UNIVERSITY OF SANTO TOMAS

Faculty of Pharmacy

Department of Biochemistry

S.Y. 2014-2015

Comparative Study on the Purity of Generic and Branded Vitamin C

Supplements sold in Metro Manila by Iodine Titration

Hajime Q. Nakaegawa

David T. Luces

Dony Lean P. Malapitan

Jennelie B. Manaog

Charisse V. Misola

Chrisha R. Montemayor

B.S. Biochemistry 1-A

Group No. 5

Mrs. Maritess D. Cation

Asst. Prof. Ma. Teresa R. Suratos

Research Advisers
 ABSTRACT

The research aims to make a comparative study on the purity of generic and branded

Vitamin C supplements sold in known pharmaceutical drugstores in Metro Manila. It will be done

through the process of Iodine Titration, using a reaction between Standard KIO3 solution and

Ascorbic Acid, titrating it with Sodium thiosulfate solution. The research requires the use of a

titration set-up and six compounds, namely Soluble Starch (C6H10O5), Potassium iodide (KI),

Sodium thiosulfate (Na2S2O3), Iodine (I2), Sodium carbonate (Na2CO3), and Sulfuric acid (H2SO4).

The researchers titrated three generic and three branded Vitamin C supplements in order to

determine its effectiveness and quality. They gathered results and determined the purity of Vitamin

C through computations using dimensional analysis. The results show that <INSERT

CONCLUSION HERE>. This shows that <INSERT INSIGHTS HERE>

 TABLE OF CONTENTS

I. Introduction

a. Background of the Study

b. Objectives

c. Hypothesis

d. Significance of the Study

e. Scope and Limitations

f. Review of Related Literature

g. Definition of Terms

II. Methodology

a. Materials

b. General Procedure

i. Preparation Phase

ii. Experimentation Phase

iii. Evaluation Phase

c. Procedure Flowchart

d. Research Design

e. Risk Assessment

f. Disposal Methods

III. Results and Discussion

IV. Conclusion

V. Recommendation

VI. Bibliography

VII. Time Table

VIII. Expenses Table

IX. Research Plan

X. Appendix
 ACKNOWLEDGEMENTS

The researchers would like to thank their research advisers, namely Mrs. Maritess D.

Cation and Asst. Prof. Ma. Teresa R. Suratos for their continuous support to guide and to give

insights to the researchers in the making of their research. Also for the provision of the necessary

apparatus and compounds needed by the researchers to make their study possible.
 INTRODUCTION

Vitamin C (Ascorbic Acid) is a common and important daily nutritional need in human’s

diet. The absence of Vitamin C inhibits the protein and collagen from forming fibers properly which

results in skin injuries and fragility of blood vessels. It also gives many benefits when it comes to

synthesis of several peptide hormones and transmitters. Although Vitamin C occurs naturally in

many fruits and vegetables, many Filipino families that belong to the financially-challenged group

take Vitamin C supplements to supplement their intake. With the presence of expensive food and

poor supply of nutritious food, these families may now take more Vitamin C from the supplements

they usually take than they can ever imagine.

Background of the Study

There had been a series of studies on the enormous nutritional benefits of Vitamin C. In

fact, man is one of the few animals that cannot synthesize Vitamin C and relies on dietary sources

to meet nutritional needs. Vitamin C is known to act as specific electrons for eight enzymes in the

body including those involved in collagen production, brain neurotransmitter synthesis, and

pituitary peptide hormone production. It also appears to have several non-enzymatic functions

based on its ability as a reducing agent (i.e., antioxidant) and to reduce free radicals thought to be

important in aging and carcinogenesis.

Many studies have tried to identify the Vitamin C intake level and tissue saturation level

at which potentially protective effect is maximized. Thus, the researchers want to take a closer look

on these studies and analyzing them for their research.

 According to Julius Goepp, MD, Vitamin C is rapidly finding new applications in

protecting against endothelial dysfunction, high blood pressure, and the blood vessel changes that

precede heart disease. Additional research is discovering that vitamin C can be helpful in preventing

asthma, protecting against cancer, and supporting healthy blood sugar levels in diabetics. While

often taken for granted, Vitamin C is a critical supplement in your program to improve cardiac

health and avoid degenerative diseases. Listed below are some important researches found about

Vitamin C:

 Vitamin C supplements reduce cellular DNA damage that is the vital first step in cancer

initiation and also reduce the inflammatory changes that allow a malignant cell to grow

into a dangerous tumor.

