College of Science

Determination of Vitamin C Concentration by Titration

(Redox Titration Using Iodine Solution) Safety
Lab coats, safety glasses and enclosed footwear must be worn at all times in the laboratory.

Solutions Needed
Iodine solution: (0.005 mol L1). Weigh 2 g of potassium 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 water and swirl for a few minutes until iodine is dissolved. Transfer iodine solution to a 1 L volumetric flask, making sure to rinse all traces of solution into the volumetric flask using distilled water. Make the solution up to the 1 L mark with distilled water.

Introduction

This method determines the vitamin C concentration in a solution by a redox titration using iodine. Vitamin C, more properly called ascorbic acid, is an essential antioxidant needed by the human body (see additional Starch indicator solution: (0.5%). Weigh 0.25 g of notes). As the iodine is added during the titration, the soluble starch and add it to 50 mL of near boiling water ascorbic acid is oxidised to dehydroascorbic acid, while in a 100 mL conical flask. Stir to dissolve and cool before (Redox Titration the iodine is reduced to iodide ions. Using Iodine Solution) using. ascorbic acid + I2 2 I + dehydroascorbic acid Introduction Thisiodine method determines the vitamin C concentration Due to this reaction, the formed is immediately 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 acid has been oxidised, iodine solution antioxidant needed by the human body (see additional notes). As the iodine added during the titration, the the excess iodine is free to react with is the starch acid is oxidised to dehydroascorbic acid, while indicator, forming theascorbic blue-black starch-iodine complex. the iodine is reduced to iodide ions. This is the endpoint of the titration.

Determination of Vitamin C Concentration by Titration

The method is suitable for with vitamin tablets, Due touse this reaction, the iodineC formed is immediately fresh or packaged fruit juices solid fruits and reduced toand iodide as long as there is any ascorbic acid present. Once all the ascorbic acid has been oxidised, vegetables.
the excess iodine is free to react with the starch

ascorbic acid + I2 2 I + dehydroascorbic acid

indicator, forming the blue-black complex. NB: This method is more straight forward thanstarch-iodine the This is the endpoint of the titration. alternative method using potassium iodate, but as method is is suitable use with vitamin the potassium iodate The solution more for stable than theC tablets, fresh or packaged fruit juices and solid fruits and iodine as a primary standard, the alternative method is vegetables. more reliable. NB: This method is more straight forward than the

Equipment Needed
burette and stand 20 mL pipette

alternative method using potassium iodate, but as the potassium iodate solution is more stable than the iodine as a primary standard, the alternative method is more reliable.
Conical flask

