Effect of Starch and Sulfuric Acid on Determination of Vitamin C in
Papaya Fruit Using Iodimetri
Khoirul Ngibad1, M. Sungging Pradana1, Ingrid Retno Y. 1

1

Department of Medical Laboratory
Technology, Faculty of Health
Science, Universitas Maarif
Hasyim Latif, Sidoarjo, Indonesia
Correspondence:
Khoirul Ngibad, Jl. Raya Ngelom
Megare No. 30, Ngelom, Taman,
Sidoarjo, East Java, Indonesia
Zip Code : 61257
Email:
khoirul_ngibad@dosen.umaha.ac.id
Received: February 11, 2019
Revised: March 16, 2019
Accepted: March 26, 2019

Abstract
Vitamin C is an antioxidant that can be used to inactivate
oxidation reactions and prevent the formation of free
radicals. Sources of vitamin C are fruits, such as papaya
fruit. The purpose of this study was to determine the effect
of the indicator volume of 1% starch and 2 N sulfuric acid
on the determination of vitamin C in papaya fruit samples.
This study used the iodimetri method with a standard iodine
solution, starch indicator and the addition of sulfuric acid.
The variations of starch indicator volume include: 0.25; 0.5,
1, 2 and 3 mL and the variations of sulfuric acid volume
include: 0, 2, 4, 6, and 8 mL. The results showed that the
optimum 1% starch indicator volume was 1 mL and the
optimum volume of 2 N sulfuric acid was 2 mL.
Keywords
Vitamin C, sulfuric acid, starch, iodimetri.

INTRODUCTION
One of the vitamins needed by humans is

also can act as a calogen–forming compound
(1).

vitamin C, also known as L–ascorbic acid.

Consumption of vitamin C that is not

Vitamin C is a chemical compound with

sufficient enough can cause the vitamin C

formula C6H8O6. Vitamin C is an antioxidant,

deficiency. In contrast, excess of vitamin C

which is a compound that can act to inactivate

intake in adolescents will also cause

oxidation reactions and prevent the formation

problems in the gastrointestinal system (2).

of free radicals. On the other hand, vitamin C

Recommended Daily Allowance (RDA) is
15

Khoirul Ngibad, et al.

the basis for adjusting the amount of adequate

with the addition of starch indicators. The

vitamins per day. The intake of at least 10 mg

accuracy in determination of vitamin C was

of vitamin C per day can prevent the

influenced

occurrence of scurvy deficiency disease.

parameters, such as: volume of starch

According to the Regulation of the Minister

indicator and sulfuric acid used for the

of Health of the Republic of Indonesia No. 75

titration

in 2013 concerning the Recommended

variations of starch volume indicators: 2

Nutritional Adequacy Rate for the Indonesian

drops (2), 3 drops (11), 2 mL (4), a few drop

Nation reveals that vitamin C is needed every

(3,12), and 3 mL (1). On the other hand, there

day at least 40 to 50 mg (infants under 1

are several studies that used the addition of

year), 40 mg (ages 1 – 3 years), 45 mg (ages

sulfuric acid in the titration process (1,3,12).

4 – 6 years), 45 to 50 mg (ages 7 –12 years),

The uncertainty in the use of volumes of

100 mg (pregnant women) and 150 mg

starch and sulfuric acid is feared to affect the

(breastfeeding mothers) (3).

accuracy in determining vitamin C levels in

by

several

process.

Some

measurement

studies

used

Sources of vitamin C are derived from

food and beverage ingredients. Thus, this

fruits such as guava fruit, raisins, kiwi,

study will know the effect of volume of the

lychees, oranges, persimmon, lime, papaya,

1% starch indicator and 2 N sulfuric acid on

strawberries, lemons, pineapples, melons,

the determination of vitamin C in samples of

mangoes, star fruit, grapes, passion fruit,

papaya fruit using iodimetri.

breadfruit,

durian,

pomegranate,

banana,

avocado,
and

jackfruit,

watermelon.

Determination of vitamin C in food and
beverage

packaging

samples

can

be

determined using the titrimetric method (4–
7) and spectrophotometry (8–10). Among the
two methods, the titrimetric method has the
advantage of not requiring standard analyte
solutions, simple equipment, easy to do and

MATERIALS AND METHODS
Material
The materials used include Carica
papaya L. (papaya fruit), 1% starch indicator,
0.01 N iodine standard solution, distilled
water, filter paper, label paper, 0.01 N
Na2S2O3, 0.01 N KIO3, 10% KI, and H2SO4.

its accuracy is high.

