College of Science

Determination of Vitamin C Concentration by Titration

(Redox Titration Using Iodine Solution)
Safety Solutions Needed
Lab coats, safety glasses and enclosed footwear must
Iodine solution: (0.005 mol L−1). Weigh 2 g of potassium
be worn at all times in the laboratory.
iodide into a 100 mL beaker. Weigh 1.3 g of iodine and
add it into the same beaker. Add a few mL of distilled
Introduction water and swirl for a few minutes until iodine is
dissolved. Transfer iodine solution to a 1 L volumetric
This method determines the vitamin C concentration
flask, making sure to rinse all traces of solution into the
in a solution by a redox titration using iodine. Vitamin
volumetric flask using distilled water. Make the solution
C, more properly called ascorbic acid, is an essential
Determination of Vitamin C Concentration
antioxidant needed by the human body (see additional
up to the 1 L mark with distilled water.
Starch indicator solution: (0.5%). Weigh 0.25 g of
by Titration
notes). As the iodine is added during the titration, the
ascorbic acid is oxidised to dehydroascorbic acid, while soluble starch and add it to 50 mL of near boiling water
(Redox
the iodine is reduced to iodide ions. Using Iodine Solution)in a 100 mL conical flask. Stir to dissolve and cool before
Titration
using.
ascorbic acid + I2 → 2 I−
+
Introduction dehydroascorbic acid
Due to this reaction, the
Thisiodine formed istheimmediately
method determines vitamin C concentration
in a solution by a redox titration using iodine. Vitamin
reduced to iodide as long as there is any ascorbic acid
C, more properly called ascorbic acid, is an essential Burette containing
present. Once all the ascorbic
antioxidantacid
neededhas been
by the humanoxidised,
body (see additional iodine solution
the excess iodine is free to react
notes). with isthe
As the iodine starch
added during the titration, the
indicator, forming theascorbic acid is oxidised to dehydroascorbic acid, while
blue-black starch-iodine complex.
the iodine is reduced to iodide ions.
This is the endpoint of the titration.
ascorbic acid + I2 → 2 I + dehydroascorbic acid
−

The method is suitableDuefortouse with vitamin

this reaction, the iodineCformed
tablets,
is immediately
fresh or packaged fruitreduced
juicestoand
iodidesolid fruits
as long and
as there is any ascorbic acid
vegetables. present. Once all the ascorbic acid has been oxidised,
the excess iodine is free to react with the starch
NB: This method is more straight
indicator, formingforward thanstarch-iodine
the blue-black the complex.
This is the endpoint of the
alternative method using potassium iodate, but as titration.

the potassium iodate The method is

solution is suitable
more for use with
stable thanvitamin
theC tablets,
fresh or packaged fruit juices and solid fruits and
iodine as a primary standard,
vegetables.the alternative method is
more reliable. NB: This method is more straight forward than the
alternative method using potassium iodate, but as
the potassium iodate solution is more stable than the
Equipment Needed iodine as a primary standard, the alternative method is
more reliable.
burette and stand
Conical flask
100 mL or 200 mL volumetric flaskNeeded
Equipment
20 mL pipette burette and stand
Vitamin C sample solution
100 mL or 200 mL volumetric flask
10 mL and 100 mL measuring cylinders
20 mL pipette
250 mL conical flasks 10 mL and 100 mL measuring cylinders
250 mL conical flasks

