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  • Spectrophotometric total reducing sugars assay based on cupric reduction.

Spectrophotometric total reducing sugars assay based on cupric reduction.

Talanta (2015-11-26)
Kevser Sözgen Başkan, Esma Tütem, Esin Akyüz, Seda Özen, Reşat Apak
ABSTRACT

As the concentration of reducing sugars (RS) is controlled by European legislation for certain specific food and beverages, a simple and sensitive spectrophotometric method for the determination of RS in various food products is proposed. The method is based on the reduction of Cu(II) to Cu(I) with reducing sugars in alkaline medium in the presence of 2,9-dimethyl-1,10-phenanthroline (neocuproine: Nc), followed by the formation of a colored Cu(I)-Nc charge-transfer complex. All simple sugars tested had the linear regression equations with almost equal slope values. The proposed method was successfully applied to fresh apple juice, commercial fruit juices, milk, honey and onion juice. Interference effect of phenolic compounds in plant samples was eliminated by a solid phase extraction (SPE) clean-up process. The method was proven to have higher sensitivity and precision than the widely used dinitrosalicylic acid (DNS) colorimetric method.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Potassium sodium tartrate tetrahydrate, BioUltra, ≥99.5% (NT)
Sigma-Aldrich
Potassium sodium tartrate tetrahydrate, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.5% (perchloric acid titration)
Sigma-Aldrich
Gallic acid monohydrate, ≥99% (HPLC)
Sigma-Aldrich
Sodium hydroxide solution, BioUltra, for molecular biology, 10 M in H2O
Sigma-Aldrich
Potassium sodium tartrate tetrahydrate, ACS reagent, 99%
Sigma-Aldrich
Copper(II) chloride dihydrate, ≥99.95% trace metals basis
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3-Ethyl-2,4-pentanedione, mixture of tautomers, 98%
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Copper(II) chloride dihydrate, 99.999%
Sigma-Aldrich
Sodium hydroxide, BioUltra, for luminescence, ≥98.0% (T), pellets
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Potassium sodium tartrate tetrahydrate, puriss., 99.0-101.0% (calc. on H2O free substance), meets analytical specification of Ph. Eur., BP, USP, FC, E 337
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Potassium disulfite, BioUltra, ≥97.0% (RT)
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Gallic acid monohydrate, ACS reagent, ≥98.0%
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3,5-Dinitrosalicylic acid, 98%
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Sodium carbonate, anhydrous, powder, 99.999% trace metals basis
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3,5-Dinitrosalicylic acid, used in colorimetric determination of reducing sugars
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Liquified Phenol, ≥89.0%
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Phenol solution, Saturated with 0.1 M citrate buffer, pH 4.3 ± 0.2, BioReagent, for molecular biology
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Phenol, puriss., ≥99.5% (GC), meets analytical specification of Ph. Eur., BP, USP, crystalline (detached)
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Sigma-Aldrich
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Sigma-Aldrich
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