Equipment Needed 100 mL or 200 mL volumetric flask

burette and stand 100 mL or 200 mL volumetric flask 20 mL pipette

Vitamin C sample solution

10 mL and 100 mL measuring cylinders

250 mL conical flasks

250 mL conical flasks 10 mL and 100 mL measuring cylinders

Solutions Needed
Iodine solution: (0.005 mol L1). Weigh 2 g of potassium

Method

4. Calculate the concentration in mol L1 of ascorbic acid in the solution obtained from fruit/vegetable/ Sample Preparation 3. Using equation the titration (below) Starch indicator solution: (0.5%). Weigh 0.25 g of juice. Also, the calculate the of concentration, in mg/100mL determine the number of moles of ascorbic soluble starch and add it to 50 mL of near boiling water or mg/100g of ascorbic acid, in the sampleacid of fruit/ For vitamin C tablets: Dissolve a single tablet in 200 mL reacting. in a 100 mL conical flask. Stir to dissolve and cool before vegetable/juice. of distilled water (in a volumetric flask if possible). using. ascorbic acid + I2 2 I + dehydroascorbic acid For fresh fruit juice: Strain the juice through Notes cheesecloth to remove seeds and pulp which may block Additional 4. Calculate the concentration in mol L1 of ascorbic Method pipettes. 1. acid Iodine stains both skin and clothing so proper care is in the solution obtained from fruit/vegetable/juice. advised. If staining does occur, alcohol may remove ForSample packaged fruit juice: This may also need to be Also, calculate the concentration, in mg/100mL or mg/ Preparation skin stains and cleaners are available for fabric stains. strained through cheesecloth if it contains a lot of pulp 100g of ascorbic acid in the sample of fruit/vegetable/ For vitamin C tablets: Dissolve a single tablet in 200 mL or seeds. 2. juice. Vitamin C, or ascorbic acid, is a water soluble of distilled water (in a volumetric flask if possible). antioxidant that plays a vital role in protecting the For fruits and vegetables: Cut a 100 g sample into small For fresh fruit in juice: Strainand the juice through body from infection and disease. It is not synthesised pieces and grind a mortar pestle. Add 10cheese mL Additional Notes cloth to remove seeds and pulp which may block by the human body and therefore must be portions of distilled water several times while grinding 1. Iodine stains bothsources skin and so proper care pipettes. acquired from dietary clothing primarily fruits and the sample, each time decanting off the liquid extract is advised. If staining does occur, alcohol may remove vegetables. The chemical structure and antioxidant into a 100 mL volumetric flask. strain the For packaged fruit juice: ThisFinally, may also need toground be skin stains and cleaners are available for fabric stains. (reducing) action of ascorbic acid are illustrated in fruit/vegetable pulp through cheesecloth, rinsing the strained through cheese cloth if it contains a lot of pulp the redox half equation below: 2. Vitamin C, or ascorbic acid, is a water soluble pulp a few 10 mL portions of water and collecting or with seeds. antioxidant that plays a vital role in protecting the body all filtrate and washings in the volumetric flask. Make For fruits and vegetables: Cut a 100 g sample into small 3. The concentration of the prepared iodine solution from infection and disease. It is not synthesised the extracted solution up to 100 mL with distilled water. can be more accurately determined by titrationby with pieces and grind in a mortar and pestle. Add 10 mL the human body and therefore must be acquired from a standard solution of ascorbic acid or a standard Alternatively the 100 g sample of fruit or vegetable portions of distilled water several times while grinding dietary sources primarily fruits and vegetables. The solution of potassium thiosulfate using a starch may be blended in atime fooddecanting processoroff together with the sample, each the liquid extract chemical structure and antioxidant (reducing) action of about mL mL of distilled water. After blending, into50 a 100 volumetric flask. Finally, strainstrain the ground indicator. This should be done if possible as iodine ascorbic acid are solutions can beillustrated unstable. in the redox half equation the pulp through cheesecloth, it with a fewthe fruit/vegetable pulp throughwashing cheese cloth, rinsing below: 10mL portions of distilled water, and makeand thecollecting extracted pulp with a few 10 mL portions of water HO HO solution up to 100 mL in a volumetric flask. flask. Make all filtrate and washings in the volumetric oxidation O O O O HO + 2H+ + 2ethe extracted solution up to 100 mL with distilled water. HO

Titration Alternatively the 100 g sample of fruit or vegetable may

be blended a food processor together with about 1. Pipette a 20in mL aliquot of the sample solution into 50 mL of distilled water. After blending, strain the a 250 mL conical flask and add about 150 mL of pulp through cloth, washing it with a few 10 distilled watercheese and 1 mL of starch indicator solution. mL portions of distilled water, and make the extracted 2. Titrate the sample with 0.005 mol L1 iodine solution. solution up to 100 mL in a volumetric flask. The endpoint of the titration is identified as the first permanent trace of a dark blue-black colour due to Titration the starch-iodine complex.