Solutions Needed
Iodine solution: (0.005 mol L−1). Weigh 2 g of potassium
Method 4. Calculate the concentration in mol L−1 of ascorbic
acid in the solution obtained from fruit/vegetable/
Sample
StarchPreparation
indicator solution: (0.5%). Weigh 0.25 g of 3.juice.Using
Also, the equation
calculate the of the titration (below)
concentration, in mg/100mL
soluble starch and add it to 50 mL of near
For vitamin C tablets: Dissolve a single tablet in 200 mL boiling water determine
or mg/100g of ascorbic acid, in the sampleacid
the number of moles of ascorbic of fruit/
in a 100 mL conical flask. Stir to dissolve
of distilled water (in a volumetric flask if possible). and cool before reacting.
vegetable/juice.
using.
For fresh fruit juice: Strain the juice through ascorbic acid + I2 → 2 I− + dehydroascorbic acid
cheesecloth to remove seeds and pulp which may block Additional Notes
4. Calculate the concentration in mol L−1 of ascorbic
Method
pipettes. 1. acid
Iodine stains
in the both skin
solution and clothing
obtained so proper care is
from fruit/vegetable/juice.
ForSample
packaged fruit juice: This may also need to be
Preparation advised.
Also, If staining
calculate does occur, alcohol
the concentration, may remove
in mg/100mL or mg/
strained through cheesecloth if it contains a lot of pulp skin stains and cleaners are available for
100g of ascorbic acid in the sample of fruit/vegetable/ fabric stains.
For
or seeds. vitamin C tablets: Dissolve a single tablet in 200 mL
of distilled water (in a volumetric flask if possible). 2. juice.
Vitamin C, or ascorbic acid, is a water soluble
For fruits and vegetables: Cut a 100 g sample into small antioxidant that plays a vital role in protecting the
For fresh fruit in
juice: Strainand
the juice through
Add 10cheese
pieces
cloth
and grind
to remove
a mortar
seeds and pulp
pestle.
which may
mL
block
Additional Notesand disease. It is not synthesised
body from infection
portions of distilled water several times while grinding by the human body and therefore must be
thepipettes.
sample, each time decanting off the liquid extract 1.acquired
Iodine stains
from bothsources
dietary skin and– clothing
primarilysofruits
properandcare
intoFora 100 mL volumetric flask. is advised. If staining does occur, alcohol
vegetables. The chemical structure and antioxidant may remove
packaged fruit juice: ThisFinally,
may alsostrain
need the
toground
be
fruit/vegetable pulp through cheesecloth, rinsing the skin stains and
(reducing) cleaners
action are available
of ascorbic acid arefor fabric stains.
illustrated in
strained through cheese cloth if it contains a lot of pulp
pulpor with
seeds. a few 10 mL portions of water and collecting the redox
2. halfC,equation
Vitamin or ascorbic below:
acid, is a water soluble
all filtrate and washings in the volumetric flask. Make antioxidant that plays a vital role in protecting the body
For fruits and vegetables: Cut a 100 g sample into small 3. The concentration of the prepared iodine solution
the extracted solution up to 100 mL with distilled water. from
can be infection and disease.
more accurately It is not synthesised
determined by titrationby with
pieces and grind in a mortar and pestle. Add 10 mL
Alternatively the 100 g sample of fruit or vegetable the
a human
standard body and
solution therefore
of ascorbic must
acid be
or acquired
a standard from
portions of distilled water several times while grinding
may dietary
solution sources – primarily
of potassium fruits andusing
thiosulfate vegetables.
a starchThe
thebesample,
blendedeach in atime
fooddecanting
processoroff together withextract
the liquid
chemical structure and antioxidant (reducing) action of
about
into50 mL mL
a 100 of distilled
volumetric water. After
flask. blending,
Finally, the ground indicator. This should be done if possible as iodine
strainstrain
thefruit/vegetable
pulp through cheesecloth, ascorbic
solutionsacidcanare
beillustrated
unstable. in the redox half equation
pulp throughwashing it with
cheese cloth, a fewthe
rinsing
10mL portions of distilled water, and makeand thecollecting
extracted below:
pulp with a few 10 mL portions of water
solution up to 100washings
mL in a volumetric flask. flask. Make HO HO
all filtrate and in the volumetric O O
oxidation O O
the extracted solution up to 100 mL with distilled water. HO HO + 2H+ + 2e-
reduction
Titration
Alternatively the 100 g sample of fruit or vegetable may HO OH O O

