Titrimetric Determination of

Vitamin C in Citrus Fruits

Presented by – Aanchal Sharma
Registration Number – 161102022
(Department of Chemistry)
Guide – Dr. Nitu Bhatnagar
Manipal University Jaipur
 Introduction:
Vitamin C (L-ascorbate)
• Ascorbate, an ion of ascorbic acid acts as an antioxidant by protecting the body against oxidative stress.
• Aids in detoxification and in improving ferrum absorption.
• Ensures the maintenance of cartilage, bone, denin and healthy blood vessel.
• Necessary in the synthesis of collagen in connective tissues, neurotransmitters, steroid hormones,
carnitine, and conversion of cholesterol to bile acid and enhances iron bio-availability
• Found to have anti-inflammatory, antioxidant, anti-tumor, anti-fungal and blood clot inhibition activities .
(Robert et al., 2000, Abeysinghe et al., 2007).
Thus, intake of Vitamin C ensures healthy lifestyle. Human beings and certain animals cannot synthesize
Vitamin C in their body, so that dietary requirements are met through food sources like fruits, vegetables
etc.
 A lack of vitamin C in the diet causes the deficiency disease
scurvy (Levine 1986), which is caused by a disruption in the
synthesis of collagen and connective tissues.

•Scurvy can have some serious signs and symptoms, including:

Slow wound healing
Bleeding gums
Easy bruising
Fatigue
Swollen gums
Weakened immune system
Dry, scaly skin
Swollen joints
Dry, splitting hair

It may also result in hemorrhages under the skin, poor

wound healing, edema and weakness (Enloe et al., 1985).
Figure : 1
 Literature Survey
• A study carried out by Tareen et al. used ………shows that among non-citrus fruits, papaya shows
higher concentration of vitamin C as compared to mango, while among citrus fruits, orange possessed
high content of vitamin C.
• Tiruwork and Ghirma, 2012 ……reported that the ascorbic acid content of freshly prepared Orange
juice and old Orange juice were 41.4mg/100 mL and 32.4mg/100 mL respectively.
• Aydogmus et al., 2002 …….shows that vitamin C content of orange juice ranged from 33 to 50 mg/100
ml .
• Nweze et al. had used iodometric titration method for the analysis of Vitamin C content in four fruits.
Result showed that pineapple and watermelon has the lowest vitamin C content and orange has the
highest vitamin C content.
• Classic (conventional) techniques are represented by volumetric methods—titration with an oxidant
solution such as dichlorophenol indophenol (DCPIP), potassium iodate, or bromate [M. Şerban, 1993,
C. Papuc, 2001, D. Balan, N. Matei, 2004]
 Objective of the Study
• To analyze Vitamin C content in Citrus Fruit juices collected from
different sources by using DCPIP
 Hypothesis
The higher the volume of fruit juices needed to decolorize DCPIP
solution, the lower the vitamin C content in the fruit juice. Fresh lime
juice has the highest content of Vitamin C among the fresh juices and
lime carton juice has the highest content of Vitamin C among carton
juices.
 Chemistry Behind the analysis –

• 2, 6-Dichlorophenolindophenol (DCPIP, DCIP or DPIP) is a chemical

compound used as a redox dye. When oxidized, DCPIP is blue with a
maximal absorption at 600 nm, when reduced, DCPIP is colorless.
• DCPIP can be used as an indicator for vitamin C. If vitamin C, which is a
good reducing agent, is present, the blue dye, which turns pink in acid
conditions, is reduced to a colorless compound by ascorbic acid. This
reaction is a redox reaction: vitamin C (ascorbic acid) is oxidized to
dehydroascorbic acid, and DCPIP is reduced to the colorless compound
DCPIPH2.
Figure 2
 Methodology –

Materials and Methods

Chemicals-
• The chemicals used for the experimental purpose in the present study includes distilled water,
ascorbic acid, and DCPIP solution.
Sample collection and preparation-
• Two ripe fruits and tetra juices were purchased from Hostel Campus mart as well as
cafeteria. The fruits include Orange (Citrus sinensi), and lemon. The fruits were squeezed
with a blender each and was filtered using a muslin cloth and made up to 100ml with
distilled water and the vitamin c determination was done by titrimetric method.
 Result of solution of vitamin C
Concentration of Volume of vitamin c need to Average volume
vitamin C (g/ml) decolorize 0.5ml of 1.0% DCPIP of vitamin c to
(ml) decolorize
DCPIP (ml)