 Vitamin C supplements enhance the health-promoting effects of exercise and reduce

exercise-induced oxidative damage.

 Vitamin C also supplements dramatically combat the oxidative damage caused by smoking

and exposure to tobacco smoke.

 In respiratory conditions, vitamin C supplements help avert or shorten the duration of

common colds and may mitigate the risk of serious respiratory conditions like asthma.

 Vitamin C supplements can speed the clearance of the stomach disease-causing bacterium

Helicobacter pylori and cut the risk of gastric cancer it causes.

Vitamin C has been an ingredient towards innovation in many fields and in a lot of food

supplements that claim to take care of one’s nutritional needs. With these amazing facts about

Vitamin C and the pressing desire to add nutrients to what Filipinos eat, the researchers came up

with this study of determining the purity of Vitamin C to directly enhance the nutrient content being

taken by the Filipino families.

Objectives

The main objective of this research is to make a comparative study on the purity of generic

and branded Vitamin C supplements sold in known pharmaceutical drugstores in Metro Manila. It

will be done through the process of Iodine Titration, using a reaction between Standard KIO 3

solution and Ascorbic Acid titrating it with Sodium thiosulfate solution. To ensure the quality and

effectiveness of these sold Vitamin C supplements whether they are worth buying or not is the main

purpose of this research. This way we can help Filipino families in their decision making towards

this matter.

Hypothesis

The hypothesis on this research is that after Iodine Titration has been done and all

computations have been made, the amount of Vitamin C found in both branded and generic

supplements will have no to very minimal difference. The researchers also hypothesize that the

branded and generic Vitamin C supplements will follow the percentage amount found on the

Nutritional Content of the supplement’s cover. This way, it is safe to prove that generic Vitamin C

supplements can be bought without hesitation even though it is cheaper than branded Vitamin C

supplements. If subjected to a high percentage error in the research, the researchers hypothesize

that this will be only due to errors in the process which can be found in the Risk Assessment of this

paper.
Significance of the Study

The research:

● will prove the direct and beneficial effect of branded and generic Vitamin C supplements

sold in pharmaceutical drugstores in Metro Manila

● will give a bird’s eye view on the difference between branded and generic supplements

● will ensure the quality and effectiveness of these sold Vitamin C supplements whether

they are worth buying or not

● may give Filipino families, especially the financially-challenged group, direct options

and decisions on the quality of these Vitamin C supplements

● may increase the demand of these Vitamin C supplements that can give direct beneficial

effect to manufacturers once proven on the effectiveness and quality of the supplements

Scope and Limitations

The research will dwell on the purity of branded and generic Vitamin C supplements sold

in known pharmaceutical drugstores across Metro Manila. The research will measure and make a

direct comparison on the differences between these two categories of Vitamin C supplements.

These will be done through one common laboratory technique known as Iodine Titration, a redox

titration where the appearance or disappearance of elementary iodine indicates the end point. It will

also tackle the effectiveness and demand for Vitamin C supplements in Metro Manila.

The research would have not enough time to further expand the proportion of analysis of

the study. It is only limited to checking on the one part of these supplements, which is the purity of

Vitamin C. The research will not be able to determine other possible topics that can be tackled such

as an analysis of the different substances present and a deeper understanding on how these Vitamin

C supplements are being manufactured.

Review of Related Literature

Nature and Sources of Vitamin C:

The structure of Vitamin or Ascorbic Acid is

shown in Figure 1. Most species are able to make

ascorbic acid from glucose but primates and some

birds lack a key enzyme (gulanolactone oxidase) on

this synthetic pathway. Ascorbic Acid is water soluble Figure 1. Structure of Vitamin C

in the solution and is readily oxidized to dehydroascorbic acid; thus it acts as an anti-oxidant.

Fruits and vegetables (including potatoes) are the classical sources of Vitamin C.

Significant amounts are also present in offal and fresh cow milk.

Functions of Vitamin C:

Vitamin C or Ascorbic Acid is necessary for the functioning of the enzyme proline

hydroxylase. This enzyme hydroxylates residues of the amino acid proline to hydroxyproline once

it has been incorporated into the structural protein collagen. Collagen is a major structural protein.

It has the capacity to form strong insoluble fibers and is the most abundant protein in mammals and

serve to hold cells together. It is a structural component of bones, skin, cartilage and blood vessels,

and indeed fulfills a structural role in most organs. Some other functions are listed below.

