PROPOSAL

ANALYSIS OF VITAMIN C (ASCORBIC ACID) IN 10

FRUITS USING UV- SPECTROPHOTOMETER
ROYAL COLLEGE OF MEDICINE PERAK (UNIKL-RCMP)
SSM MODULE YEARR 2 2012
GROUP 33
RESEARCHERS
NURUL ATIQAH MAHMUD

57260211083

NUR HIDAYU FHATIHAH GHADZALI @GHAZALI

57260211062

NUR SYUHAINA ABU BAKAR

57260211126

MAZRIA AMANI MAZLAN

57260211130

SITI RASHIDAH ABU BAKAR

57260211051

SUPERVISOR
Checked by:

Approved by:

1.

INTRODUCTION

Vitamin C or ascorbic acid is a six-carbon lactone, which is synthesized from glucose

by many animals. Vitamin C is synthesized in the liver in some mammals and in the
kidney in birds and reptiles. However, this is not synthesized in a human body, which
makes it an essential nutrient which human obtained through dietary. Due to
deficiency of vitamin C, human suffer from the potentially lethal disease scurvy.
Vitamin C is largely distributed in fruits and plants that make it accessible to be
obtained. It is abundantly found in watermelon, papaya, oranges, apples, strawberries,
pineapple, mango and guava. It is also found in green leafy vegetables, cabbages,
cauliflower, broccoli and tomatoes.
Vitamin C is one of the important water-soluble vitamins. It is essential for
collagen, carnitine as well as neurotransmitter biosynthesis. Hence, vitamin C is
undeniably an important nutrient for metabolic processes in human. The current US
recommended daily allowance (RDA) for ascorbic acid ranges 100-120-mg/per day
for adults. Many health benefits have been attributed such as being a powerful
antioxidant, anti-atherogenic, anti-carcinogenic, immune-modulator and prevent cold
(K.A Naidu et. al, 2003).
Lately there has been mast production of synthetic or nutraceutical ascorbic
acid in the market to keep up with the demand of vitamin C supplement by the
consumers. A study showed that adults taking synthetic vitamin C supplementation
had serious effect. The result shows that synthetic vitamin C supplementation
decreases the exercise or training efficiency. This is because it depreciates the cellular
adaptations of exercise in the mitochondrial level.
Therefore, it is always best to consume natural vitamin C. Due to that, a
research to measure the content of vitamin C in five local fruits in Malaysia is carried

out to compare the ascorbic acid content between these fruits.

2.

LITERATURE REVIEW

A research study had been done regarding iodimetric determination of ascorbic acid
(vitamin C) in citrus fruits in year 2007. The study is conducted to determine the
ascorbic acid content of the juices from four different types of citrus fruits which are
orange, tangerine, grapefruit and lime. The study showed that orange has the highest
value of ascorbic acid (600 g /mL) followed by grape (446 g /mL) and then
tangerine (415 g /mL). Lime had the least value (306 g /mL). It is concluded that
orange would supply more ascorbic acid per milliliter for body need. From this
research, it is also concluded that the juices of oranges and tangerine are hereby
recommended, preferably, to be taken daily during meal at quantities sufficient to
meet daily vitamin C need (Izuagie et al. , 2007).
Besides, another research is carried out to quantify the total phenolics,
flavonoids and carotenoides, vitamin C contents, reported as antioxidants, in the
extracts of four locally available varieties of durian. In the research, the fruits are
extracted using dichloromethane: pentane (1:1 v/v) and subjected to study the various
quality parameters. The result showed the total of vitamin C contents.
This research also proved that caffeic acid and quercetin are the dominant
antioxidant substances found in durian. From this research, we know that the
bioactivity of ripe durian is high and the total polyphenols are the main contributors to
the overall antioxidant capacity. It is thus recommended to consume durian fruit for
uptake of antioxidant phytochemicals and vitamin C (Ashraf et al. , 2011).

3.

