Ascorbic Acid Determination in Natural Orange Juice

As a Teaching Tool of Coulometry and Polarography

Mauro Bertotti, Jorge Moreira Vaz, and Rogerio Telles
lnstituto de Quimica, USP, CP 26077, CEP 05599-970, SBo Paulo, SP, Brazil
Electroanalytical techniques have-been defined a s the
application of electrical measurements for analytical purposes ( I ) , and they have undergone a large development in
the last two decades due to the improvements and simplifications of instrumentation (2.3). In this wav, the use of
operational amplifiws led to more versatile elertrorhemical i n s t ~ m e n t sThe
. interest in thc detcrminatim of metals and organlr compound< at lower conccntratmns, esperiallv in envwonmenrnl nnaI\r.+es,ha$ hroughr about the
application of some electroanalytical methods in both academic and industrial contexts. I n spite of these important
aspects, the teaching of electroanalytical methods is a difficult task mainly due to the apparent aversion of students
relating to electrochemistry ( 4 ) .I n this sense, we have proposed a n attractive experiment involving coulometry and
polarography in order to determine ascorbic acid concentrations in natural orange juice. This experiment has been
performed by undergraduate students of a pharmacy
course. Our expectation is to present to these students a
practical experiment involving a common substance to allow understanding of the principles of operation of the coulometer and polaromaph, besides pointing out the connection between the measured phykcal p;operty and the
concentration of a substance. Important concepts as Faraday's law. diffusive transport, and the ronnrction hrtwwn
the electrical quant~tiesof interest in clcctrorhcmistry
(i.e., potential a i d current) are emphasized strongly.
A usual method for coulometric determination of ascorbic acid consists in the reaction of this substance with electrogenerated bromine, using constant current (coulometric
titration) (5,6).The endpoint in these titrations has been
determined by methods such a s amperometry or potentiometrv. With the aim to present to the students another
differek e l e ~ t r o ~ e n e r a t esubstance
d
able to oxidize the
ascorbic acid. the experiments also are performed using 12.
The two oxidant spkcies are generatea separately a t t h e
anode from the respective reduced forms by the application of constant current, and the detection of endpoint in
these different coulometric titrations is made by use of visual indicators. The reaction between ascorbic acid and iodine or bromine is shown below

ascorbic acid

ascorbic acid concentrations are recorded and the calibration curve is plotted in order to verify the linearity between
limiting current and ascorbic acid concentration (Ilkovic
equation). The standard addition method is used in the determination of the substance in orange juice. This analytical procedure minimizes possible interferences of the matrix (8).
Experimental Section
Reagents
The supporting electrolyte for the coulometric titrations
was 1.0 M HpSO4. The iodide is oxidized easily by air a t
sufiicientlv acidic solutions: therefore. for the iodometric
M) buffer was
titration a n acetatelacetic acid (0.1 k0.1
em~lovedas electrolvtic solvent. The salts used a s startine
mateAals were ~ 1 a n KBr,
d
both analytical grade reagents. The indicators were 2% starch solution and 1%
methyl orange solution; this latter for the bromine titration. An ascorbic acid solution was prepared a t approxiademately 0.1 M concentration from the dissolution of
quate quantity of the solid substance in a volumetric flask.
%s solution was stored in a piston buret to limit the contact with air and to deliver more precise volumes to the
coulometric and polarographic cells. The solution was
standardized by a usual method of titration with ascorbic
acid of the chemicallv eenerated iodine after oxidation of
iodide with a known hcantity of iodate (9).The solution of
ascorbic acid holds this concentration for one dav in the
mentioned storage conditions (10).

an

Coulometric Titrations
AMetrohm Herisau coulometer E211 A was used in the
coulometric titrations and the constant current was adjusted a t 20 mA. This current value was calibrated by use
of a usual method, i.e., titration of a defined volume of
standardized AsOi solution with electrogenerated iodine
(starch a s indicator) in Na?HPOa medium (8).
. . The olatinum generator electrodes were a gauze cylinder and a wire
spiral. These electrodes were resnectivelv the anode and
the cathode when iodine and broniine were generated. The
wire spiral was placed into a glass isolation tube containing a Na2S04saline bridge in order to avoid contamination
of the solution due to electrogenerated species. The solution in the cell was continuouily stirred ;sing a magnetic
bar.
Pure Ascorbic Acid

dehydroascorbic acid
The determination of ascorbic acid in orange juice by polarography is connected with the measurement of the anodic limiting current originated from the oxidation of this
substance to dehydroascorbic acid a t the dropping mercury
electrode (7). The polarograms of solutions a t different