1st reading 2nd reading

0.125/100 0.75 0.80 0.77
0.250/100 0.45 0.47 0.46
0.500/100 0.50 0.70 0.60
0.750/100 0.40 0.35 0.37
1.000/100 0.25 0.30 0.27
 Curve of Ascorbic Acid
Graph 2
1
0.8
0.6
0.4 Volume of ascorbic acid
0.2 need to decolorize DCPIP
0
0 0.2 0.4 0.6 0.8 1 1.2
Concentration of ascorbic acid
Juice Volume of vitamin c required to C= Concentr
Samples decolourise 0.5ml of 1.0 % of DCPIP (ml) 0.192/V( ation of
ml) vitamin
C (g
mol^-)
Category : 1st reading 2nd 3rd reading Avera
Orange Juice reading ge
Fresh juice 3.3 ml 3.5ml 4.0ml 0.192/3.6 0.053
3.6ml
Health Bar 0.192/13. 0.01
Juice
10ml 12ml 18ml 3
13.3ml 0.768
Tetra Pack 0.192/25
juice 20 ml 25ml 30ml
25ml
• Fresh 4.0ml 4.5ml 5.0ml 0.192/3.5 0.054
lemon juice 3.5ml
0.192/28. 0.678
•Tetra Pack 25ml 30ml 30 ml 3
juice 28.3ml
UV Spectrum of Orange Juice
Sample 1 –Market Orange Juice

The absorption maxima for fresh

orange juice samples were found to be
282 nm. The first absorption maxima
was due to the naturally occurring
compounds which would be beneficially
for human health.
UV Spectrum of Carton orange juice

Sample 2 – Tetra pack of orange juice

The absorption maxima for Carton

orange juice samples were found to be
around 260nm.
UV Spectrum of Carton of lemon juice

Sample 3 –Tetra Pack of lemon juice

The absorption maxima for Carton

lemon juice samples were found to be
around 260-280nm.
UV Spectrum of DCPIP Solution

Sample 4 – DCPIP Solution

Dichlorophenol indophenol (DCPIP)

absorbs within the visible region of
the spectrum with a lambda(max)
approximately 600 nm. When reduced,
the molar absorption coefficient at this
wavelength decreases significantly and
DCPIP effectively becomes colorless.
UV Spectrum of Ascorbic Acid

Sample 5 – Ascorbic Acid Solution

[4]
 Conclusion -
• Every fruit group has different concentration of vitamin C. the smaller the
volume of fruit juice needed to decolorize DCPIP solution, the higher the
content of vitamin C in fruit juice.
• Fresh juice contain more vitamin C content than carton fruit juices.
• Fresh lime juice has the highest vitamin C content and orange juice from
health bar has the lowest vitamin C content among fresh fruit juices while
for carton fruit juice, lime has highest and orange has the lowest content of
vitamin C.
 References:
[1] C. C., Abdulganiyu M. G., & Erhabor, O. G. (2015).Comparative analysis of Vitamin C in fresh Juice of Malusdomestica,
Citrus sinensi, Ananascomosus and Citrulluslanatus by idometric titration. International Journal of Science,
Environment and Technology, 4(1): 17 – 22.
[2] Tareen Huma, Mengal Fariha, Zubia Masood and Zainab Nawaz. (2015). Determination of Vitamin C content in Citrus
Fruits and in Non-Citrus Fruits by Titrimetric method, with special reference to their nutritional importance in Human
diet. Biological Forum – An International Journal 7(2): 367-36
[3] R.H. Larsen. Vitamin C: Your ultimate health insurance.(1997) International Journal of Alternative and Complementary
Medicine. 15(8) 19-2
[4] Vallapudas Hima, S. Rubesh Kumar, N. Duganath and N. Devanna .(2013) Quantization of ascorbic acid in ayurvedic
amla capsule by various analytical techniques. Scholars Research Library Der Pharma Chemica, 5(3):8-17.
[5] Tiruwork M. and Ghirma M. (2012). All-Solid-State Iodide Selective Electrode for Iodimetry of Iodized Salts and
Vitamin C. Orient. J. Chem., 28(4): 1547-1555.
Acknowledgement