STUDY RATIONAL

Diets rich in fresh fruits and vegetables can protect us against chronic and
degenerative diseases (Joshipura et al. , 1999). The significant of this study is to study
the content of vitamin C in fresh fruits highly available in the market. Due to the side
effects of the synthetic supplementation of vitamin C, this study is carried out to find
the content of naturally available vitamin C in fresh fruits. This is significant to
educate the public that vitamin C supplementation can be taken naturally from fruits.
It is without a doubt that synthetic, nutraceutical vitamin C capsule available in the
market is modified chemically to prolong their time of consume. With this study, we
can help educate the public about the significant of eating fresh fruits that is readily
available in the market and therefore create awareness to take natural
supplementation, which is cheaper and healthier.

4.

OBJECTIVES

4.1.

GENERAL OBJECTIVES

4.1.1.

To determine the content of vitamin C in nine local fruits and an import fruit
in comparison with supplement vitamin C.

4.2.

SPECIFIC OBJECTIVES

4.2.1. To study the concentration of vitamin C using UV spectrophotometer method

with sodium oxalate acts as stabilizer.
4.2.2. To compare the ascorbic acid content between Orange and nine local fruits in
Malaysia which are:
4.2.2.1.
4.2.2.2.
4.2.2.3.
4.2.2.4.
4.2.2.5.
4.2.2.6.
4.2.2.7.
4.2.2.8.
4.2.2.9.

Guava
Mango
Durian
Pamelo
Banana
Pineapple
Star fruit
Papaya
Water apple/ bell fruit

4.2.3. To compare all the 10 fruits with the supplement vitamin C available in the
market.
4.2.4. To determine which fruits have the highest concentration of vitamin C.
4.2.5. To determine the pH value in each fruits used in the experiment.
4.2.6. To educate the public about the importance of vitamin C in daily diet.
4.2.7. To provide alternative for the public who want cheaper and healthier supply of
vitamin C.

5.0.

METHODOLOGY

There are many analytical methods have been reported to determine vitamin C
concentration such as titrimetry (Gunjan, 2012), electrochemical (Kim, 1989) and
potentiometric method (Leslie et al. , 1941). Some methods are available for
determination of ascorbic acid but only few are found to be used to determine both
forms of vitamin C (L-ascorbic acid and dehydroascorbic acid). It is very difficult to
choose a suitable method to determine the vitamin C contents in fruit juice. All the
methods have their own limitation for different purpose. (Mohammad et al. , 2007).
In this study, the method chosen is direct spectrophotometric method. It is
simple and highly sensitive method for determination of L-ascorbic acid in pure form.
This method used sodium oxalate (0.0056 mol/dm 3) as a stabilizer and molar
absorptivity (which does not require an extraction procedure) was 1.42x104 dm3mol1

cm-1 at 266nm. Beers law is obeyed in concentration range of 0.857-12.0 g ascorbic

acid/cm3 (Amra et al. , 2011).

5.1.

INSTRUMENTS

UV-Visible spectrophotometer using 1cm path length is used to measure the content
of vitamin C in each sample solutions. Digital pH meter is used to determine the pH
value in each sample solutions.

5.2.

REAGENTS

All reagents used are of analytical-reagent grade. Buffer solution (pH = 5.4), Sodium
oxalate solution (0.0056 mol/dm3) and L-ascorbic acid solution (1.13x10-3 mol/dm3).

5.3.

REAGENT PREPARATION

Buffer solution (pH = 5.4)

A mixture of potassium dihydrogenphosphate (0.03 mol/dm 3) and disodium
hydrogenphosphate (8.99x10-4 mol/dm3) is prepared by dissolving 4.08 g of KH 2PO4
(Fluka) and 0.16 g of Na2HPO4.2H2O (Merck) in 100 cm3 of distilled water.

Sodium oxalate solution (0.0056 mol/dm3)

This solution is prepared by dissolving 0.75 g of sodium oxalate (Sigma) in 1000 cm 3
of the buffer solution.

L-Ascorbic acid solution (1.13x10-3 mol/dm3)

A 0.05 g amount of L-ascorbic acid (Riedel-de Han) was dissolved in 250 cm 3 of the
sodium oxalate solution.

5.4.

SAMPLE

10 raw material of fresh fruits used in this study are purchased from a local
supermarket on the day of analysis. Guava (Psidium guajava), mango (Mangifera
indica), durian (Durio zibethinus), pamelo (Citrus grandis), banana (Musa spp),
pineapple (Ananas comosus), star fruit (Averrhoa carambola), papaya (Carica
papaya), water apple (Syzygium samarangense). The fruits chosen are in ripest state
(according to assumption).