HO

OH

reduction

ascorbic acid

dehydroascorbic acid

3. The concentration of the prepared iodine solution 4. The average titre volume should ideally be in the can be more accurately determined by titration with a range of 10 30 mL. If the titre required for a 20 mL standard solution of ascorbic acid or a standard solution aliquot of sample solution is well outside this range of potassium thiosulfate using a starch indicator. This then a larger or smaller aliquot volume should be should be done if possible as iodine solutions can be 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 further aliquots sampleinto 4. 1. Pipette a 20 mLwith aliquot of the sampleof solution The average titre volume should ideally be in the sample. solution you obtain results (titres a 250 mLuntil conical flask andconcordant add about 150 mL of distilled the range of 10 30 mL. If the titre required for a 20 5. mL Ascorbic acid is susceptible tois oxidation by this agreeing within 0.1 mL). indicator solution. water and 1 mL of starch aliquot of sample solution well outside atmospheric oxygen over time. For this reason, the range then a larger or smaller aliquot volume should 2. Titrate the sample with 0.005 mol L1 iodine samples should be prepared immediately before be chosen. If the volume of the titre is too low, dilute Calculations solution. The endpoint of the titration is identified as the titrations. However, if the samples have to bethe the standard. If the titre volume is too high, dilute the first permanent trace of a dark blue-black colour due prepared several hours earlier, oxidation can be 1. Calculate the average volume of iodine solution used sample. to the starch-iodine complex. minimised by the addition of a small amount of from your concordant titres. 5. Ascorbic acid is susceptible oxidation by acid (eg 1 g oxalic acid perto 100 mL of sample 3. Repeat the titration with reacting. further aliquots of sample oxalic 2. Calculate the moles of iodine atmospheric oxygen over time. For this reason, the solution). solution until you obtain concordant results (titres 3. Using thewithin equation of the titration (below) determine samples should be prepared immediately before the agreeing 0.1 mL). 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 sample solution used. If the solutions are colourless ascorbic acid + I2 2 I + dehydroascorbic acid Calculations addition of a small amount of oxalic acid (eg 1 g oxalic or are pale in colour, there is no problem identifying acid per 100 mL of sample solution). 1. Calculate the average volume of iodine solution used from your concordant titres. 6. Identification of the endpoint in this titration is significantly affected by the colouration of the sample 2. Calculate the moles of iodine reacting. 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 endpoint. For coloured juices there can the and it strongly is advised to carry out a rough be a problem with the endpoint and it is advised titration in order to become familiar with any distinct in colour, there is no problem identifying the endpoint. to carry out awhich rough titration in order to(it become colour change occurs at the endpoint may For strongly coloured there a problem with just be a juices darkening ofbe the colour) This will also help by familiar with anycan distinct colour change which the endpoint and it is advised to carry out a rough establishing anendpoint, approximate volume of iodate solution Figure 1 Vitamin C tablet. Left photo: before endpoint, added iodine reacts occurs at the (itdistinct may just be a darkening Figure 1 Vitamin C tablet. Left photo: before endpoint, added iodinetitration in order to become familiar with any with ascorbic acid leaving the solution colourless. Centre photo: At the required. of the colour) This will also help by establishing an reacts with ascorbic acid leaving the solution colourless. colour change which occurs at the endpoint (it may
titration endpoint all the ascorbic acid has reacted and the excess iodine

