Determinação Colorimétrica de Glicose

Medição do análogo de glicose sanguínea usando o método DNS

Colorimetry Intermediate (High School) 60 min ~$10,00

Objetivo

Determinar a concentração de glicose usando o método colorimétrico do ácido dinitrossalicílico (DNS) e entender a química do açúcar redutor.

Contexto

The DNS method is a classic colorimetric assay for reducing sugars. 3,5-dinitrosalicylic acid (yellow) is reduced by the free aldehyde group of glucose to 3-amino-5-nitrosalicylic acid (orange-red). The color change occurs upon heating, and the intensity at 540 nm is proportional to the reducing sugar concentration. This method is widely used in food science and fermentation monitoring.

Avisos de segurança

  • DNS reagent contains NaOH — corrosive to skin
  • Boiling water bath — handle tubes with tongs
  • Cool tubes before opening to prevent splattering
  • DNS reagent stains skin yellow

EPI necessário

goggles gloves lab_coat

Materiais

  • DNS reagent (100 mL)
    1% DNS in NaOH with sodium potassium tartrate
  • Glucose stock solution (100 mL)
    10 mg/mL in water
  • Unknown sugar solution (20 mL)
  • Distilled water (200 mL)

Equipamentos

Spectrophotometer or colorimeter Cuvettes Boiling water bath Test tubes (10) with caps Pipettes (1 mL, 5 mL) Ice bath Timer

Procedimento

1

Prepare glucose standards: 0, 1, 2, 4, 6, 8, 10 mg/mL by diluting the stock solution.

10 min
2

Add 1 mL of each standard and the unknown to labeled test tubes. Add 1 mL of DNS reagent to each tube.

5 min DNS contains NaOH
3

Place all tubes in a boiling water bath for exactly 5 minutes. The color changes from yellow to orange-red.

5 min Use tongs for hot tubes
4

Transfer tubes immediately to an ice bath to stop the reaction. Cool for 2 minutes.

2 min
5

Add 8 mL of distilled water to each tube and mix well.

5 min
6

Measure absorbance at 540 nm. Zero with the blank (0 mg/mL glucose).

15 min
7

Plot the calibration curve and determine the glucose concentration of the unknown.

18 min

Resultados esperados

A linear calibration curve from 1–10 mg/mL glucose. The color intensifies from pale yellow (blank) to deep orange-red (10 mg/mL). Non-reducing sugars like sucrose give no color change.

Limpeza

Neutralize DNS waste with dilute HCl before disposal. Clean test tubes immediately as dried DNS residue is difficult to remove.

Detalhes

Categoria
Colorimetry
Dificuldade
Intermediate (High School)
Duração
60 min
Custo estimado
$10,00
Passos
7
Materiais
4

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Frequently Asked Questions

What is the objective of Determinação Colorimétrica de Glicose?
Determinar a concentração de glicose usando o método colorimétrico do ácido dinitrossalicílico (DNS) e entender a química do açúcar redutor.
How difficult is Determinação Colorimétrica de Glicose?
This experiment is rated as Intermediate (High School). It takes approximately 60 minutes to complete.
What safety precautions are needed for Determinação Colorimétrica de Glicose?
Key safety precautions include: DNS reagent contains NaOH — corrosive to skin; Boiling water bath — handle tubes with tongs; Cool tubes before opening to prevent splattering.
What materials are needed for Determinação Colorimétrica de Glicose?
The main materials required are: DNS reagent, Glucose stock solution, Unknown sugar solution, Distilled water.
What results should I expect from Determinação Colorimétrica de Glicose?
A linear calibration curve from 1–10 mg/mL glucose. The color intensifies from pale yellow (blank) to deep orange-red (10 mg/mL). Non-reducing sugars like sucrose give no color change.