymers
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The molecular weight of HA and HA-C12 was determined by SEC-MALLS on a Waters Alliance liquid chromatography (Model e2695), equipped with a refractive index detector (Model 2414, Waters, Santa Clara, CA, USA). The chromatograph is provided by a mini DAWN TREOS multiangle laser light scattering photometer (Wyatt Technology Corporation, Dernbach, Germany). The injection volume was 100 µL of HA (0.02%). The separation was carried out using two columns thermostated at 40 °C: PL aquagel-OH 60 (7.5 × 300 mm, 8 µm) and PL aquagel-OH mixed-H (7.5 × 300 mm, 8 µm). Each sample was filtered across an acrodisc syringe filter (0.22 µm, 13 mm diameter) with the support membrane (Pall corporation, New York, NY, USA). An aqueous solution of NaH2PO4 (50 mM, pH adjusted to 7.5) was filtered through a Nylaflo® (nylon) membrane filter (0.2 µm, 47 mm diameter, Pall corporation, New York, NY, USA) was used as a mobile phase. Data acquisition and molecular weight calculations were performed using the ASTRA V software (Wyatt Technology Corporation). A specific refractive index increment dn/dc = 0.155 mL/g was used for HA [18 (link),19 (link)].
5
Monosaccharide Composition Analysis of Rice Straw Prehydrolyzates
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The monosaccharide
composition of the lyophilized rice straw prehydrolyzates was determined
using a Waters Alliance HPLC system (Model 2695, Waters Corporation,
Milford, MA) equipped with a SP-G precolumn, SP0810 analytical column
(Shodex, Kawasaki, Japan), and refractive index detector (Model 2414,
Waters Corporation, Milford, MA). The xylo-oligosaccharide concentration
was determined by equipping the HPLC described above with a Bio-Rad
Aminex-HPX 42A analytical column (Bio-Rad, Hercules, CA) and a Micro-Guard
deashing precolumn. Calibration curves for the xylo-oligosaccharides
(DP 2 to DP 6) were determined using pure (>95.0%) reference compounds.
The analytical columns and the detector were maintained at 85 and
50 °C, respectively. Millipore water was used as eluent at a
flow rate of 0.2 mL min–1, and the sugars were quantified
using in-house calibration curves.
Analyses of formic acid,
acetic acid, HMF, and furfural were performed using the Waters Alliance
HPLC system fitted with a Bio-Rad Aminex HPX-87H ion exclusion analytical
column (Life Sciences Research, Hercules, CA) and photodiode array
detector (Model 2996, Waters Corporation, Milford, MA). The samples
were eluted with a 5 mM sulfuric acid at a flow rate of 0.6 mL min–1 and were detected at 280 nm.23
composition of the lyophilized rice straw prehydrolyzates was determined
using a Waters Alliance HPLC system (Model 2695, Waters Corporation,
Milford, MA) equipped with a SP-G precolumn, SP0810 analytical column
(Shodex, Kawasaki, Japan), and refractive index detector (Model 2414,
Waters Corporation, Milford, MA). The xylo-oligosaccharide concentration
was determined by equipping the HPLC described above with a Bio-Rad
Aminex-HPX 42A analytical column (Bio-Rad, Hercules, CA) and a Micro-Guard
deashing precolumn. Calibration curves for the xylo-oligosaccharides
(DP 2 to DP 6) were determined using pure (>95.0%) reference compounds.
The analytical columns and the detector were maintained at 85 and
50 °C, respectively. Millipore water was used as eluent at a
flow rate of 0.2 mL min–1, and the sugars were quantified
using in-house calibration curves.
Analyses of formic acid,
acetic acid, HMF, and furfural were performed using the Waters Alliance
HPLC system fitted with a Bio-Rad Aminex HPX-87H ion exclusion analytical
column (Life Sciences Research, Hercules, CA) and photodiode array
detector (Model 2996, Waters Corporation, Milford, MA). The samples
were eluted with a 5 mM sulfuric acid at a flow rate of 0.6 mL min–1 and were detected at 280 nm.23
6
Quantifying Soluble Sugars in Powdered Samples
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RFOs evaluation was performed in samples of 150 mg of powdered frozen material, following the HPLC method for soluble sugars described in Ramalho et al. (2013b (link)), with some modifications. To overcome the presence of non-pure peaks the separation of sugars was performed using a Sugar-Pak 1 column (300 × 6.5 mm, Waters) at 90°C, with H2O as the eluent (containing 50 mg EDTA-Ca L−1 H2O) and a flow rate of 0.5 mL min−1. Another 20 μL aliquot of each sample was injected through a DionexCarboPac PA1 analytical column (4 × 250 mm, Thermo Scientific, USA) coupled to a DionexCarboPac PA1 Guard (4 × 50 mm) at 20°C. Ultrapure water and 300 mM NaOH were used as eluents (water from 0 to 50 min; NaOH from 50 to 65 min; and water from 65 to 80 min for re-equilibration) at a 1 mL min−1 flow rate. A refractive index detector (Model 2414, Waters, USA) was used for sugar detection. Sugars were quantified using specific standard curves.