injust colour, is no problem identifying the endpoint. Centre photo: Atthe the titration endpoint all the ascorbic acid hasphoto: darkening of the colour) This will also help by reacts with starch indicator to give a pale blue colour. Right If be athere approximate volume of iodine solution required. 7. The above method may be used to carry out a For strongly coloured juices there can be a problem with addition of1iodine continued after the the endpoint, further iodine-starch establishing an approximate volume of iodate solution reacted and the excess iodine reacts with starch indicator to give Figure Vitaminis C tablet. Left photo: before endpoint, added iodine reacts in colour, there is no problem identifying the endpoint. number of interesting investigations regarding the endpoint and it is advised to carry out a rough is formed. in each of these images a flask showing pale ascorbic acidNB: leaving theIf solution colourless. Centre photo: At the the 7. The above method may be used to carry out the a required. a palecomplex bluewith colour. Right photo: addition of iodine is continued after For strongly coloured juices there can be a problem with titrationof endpoint all the ascorbic acid has reacted and the excess iodine blue colour the endpoint is shown for comparison. concentration of vitamin C in various foods and drinks: titration in order to become familiar with any distinct the endpoint, further iodine-starch complex is formed. NB: in each of reacts with the starch indicator to give a pale blue colour. Right photo: If number interesting investigations regarding the 7. The above method may used to carry out a the endpoint and it isof advised to carry out rough colour change which occurs at be the endpoint (it a may addition of iodine is continued after theblue endpoint, further iodine-starch these images a flask showing the pale colour of the endpoint is Vitamin C content of different types of fruits/ number of interesting investigations regarding the titration in order to become familiar with any distinct concentration of vitamin C in various foods and complex is formed. NB: in each of these images a flask showing the pale just be a darkening of the colour) This will also help by shown for blue comparison. colour of the endpoint is shown for comparison. concentration of vitamin C in various foods and drinks: vegetables/juices. colour change which occurs at the endpoint (it may establishing an approximate volume of iodate solution Figure 1 Vitamin C tablet. Left photo: before endpoint, added iodine reacts drinks: with ascorbic acid leaving the solution colourless. Centre photo: At the just be aC darkening of colour) This will also help by required. Vitamin content of different types ofdifferent fruits/ Vitamin Cthe content of types/brands of vitamin titration endpoint all the ascorbic acid has reacted and the excess iodine Vitamin C content of different types of fruits establishing an approximate volume of iodate solution Figure Vitamin C tablet. Left photo: endpoint, added iodine If reacts vegetables/juices. reacts1with the starch indicator to givebefore a pale blue colour. Right photo: 7. The above method may be used to carry out a tablets. with ascorbic acid leaving the solution Centreiodine-starch photo: At the addition of iodine is continued after thecolourless. endpoint, further required. vegetables/juices. number of interesting investigations regarding of the Vitamin C content of different types/brands vitamin titration endpoint all the acid has reacted and the excess iodine complex is formed. NB: inascorbic each of these images a flask showing the pale Vitamin C of tablets or food/drink in the reacts with the starch indicator to give pale blue colour. Right photo: If blue colour of the endpoint is shown foracomparison. concentration of vitamin C content in various foods and drinks: tablets. 