16

Determination of vitamin C used the

Equipment

titrimetric method based on iodimetri,

The equipments used include beaker

namely vitamin C will be oxidized by iodine

glass, Erlenmeyer flask, volumetric flask,

standard solution and the endpoint of titration

burette,

was indicated by the formation of blue color

measuring pipette, scissors, knife, blender,

statif,

clamp,

volume

pipette,

spatula, and analytic balance.
Ina J Med Lab Sci Tech 2019; 1(1): 15-21

Khoirul Ngibad, et al.

Procedure

Effect of 1% starch indicator volume on

Na2S2O3 standard

the determination of vitamin C in papaya

As much as 10 mL of 0.1 N KIO3

fruit

solution was taken and then was put into the

As much as 25 mL of papaya sample

Erlenmeyer flask. Then, as much 5 mL of

solution was taken and put into Erlenmeyer

10% KI solution and 2 mL of H2SO4 was

flask. Then, solution was added by indicator

added into the flask. Then, solution was

solution of 1% starch with volume variation

titrated using the Na2S2O3 solution until a

of 0.25; 0.5, 1, 2 and 3 mL. Then, mixtures

light yellow color was formed. Next,

was titrated with I2 standard solution until a

mixtured was added with 0.5 mL of 1%

blue color was formed. From this treatment,

starch indicator and was titrated with

the optimum volume of starch indicator will

Na2S2O3

be obtained.

solution

until

a

blue

color

disappears (12).

Effect of addition of 2 N sulfuric acid on

I2 standard

determination of vitamin C in papaya fruit

As much as 10 mL of I2 solution was

As much as 25 mL of papaya sample

taken into the Erlenmeyer flask. Furthermore,

solution were taken and put into Erlenmeyer

solutions was titrated using Na2S2O3 solution

flask. Then, as much as 1 mL of 1% starch

to form a light yellow color. Next, mixtured

indicator solution was added. Then, as much

was added with 0.5 mL of 1% starch

as 2 N H2SO4 solution were added with

indicator. Then, mixtures was titrated with a

volume variations include 0, 2, 4, 6, and 8

Na2S2O3

mL. Then, mixtures were titrated with I2

solution

until

a

blue

color

disappears (12).

solution standard until a blue color was

Preparation of Sample

formed. From this treatment, the optimum

Papaya fruit was shelled and the seed was

volume of sulfuric acid will be obtained.

removed. Then, it was cut into small pieces
and was weighed as much as 100 grams.

RESULTS

Furthermore, was blended until it resembles

Effect of 1% starch Vitamin C levels in

a slurry (juice). Then, mixtures was filtered

papaya fruit are tabulated in Table 1 and

with a filter cloth to separate between residue

Fig.2. The obtained results revealed that

and filtrate. The residue was removed and the

papaya fruit contain maximum amount of

filtrate was put into 1000 mL volumetric

vitamin C that is 12.91 mg/mL.

flask and added with distilled water to the

to be measured using the iodimetri method.
Ina J Med Lab Sci Tech 2019; 1(1): 15-7 21

17

boundary mark. The filtrate obtained is ready

Khoirul Ngibad, et al.

Table 1. Effect of 1% starch indicator volume on determination the vitamin C levels in papaya
fruit
Titrant volume (mL)

Vitamin C levels (mg)

Indicator
volume
(mL)

n1

n2

n3

Average

Standard
deviation

n1

n2

n3

Average

Standard
deviation

0,25

1.1

0.9 1.1

1.03

0.12

9.68

7.92 9.68

9.09

1.02

0,5

0.6

1.1 0.9

0.87

0.25

5.28

9.68 7.92

7.63

2.21

1

1.4

1.5 1.5

1.47

0.06

12.32 13.2 13.2

12.91

0.51

2

0.9

1

1

0.97

0.06

7.92

8.8

8.8

8.51

0.51

3

1.4

1

0.5

0.97

0.45

12.32

8.8

4.4

8.51

3.97

1,5

2

2,5

Vitamin C levels (mg)

Description: n1 = test 1, n2 = test 2, n3 = test 3
16
14
12
10
8
6
4
2
0
0

0,5

1

3

3,5

1% starch indicator volume (mL)

Fig 1. Effect of 1% starch indicator volume on determination the vitamin C levels in papaya

In order to established methods for determination of vitamin C in papaya fruit, sulfuric acid
was used in treatment (Fig.2 and Table 2). After incubation with sulfuric acid, a colored product

18

Vitamin C Levels (mg)

is formed which is in a blue color.

Sulfuric Acid Volume 2N (mL)

Fig 2. Effect of sulfuric acid addition on the determination of vitamin C levels in papaya
Ina J Med Lab Sci Tech 2019; 1(1): 15-21

Khoirul Ngibad, et al.