be blended ascorbic acid dehydroascorbic acid

1. Pipette a 20inmLa food processor
aliquot togethersolution
of the sample with about
into
50 mL of distilled water. After blending, strain
a 250 mL conical flask and add about 150 mL of the
3. The concentration of the prepared iodine solution
pulp through
distilled watercheese cloth,
and 1 mL washing
of starch it with asolution.
indicator few 10 4. The average titre volume should ideally be in the
can be more accurately determined by titration with a
mL portions of distilled water, and make the extracted range of 10 – 30 mL. If the titre required for a 20 mL
2. Titrate the sample with 0.005 mol L−1 iodine solution. standard solution of ascorbic acid or a standard solution
solution up to 100 mL in a volumetric flask. aliquot of sample solution is well outside this range
The endpoint of the titration is identified as the first of potassium thiosulfate using a starch indicator. This
permanent trace of a dark blue-black colour due to then a larger or smaller aliquot volume
should be done if possible as iodine solutions can beshould be
Titration
the starch-iodine complex. chosen. If the volume of the titre is too low, dilute
unstable.
the standard. If the titre volume is too high, dilute
3. Repeat the titration
1. Pipette a 20 mLwith further
aliquot aliquots
of the sampleofsolution
sampleinto 4. The average titre volume should ideally be in
the sample.
solution
a 250 mLuntil youflask
conical obtain
andconcordant results
add about 150 (titres
mL of distilled the range of 10 – 30 mL. If the titre required for a 20
agreeing
water andwithin
1 mL of0.1starch
mL). indicator solution. 5. mL
Ascorbic
aliquotacid is susceptible
of sample solutiontoisoxidation by this
well outside
atmospheric oxygen over time. For this
range then a larger or smaller aliquot volume shouldreason, the
2. Titrate the sample with 0.005 mol L−1 iodine samples
Calculations
solution. The endpoint of the titration is identified as be chosen.should be prepared
If the volume of the immediately before
titre is too low, dilute
the
the titrations.
standard. IfHowever,
the titre if the
volume samples
is too have
high, to bethe
dilute
the first permanent
1. Calculate the average trace of a dark
volume blue-black
of iodine colour
solution used due prepared several hours earlier, oxidation can be
to the starch-iodine complex. sample.
from your concordant titres. minimised by the addition of a small amount of
3. Repeat
2. Calculate thethe titration
moles further aliquots of sample oxalicAscorbic
with reacting.
of iodine
5. acid (egacid is susceptible
1 g oxalic acid perto oxidation
100 by
mL of sample
solution until you obtain concordant results (titres atmospheric
solution). oxygen over time. For this reason, the
3. Using thewithin
agreeing equation of the titration (below) determine
0.1 mL). samples should be prepared immediately before the
6. Identification of the endpoint in this titration is
the number of moles of ascorbic acid reacting. titrations. However, if the samples have to be prepared
significantly affected by the colouration of the
several hours earlier, oxidation can be minimised by the
ascorbic acid + I2 → 2 I + dehydroascorbic acid
Calculations
− sample solution used. If the solutions are colourless
addition of a small amount of oxalic acid (eg 1 g oxalic
or are pale in colour, there is no problem identifying
1. Calculate the average volume of iodine solution acid per 100 mL of sample solution).
used from your concordant titres. 6. Identification of the endpoint in this titration is
2. Calculate the moles of iodine reacting. significantly affected by the colouration of the sample
solution used. If the solutions are colourless or are pale
 in colour, there is no problem identifying the endpoint.
For strongly coloured juices there can be a problem with
theendpoint
the endpoint. andForitstrongly
is advisedcoloured juices
to carry out there can
a “rough”
be a problem with the endpoint and it
titration in order to become familiar with any distinct is advised
to carry
in colour, there out awhich
is nochange
colour problem “rough”
identifying titration
the
occurs at theinendpoint
endpoint. order to(itbecome
may
For stronglyjust
coloured
be a juices
familiar withthere
anycan
darkening ofbethe
a problem
distinct colour
colour) withchange
This which
will also help by
the endpoint and it is advised to carry out a “rough”
FigureFigure 1 Vitamin C tablet. Left photo: before endpoint, added iodine reacts
1 Vitamin C tablet. Left photo: before endpoint, added iodinetitration in order
establishing
occurs anendpoint,
at the approximate volume
(itdistinct
may justofbeiodate solution
a darkening
with ascorbic acid leaving the solution colourless. Centre photo: At the
to become familiar with any
reacts with ascorbic acid leaving the solution colourless. required.
of the colour) This will also
colour change which occurs at the endpoint (it may help by establishing an
titration endpoint all the ascorbic acid has reacted and the excess iodine
Centrereacts
photo: Atthethestarch
titration endpoint to givealla palethe ascorbic acid injust
hasphoto: colour,
If be athere is no problem identifying thealsoendpoint.
with indicator blue colour. Right
For strongly 7. approximate
darkening
coloured Theof the
above
juices there
volume
colour) This will
method
can be a
ofmay iodine
problem
help
bewithbysolution
used required.
to carry out a
reacted andFigure
addition the excess
VitaminisCiodine
of1iodine continued
tablet. Leftreacts after
photo: with
the the
before starch
endpoint,
endpoint, added indicator
further to give
iodineiodine-starch
reacts establishing
in colour, an
there approximate
is no problem volume of iodate
identifying thesolution
endpoint.
the endpoint number
7. Theand itabove of
is advised interesting
method
to carry out investigations
may be
a “rough” used to regarding
carry out thea
a palecomplex
bluewith is
colour. formed.
ascorbic acidNB:
Right leavingin each
photo: theIf of these
solution
addition images
colourless.
of iodine a flask
Centre photo: showing
At the the
is continued pale
after
required.
For strongly coloured juices there can be a problem with
titrationof
blue colour endpoint all the ascorbic
the endpoint is shown acid has forreacted and the excess iodine
comparison. titration concentration of vitamin
in order to become familiar with any distinct C in various foods and drinks:
the endpoint, further iodine-starch complex is formed.
reacts with the starch indicator to give a pale blue colour. Right photo: If NB: in each of
7.the endpoint
The abovenumber method
and it isofmay interesting
at be
advised theused
to investigations
to carry
carry out out
(it amay a
“rough” regarding the
these images a flask
addition showing
of iodine is continued theafter pale theblue
endpoint,colour
further the endpointcolour
ofiodine-starch isnumber change
•
which
Vitamin
occurs
C content
endpoint
of different types of fruits/
complex is formed. NB: in each of these images a flask showing the pale titration ofin concentration
interesting
order to investigations
become of vitamin
familiar
just be a darkening of the colour) This will also help by regarding
with C
any in
the various
distinct foods and
shown for blue comparison.
colour of the endpoint is shown for comparison. concentration vegetables/juices.
of vitamin C in various foods and drinks:
Figure 1 Vitamin C tablet. Left photo: before endpoint, added iodine reacts colour
establishing anchange drinks: which occurs at the endpoint
approximate volume of iodate solution (it may
just be aCdarkening ofofdifferent
Cthe colour) This will also help by
with ascorbic acid leaving the solution colourless. Centre photo: At the required.
• Vitamin • content
Vitamin content typesof ofdifferent
fruits/ types/brands of vitamin
titration endpoint all the ascorbic acid has reacted and the excess iodine
Figure
reacts1withVitamin C tablet.
the starch Left photo:
indicator to givebefore endpoint,
a pale blue colour.added
Right photo: 7. The