5.5.

SAMPLE PREPARATION

All the 10 sample of fruits are washed thoroughly with water and the juices are
extracted manually using juice squeezer. All samples are filtered to remove pulp and
seed. After that they are stored in already labelled plastic containers. A small amount
of fruit juice is sufficient for the determination of ascorbic acid in these samples.

5.6.

GENERAL PROCEDURE

Transfer a portion of the sample solution containing 60 300 g of L-ascorbic acid to

a 25 cm3 standard flask. Dilute to the mark with the (0.0056 mol/dm 3) sodium oxalate
solution and measure the absorbance at 266 nm against the sodium oxalate solution as
a blank.

5.7.

DETERMINATION OF L-ASCORBIC ACID IN FRUIT SAMPLES

Transfer an accurately weighed amount of powder obtained from several tablets into a
100 cm3 volumetric flask, dissolve and make up to the mark with the (0.0056
mol/dm3) sodium oxalate solution. Filter and dilute a suitable aliquot of the filtrate to
50 cm3 with the stabilizer solution. Take an aliquot of the final solution and determine
the ascorbic acid content as described under general procedure.

5.8.

DETERMINATION pH IN FRUIT SAMPLES

Set the digital pH meter to set mode and insert the electrode into the solution samples.
Wait the reading to stabilize before record the value appeared on the display. Always
rinse the electrode before use it to test other sample solutions.

6.0.

RESULTS

Plan for data analysis and interpretation

Table 1: Biochemical characteristics of different types of fruits in Malaysia

Types of fruits
Vitamin C Contents(mg/100g)
pH
Orange
Bell fruit
Guava
Banana
Mango
Papaya
Starfruit
Pineapple
Durian
Pomelo

Figure 1: Experimental and literature values of vitamin C content for fresh

tropical fruits

7.0.

REFERENCES

K.A Naidu (2003) Vitamin C in human health and disease is still a mystery? An
overview, Nutrition Journal, 2(7), 1.
Joshipura, Alberto. A, JoAnn E.M, Meir J. S, Eric B.R, Frank E. S, Charles H.H,
Donna S, Walter C.W (1999) Fruits and vegetables Intake in Relation to Risk
of Ischemic Stroke, The Journal of American Medical Association (JAMA),
282(13), 1237-1239.
Ashraf, M. A., Maah, M. J., Yusoff, I., Mahmood, K., & Wajid, A. (2011). Study of
Antioxidant Potential of Tropical Fruit. International Journal of Bioscience,
Biochemistry and Bioinformatics, 1(1), 53-57.
Izuagie, A. A., & Izuagie, F. O. (2007). Iodimetric Determination of Ascorbic Acid
(Vitamin C) in Citrus Fruits. Research Journal of Agriculture and Biological
Sciences, 3(5), 367-369).
Gunjan Kashyap & Mangla Dave Gautam (2012). Analysis of Vitamin C in
Commercial and Naturals substances by Iodometric Titration found in Nimar
and Malwa region. Journal of Scientific Research in Pharmacy, 2277-9469.
Kim HJ. (1989). Determination of total vitamin C by ion exclusion chromatography
with electrochemical detection. US National Library of Medicine National
Institutes of Health Journal, Jul-Aug; 72(4): 681-6.
Leslie J. Harris, L. W. Mapson & Y. L. Wang (1941). A Simple Potentiometric
Method for Determining Ascorbic Acid, Suitable for Use with Coloured
Extracts. Nutritional Laboratory, University of Cambridge and Medical
Research Council.
Mohammad Mizanur Rahman Khan & Mohammad Mazedul Hosain (2007). Analysis
of Vitamin C (ascorbic acid) Contents in Various Fruits and Vegetables by
UV-spectrophotometry. Bangladesh J. Sci. Ind. Res. 42(4), 417-424, 2007.
Amra Selimovi, Mirsad Salki & Amel Selimovi (2011). Direct Spectrophotometric
Determination of L-ascorbic acid in Pharmaceutical Preparations Using
Sodium oxalate as a Stabilizer. International Journal of Basic & Applied
Sciences IJBAS-IJENS Vol: 11 No: 02.