● It is a cofactor in the synthesis of the important nerve transmitter noradrenalin (norepinephrine)

● It is a cofactor in the synthesis of carnitine, which is necessary for the transfer of long-chain fatty

acids into the mitochondrion where β–oxidation occurs.

● Large doses of Vitamin C increase the absorption of non- harm iron from the gut
Deficiency states

Scurvy is a disease caused by lack of Vitamin C. According to the World Health

Organization, intake of Vitamin C differ in different places. In the Philippines, the intake of

Vitamin C should at least be 75mg/day for moderately active adult males. Studies with radio

labelled Ascorbic acid have shown that when doses of <100mg of Ascorbic acid are ingested by

humans, the absorption efficiency may be as high as 98% (Olson & Hodges, 1987). When larger

amounts are ingested, the absorption efficiency decreases to 70% at a dose of 180 mg, to 50% at

1500 mg, and to 16% at 12000 mg. Vitamin C absorption is increased significantly when given

with food, perhaps because of a slowing of intestinal transit time. Thus, the absorption efficiency

seems to be 90% or more over a range of 20-120 mg per day of usual intakes in food. Table 1 shows

the daily intakes of vitamin C recommended by the joint FAO/WHO expert group (1970). It was

stated that heavy physical activity may increase the need for Vitamin C, but that there was no

definite evidence for this.

Table 1. FAO/WHO recommended daily allowances for vitamin C Vitamin C (mg/day)

Infants (0–6 months) Breast-feeding

Infants (6–12 months) 20

Children > 13 years and adults 30

Pregnant women (2nd and 3rd trimester) 50

Lactating women 50
Generic and branded Vitamin C supplements:

According to the 2013 OTC/DTC Study, about 16% of adults (37 million) agree that it’s

worth paying more for branded prescription medications rather than getting generic products. In

contrast, 45% of adults (106 million) disagree with that statement. Nearly 40 million of them

disagree while still choosing to treat a health condition with a branded prescription drug. Adults

with HH annual incomes between $50-75k are 32% less likely to agree that it’s worth paying more

for branded Rx vs. generics. On the opposite side of the spectrum, they found that adults with $250k

or more in annual HH income are 40% more likely to agree that it’s worth paying more for branded

Rx. Graph 1 shows the number of millions of people who agree or disagree that is worth paying for

more branded than generic products.

Graph 1. Generic Vs. Branded Prescription Drugs: Perceptions and Actions

45
40
35
30 Treating a Health Condition
with a Generic Description
In Millions

25 Product
20 Treating a Health Condition
with a Branded Prescription
15
10
5
0
Agree that its worth Disagree that its worth
paying more for paying more for
Branded Prescription Branded Presciption
than buying Generic than buying Generic
Source: Kantar Media's 2013 MARS OTC/ DTC Study
Potency of Sales of Vitamin C in the Philippines:

According to a food consumption survey conducted by the Food and Nutrition Research

Institute of the Department of Science and Technology (FNRI-DOST), Filipinos mostly rely on

vegetables for their Vitamin C requirements. However, no more than 30 percent of the population

meets the estimated average requirement (EAR) which is 80 percent of the recommended intake of

Vitamin C. With potential sales of at least P150 billion annually, the local vitamins and supplements

industry is robust and continues to rise with encouraging numbers. The dietary and nutritional

supplement market generated $50B sales worldwide in 2010. Multivitamins make up a major

portion of this market and grew almost 7.9 %

Manufacturing Process of Supplements:

The synthesis of Ascorbic acid was achieved by Reichstein in 1933, followed by industrial

production of Ascorbic acid two years later by Roche. Today, vitamin C identical to that occurring

in nature is produced on a very large industrial scale. Figure 2 shows an outline on how supplements

are being made.

Figure 2. Manufacturing Process of Supplements

 For vitamin supplements, particle size is extremely important in determining how well the

formula will run through the tabletting machine. In some cases, the raw vitamins arrive from the

distributor milled to the appropriate size for tabletting. In other cases, a wet granulation step is

necessary. In wet granulation, the fine vitamin powder is mixed with a variety of cellulose particles,

then wetted. The finished vitamin mixture can be compressed into supplements, sometimes with a

coating, or encapsulated in preformed gelatin capsules. A manufacturer may coat in two different

colors supplements that are the same size and shape, for identification. Supplements may also be

given an enteric coating—a pH sensitive chemical coating that resists gastric acid.