7. The above method may be used to carry out a types/brands Vitamin C content of different of addition of iodine is continued after the endpoint, further iodine-starch presence and absence of added oxalic acid or metal ions interesting investigations regarding Vitamin Cof content of different types of fruits/ complex is formed. NB: in each of these images a flask showing the pale number Vitamin C content of tablets or food/drink in the the vitamin tablets. blue colour of the endpoint is shown for comparison. over various periods of time. concentration of vitamin C inoxalic various foods and drinks: vegetables/juices. presence and absence of added acid or metal ions Vitamin C content of tablets or food/drink in the over various periods of time. Vitamin C content of different types of fruits/ Vitamin C content of different types/brands of vitamin Vitamin C content of food/drink before and after Figure 2 Commercial fruit juice. Left flask: before the endpoint, the colour presence and absence of added oxalic acid or vegetables/juices. tablets. Vitamin content of food/drink before and after subjection to cooking conditions. of the solution reflectsfruit thejuice. paleLeft yellow ofendpoint, the fruit juice. Centre flask: C Figure 2 Commercial flask: colour before the the colour Figure At the titration endpoint all the ascorbic has reacted and any excess subjection to cooking conditions. of the solution reflects the pale yellow colouracid of the fruit juice. Centre flask: content metal ionsor over various periods of time. Vitamin C content of tablets food/drink in the Vitamin of types/brands of vitamin 2 Commercial fruit juice. Leftall flask: before the endpoint, the colour of Atreacts the titration the ascorbic acid has reacted and any excess complex. CVitamin Cdifferent content of fruits/vegetables at different iodine withendpoint the starch indicator to form a dark blue-black presence and absence of added oxalic acid or metal ions Vitamin C content of fruits/vegetables at different tablets. the solution reflects the pale yellow colour of the fruit juice. iodine reacts with the starch indicator to form a dark blue-black complex. In this case the result is a darkening of the solutions colour from yellow Vitamin C content of food/drink before and after stages of ripeness. In this result is aendpoint darkening of all the solutions colour from yellow over periods of time. stages Centre flask: At case thethe titration the ascorbic acid hasjust a to brown-grey. Right flask: This illustrates the effect of adding mL various or of ripeness. to brown-grey. Right flask: This illustrates the effect of adding just a mL or Vitamin C content of tablets or food/drink in the subjection tobefore cooking conditions. two more ofexcess iodine solution after the endpoint is reached, resulting the reacted and any iodine with the starch indicator to in two more of iodine solutionreacts after the endpoint is reached, resulting in the Vitamin C content of food/drink and after presence and absence of added oxalic acid or metal ions Figure 2 of Commercial fruit juice. Left flask: before the endpoint, the colour formation further iodine-starch complex. formation of further iodine-starch complex. form a dark blue-black complex. In this case the result is a darkening subjection to cooking conditions. of the solution reflects the pale yellow colour of the fruit juice. Centre flask: Contact Us Contact Us Vitamin C content of fruits/vegetables at different over various periods of time. At the titration endpoint all the ascorbic has reacted and any excess of the solutions colour from yellow toacid brown-grey. Right flask: Vitamin C content of fruits/vegetables at different iodine reacts with the starch indicator to form a dark blue-black complex. you stages If have any questions orripeness. comments relating to this If have any questions or comments relating to this you Vitamin C content of of food/drink before and after This illustrates the effectis of adding just a mL the orcolour two more of iodine Figure Commercial juice. Leftof flask: before endpoint, colour In this2case the resultfruit a darkening the solutions from the yellow stages of ripeness. experiment, please contact us: subjection to cooking conditions. the solution reflects the pale yellow colour of the fruit juice. Centre flask: solutionof after the endpoint is reached, resulting in the formation of to brown-grey. Right flask: This illustrates the effect of adding just a mL or experiment, please contact us:
two more of iodine solution after the endpoint is reached, the At the titration endpoint all the ascorbic acid has reactedresulting and any in excess further iodine-starch complex. formation of further iodine-starch complex.