establishing
iodine Ifreacts vegetables/juices.
above
• anVitamin
approximate
method may
C be
content
volume
used to
of
of
carry
different
iodate
out a
types of fruits
solution
tablets.
C content vegetables/juices.
with ascorbic
addition acid leaving
of iodine the solution
is continued after thecolourless. Centreiodine-starch
endpoint, further photo: At the required.
titration
complexendpoint
is formed.allNB: theinascorbic
each of these acid has
images reacted and
a flask the excess
showing iodine
the pale number
• Vitamin of interesting ofinvestigations regarding of
different types/brands thevitamin
reacts with the
blue colour starch
of the indicator
endpoint to give
is shown foracomparison.
pale blue colour. Right photo: If concentration
7. The
tablets. • Vitamin
above of vitamin
method CC content
inmay
various
be of
foods
used tablets
to and
carry or
outfood/drink
drinks: in the
a types/brands
addition of iodine is continued after the endpoint, further iodine-starch • Vitamin C content of different of
•number
VitaminCof
• Vitamin presence
interesting
Ccontent
content
and absence
investigations
ofofdifferent
tablets types
or
of added
regarding
of fruits/
food/drink in the the
oxalic acid or metal ions
complex is formed. NB: in each of these images a flask showing the pale
blue colour of the endpoint is shown for comparison. concentration