Definition of Terms

● Vitamin C – also known Ascorbic Acid, with the chemical formula C6H8O6

● Generic drug- is a pharmaceutical product that is manufactured without a license from

the innovator company and marketed after the expiry date of the patent or other exclusive

rights (World Health Organization)

● Branded drug - a drug that has a trade name and is protected by a patent (can be produced

and sold only by the company holding the patent)

● Iodine Titration – also known as Iodometry, is a method of volumetric chemical analysis,

a redox titration where the appearance or disappearance of elementary iodine indicates the

end point.

● RDA – recommended dietary allowance; are quantities of nutrients in the diet that are

required to maintain good health in people.

 METHODOLOGY

A. Materials

The materials used by the researchers includes the titration set-up and six compounds

which were used for experimentation. The titration setup includes the 50mL burette with stopcock,

iron stand, burette clamp, analytical balance, and a wash bottle. In addition to this, glassware

apparatus were also used. This would include the 250mL Erlenmeyer flask, 250L volumetric flask,

25mL volumetric pipette, distilled water, 1L amber bottles and some extra glassware to hold the

necessary reagents and compounds. The six compounds used by the researchers are namely Soluble

Starch (C6H10O5), Potassium iodide (KI), Sodium thiosulfate (Na2S2O3), Iodine (I2), Sodium

carbonate (Na2CO3), and Sulfuric acid (H2SO4)

B. General Procedure

The general procedure is divided into three phases namely: the Preparation Phase, where

they would setup all the necessary solutions needed for the experiment; the Experimentation Phase,

where they would do the Iodine Titration of Vitamin C supplements; and lastly the Evaluation

Phase, where they would actually analyze, compute, and make conclusions toward the experiment.

Preparation Phase

I. Preparation of Starch Indicator

The researchers fill a 100mL beaker to the 100-mL mark with distilled water and place it

on a hot plate until boiling. They weigh out 1g of soluble starch (C6H10O5) and slowly add to the

beaker of boiling water with stirring until the solution is clear.

 II. Preparation of Sodium Thiosulfate Solution

The researchers fill a 500mL beaker to the 500-mL mark with distilled water and place it

on a hot plate. They boil the water for 5 minutes to expel dissolved CO2 gas. After allowing the

distilled water to cool, they weigh out 0.05g of Na2CO3 and place into the 500-mL beaker of boiling

water. They again weigh out 8.7 grams of Na2S2O35H2O (0.07 M) and dissolve it into the 500-

mL beaker of boiling water buffered with Na2CO3. The researchers then place the solution in a 1L

amber bottle properly labelled with “Sodium Thiosulfate Solution”.

III. Preparation of Standard Iodate Solution

The researchers fill a 500mL volumetric flask with 400mL of distilled water. They

accurately weigh out 1g of KIO3 using the analytical electronic balance and record the mass up to

four decimal places. Dissolve the KIO3 into the distilled water contained in the 500-mL volumetric

flask and stir until all solid particles have been dissolved. Fill the volumetric flask to the mark with

distilled water. They determine the exact molarity of this solution using the following molecular

masses: K: 39.0983 I: 126.90447 O: 15.9994. The researchers then placed the solution in a 1L

amber bottle properly labelled with “Standard KIO3 Solution”.

IV. Standardization of Sodium Thiosulfate Solution

The researchers prepared the setup to be used for titration. Then they fill the 50mL burette

with the Sodium Thiosulfate solution. Accurately pipet 50.00 mL of standard KIO 3 solution into

the 250mL Erlenmeyer flask. Then they weigh 2 grams of KI and place it into the flask. Add 10mL

of 0.5 M H2SO4 to the flask. The solution start out as a “red-ish” solution due to the presence of the

triiodide. Titrate the solution with the Sodium Thiosulfate until the solution has lost most of the

red-ish color (should be at least pale yellow). At this point add 2 mL of the starch indicator (may

turn smoky-blue or remain yellowish). Carefully add Sodium Thiosulfate until the solution turns

colorless (may be a milky-white). Record this volume as the end point. The indicator is not added
until before the end point as the triiodide/starch complex can “hold on to” the triiodide in

concentrated solutions and prevent it from reacting with the Sodium Thiosulfate. The researchers

repeat this three times to get an accurate and precise result. Using the known stoichiometry, the

molarity of the Sodium Thiosulfate solution can now be calculated.

Experimentation Phase

I. Reaction of Triiodide with Ascorbic Acid

After preparing and knowing the molarity of the Sodium Thiosulfate solution and preparing

the Standard KIO3 Solution, three branded and three generic Vitamin C supplements are to be used.

