International Starch Institute

Science Park Aarhus, Denmark

ISI 27-1e Determination of Reducing Power and DE by Lane and Eynon's method.

1. Scope The method is applicable to all starch hydrolysis LT 7/12 1966

products. Rev.: LT 3. May 1999

2. Principle The Lane and Eynon constant titre method. Mixed

Fehling's solution is titrated with sample using
methylene blue as indicator.

3. Reagents 3.1 Fehling stock solution A. Dissolve 69.3 g copper

(II) sulphate pentahydrate (CuSO4, 5H2O) in distilled
water to 1000.0 ml.

3.2 Fehling stock solution B. Dissolve 346 g

potassium sodium tartrate tetrahydrate (KNaC4H4O6,
4H2O) and 100.0 g sodium hydroxide (NaOH) in
distilled water to 1000,0 ml. Decant solution from
sediment if necessary before use.

3.3 Methylene blue (C16H18CIN3S, 2H2O) indicator,

1% aqueous solution.

3.4 Anhydrous D-glucose pro analyse. Dry two hours

at 100 oC and cool in dessicator before use.

3.5 Standard glucose solution. Weigh to the nearest

0.1 mg 1.25 g dried anhydrous D-glucose (3.4) and
dissolve with distilled water to 250 ml.

4. Apparatus 4.1 Automatic burette.

4.2 Burette, 50 ml graduated to 0.1 ml

4.3 Bunsen burner

4.4 Desiccator with dry drying agent

4.5 Analytical scale 0.1 mg

5. Procedure Prepare mixed Fehling's solution and standardise.
Mixed Fehling's solution does not keep.
A. Make mixed Fehling's solution by transferring first Use of automatic
Standardisation 12.5 ml of solution A (3.1) and then 12.5 ml of burette for solution A
solution B (3.2) to a 250 ml conical flask and mix is strongly
well. recommended.
Fill 50 ml burette (4.2) with standard glucose solution
(3.5). Add from burette about 23.5 ml of the standard
glucose solution (3.5) to mixed Fehling's solution.
Add a few anti-bumping granules and place on wire
gauze with clean asbestos mat. Heat over Bunsen
burner. Boil for two minutes and add three drops of
methylene blue indicator (3.3).
Add - without interrupting boiling - 2-3 drops of Detect colour change
dextrose solution at about 10 seconds intervals until in the clear
the blue colour completely disappear. supernatant liquid
from light blue to
colourless
Artificial indirect lighting is strongly recommended. Diffuse daylight is not
suitable for detecting
the end-point.
Titration should be completed in less than a minute.
Repeat titration twice.
Exactly 24.1 ml of standard glucose solution should
be used for the titration - otherwise adjust Fehling's
stock solution A appropriately and repeat
standardisation.
B. Weigh accurately into a beaker (g) g sample, where The test solution
Determination approximate sample weight (g) = 12500 / (expected should contain
DE x dry substance). Dilute sample with distilled approximately 1.25 g
water to 250 ml dextrose in 250 ml.
Proceed as described under A. Standardisation using
sample solution in stead of standard glucose solution
for the titration. Note b = ml test solution used for
titration.

6. Calculation DE (average results of two samples to one decimal)

= (250 x a x 10) / (b x g x d) ;

where

a = factor from table below

b = ml test solution for titration
g = gram sample dissolved to 250 ml
d = % dry solids in sample
7. Notes Alternative method: Calculation of DE from a sugar
distribution obtained by HPLC.

8. Reference International Standard: ISO 5377