HPLC system coupled to a refraction index detector (RI detector Knauer Smartline 2300) was used to determine the composition of mesquite flour sugar, as previously described in Barros et al. [15 (link)]. Sugars were identified by comparing the relative retention times. The equipment consisted of an integrated system with a pump (Knauer, Smartline system 1000, Berlin, Germany), degasser system (Smartline manager 5000), autosampler (AS-2057 Jasco), and an RI detector (Knauer Smartline 2300). Data were analyzed using Clarity 2.4 Software (DataApex, Prague, Czech Republic). The chromatographic separation was achieved with a Eurospher 100-5 NH2 column (5 µm, 250 mm × 4.6 mm i.d., Knauer) operating at 35 °C (7971 R Grace oven). The mobile phase was acetonitrile/deionized water, 70:30 (v/v) at a flow rate of 1 mL/min. Quantification was achieved using the internal standard (IS, melezitose, Sigma-Aldrich, St. Louis, MO, USA) method and by calibration curves from commercial standards. Analyses were carried out in triplicate, and results were expressed in g per 100 g of dry weight.
Smartline 2300
Manufactured by Knauer
Sourced in Germany, United States, Czechia
The Smartline 2300 is a high-performance liquid chromatography (HPLC) system designed for a wide range of analytical applications. It features a modular design, allowing for customization to meet specific user requirements. The system includes a solvent delivery unit, auto-sampler, column oven, and a variety of detection options. The Smartline 2300 is capable of performing precise and reliable HPLC analyses.
Automatically generated - may contain errors
Lab products found in correlation
Eurospher 100 5 nh2 column, by Knauer (12 mentions)
As 2057, by Jasco (9 mentions)
Smartline system 1000, by Knauer (7 mentions)
Smartline manager 5000, by Jasco (6 mentions)
Smartline 1000, by Knauer (5 mentions)
Melezitose, by Merck Group (4 mentions)
20a series, by Shimadzu (2 mentions)
D trehalose, by Merck Group (2 mentions)
D sucrose, by Merck Group (2 mentions)
D fructose, by Merck Group (2 mentions)
D glucose, by Merck Group (2 mentions)
Melezitose as is, by Merck Group (1 mentions)
Hplc grade acetonitrile, by RCI Labscan (1 mentions)
D xylose, by Merck Group (1 mentions)
D raffinose pentahydrate, by Merck Group (1 mentions)
28 protocols using smartline 2300
1
Quantifying Mesquite Flour Sugars by HPLC
2
HPLC Analysis of Free Sugars
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The content in free sugars was determined by High-Performance Liquid Chromatography (HPLC, Knauer Smartline 2300, Knauer, Berlin, Germany), coupled to a refractive index detector (RI detector, Knauer Smartline 2300, Knauer, Berlin, Germany), according to the procedure previously described by Dias et al. [37] (link). The identification and quantification of free sugars were performed using the Clarity 2.
3
Quantifying Free Sugars in Cardoon Seeds
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To the determination of the free sugars of cardoon seeds, the samples were extracted with a aqueous ethanol (EtOH:H 2 O, 80:20, v/v, 40 mL) and the internal standard melezitose (IS, 5 mg/mL) at 80 • C for 90 min. The solution was centrifuged at 15,000 g for 10 min and the supernatant was concentrated under reduced pressure (rotary evaporator Büchi R-210, Flawil, Switzerland). The obtained solution was defatted with ethyl ether (10 mL, three times) and the solid residues were dissolved in water to a final volume of 5 mL. The free sugars content was analysed by High-Performance Liquid Chromatography (HPLC, Knauer Smartline 2300, Knauer, Berlin, Germany), coupled to a refractive index detector (RI detector, Knauer Smartline 2300, Knauer, Berlin, Germany) following the procedure previously described (Dias et al., 2015) (link). The identification was performed through the comparison of the retention times with the commercial standards (D-(-)-fructose, D-(+)-sucrose, D-(+)-glucose, D-(+)-trehalose, and D-(+)-raffinose pentahydrate purchased from Sigma-Aldrich (St. Louis, MO, USA). For the quantification, the areas of the peaks were compared with the calibration curves of the commercial standards. The results were analysed using the Clarity 2.4 software (DataApex, Prague, Czech Republic) and were expressed in g per 100 g of dry weight.
4
Molecular Weight Determination by SEC
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The average molecular weight (Mw), molecular number (Mn), and polydispersity index (PDI = Mw/Mn) were determined by size exclusion chromatography (SEC). The samples were dissolved in deionized water (2–4 mg/mL) and injected in a size exclusion-high-performance liquid chromatography (SE-HPLC) system (Knauer, Smartline system 1000, Berlin, Germany), equipped with a Phenomenex Phenogel Linear LC Column (300 mm × 7.8 mm, Knauer) coupled to a refractive index detector (RI detector, Knauer Smartline 2300). The analysis was performed at 25 °C with LiNO3 0.1 M as the eluent, at a flow rate of 0.6 mL/min and an injection volume of 50 µL. The Mw was estimated after a universal calibration with the pullulans (≤600 kDa).