The researchers place 60 mL of 0.3 M H2SO4 into the 250mL Erlenmeyer flask. Add 2 Vitamin C

supplements to the flask and dissolve it in the sulfuric acid. Make sure it is completely dissolved.

Then accurately pipet 50.00 mL of standard KIO3 solution into the 250mL Erlenmeyer flask. 24.

Weigh out around 2 grams of KI and place into the flask. Swirl the flask for 1 minute to insure

complete formation of the triiodide complex and complete reaction between the triiodide and the

Vitamin C.

II. Titration of Solution with Sodium Thiosulfate

The solution should start out as a “red-ish” solution due to the presence of the triiodide.

The researchers titrate the solution with the Sodium Thiosulfate until the solution has lost most of

the red-ish color (should be a pale yellow). Then, add 2 mL of the starch indicator (may turn smoky-

blue or remain yellowish). Carefully add Sodium Thiosulfate until the solution turns colorless (may

be a milky-white). Record this volume as the end point. The indicator is not added until before the

end point as the triiodide/starch complex can “hold on to” the triiodide in concentrated solutions

and prevent it from reacting with the Sodium Thiosulfate. The researchers made two trials and it

was done repeatedly to the three generic and three branded Vitamin C supplements.
Evaluation Phase

The researchers are to compute for the amount of Vitamin C present using the known

stoichiometry of the Sodium Thiosulfate/IO3 - and Ascorbic Acid/IO3 - reactions, and then calculate

the average number of moles of Ascorbic Acid contained in each Vitamin C supplement. Using the

formula weight of Vitamin C (C6H8O6, MW = 176.13), they calculate the average number of

milligrams of Vitamin C contained in each supplement along with the standard deviation.

Presentation of data and computations is shown in the Results and Discussion of the research paper.

Procedure Flowchart

Preparation of Different Solutions

Preparation of Preparation of Standardization of
Preparation of
Sodium Standard Sodium
Starch Indicator
Thiosulfate Iodate Solution Thiosulfate

Experimentation Proper
Repetition of
Reaction of Titration of
Titration Process
Triiodide Solution with
for remaining
with Ascorbic Acid Sodium Thiosulfate Vitamin C Supplements

Computation and Analysis

Calculation on the
Calculation on the
Computation on the Average Number of
Number of Moles of
Amount of mg of Vitamin C
Vitamin C Consumed
Vitamin C Present with
in Each Supplement Standard Deviation
Research Design

The Sodium Thiosulfate Solution serve as the independent variable of the research which

was titrated with the Ascorbic Acid Solution. The Ascorbic Acid solution, which contains the two

supplements of Vitamin C, distilled water, and starch indicator, serve as the dependent variable as

the researchers hypothesize no to very minimal difference between the two categories of Vitamin

C supplements.

The Nutritional Content found on the label of the Vitamin C supplement serve as the

constant variable in the setup since it is the number that cannot be changed once it has been assigned

a value. The difference between the dependent variable and the constant variable will be put under

analysis through computations in the Evaluation Phase of the research.

Risk Assessment

Many risks are present in the process of the research. These would include the availability

of the compounds and apparatus needed; and the presence of human error. The researchers take

note on the risks that may lead to the failure of the research. This would include the following:

● Standard KIO3 Solution should be stored in a cabinet and must be out of direct sunlight. It may

not be possible to completely dissolve the I2. If some undissolved I2 is present, decantation of the

solution is necessary and diluting it up to the 1L mark with distilled water. Standardization of the

solution should be repeated.

● I2 sublimes at room temperature. This vapor form is caustic and a potential health threat.

Weighing and mixing with the presence of this element must be done at the fume hood.

● The Sodium Thiosulfate Solution must be used within 24 hours. Using of this solution after 24

hours might lead to an error in the results.

● Vitamin C spontaneously oxidizes when exposed to air. If not kept tightly closed, the potency of

these Vitamin C supplements will slowly be lost. The solution must be used within 24 hours. Do

not store the solution overnight.

