5/30/23, 4:39 PM Ethanol Assay
EXPERIMENT NO. 9E
ETHANOL ASSAY
BY DICHROMATE COLORIMETRIC METHOD
Prepared by
Nam Sun Wang
Department of Chemical & Biomolecular Engineering
University of Maryland
College Park, MD 20742-2111
ENCH485
Table of Contents
Method
List of Reagents and Instruments
Procedures
Questions
References
Comments
Method
Most of the chemical oxidation methods are based on the complete oxidation of ethanol by dichromate in the
presence of sulfuric acid with the formation of acetic acid. This reaction is popular because potassium
dichromate is easily available in high purity and the solution is indefinitely stable in air. The theoretical
reaction stoichiometry is shown below:
2Cr2O7-- + 3C2H5OH + 16H+ -----> 4Cr+++ + 3CH3COOH + 11H2O
Dichromate (Cr2O7--, Cr(VI)) is yellowish in color and the reduced chromic product (Cr+++, Cr(III)) is
intensely green. Because the absorption spectra of dichromate and chromic ions overlap significantly, Beer's
law is not obeyed. Instead, the spectra of the solution of interest must be analyzed at multiple wavelengths to
calculate the individual concentrations of dichromate and chromic ions in a mixture subject to the material
balance that the total number of chromium atoms must be conserved.
Other methods of determination primarily based on the above reaction are commonly used. In these methods,
any one of the reactants or products participating in Reaction (1) can be analyzed through another separate
reaction. For example, the excess dichromate remaining in the solution can be further reduced by titration
with other oxidizing reagents such as ferrous ammonium sulfate:
Cr2O7-- + 6Fe++ + 14H+ -----> 2Cr+++ + 6Fe+++ + 7H2O
To enhance the visualizatio of titration endpoint, organic indicators such as sodium diphenylamine sulfonate
and 1,10-o-phenanthroline are added. Another similar method is based on iodometric titration.
Proper concentration of sulfuric acid in the surronding solution will direct the oxidation of ethanol toward
acetic acid instead of acetaldehyde.
Or the excess dichromate can be analyzed.
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5/30/23, 4:39 PM Ethanol Assay
List of Reagents and Instruments
A. Equipment
Test tubes
Pipets
Spectrophotometer
B. Reagents
Potassium Dichromate Reagent Solution
Potassium dichromate: 1 g
Add concentrated (6N) sulfuric acid to: 100 ml
Shake to dissolve; heat slightly if necessary
Saturated s-Diphenylcarbazide solution in 95% ethanol
s-Diphenylcarbazide: ? g to saturate
95% ethanol: ? ml
Collect supernatant
Procedures
Prepare the dichromate reagent solution and the secondary s-Diphenylcarbazide solution.
1. Pipet 1 ml of ethanol solution to be analyzed into of glucose sample in a lightly capped test tube. (To
avoid the loss of liquid due to evaporation, cover the test tube with a piece of paraffin film if a plain
test tube is used.)
2. Heat the mixture at 90ºC for 5-15 minutes to develop the red-brown color.
3. Add 1 ml of a 40% potassium sodium tartrate (Rochelle salt) solution to stabilize the color.
4. After cooling to room temperature in a cold water bath, record the absorbance with a
spectrophotometer at 575 nm.
3ml 1ml O.D.
reagent --->----+ Rochelle soln --->----++-------> at 575nm
| | | | || |
|-+--| |-++-|
| | heat | |
| | --------> | |
| | | |
| | | |
|____| |____|
3ml sample soln
Notes
1. Phenol, up to 2g/l, intensifies the color density. It changes the slope of the calibration curve of
absorbance versus glucose concentration but does not affect the linearity. The above procedure yields
an absorbance of 1 for 1 g/l of glucose in the original sample in the absence of phenol in the reagent,
as opposed to an absorbance of 2.5 for 1 g/l of glucose in 2 g/l of phenol. This property can be
exploited to achieve the maximum sensitivity for dilute samples.
Questions
1. How much time was needed for the complete color development? Justify your answer with a plot of
color intensity as a function of time.
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5/30/23, 4:39 PM Ethanol Assay
2. Obtain an absorption spectrum over wavelengths in the visible range (i,e. 400-700 nm). Justify the use
of 575 nm chosen in the Procedure.
3. Find one other method commonly used to determine ethanol concentration in aqueous solutions. What
are the claimed advantages and disadvantages of this method.
References
1. Williams, M. B. and Reese, Darwin, Colorimetric determination of ethyl alcohol, Anal. Chem., 22,
1556, 1950.
Return to Prof. Nam Sun Wang's Home Page
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Ethanol Assay by Dichromate Colorimetric Method
Forward comments to:
Nam Sun Wang
Department of Chemical & Biomolecular Engineering
University of Maryland
College Park, MD 20742-2111
301-405-1910 (voice)
301-314-9126 (FAX)
e-mail: nsw@umd.edu
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