over vitamin
various
of vitamin
tablets.
periods
C inoxalic ofacid
various time.
foods and drinks:
vegetables/juices.
presence and absence of added or metal ions
over
• Vitamin
various
• Vitamin C content C • Vitamin
periods
content
• Vitamin ofoftime. C content
different
C content
of different types of
of tablets
of vitaminor
fruits/
of food/drink
types/brands food/drink
before and afterin the
Figure 2 Commercial fruit juice. Left flask: before the endpoint, the colour
of theFigure
solution reflectsfruit
2 Commercial thejuice.
paleLeft yellow
flask: colour
before the ofendpoint,
the fruitthejuice.
colourFigure

vegetables/juices.
tablets.
flask: Csubjection
Centre• Vitamin presence
content of food/drink and
to cooking absence
beforeconditions.of
and after added oxalic acid or
At theoftitration
2 Commercial the solution endpoint
reflects the allpale
theyellow
ascorbic
colouracidof thehasfruitreacted andflask:
juice. Centre subjection
any excess
the colour• Vitamin • Vitamin to content
C content metal
cooking ofCions
conditions.
of tablets orover various
food/drink in the periods of time.
iodineAtreacts
fruitwithjuice.
the titration
Leftallflask:
the starch
endpoint the
beforetothe
indicator
ascorbic acid form
endpoint,
a dark
has reacted andblue-black
any excess complex.
of •CVitamin different
content types/brands
of fruits/vegetables of vitamin at different
the solution reflects
iodine reacts the
with pale
the starchyellow
indicator colour
to form of
a the
dark fruit
blue-black juice.
complex. presence
• tablets.
Vitamin and
C absence
content of
of added oxalic
fruits/vegetables acid ator metal
different ions
In this case the result is a darkening of the solution’s colour from yellow •
stages Vitamin
of C
ripeness. content of food/drink before and after
Centretoflask: In this
At case
brown-grey. thethe result is aendpoint
titration
Right flask:darkening of all
This illustrates the solution’s
thethe colour from
ascorbic
effect ofacid
addinghasjust aover
yellow
mL various
stages periods of time.
or of ripeness.
two
to brown-grey. Right flask: This illustrates the effect of adding just a mL or
more ofexcess
iodine
• Vitamin C content
subjection of tablets or food/drink
tobefore
cooking in the
conditions.
reacted and any of iodinesolution
iodine
solutionreactsafterthethe with endpoint is reached,
theis reached,
starch indicatorresulting
in theto • in the
Vitamin C content of food/drink
two
Figure
more
2 of
Commercial fruit
after
juice. Left flask:
endpoint resulting
before the endpoint, the colour presence and absence of added oxalicand acidafter
or metal ions
form aformation
dark blue-black
formation further
of further iodine-starch
complex.
iodine-starch In complex.
this
complex. case the result
of the solution reflects the pale yellow colour of the fruit juice. Centre flask: is a darkening Contact
subjection toContact
Us• Vitamin
cooking Us C
conditions. content of fruits/vegetables at different
over various periods of time.
of the solution’s colour
At the titration endpointfrom yellow
all the ascorbic toacid
brown-grey.
has reacted and Right flask:
any excess
iodine reacts with the starch indicator to form a dark blue-black complex.
This illustrates thetheeffect of adding just abefore
mL the orcolour
two more of iodine• Vitamin
If• you have
Vitamin you
C content
IfCany stages
questions
contenthave of of orripeness.
of fruits/vegetables
any comments
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andcommentsthis
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Figure
In this2case
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is a darkening flask:
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yellow stages of ripeness.
solutionofafter
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palereached,
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colour of the in
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to brown-grey. Right flask: This illustrates the effect of adding just a mL or the
juice. formation
Centre flask: of experiment,
subjection please contact us:
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to cooking conditions.
Attwo
themore
titration
further iodine-starch endpoint
of iodine all the
solution
complex. afterascorbic acid has
the endpoint reactedresulting
is reached, and any in
excess
the
Outreach
formation of further iodine-starch complex.
iodine reacts with the starch indicator to form a dark blue-black complex. • Vitamin
Contact
College ofContact
C content ofUs
Outreach
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In this case the result is a darkening of the solution’s colour from yellow stages ofScience
ripeness.
College of Science
to brown-grey. Right flask: This illustrates the effect of adding just a mL or
two more of iodine solution after the endpoint is reached, resulting in the
IfUniversity
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ContactPrivate
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Outreach service
New Zealand is
Christchurchfor senior school chemistry students in
Figure 3 Freshly squeezed orange juice. Left flask: before the endpoint, the College If youofhave any questions or comments relating to this
Science
New Zealand only. We regret we are unable to respond