Human error will include the following:

● Error in measuring and reading values (parallax error) while in the process of research

● Use of Contaminated Compounds

● Misuse or use of broken/contaminated apparatus in the laboratory

Disposal Methods

The researchers consulted the Material Safety Data Sheet (MSDS) of the used compounds

in order to determine the proper method of disposal for each compound. Each compound, namely

Soluble Starch (C6H10O5), Potassium iodide (KI), Sodium thiosulfate (Na2S2O3), Iodine (I2),

Sodium carbonate (Na2CO3), and Sulfuric acid (H2SO4), must be placed in a sealed black plastic

bag or container before placing it in a trash bin, as designated color for chemical wastes. It is

important to ensure that these bags are perfectly sealed without leaks or damage. Sulfuric acid may

be placed in sealed container or absorbed in vermiculite, dry sand, earth, or a similar material. It

may also be diluted and neutralized.

Dry materials contaminated with chemicals (paper, rags, towels, wipes, or spill

contaminated materials) must be double bagged in heavy-duty plastic bags. Tape all containers of

chemically-contaminated dry materials securely shut and label the outside of any container clearly

with descriptions of its contents and the chemical contaminants. Biologically contaminated

glassware and sharps should be placed into appropriate sharps containers.

RESULTS AND DISCUSSION
CONCLUSION
 RECOMMENDATION

The researchers recommend anyone who will do this research to:

● Search for a broader and wider discussion in Vitamin C. It is recommended that anyone who will

do this research will tackle other important food good for human’s diet such as fruits and

vegetables, or even fruit juices being sold across different stores.

● Tackle health statuses in rural places with relation to Vitamin C. Look at their sources of these

vitamins and compare if it can level the supplementation that Vitamin C supplements can provide

● Try a different technique different from Iodine Titration. Something that can give a different

result or analysis towards Vitamin C.

 BIBLIOGRAPHY

A. Books:

Webb, Geoff. (2012). Nutrition, Fourth Edition, Maintaining and Improving Health. ISBN

9781444142464. CRS Press. (P.390-392)

La Salle University, Philadelphia. Chemistry Laboratory Manual. CHL 212, Quantitative Analysis.

Retrieved from: http://www.lasalle.edu/~prushan/Experiment8-redox%20titration.pdf

University of Canterbury, New Zealand. Outreach, College of Science. Retrieved from:

http://www.outreach.canterbury.ac.nz/chemistry/documents/vitaminc_iodine.pdf3

Life Extension Magazine. Foundation for Longer Life. Newly Discovered Health Benefits of

Vitamin C. Goepp, Julius, MD. April 2008 Issue. Retrieved from: http://www.lef.org/.

B. Journals

The American Journal of Clinical Division (1995). Determination of optimal vitamin C

requirements in humans. (pp. 1347s-1356s)

World Health Organization & Nutrition on Health and Development. SCURVY and its prevention

and control in major emergencies (1999).

Deutschman, M. (2013). Generic vs. prescription drugs: perceptions and actions. Kantar Media:

Healthcare Research Insights. Retrieved March 30, 2015 from http://www.kantarmedia-

healthcare.com/generic-vs-prescription-drugs-perceptions-and-actions

Nutrition Review (1999). Brief Critical Reviews: Criteria and Recommendations for Vitamin C

Intake. Ausman L. & Mayer J. (pp. 222-224)

Philstar. (2011). Market is wide enough for new players in vitamins and supplements industry,

March 30, 2015. Retrieved from http://www.philstar.com/business-usual/697514/market-wide-

enough-new-players-vitamins-and-supplements-industry

C. Internet

DSM Home (Dutch-based multinational life sciences and materials sciences Company). Industrial

production. Retrieved from: http://www.dsm.com/products/quali-c/en_US/vitamin-c/industrial-

production.html.
TIME TABLE
EXPENSES TABLE
 RESEARCH PLAN

A. Objectives

The main purpose of this research is to make a comparative study on the purity of generic

and branded Vitamin C supplements sold in known pharmaceutical drugstores in Metro Manila. It

will be done through the process of Iodine Titration, using a reaction between Standard KIO 3

solution and Ascorbic Acid titrating it with Sodium thiosulfate solution. To ensure the quality and

effectiveness of these sold Vitamin C supplements whether they are worth buying or not is the main

objective of this research. This way we can help Filipino families in their decision making towards

this matter.

