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 2!"2#2$$$
E%C&"'(
Table of Conen!
Method
)ist of *eagents and +nstruments
Procedures
,uestions
*eferences
Comments
Me"o#
Most of the chemical o-idation methods are .ased on the complete o-idation of ethanol
.y dichromate in the presence of sulfuric acid /ith the formation of acetic acid0 1his
reaction is popular .ecause potassium dichromate is easily availa.le in high purity and
the solution is indefinitely sta.le in air0 1he theoretical reaction stoichiometry is sho/n
.elo/2
2Cr
2
O
7
--
+ 3C
2
H
5
OH + 16H
+
-----> 4Cr
+++
+ 3CH
3
COOH + 11H
2
O
Dichromate 3Cr24!
##
, Cr35+66 is yello/ish in color and the reduced chromic product 3Cr
77
7
, Cr3+++66 is intensely green0 Because the a.sorption spectra of dichromate and chromic
ions overlap significantly, Beer8s la/ is not o.eyed0 +nstead, the spectra of the solution of
interest must .e analy9ed at multiple /avelengths to calculate the individual
concentrations of dichromate and chromic ions in a mi-ture su.:ect to the material
.alance that the total num.er of chromium atoms must .e conserved0
4ther methods of determination primarily .ased on the a.ove reaction are commonly
used0 +n these methods, any one of the reactants or products participating in *eaction 3$6
can .e analy9ed through another separate reaction0 ;or e-ample, the e-cess dichromate
remaining in the solution can .e further reduced .y titration /ith other o-idi9ing reagents
such as ferrous ammonium sulfate2
Cr
2
O
7
--
+ 6Fe
++
+ 14H
+
-----> 2Cr
+++
+ 6Fe
+++
+ 7H
2
O
1o enhance the visuali9atio of titration endpoint, organic indicators such as sodium
diphenylamine sulfonate and $,$#o#phenanthroline are added0 <nother similar method is
.ased on iodometric titration0
Proper concentration of sulfuric acid in the surronding solution /ill direct the o-idation
of ethanol to/ard acetic acid instead of acetaldehyde0
4r the e-cess dichromate can .e analy9ed0
L$! of Reagen! an# In!%umen!
A. E&u$'men
1est tu.es
Pipets
=pectrophotometer
B. Reagen!
Potassium Dichromate *eagent =olution
o Potassium dichromate2 $ g
o <dd concentrated 3>%6 sulfuric acid to2 $ ml
o =hake to dissolve? heat slightly if necessary
=aturated s#Diphenylcar.a9ide solution in @(A ethanol
o s#Diphenylcar.a9ide2 B g to saturate
o @(A ethanol2 B ml
o Collect supernatant
P%o(e#u%e!
Prepare the dichromate reagent solution and the secondary s#Diphenylcar.a9ide solution0
$0 Pipet $ ml of ethanol solution to .e analy9ed into of glucose sample in a lightly
capped test tu.e0 31o avoid the loss of liCuid due to evaporation, cover the test
tu.e /ith a piece of paraffin film if a plain test tu.e is used06
20 &eat the mi-ture at @DC for (#$( minutes to develop the red#.ro/n color0
E0 <dd $ ml of a "A potassium sodium tartrate 3*ochelle salt6 solution to sta.ili9e
the color0
"0 <fter cooling to room temperature in a cold /ater .ath, record the a.sor.ance
/ith a spectrophotometer at (!( nm0
5. 3ml 1ml O.D.
6. reagent --->----+ Rochelle soln --->----++-------> at
575nm
7. | | | | || |
. |-+--| |-++-|
!. | | heat | |
1". | | --------> | |
11. | | | |
12. | | | |
13. |####| |####|
14. 3ml sam$le soln
Noe!
$0 Phenol, up to 2gFl, intensifies the color density0 +t changes the slope of the
cali.ration curve of a.sor.ance versus glucose concentration .ut does not affect
the linearity0 1he a.ove procedure yields an a.sor.ance of $ for $ gFl of glucose in
the original sample in the a.sence of phenol in the reagent, as opposed to an
a.sor.ance of 20( for $ gFl of glucose in 2 gFl of phenol0 1his property can .e
e-ploited to achieve the ma-imum sensitivity for dilute samples0
)ue!$on!
$0 &o/ much time /as needed for the complete color developmentB Gustify your
ans/er /ith a plot of color intensity as a function of time0
20 4.tain an a.sorption spectrum over /avelengths in the visi.le range 3i,e0 "#!
nm60 Gustify the use of (!( nm chosen in the Procedure0
E0 ;ind one other method commonly used to determine ethanol concentration in
aCueous solutions0 Hhat are the claimed advantages and disadvantages of this
method0
Refe%en(e!
$0 Hilliams, M0 B0 and *eese, Dar/in, Colorimetric determination of ethyl alcohol,
Anal. Chem., **+ $((>, $@(0
Return to Prof. Nam Sun Wang's Home Page
Return to Biohemial !ngineering "aboratory #!NCH$%&'
Ethanol Assay by Dichromate Colorimetric Method
(or)ard omments to*
Nam Sun Wang
+epartment of Chemial , Biomoleular !ngineering
-ni.ersity of /aryland
College Par0, /+ 123$141555
6254$2&45752 #.oie'
625465$47518 #(A9'
e4mail* ns):umd.edu