I z a s a n Oxidant The electrolytic solution contained

100 mL of acetatel acetic acid buffer and 1 mL of starch
solution. The solution was 0.1 M in iodide. A 0.125-mL
s a m ~ l of
e standard ascorbic acid solution was added to the
cell (a 200-mL beaker) and the constant current was applied until the appearance of blue color. Previous experiments were made with electrolyte in the absence of a
reducing agent to verify the time required for the visualiVolume 72 Number 5 May 1995

445

Table 1. Results of Coulometric Titrations of Pure Ascorbic Acid

Using Bromine and Iodine as Electrogenerated Oxidants (I = 19.90

Natural Orange Juice

T h e juice was obtained by
squeezing fresh oranges and filEleclroanalytical
Ascorbic
Time (s)
Ascorbic
Recovery
Trials
tering to remove solids. The re(%)
technique
acid added
acid found
sulting solution was stocked in a
(mM)
(mM)
closed flask under nitrogen atCoulometry 12 Method
102.5
126.0f0.2 104.0f0.1
101.5
3
mosphere. Five milliliters of this
Br2 Method
102.5
125.8f 0.8 104.7f 0.6
102.1
3
juice were added to the coulometric cell for the analysis with ioPolarography Standard addition
0.91
0.92f 0.01
101.1
3
dine. In spite of the presence of
t h e juice, the excess of iodine
(blue color) was easily observed.
Table 2. Comparison of the Results of Ascorbic Acid Determinations
I n t h e coulometric titrations
in Natural Orange Juice by Coulometric (with iodine and bromine)
with bromine the added volume
and Polarographic (standard addition procedure) ~ e t h o d s ~
of orange juice was 2 mL in order
to diminish the interference of
Eleclroanalytical
Ascorbic acid found (mM)
Trials
the juice color in the detection of
technique
the endpoint.
Polarography. The
polaro12 Method
2.46M.02
3
Coulometry
m h i c determinations were carRrg Method
6R7+004
3
riea out with a Sargent Welch
3
Polarography
Standard addition
2.43M.07
Polarograph Model XVI using a
conventional H-cell and a dropping
T h e percent recovery for polarography was 98G %.
mercury electrode (SCE). All the
potentials were measured against
a saturated calomel electrode. The
zation of the blue color of the iodine-starch complex (blank
solutions were bubbled adequately with nitrogen. Chloride
correction).
from the SCE gives a n anodic wave in the region of ascorBrz a s a n Oxidant The detection of endpoint i n these
bic acid electroxidation (anticipation of the mercury oxidacoulometric titrations was based on the reaction of the
tion); therefore, the working electrode compartment was
separated from the reference electrode by a sintered glass
electrogenerated bromine with methyl orange (11). Bedisk and 4% agar-saturated potassium nitrate saline
cause ascorbic acid is first oxidized by bromine, the endbridge. This procedure minimizes the migration of chloride
point can be found by the reaction of the excess of the
ions
from the reference electrode to the test solution.
oxidant with the indicator, the red color of the solution
Calibration curves were constructed from the polaro(acidic medium) is changed to colorless. Initially, three
grams recorded after the addition of known volumes of
drops of methyl orange were added to the solution constandard ascorbic acid solution (0.100 to 0.400 mL) to 10.0
tained in the cell (0.1 M H2S04 and 0.1 M B r (100 mL))
mL of 0.5 M KN03 contained in the polarographic cell. The
and the current was applied in order to determine the time
polarograms of the blank and the solutions were recorded
from -0.1 V going in the positive direction, until the aprequired for the indicator oxidation (blank correction).
pearance of the anodic wave corresponding to the mercury
Then, three new drops of methyl orange and a 0.125-mL
oxidation. The polarographic determination of ascorbic
sample of standard ascorbic acid were added to the cell for
acid in the juice was performed by use of the standard adthe titration.
dition method. A0.5 M KNOBsolution was used a s supporting
electrolyte (8.0 mL). Then, a 2.0mL sample of the filtered natural
juice was added to t h e polarographic cell and the polarogram of the resulting solution
was recorded. Afterwards, four
aliquots (0.050 mL each) of the
standard ascorbic acid contained
in the piston buret were added to
the cell and the respective polarograms were obtained.