B. Hypothesis

The hypothesis on this research is that after Iodine Titration has been done and all

computations have been made, the amount of Vitamin C found in both branded and generic

supplements will have no to very minimal difference. The researchers also hypothesize that the

branded and generic Vitamin C supplements will follow the percentage amount found on the

Nutritional Content of the supplement’s cover. This way, it is safe to prove that generic Vitamin C

supplements can be bought without hesitation even though it is cheaper than branded Vitamin C

supplements. If subjected to a high percentage error in the research, the researchers hypothesize

that this will be only due to errors in the process which can be found in the Risk Assessment of this

paper.
C. Methodology

1. Materials

The materials used by the researchers includes the titration set-up and six compounds which were

used for experimentation. The titration setup includes the 50mL burette with stopcock, iron stand,

burette clamp, analytical balance, and a wash bottle. In addition to this, glassware apparatus were

also used. This would include the 250mL Erlenmeyer flask, 250L volumetric flask, 25mL

volumetric pipette, distilled water, 1L amber bottles and some extra glassware to hold the necessary

reagents and compounds. The six compounds used by the researchers are namely Soluble Starch

(C6H10O5), Potassium iodide (KI), Sodium thiosulfate (Na2S2O3), Iodine (I2), Sodium carbonate

(Na2CO3), and Sulfuric acid (H2SO4)

2. General Procedure

The general procedure is divided into three phases namely: the Preparation Phase, where

they would setup all the necessary solutions needed for the experiment; the Experimentation Phase,

where they would do the Iodine Titration of Vitamin C supplements; and lastly the Evaluation

Phase, where they would actually analyze, compute, and make conclusions toward the experiment.

Preparation Phase

I. Preparation of Starch Indicator

The researchers fill a 100mL beaker to the 100-mL mark with distilled water and place it

on a hot plate until boiling. They weigh out 1g of soluble starch (C6H10O5) and slowly add to the

beaker of boiling water with stirring until the solution is clear.

 II. Preparation of Sodium Thiosulfate Solution

The researchers fill a 500mL beaker to the 500-mL mark with distilled water and place it

on a hot plate. They boil the water for 5 minutes to expel dissolved CO2 gas. After allowing the

distilled water to cool, they weigh out 0.05g of Na2CO3 and place into the 500-mL beaker of boiling

water. They again weigh out 8.7 grams of Na2S2O35H2O (0.07 M) and dissolve it into the 500-

mL beaker of boiling water buffered with Na2CO3. The researchers then place the solution in a 1L

amber bottle properly labelled with “Sodium Thiosulfate Solution”.

III. Preparation of Standard Iodate Solution

The researchers fill a 500mL volumetric flask with 400mL of distilled water. They

accurately weigh out 1g of KIO3 using the analytical electronic balance and record the mass up to

four decimal places. Dissolve the KIO3 into the distilled water contained in the 500-mL volumetric

flask and stir until all solid particles have been dissolved. Fill the volumetric flask to the mark with

distilled water. They determine the exact molarity of this solution using the following molecular

masses: K: 39.0983 I: 126.90447 O: 15.9994. The researchers then placed the solution in a 1L

amber bottle properly labelled with “Standard KIO3 Solution”.

IV. Standardization of Sodium Thiosulfate Solution

The researchers prepared the setup to be used for titration. Then they fill the 50mL burette

with the Sodium Thiosulfate solution. Accurately pipet 50.00 mL of standard KIO3 solution into

the 250mL Erlenmeyer flask. Then they weigh 2 grams of KI and place it into the flask. Add 10mL

of 0.5 M H2SO4 to the flask. The solution start out as a “red-ish” solution due to the presence of the

triiodide. Titrate the solution with the Sodium Thiosulfate until the solution has lost most of the

red-ish color (should be at least pale yellow). At this point add 2 mL of the starch indicator (may

turn smoky-blue or remain yellowish). Carefully add Sodium Thiosulfate until the solution turns

colorless (may be a milky-white). Record this volume as the end point. The indicator is not added
until before the end point as the triiodide/starch complex can “hold on to” the triiodide in

concentrated solutions and prevent it from reacting with the Sodium Thiosulfate. The researchers

repeat this three times to get an accurate and precise result. Using the known stoichiometry, the

molarity of the Sodium Thiosulfate solution can now be calculated.

Experimentation Phase

I. Reaction of Triiodide with Ascorbic Acid

After preparing and knowing the molarity of the Sodium Thiosulfate solution and preparing

the Standard KIO3 Solution, three branded and three generic Vitamin C supplements are to be used.

The researchers place 60 mL of 0.3 M H2SO4 into the 250mL Erlenmeyer flask. Add 2 Vitamin C

supplements to the flask and dissolve it in the sulfuric acid. Make sure it is completely dissolved.

Then accurately pipet 50.00 mL of standard KIO3 solution into the 250mL Erlenmeyer flask. 24.

