Detection and quantification of the respective glyco-coat of 6-CD, 3-CD and Lac-CD was performed using anthrone reagent as described.51 (link) In brief a stock solution of ACS (97%) anthrone (Sigma-Aldrich) was prepared by dissolving 20 mg of anthrone into 10 mL concentrated ACS (98%) sulphuric acid (Sigma-Aldrich) and protected from light. Lactose disaccharides were used as a standard to quantitate the carbohydrate concentration of each CD. In glass vials, 400 μL of anthrone reagent was added to 200 μL of each standard or test compound, heated in a boiling water bath for 15 min and cooled in an ice bath. 200 μL of each sample was aliquoted into a 96 well plate and absorbance measured at 622 nm using an Infinite M200 Pro Multimodal Plate reader (Tecan). A linear calibration curve (y = 0.001x + 0.0386, R2 = 0.9969) was obtained by plotting absorbance against concentration of lactose on Microsoft EXCEL. The extent of the CD glyco-coat functionalization was determined using the above calibration curve. The molar concentration was calculated based on the respective concentrations of glycans in each carbon dot sample and subsequently used to compare monovalent glycan concentrations to multivalent glyco-coated carbon dot concentrations.
Anthrone
Manufactured by Merck Group
Sourced in United States, Germany, China, India
Anthrone is a chemical compound primarily used in analytical chemistry and biochemistry. It is a cyclic hydrocarbon with the chemical formula C₁₄H₁₀O. Anthrone is commonly used as a reagent for the detection and quantification of carbohydrates, particularly in colorimetric assays.
Automatically generated - may contain errors
Lab products found in correlation
Gallic acid, by Merck Group (7 mentions)
Sodium bicarbonate, by Thermo Fisher Scientific (6 mentions)
Bovine serum albumin (bsa), by Merck Group (5 mentions)
Ferric chloride, by Sinopharm (4 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (4 mentions)
Multiskan fc, by Thermo Fisher Scientific (4 mentions)
Chloroform, by Merck Group (4 mentions)
Methanol, by Tedia (3 mentions)
Acetonitrile, by Tedia (3 mentions)
Glucose 6 phosphate (g6p), by Merck Group (2 mentions)
Anthrone reagent, by Merck Group (2 mentions)
Orcinol, by Merck Group (2 mentions)
D xylose, by Merck Group (2 mentions)
D glucose, by Merck Group (2 mentions)
Vanillic acid, by Merck Group (2 mentions)
Syringic acid, by Merck Group (2 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (2 mentions)
Avicel cellulose, by Merck Group (2 mentions)
Sodium carbonate, by Merck Group (2 mentions)
Formic acid, by Merck Group (2 mentions)
52 protocols using anthrone
1
Quantification of Carbon Dot Glyco-Coats
2
Quantifying Polysaccharide Content via Anthrone and VaxArray
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Anthrone reagent was prepared fresh by dissolving 1 g Anthrone (Sigma-Aldrich, St. Louis, MO, USA) in 500 mL concentrated sulfuric acid. A 12-point dilution series of the PS of interest was prepared, and the assay was performed in 96-well plate format [30 (link)]. Briefly, a 100 µL sample was mixed with 200 µL Anthrone reagent, with each dilution analyzed in triplicate. The plate was incubated at 95 °C for 20 min and scanned at 630 nm on a FLUOstar OPTIMA microplate reader (BMG LABTECH, Ortenberg, Germany). Because VaxArray has improved sensitivity over the Anthrone assay, serial dilutions prepared for Anthrone assay analysis were diluted an additional 100× or 200× prior to VaxArray analysis. The samples were assessed by VaxArray in triplicate, as described above, using individual serotype-specific Primary Detection Labels. Correlations were generated by plotting the Anthrone assay blank-corrected against the VaxArray signal response (median RFU minus background) for each sample.
3
Quantification of Total Soluble Sugars
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Total soluble sugar was measured spectrophotometrically using the method of Ibrahim and Jaafar [50 (link)]. Samples (0.5 g) were placed in 15 mL conical tubes, and distilled water was added to make up the volume to 10 mL. The mixture was then vortexed and later incubated for 10 min. Anthrone reagent was prepared by dissolving Anthrone (Sigma Aldrich, St Louis, MO, USA, 0.1 g) in 95% sulphuric acid (Fisher Scientific, USA, 50 mL). Sucrose was used as a standard stock solution to prepare the standard curve for the quantification of sucrose in the sample. The mixed sample of ground dry sample and distilled water was centrifuged at a speed of 3400 rpm for 10 min and then filtered to get the supernatant. A sample (4 mL) was mixed with Anthrone reagent (8 mL) and then placed in a water bath set at 100°C for 5 min before the sample was measured at an absorbance of 620 nm using a spectrophotometer (model UV160U; Shimadzu Scientific, Kyoto, Japan). The total soluble sugar in the sample was expressed as mg/g sucrose dry sample.