~
1
.
'

Results
Table 1presents the electrolysis times for the coulometric titrations of the standard ascorbic
acid samples using the two proposed methods, and the amount
2
4
of ascorbic acid determined from
each method. The results found,
C ASCORBIC ACID/ m M
using both iodine and bromine as
oxidants for pure ascorbic acid,
are reproducible and a difference
than 2% was obtained in
Figure 1.Polarograms of 0.5M KN03 solution (0)and after addition of pure ascorbic acid aliquots (1-4).
comparison w i t h iodometric
The resulting calibration curve is also depicted.

446

Journal of Chemical Education

standardization (9).A partial explanation for the higher results

of the coulometric analysis may
be related to the presence of impurities, so that the current efficiency was not 100%. The data
obtained i n the coulometric titrations of t h e n a t u r a l juice a r e
shown in Table 2, where the discrepancies between the titration
with bromine and iodine are evident. The explanation for these
results is related to the greater
oxidizing power of the bromine,
which can oxidize other organic
I
c o m ~ o n n d sof the natural iuice
w
0
1
2
besides ascorbic acid. This exoeriment shows the higher selecC ASCORBIC ACID ADDED I m M
tivity of ascorbic acid-oxidation
bv iodine. The ~ o l a r o m a m sand
calibration curve obiained for Figure 2. Polarograms of natural orange juice (0) and after the addition of pure ascorbic acid aliquots
the analysis of pure ascorbic acid ( 1 4 ) (0.5 M K N 0 3 ) . The extrapolation of the straight line to x-axis represents the ascorbic acid concenare shown in Figure 1.The linear tration in the juice contained in the polarographic cell.
plot was perfectly. adiusted
to the
.
experimental points, also passing through the origin. So,
Acknowledgment
the linear dependence of the limiting current with respect
We thank Lniz Roberto de Moraes Pitombo for helpful
to ascorbic acid concentration was proved. Figure 2 presuggestions and Paulo Celso Isolani for correcting the Engsents the polarogram of natural juice i n KN08, as well a s
lish.
the resulting polarograms after the addition of known
Literature Clted
amounts of standard ascorbic acid. The anomalous de1. Kolthaff, I, M. J. Eledroehrm. Soc. 1971,118, 5C4C.
crease of the current before the mercury oxidation may be
2. Bond, A . M . Modem Polomgmgmgphie Methods in Analytical ChamLslry; Marcel Dekter ~.
New Ynrt.,.19RO
associated with unknown compounds present in the or. ~,..
3. Bard, A. J.; Faulkne~L. R. E l ~ f r n c h ~ m i Methods:
ml
Wiiey: New York, 1980.
ange juice, because this unexpected effect was not oh4. Chambers, J. Q. J. Chem Educ 1983.60.259-262,
served i n the polarograms of Figure 1(pure ascorbic acid).
5. Marsh, D. J.;Jacobs. D. L.:Veening H. J. Chem. Educ. 1973.50,626628.
, D.; Burch6eld, D. E.: Veening, H. J. Chem. Educ 1985.62.688690.
6. G ~ e n s p a nP
However, this anomaly does not interfere in the analytical
7. Milner, G. W C. The Princ~plesand Applications ofPoiampmphy; Lonprmans: Lonapplication of polarography for this study. The results
don, 1966. p 602.
8. Willard, H.H.:Mertitt, L. L., Jr;Dean. J.A. Insbumntol Methalr ~ f A n a i ~ ~ i ~ : V ~
found from the standard addition method (Fig. 2) are in
Nostrand: New York, 1965.
excellent agreement with the ones obtained using electro9. KolthofC I. M. VolumelricAnolysis; Interscience: New York, 1957. p 626.
10. Erdey, L.; Bodor, E. A n d Cham. 1952,24,418420.
generated iodine in the constant current coulometric exn. A ; west, M w F ~ L ~ o , f~~ n a~i y t i~mchemistry;
~P
~ saunaers:
~
~
I mila~
11. skoog,
periment, a s shown in Table 2.
delphia, 1982, p379.
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Number 5

May 1995

447