Weigh out around 2 grams of KI and place into the flask. Swirl the flask for 1 minute to insure

complete formation of the triiodide complex and complete reaction between the triiodide and the

Vitamin C.

II. Titration of Solution with Sodium Thiosulfate

The solution should start out as a “red-ish” solution due to the presence of the triiodide.

The researchers titrate the solution with the Sodium Thiosulfate until the solution has lost most of

the red-ish color (should be a pale yellow). Then, add 2 mL of the starch indicator (may turn smoky-

blue or remain yellowish). Carefully add Sodium Thiosulfate until the solution turns colorless (may

be a milky-white). Record this volume as the end point. The indicator is not added until before the

end point as the triiodide/starch complex can “hold on to” the triiodide in concentrated solutions

and prevent it from reacting with the Sodium Thiosulfate. The researchers made two trials and it

was done repeatedly to the three generic and three branded Vitamin C supplements.
Evaluation Phase

The researchers are to compute for the amount of Vitamin C present using the known

stoichiometry of the Sodium Thiosulfate/IO3 - and Ascorbic Acid/IO3 - reactions, and then calculate

the average number of moles of Ascorbic Acid contained in each Vitamin C supplement. Using the

formula weight of Vitamin C (C6H8O6, MW = 176.13), they calculate the average number of

milligrams of Vitamin C contained in each supplement along with the standard deviation.

Presentation of data and computations is shown in the Results and Discussion of the research paper.

3. Risk Assessment

Many risks are present in the process of the research. These would include the availability

of the compounds and apparatus needed; and the presence of human error. The researchers take

note on the risks that may lead to the failure of the research. This would include the following:

● Standard KIO3 Solution should be stored in a cabinet and must be out of direct sunlight. It may

not be possible to completely dissolve the I2. If some undissolved I2 is present, decantation of the

solution is necessary and diluting it up to the 1L mark with distilled water. Standardization of the

solution should be repeated.

● I2 sublimes at room temperature. This vapor form is caustic and a potential health threat.

Weighing and mixing with the presence of this element must be done at the fume hood.

● The Sodium Thiosulfate Solution must be used within 24 hours. Using of this solution after 24

hours might lead to an error in the results.

● Vitamin C spontaneously oxidizes when exposed to air. If not kept tightly closed, the potency of

these Vitamin C supplements will slowly be lost. The solution must be used within 24 hours. Do

not store the solution overnight.

Human error will include the following:

● Error in measuring and reading values (parallax error) while in the process of research

● Use of Contaminated Compounds

● Misuse or use of broken/contaminated apparatus in the laboratory

4. Disposal Methods

The researchers consulted the Material Safety Data Sheet (MSDS) of the used compounds

in order to determine the proper method of disposal for each compound. Each compound, namely

Soluble Starch (C6H10O5), Potassium iodide (KI), Sodium thiosulfate (Na2S2O3), Iodine (I2),

Sodium carbonate (Na2CO3), and Sulfuric acid (H2SO4), must be placed in a sealed black plastic

bag or container before placing it in a trash bin, as designated color for chemical wastes. It is

important to ensure that these bags are perfectly sealed without leaks or damage. Sulfuric acid may

be placed in sealed container or absorbed in vermiculite, dry sand, earth, or a similar material. It

may also be diluted and neutralized.

Dry materials contaminated with chemicals (paper, rags, towels, wipes, or spill

contaminated materials) must be double bagged in heavy-duty plastic bags. Tape all containers of

chemically-contaminated dry materials securely shut and label the outside of any container clearly

with descriptions of its contents and the chemical contaminants. Biologically contaminated

glassware and sharps should be placed into appropriate sharps containers.

 APPENDIX

Appendix A: Important Details on Compounds involved in Research

Name of Chemical Mol. Molecular Structure Melting Boiling

Compound Formula Weight Point Point
(g/mol)

Ascorbic C6H8O6 176.13 190 °C 553 °C

Acid

Soluble C6H10O5 150.15 0.8 °C 256°C

Starch

Potassium KI 166.00 K-I 681 °C 1,330 °C

Iodide

Sodium Na2S2O3 *
Thiosulfate 5H2O 248.17 48.3 °C 100 °C

Iodine I2 253.81 I-I 113.7 °C 184.3 °C

Sodium Na2CO3 105.99 851 °C 1,600 °C

Carbonate

Sulfuric H2SO4 98.08 10 °C 337 °C

Acid