5
Quantification of Free Sugars by HPLC
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Free sugars were determined by HPLC coupled to a refraction index (RI) detector as described previously [8 ]. The equipment consisted of a pump (Knauer, Smartline System 1000, Berlin, Germany), a degasser (Smartline Manager 5000), an autosampler (AS-2057 Jasco, Easton, MD, USA), and a RI detector (Knauer Smartline 2300). The identification was achieved by comparing the relative retention times of sample peaks with standards. Quantification was made by the internal standard method, and the results are expressed in mg per g of lyophilized decoction or infusion.
6
HPLC Analysis of Sugars in Lyophilized Fruit
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These sugars were extracted from the lyophilized fruit and were further injected in HPLC equipment (Knauer, Smartline system 1000, Berlin, Germany) coupled to a refractive index detector (RI detector, Knauer Smartline 2300) as described by Barros et al. [8 (link)]. Separation was achieved with a Eurospher 100-5 NH2 column (250 mm × 4.6 mm, 5 µm, Knauer), with an isocratic elution using acetonitrile/deionized water (70:30, v/v) at a flow rate of 1 mL/min, operating at 35 °C. The internal standard method was applied, and the results were expressed in g per 100 g of fresh fruit.
7
Quantification of Free Sugars in Dry Samples
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The extraction of free sugars from the dry samples was carried out according to Barros et al. [68 (link)] The compounds were identified by high performance liquid chromatography with a refraction index detector (HPLC-RI; Knauer, Smartline 1000 and Smartline 2300 systems, respectively) operating as previously described by the authors. Peaks identification was performed by comparisons of their relative retention time (Rt) with authentic standards. Quantification was completed using melezitose as IS, (Sigma-Aldrich, St. Louis, MO, USA). Results were processed in a Clarity Software (Data Apex, Prague, Czech Republic) and expressed in g per 100 g dw.
8
Quantifying Soluble Sugars in Samples
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Soluble sugars were determined using a High-Performance Liquid Chromatography (HPLC) system coupled to a refraction index (RI detector Knauer Smartline 2300, Berlin, Germany) detector. Chromatographic separation was achieved with a Eurospher100-5 NH2 column (5 μm, 4.6 × 250 mm, Knauer) as comprehensively described by Reis, Barros, Martins, Vasconcelos, Morales and Ferreira [20 (link)]. The samples (1 g) were extracted in a hydroethanolic (80:20 v/v) solution during 30 min at 80 °C. Melezitose was added as an internal standard (IS, Matreya, State College, PA, USA). The extracts were centrifuged (15,000× g, 10 min) and the supernatants were collected and concentrated under reduced pressure in a rotatory evaporator. Finally, the samples were defatted in ethyl ether (three times) and filtered using 0.2 µm Whatman’s nylon filters for HPLC analysis. The quantification was based on the RI signal response of each standard, depending on the IS method, and using calibration curves from commercial standards of each compound. The results were expressed in g 100 g−¹ of DW.
9
Quantification of Free Sugars by HPLC-RI
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Free sugars were determined by high performance liquid chromatography coupled to a refraction index detector (HPLC-RI), after an extraction procedure previously described by the authors [28 (link)] using melezitoze as internal standard (IS). The equipment consisted of an integrated system with a pump (Knauer, Smartline system 1000, Berlin, Germany), degasser system (Smartline manager 5000), auto-sampler (AS-2057 Jasco, Easton, MD, USA), and an RI detector (Knauer Smartline 2300). Data were analyzed using Clarity 2.4 Software (DataApex, Podohradska, Czech Republic). The chromatographic separation was achieved with a Eurospher 100-5 NH2 column (4.6 × 250 mm, 5 µm, Knauer), operating at 30 °C (7971 R Grace oven). The mobile phase was acetonitrile/deionized water, 70:30 (v/v) at a flow rate of 1 mL/min. The compounds were identified by chromatographic comparisons with authentic standards. Quantification was performed using the internal standard method and sugar contents were further expressed in g per 100 g of dry weight (dw).
10
Quantification of Free Sugars in Samples
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Free sugars were determined by high performance liquid chromatography coupled to a refraction index detector (HPLC-RI), after an extraction procedure previously described [14 (link)]. Analysis was performed by HPLC (equipment described above) using an RI detector (Knauer Smartline 2300, Berlin, Germany). Data were analyzed using Clarity 2.4 Software (DataApex). The chromatographic separation was achieved with a Eurospher 100-5 NH2 column (4.6 × 250 mm, 5 mm, Knauer, Berlin, Germany) operating at 30°C. The mobile phase was acetonitrile/deionized water, 70 : 30 (v/v) at a flow rate of 1 mL/min. The compounds were identified by chromatographic comparisons with authentic standards. Quantification was performed using the internal standard method; melezitose was used as IS. The results were expressed in mg per g of dry weight.
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