Table 2. Effect of volume of 2 N sulfuric acid on determination of vitamin C levels in papaya
Titrant volume (mL)

Levels of vitamin C (mg)

Volume of
sulfuric
acid (mL)

n1

n2

n3

Ave
rage

Standard
deviation

n1

n2

n3

Ave
rage

Standard
deviation

0

0.8

1

1

0.93

0.12

7.04

8.8

8.8

8.21

1.02

2

0.8

0.9

0.9

0.87

0.06

7.04

7.92

7.92

7.63

0.51

4

0.8

0.6

0.8

0.73

0.12

7.04

5.28

7.04

6.45

1.02

6

0.8

0.8

0.8

0.8

0

7.04

7.04

7.04

7.04

0

0

7.04

7.04

7.04

7.04

0

8
0.8
0.8
0.8
0.8
Description: n1 = test 1, n2 = test 2, n3 = test 3

DISCUSSION
The principle of determination the
vitamin C contents using the iodimetri
method is based on the reaction between
vitamin C which is a strong reducing agent
and standard solution of iodine which is an
oxidizer with starch indicator. In this study,

iodine standard solution so that an addition

Fig 3. The reaction between vitamin C in
papaya fruit with iodine produced
the dehydroascorbic acid

reaction occurs between iodine and double

Effect of 1% starch indicator volume on

bonds of vitamin C on number 2 and 3 of

determination of vitamin C in papaya

vitamin C in papaya fruit samples react with

carbon atoms become a single bond. If the

In this study, the effect of 1% starch

whole vitamin C has been supplemented by

indicator volume on the determination of

standard iodine solution, then a reaction

vitamin C levels in papaya fruit was studied

occurs between the droplets of the next iodine

using iodimetric titration. In the process of

solution and the starch indicator solution so

titration, the addition of an indicator serves to

that iod–amilum was formed. The end point

show that the endpoint of the titration has

of the titration was indicated by the presence

occurred so that the titration process must be

of blue color indicating that the titration has

stopped immediately.

been completed because all vitamin C has

incorrect volume indicators is feared to affect

been supplemented by iodine (5). Reactions

the accuracy of the measurement of vitamin

that occur can be seen in Figure 3.

C levels in a sample.

of

19

Ina J Med Lab Sci Tech 2019; 1(1): 15-7 21

The addition

Khoirul Ngibad, et al.

Table 1 and Figure 1 showed that the

sulfuric acid volume produced the optimum

addition of the indicator volume by 1 mL

vitamin C levels of 7.63 mg with standard

resulted the optimum vitamin C levels of

deviation of 0.51. On the other hand, the

12.91 mg with the smallest standard

more volume of 2 N sulfuric acid added

deviation of 0.51. On the other hand, the

caused slight decrease in fluctuating of

addition of volume indicators of 1% starch as

vitamin C levels. In general, the standard

much as 2 and 3 mL decreased the vitamin C

deviation value in the study of the effect of

levels and increased the standard deviation

sulfuric acid volume on determination of

value for addition the volume indicator of 1%

vitamin C levels in papaya fruit was smaller

starch by 3 mL. This is probably caused by

than the study of the effect of 1% starch

the excess volume of 1% starch indicator, the

indicator volume on the determination the

less the volume of iodine standard solution

vitamin C levels in papaya. It was also

needed so that the vitamin C levels in the

supported by visual observations in the

sample reduced. Thus, the optimum volume

laboratory proving that the color yield of the

of 1% starch indicator in determination the

titration end point of all sulfuric acid addition

vitamin C in papaya fruit was 1 mL.

treatments produced more stable color. This

The effect of addition of 2 N sulfuric acid

shows that at low pH vitamin C is more stable

on determination the vitamin C levels in

than high pH. Thus, the optimum volume of

papaya

2 N sulfuric acid on determination of vitamin

This study also was studied the effect of

C levels in papaya fruit was 2 mL.

addition of 2 N sulfuric acid on determination
of vitamin C levels in fruit Papaya using
iodimetri. In the process of titration, the
addition of sulfuric acid give an acidic
condition in the solution of vitamin C when
the titration process occurs with iodine
standard solution. The effect of 2 N sulfuric
acid on determination of vitamin C in papaya

CONCLUSIONS
The addition of 1% starch indicator and 2
N sulfuric acid volume had an effect on the
determination of vitamin C levels in papaya
fruit. The optimum 1% starch indicator
volume obtained was 1 mL and the optimum
volume of sulfuric acid 2 N was 2 mL.

fruit was studied with 2 N sulfuric acid
volume variations of 0, 2, 4, 6 and 8 mL.
Based on Table 2 and Figure 3, we can

No potential conflict of interest.

20

calculated that the addition of 2 mL of

CONFLICT OF INTEREST

Ina J Med Lab Sci Tech 2019; 1(1): 15-21

Khoirul Ngibad, et al.

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