Free sugars were determined with the implementation of a high-performance liquid chromatography (HPLC) system coupled to a refractive index detector and by applying the internal standard method (IS, melezitose; Sigma, St. Louis, MO, USA) [26 (link)]. Data were obtained and evaluated with Clarity 2.4 software (Informer Technologies, Inc., Solihull, UK).
Melezitose
Manufactured by Merck Group
Sourced in United States
Melezitose is a trisaccharide found in nature. It is used as a laboratory reagent in various analytical and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Sucrose, by Merck Group (6 mentions)
Eurospher 100 5 nh2 column, by Knauer (5 mentions)
Hplc grade acetonitrile, by RCI Labscan (4 mentions)
Smartline system 1000, by Merck Group (4 mentions)
Smartline 2300, by Knauer (4 mentions)
Glucose, by Merck Group (4 mentions)
Fructose, by Merck Group (4 mentions)
Trehalose, by Merck Group (3 mentions)
Maltose, by Merck Group (3 mentions)
Turanose, by Merck Group (2 mentions)
Erlose, by Merck Group (2 mentions)
Methanol, by Merck Group (2 mentions)
Standard 47885, by Merck Group (2 mentions)
Smartline 1000, by Knauer (2 mentions)
Clarity 2, by Informer Technologies (1 mentions)
Smartline manager 5000, by Jasco (1 mentions)
As 2057, by Jasco (1 mentions)
Chromatographic system, by Shimadzu (1 mentions)
Melibiose, by Merck Group (1 mentions)
Xylose, by Merck Group (1 mentions)
21 protocols using melezitose
1
Quantitative Analysis of Free Sugars
2
HPLC Analysis of Soluble Sugars
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Soluble sugars were assessed via HPLC coupled to an RI detector (Knauer, Smartline system 1000, Berlin, Germany) applying the internal standard (IS, melezitose, Sigma-Aldrich, St. Louis, MO, USA) method, as formerly described by the authors [36 (link)]. Mobile phase was composed by an acetonitrile: water mixture (70:30 v/v, acetonitrile HPLC-grade, Lab-Scan, Lisbon, Portugal), whereas separation was completed using a Eurospher 100-5 NH2 column (4.6 × 250 mm, 5 µm, Knauer). The results were recorded and treated using Clarity 2.4 software (DataApex, Prague, Czech Republic).
3
Quantifying Mesquite Flour Sugars by HPLC
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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.
4
Honey Sugars Profiling by HPLC
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Sugar profiles of 11 honey samples were analyzed by HPLC using the Shimadzu chromatographic system (Kyoto, Japan) with the RID-10A refractive index detector. The mobile phase (Milli-Q water obtained using the Elix® Essential 3 Water Purification System with Synergy® UV Water Purification System, Merck Millipore, Darmstadt, Germany) was run at a flow rate of 0.6 mL/min at 75 °C through the REZEX RPM-Monosaccharide Pb2+ column (300 × 7.8 mm, Phenomenex, Torrence, USA). The column was calibrated using sixteen carbohydrate standards, including mono-, di- and trisaccharides. Standard solutions of mono-, di- and trisaccharides: glucose, fructose, galactose, rhamnose, xylose, mannose, sucrose, turanose, maltose, celobiose, fucose, trehalose, melibiose, erlose, melezitose and raffinose (Sigma-Aldrich, Saint Louis, MO, USA) were used for interpretation and quantification of sugars in the honey samples. Sugar concentrations were expressed in g/100 g honey.
5
Characterization of Carbohydrate Standards
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The following compounds were purchased from Sigma-Aldrich: D-glucose ≥ 99.5%, D (−) fructose ≥ 99%, D (+) Turanos ≥ 98%, erlose ≥ 94%, isomaltotriose, D (+) melibiose ≥ 99,0%, D (+), raffinose pentahydrated ≥ 98.0%, palatinose hydrate ≥ 99%, sucrose ≥ 99.5%, D (+) maltose monohydrate min 98%, melezitose ≥ 99.0%, trehalose dihydrate (Certified Reference Materials), D-panose ≥ 88 97%, maltotriose, isomaltose 98%,ascorbic acid, gallic acid, 2,2-diphenyl-1 picrylhydrazyl (DPPH), Folin–Ciocalteu’s reagent, hexamethyldisilazane ≥ 99%, trifluoroacetic acid, hydroxylamine hydrochloride, acetic anhydride, pyridine, potassium hexacyanoferrate (II) trihydrate, and zinc acetate dihydrate. Methanol, sulfuric acid, ethanol, and acetonitrile were purchased from Merck (Darmstadt, Germany). Aluminum chloride, sodium carbonate, sodium hydroxide, and sodium nitrite were purchased from Merck (Darmstadt, Germany). All chemicals used were of analytical grade.