4
Detecting Sand Fly Plant Feeding
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Each sand fly abdomen containing the crop was macerated in 50 μL absolute ethanol using a sterilized pestle. An aliquot (25 μL) of the sand fly homogenate from each individual was placed in the wells of a flat-bottomed 96-well microplate, and tested for presence of fructose as evidence of recent plant feeding by the cold anthrone test [21 ] with modifications by Matheson et al [22 (link)]. Briefly, 200 μL anthrone solution (0.15% anthrone (Sigma -Aldrich) w/v in 71.7% sulphuric acid) was added to each well containing the homogenate and incubated for 60 min at room temperature (25°C). A change in color from yellow to green or blue was deemed indicative of a positive test [21 ].
5
Colorimetric Trehalose Quantification
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Trehalose extracts were subjected to the reaction with anthrone (Sigma, St. Louis, Missouri, USA) and colorimetric measurement at 630 nm, according to the anthrone method (Brin 1966; Ferreira et al. 1997 ). Trehalose served as external standard for quantification. Each treatment was performed in triplicate.
6
Purification and Characterization of Organic Compounds
7
Streptozotocin-Induced Diabetes Assays
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NA, STZ [2-deoxy-2-(3-methyl-3nitrosoureido)-D-glycopyranoside], glucose-6-phosphate, glucose-1-phosphate, anthrone, reduced glutathione (GSH), malondialdehyde (MDA), and 1-Chloro-2,4-dinitrobenzene were purchased from Sigma-Aldrich Chemical Co., St Louis, MO, USA. All other chemicals were of analytical grade and were obtained from standard commercial supplies.
8
Spectrophotometric Quantification of Rhizome Sugars
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Sugars were assayed spectrophotometrically with the anthrone method [84 (link)]. Rhizome samples (n = 3) with FW of 250 mg were homogenized in a mortar in 10 mL of distilled water, then heated for 15 min in a water bath at 90 °C and centrifuged for 10 min at 5000 rpm (5430R; Eppendorf, Hamburg, Germany). anthrone reagent (2 mL; 1 mg of anthrone; Sigma-Aldrich, Darmstadt, Germany) per 100 mL of concentrated H2SO4, analytical grade, (ChemPur, Piekary Śląskie, Poland) was added to a cooled and 10-fold-diluted sample. The absorbance of the complex compound was measured at the wavelength of 620 nm (Ultrospec 2100; Amersham, UK). The results were referenced to the calibration curve obtained for glucose (Sigma-Aldrich) at the concentration 0.0039–0.0312 mg per 1 mL. The control was a sample of 1 mL distilled water.
9
Quantitative Analysis of Hexoses and Pentoses
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A UV—vis opectrophotometer (756 PC, Shanghai Hongji Instruments Co., Ltd., Shanghai, China) was used for absorbance reading values. The Anthrone/H2SO4 method [48 ] was used to determine hexoses and the orcinol/HCl method [49 ] was used to determine pentoses. The details of the determination method were carried out according to Xu et al [50 (link)]. Anthrone and orcinol were purchased from Sigma-Aldrich Co., LLC and ferric chloride was obtained from Sinopharm Chemical Reagent Co., Ltd. D-glucose and D-xylose purchased from Sigma-Aldrich Co., LLC were used as standards for creating standard curves, with R2 higher than 0.9990, of hexoses and pentoses.
10
Phytochemical Characterization of Olive Derivatives
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For this work, 3,4-dihydroxyphenylglycol, syringic acid, verbascoside and luteonin-7-O-glucoside were obtained from Sigma-Aldrich (Deisenhofer, Germany). Tyrosol was obtained from Fluka (Buchs, Switzerland) and hydroxytyrosol was obtained from Extrasynthese (Lyon Nord, Geney, France). Gallic acid, trifluoroacetic acid, anthrone, glucose, Folin–Ciocâlteu phenol reagent, vanillic acid, oleocanthal, oleacein and 2,2-diphenyl-1-picrylhydrazyl were obtained from Sigma-Aldrich (Madrid, Spain). Sodium bicarbonate (Na2CO3), acetonitrile (HPLC grade), hexane and sulfuric acid (H2SO4) were purchased from Panreac Quimica S.A. (Barcelona, Spain).
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