colour of the solution reflects the bright orange colour of fresh orange juice University Phone: ofNew
+64
experiment, 3 364 Zealand
2178contact
please
Canterbury us:
Figureand is unaffected
3 Freshly by addition
squeezed of iodine.
orange Centre
juice. Leftflask: Once
flask: all the the
before ascorbic
endpoint,Fax:
the+64 to
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364 2490 from overseas.
oxidised, a slight excess of added iodine complexes with the Private Bag 4800
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colouracidof hasthebeen
solution reflects the bright orange colour of fresh orange juiceoutreach@canterbury.ac.nz
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starch indicator, giving the solution a green colour in this case. This is the Christchurch
and isendpoint
unaffected of the by addition
titration. Rightof iodine.
flask: Centre
If further iodineflask:
solution Once
were all
to be CollegeOutreach
the ascorbic ofFax:
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acid has been the oxidised, a slight
colourexcess of added iodine complexes with New Zealand
www.outreach.canterbury.ac.nz
the College of Science
added,
Figure
solution’s green
3 Freshly squeezed orange
would become
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as shown.
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colour of the
endpoint
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solution reflects
of the titration.
solution
Right
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this case.
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the colour
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and
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solution of iodine.
reflects Centre
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acid the solution’s
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excess of added become darkerwith
iodine complexes shown.
the
orange juicestarchand is unaffected by addition of iodine. Centre flask: Email:
Newoutreach@canterbury.ac.nz
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endpoint
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the
indicator,
ascorbic
3 Freshly
giving the solution
of thesqueezed
titration.
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Left flask:
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Christchurch
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green colour
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case. This isexcess
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of added iodine of the titration.
complexes with the
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starch
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furthergiving iodine thesolution
solution a greenwerecolour
to beinadded,
this case.the
endpoint of the titration. Right flask: If further iodine solution were to be
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green colour
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colour would as shown.
become darker as shown. www.outreach.canterbury.ac.nz
Email: outreach@canterbury.ac.nz
Figure 4 Red capsicum. Left flask: Before the endpoint, the solution www.outreach.canterbury.ac.nz
retains its original colour. Centre flask: Once all the ascorbic acid has been
oxidised, a slight excess of added iodine forms a dark complex with starch
indicator giving a purple colour. This is the titration endpoint. Right flask:
If a further mL or two of iodine were to be added after the endpoint, the
solution would develop the dark purple colour shown here.
Figure 4 Red capsicum. Left flask: Before the endpoint, the solution
3Figureits4original
retains Red capsicum. Left Centre
colour. flask: Before theOnce
flask: endpoint,
all the
thesolution
ascorbic acid has been
retains its original colour. Centre flask: Once all the ascorbic acid has been
oxidised, a slight excess of added iodine forms a dark complex with starch
oxidised, a slight excess of added iodine forms a dark complex with starch
indicator
indicatorgiving
givingaa purple colour.
purple colour. ThisThis
is theistitration
the titration
endpoint.endpoint.
Right flask: Right flask:
If a further mL
If a further mLoror two
two ofofiodine
iodine
werewere
to beto be added
added after theafter the the
endpoint, endpoint, the
solution
solutionwould
woulddevelop
develop thethe dark
dark purple
purple colourcolour shown here.
shown here.
Figure 4 Red capsicum. Left flask: Before the endpoint, the solution
3 43retains
Figure Red capsicum. LeftCentre
its original colour. flask:flask:
Before
Oncethe endpoint,
all the the
ascorbic acid hassolution
been
retains its original
oxidised, colour.
a slight excessCentre
of addedflask: Oncea all
iodine forms darkthe ascorbic
complex acid has
with starch
indicator giving a purple colour. This is the titration endpoint. Right flask:
been oxidised, a mL
If a further slight excess
or two ofwere
of iodine addedto beiodine forms
added after a dark complex
the endpoint, the
with starch indicator
solution giving
would develop theadark
purple
purplecolour. This here.
colour shown is the titration
endpoint.
3 Right flask: If a further mL or two of iodine were to be
added after the endpoint, the solution would develop the dark purple
colour shown here.