6
Analytical Methods for Honey Characterization
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All chemical standards were HPLC-grade pure. Folin-Ciocalteu reagent, gallic acid, aluminum chloride, sodium carbonate, potassium iodide, and bisulfite were purchased from Panreac (Barcelona, Spain). Quercetin and 2,2-diphenyl-1-picrilhidrazil (DPPH) were purchased from Alfa Aesar (Massachusetts, USA) and methanol was obtained from Merck (Darmstandt, Germany). Hydrolyzed starch for diastase determination was purchased from Carlo Erba (Barcelona, Spain). The calibration of the HANNA Honey Color C221 colorimeter was with glycerin was provided by Glycerol HANNA instruments (Woonsocket, Rhode Island, USA). Standards of glucose, fructose, sucrose, maltose, trehalose, turanose and melezitose were obtained from Sigma–Aldrich (Madrid, Spain).
7
Synthesis of Cholesterol-Based Lipid Conjugates
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Cholesteryl (3-((2-hydroxyethyl)amino)propyl)carbamate hydroiodide (HAPC-Chol) was synthesized as described previously (13 (link)). N-(2-(2-Hydroxyethylamino)ethyl)cholesteryl- 3-carboxamide (OH-Chol) and cholesteryl (2-((2-hydroxyethyl)amino)ethyl)carbamate (OH-C-Chol) were synthesized as described previously (14 (link)). 1,2-Dioleoyl- 3- trimethylammonium-propane methyl sulfate salt (DOTAP) was obtained from Avanti Polar Lipids Inc. Dimethyldioctadecylammonium bromide (DDAB, product name: DC-1-18) and 11- ((1,3- bis(dodecanoyloxy)- 2- ((dodecanoyloxy)methyl)propan- 2- yl) amino)- N,N,N- trimethyl-11- oxoundecan-1- aminium bromide (product name: TC-1-12) were obtained from Sogo Pharmaceutical Co., Ltd.. 1,2-Dioleoyl- sn- glycero- 3-phosphoethanolamine (DOPE, COATSOME ME-8181) was obtained from NOF Co., Ltd.. Glucose, fructose, galactose, mannose, sucrose, trehalose dihydrate, maltose monohydrate, lactose monohydrate, lactulose, cellobiose, and raffinose pentahydrate were obtained from Wako Pure Chemical Industries, Ltd.. Melezitose was purchased form Sigma-Aldrich Co. LLC. All other chemicals were of the highest grade available.
8
Donut Composition Determination
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The chemical composition of the donut samples was determined by assaying the sugars and fatty acids according to methodologies previously described by Barros et al. [39 (link)], and analyzed through chromatographic systems, namely, HPLC-RI and GC-FID, respectively. The compounds were characterized by comparison with available standards (standard 47885, Sigma-Aldrich, St. Louis, MO, USA). The content in sugars was expressed in g/100 g of fresh weight, and melezitose (Sigma Chemical Co.; Saint Louis, MO, USA) was used as internal standard in sugars evaluation. The fatty acids concentration was expressed as relative percentages (%) of each fatty acid.
9
Quantification of Phytochemicals in Samples
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HPLC-grade acetonitrile was obtained from Merck KgaA (Darmstadt, Germany). Standards of melezitose, fatty acids methyl ester (FAME, reference standard mixture 3747885-U), Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), and 2,2′-azobis(2-methylpropionamidine) dihydrochloride (AAPH) were purchased from Sigma Chemical Co. (St. Louis, MO, USA); tocol standard (50 mg/mL) was acquired from Matreya, Pleasant Gap (PA, USA), and the phenolic compound standards were acquired from Extrasynthese (Genay, France). Ethanol and all other chemicals and solvents were of analytical grade and purchased from common sources. Water was treated in a Milli-Q water purification system (TGI Pure Water Systems, Greenville, SC, USA)
10
Synthesis and Characterization of Fructo-oligosaccharides
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Glucose, fructose, and sucrose were from Merck (Darmstadt, Germany). Melezitose, kanamycin sulphate, isopropyl-β-D-1-thiogalactopyranoside (IPTG), 3,5-dinitrosalicylic acid (DNS) and beef extract were from Sigma-Aldrich (Saint Lois, MO, USA). Polysorbate 80 was from Santa Cruz Biotechnology (Dallas, TX, USA) and 3-(N-morpholino)-propanesulfonic acid (MOPS) from NzyTech (Lisbon, Portugal). Sodium alginate Protanal LF 120 LS was from FMC Biopolymer (Grivan, Ayrshire, UK). Beghin Meiji SA (Neuilly-sur-Seine, France) kindly supplied the commercial fructo-oligosaccharides mixture “Actilight”. Yeast extract, peptone and tryptone were from Laboratorios Conda S.A. (Madrid, Spain).
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