Vitamin C content ofUs fruits/vegetables at different Contact Outreach Contact Us College of stages ofScience ripeness. College of Science of Canterbury IfUniversity you have questions or comments relating to this Ifany you have any questions or comments relating to this University of Canterbury Private Bag 4800 experiment, please contact us: experiment, please contact us. Please note that this Christchurch Contact Us Bag 4800 Private Outreach service is for senior school chemistry students in New Zealand Christchurch If youof have any questions or comments relating to this Science Figure 3 Freshly squeezed orange juice. Left flask: before the endpoint, the College New Zealand only. We regret we are unable to respond Phone: +64 3 364 2178contact New Zealand colour of the solution reflects the bright orange colour of fresh orange juice University experiment, please us: of Canterbury and is unaffected by addition of iodine. Centre flask: Once all the the ascorbic to queries from overseas. Fax: +64 3 364 2490 Figure 3 Freshly squeezed orange juice. Left flask: before endpoint, the Bag 4800 acid has been oxidised, a slight excess of added iodine complexes with the Private Outreach Phone: +64 3 364 2178 colour of the solution reflects the bright orange colour of fresh orange juiceoutreach@canterbury.ac.nz Email: starch indicator, giving the solution a green colour in this case. This is the Christchurch CollegeOutreach of Science and is unaffected addition iodine. Centre Once the ascorbic Fax: +64 3 364 2490 endpoint of the by titration. Rightof flask: If further iodineflask: solution were all to be New Zealand www.outreach.canterbury.ac.nz acid has been a slight excess of added iodine complexes with the added, the oxidised, solutions green colour would become darker as shown. College of Science University of Canterbury Email: outreach@canterbury.ac.nz Figure 3 Freshly squeezed orange juice. Left flask: before thein endpoint, the This is the starch indicator, giving the solution a green colour this case. Phone: +64 3 364 2178 Private Bag 4800 colour of the solution reflects the bright orange colour ofbefore fresh orange juice Figure 3 Freshly squeezed orange juice. Left flask: the endpoint, University of Canterbury endpoint of the titration. Right flask: If further iodine solution were to be and isthe unaffected by addition of iodine. Centre flask: Once allcolour the ascorbic Fax: +64 3 364 2490 www.outreach.canterbury.ac.nz Christchurch the colour of solution reflects thewould bright orange fresh added, solutions green colour darker asof shown. Private Bag 4800 acid the has been oxidised, a slight excess of added become iodine complexes with the orange juice and is unaffected by addition of iodine. Centre flask: Email: Newoutreach@canterbury.ac.nz Zealand starch indicator, giving the solution a green colour in this case. This is the Christchurch endpoint of thesqueezed titration. Right flask: If further iodine solution were to be 3 Freshly orange juice. Left flask: before theexcess endpoint, the Once allFigure the ascorbic acid has been oxidised, a slight of added www.outreach.canterbury.ac.nz +64 3 364 2178 added, the solutions colour would orange becomecolour darker of as fresh shown. colour of the solution green reflects the bright orange juice Phone:New Zealand iodine complexes with the starch indicator, giving the solution a and is unaffected by addition of iodine. Centre flask: Once all the ascorbic Fax: +64 3 364 2490 Phone: +64 3 364 2178 green colour inbeen this case. This isexcess the endpoint of the titration. acid has oxidised, a slight of added iodine complexes with the Email: outreach@canterbury.ac.nz starch thesolution solution a green inadded, this case.the This is the Right flask: If indicator, furthergiving iodine werecolour to be solutions Fax: +64 3 364 2490 endpoint of the titration. Right flask: If further iodine solution were to be www.outreach.canterbury.ac.nz green colour become as shown. Email: outreach@canterbury.ac.nz added,would the solutions greendarker colour would become darker as shown. www.outreach.canterbury.ac.nz Figure 4 Red capsicum. Left flask: Before the endpoint, the solution
iodine reacts with the starch indicator to form a dark blue-black complex. In this case the result is a darkening of the solutions colour from yellow 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 formation of further iodine-starch complex.
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.

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Figure 4 Red capsicum. Left flask: Before the endpoint, the solution Figure 4original Red capsicum. Left Centre flask: Before theOnce endpoint, solution 3 retains its colour. flask: all the the 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 giving colour. is the titration endpoint. indicator givinga a purple purple colour. ThisThis is the titration endpoint. Right flask: Right flask: If a further mL of iodine were be added the the endpoint, the If a further mLor or two two of iodine were to beto added after theafter endpoint, solution woulddevelop develop the dark purple colour shown here. solution would the dark purple colour shown here.

3retains Figure Red capsicum. LeftCentre flask:flask: Before the endpoint, the solution 3 4 its original colour. Once all the ascorbic acid has been oxidised, a slight excessCentre of addedflask: iodine forms a all dark complex with starch retains its original colour. Once the ascorbic acid has indicator giving a purple colour. This is the titration endpoint. Right flask: been oxidised, a mL slight excess ofwere added forms a dark complex If a further or two of iodine to beiodine added after the endpoint, the with starch indicator giving adark purple colour. This here. is the titration solution would develop the purple colour shown endpoint. Right flask: If a further mL or two of iodine were to be 3 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