7
Determining Degraded LJP Molecular Weight
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The molecular weight of the degraded LJP was determined by high-performance gel-permeation chromatography (Model 2414, Waters Co., Milford, MA, USA) as previously described [23 (link)]. A TSK G6000 PWXL column (7.8 × 300 mm i.d., 13 μm, Tosoh Co., Ltd., Tokyo, Japan) and a TSK G3000 PWXL column (7.8 × 300 mm i.d., 7 μm, Tosoh Co., Ltd.) were connected in series. Dextran with molecular weights of 4.320, 12.6, 126, 289, and 496 kDa, respectively, were used as the standards. Both the standards and samples (30 μL each run) were eluted with 0.02 M monopotassium phosphate (0.5 mL/min) at 35 ± 0.1 °C.
8
Gel Permeation Chromatography of Lignins
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The molecular weights of lignins were characterized by a gel permeation chromatography instrument (Waters Breeze 2) equipped with a refractive index detector (Waters model 2414) and two Waters Styragel columns (models: HR 0.5 and HR 2; dimensions: 7.8 × 300 mm)22 (link). Polystyrenes with different molecular weights were used as standard compounds. Tetrahydrofuran was used as the mobile phase at a flow rate of 1.0 ml min−1. The column compartment and the refractive index detector temperature were set at 40 °C. To facilitate dissolution of lignins in tetrahydrofuran, hydrogenolysis of lignins and hot-pressed lignins was conducted before gel permeation chromatography analysis.
9
Quantitative Soluble Grain Sugar Analysis
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Soluble grain sugars were determined in approximately 400 mg of ground grain, based on the method of [46 (link)]. Briefly, the samples were homogenized in 10 mL of cold H2O, left to extract for 30 min on ice at 100 rpm, followed by 5 min in an ultrasonic bath and centrifuged (15,000× g, 20 min, 4 °C). The supernatant was collected, and the extraction procedure was repeated with the pellet. Both supernatants were combined and cleared with nylon syringe filters (0.45 µm) before injection. Sugars separation was performed with an HPLC system coupled to a refractive index detector (Model 2414, Waters, MA, USA), using a SugarPak 1 column (300 mm length × 6.5 mm diameter, Waters) at 90 °C, with H2O as eluent (containing 50 mg EDTA-Ca L−1 H2O) and a flow rate of 0.5 mL min−1. Standard curves were used for the quantification of each sugar.
10
Characterization of High-G/M and Mid-G/M Alginates
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Sodium alginate with molecular weight (Mw) of 520 kDa and G/M ratio of 64/36 was designated High-G/M alginate, and sodium alginate with Mw of 532 kDa and G/M ratio of 34/66 was designated Mid-G/M alginate. Both were purchased from Qingdao Mingyue Seaweed Group Co., Ltd. (Qingdao, China). The detailed information of both purchased alginates was characterized by GPC and 1HNMR spectroscopy (Table 1 ).
The G/M ratio of the respective alginates was determined at 80°C by 1HNMR spectroscopy using an AVANCE NB-360 spectrometer (Bruker, Germany). The Mw and molecular weight distribution (Mw/Mn) of alginate samples were analyzed via gel permeation chromatography (GPC) using a system equipped with two TSK-gel size-exclusion columns (G4000PWXL and G3000PWXL), a refractive index detector (Waters, model 2414, Milford, MA), and an HPLC pump (Waters, model 515).
Collagen, acquired from rat tails, and was obtained by our laboratory. Anhydrous calcium chloride (CaCl2) was purchased from Sigma-Aldrich (Merck Life Science (Shanghai) Co., Ltd.). All other chemical reagents were analytical grade and used as received.
The G/M ratio of the respective alginates was determined at 80°C by 1HNMR spectroscopy using an AVANCE NB-360 spectrometer (Bruker, Germany). The Mw and molecular weight distribution (Mw/Mn) of alginate samples were analyzed via gel permeation chromatography (GPC) using a system equipped with two TSK-gel size-exclusion columns (G4000PWXL and G3000PWXL), a refractive index detector (Waters, model 2414, Milford, MA), and an HPLC pump (Waters, model 515).
Collagen, acquired from rat tails, and was obtained by our laboratory. Anhydrous calcium chloride (CaCl2) was purchased from Sigma-Aldrich (Merck Life Science (Shanghai) Co., Ltd.). All other chemical reagents were analytical grade and used as received.
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