This slide was prepared by dawarayamajhi@gmail.com and presented at the International symposium on himalayan biodiversity and bio-resources, 2018.

1. Utilization of Stevia rebaudiana Bertoni
(chini biruwa) as replacement of sucrose in
yoghurt
International Symposium on Himalayan Biodiversity and Bioresources
February 14-15, 2018, Pokhara, Nepal
Alisha Karmacharya1 Dawa Rayamajhi Sherpa*1 Laxmi Ghimire2
and Pravin Ojha3
1 ,2National College of Food Science and Technology
3Nepal Agriculture Research Council
*Corresponding Author: dawarayamajhi@gmail.com

2. Contents
• Introduction
• Materials and Methods
• Result and Discussion
• Conclusion and Recommendation
• Acknowledgment

3. Introduction
• Nepal is a landlocked country located in the central part
of main Himalayan range and is floristically rich and
diversified
• Nepal possesses 2.80 percent of the total number of
plants species found globally
• The Himalayas are famous for many medicinal plants
and are used in traditional healing systems like
Ayurvedic, Homoeopathic, Amchi, etc,
• Some of these plants are even used for allopathic
medicine

4. Introduction continues…
• Examples of medicinal important plants are:
Fig a: Aloe vera
Fig c: Yarsagumba (fungi)
Fig b: Lavender flower
Fig d: Stevia plant

5. • Stevia rebaudiana is commonly known as sweet leaf, honey leaf, simply
stevia or as (chini biruwa in Nepal) is widely grown for its sweet leaves
• Native to subtropical and tropical regions from western North America to
South America
• Today its cultivation has spread to other regions of the world, including
Canada and some parts of Asia and Europe
Introduction continues…
Fig e: Stevia plant and sugar grains

6. • The sweet compounds in the leaves are called
stevioside and rebaudioside and they can be more
than 200 times sweeter than sucrose and its leaf by
itself is about 20 to 30 times sweeter than sucrose.
Introduction continues…

7. • Yoghurt is one of the most popular fermented dairy products widely consumed
all over the world and considered a healthy food due to high digestibility and
bioavailability of its protein, calcium and energy
• The sugar present in it is milk-sugar i.e. lactose and table sugar where table sugar
(sucrose) is incorporated to enhance the sweet taste
• Sucrose is highly avoided by diabetic and health concerned individuals hence
decreasing the consumption of nutritious yoghurt by significant amount.
Introduction continues…
Fig f: Yoghurt

8. Objective
• To utilize the water-extract of Stevia rebaudiana Bertoni (chini biruwa) as
the replacement of sucrose in yoghurt.
• The specific objectives are:
1. to calculate the proximate composition of Stevia leaf powder
2. to extract the sweet glycosides or steviol glycosides from Stevia powder
3. to optimize the stevia extract with sugar solution of different
concentration by comparing sweetness of stevia extract with sugar
solution
5. to optimize the Stevia concentration in yoghurt on the basis of sensory
evaluation and
6. to carry out physicochemical analysis of the formulated yoghurt.

9. Statement of Problem
• Consumers have more inclinated
towards products that are
claimed to be ‘All Natural’ and
‘Low Carbs’
• Large amount of cane-sugar is
used in products like biscuits,
jam, chocolates, ice-creams, soft
drinks, candies, yoghurt, etc,
• Yoghurt available for diabetic
patients are compromised with
sweet taste
Fig g: Sweet-less yoghurt

10. Materials
Fig h: Stevia Leaf powder Fig i: Milk
Fig j: Skim milk powder Fig k: Yoghurt culture packet

11. Research Methodology
Fig l: Research Methodology

12. Analysis Methodology
• Proximate calculation as per (AOAC, 2005)
• Extraction of sweet glycosides from stevia powder was done
by boiling in water at different ratios (Abou-Arab et al., 2011)
• Optimization of extract and comparison of sweetness of
stevia extract with sucrose solution was done by sensory
method (Rangana, 2007)
• Optimizations of the yoghurt formulation with stevia extract
(15°Bx) of different concentrations were carried out.

13. Result and Discussion
Parameters Stevia Powder
Moisture (%) 8.06±1.2
Ash (g/100g) 8.34±1.7
Fat 5.72±2.4
Protein 2.4±1.3
Reducing Sugar 1.5±0.2
Crude fiber 15.21±1.3
Carbohydrate 68.28±6.8
Table 1. Proximate analysis of stevia powder
The values are Mean ±S. Deviation for triplicate data.

14. Table 2. Extraction of steviol from stevia leaf powder
Sample No.
Extraction ratio
(powder/water)
Extract from 5 grams TSS °Bx
1. 1:15 2.51a±0.0 3.2±0.0
2. 1:20 2.47a±0.01 2.4±0.01
3. 1:25 2.28b±0.01 1.8±0.01
4. 1:30 2.0c±0.1 1.3±0.1
5. 1:35 1.76d±0.0 1.0±0.0
6. 1:40 1.22e±0.01 0.6±0.01
LSD at 5% 0.075
Superscript with different alphabets in the column differs significantly

15. 6.70%
10%
16.70%
56.70%
6.70%
3.30%
0%
10%
20%
30%
40%
50%
60%
1;15 1;20 1;25 1;30 1;35 1;40
PanelistScore
Extraction Ratio
Fig m: Preference (optimization) chart of extract by sensory
method from 30 panelist

16. 6.70%
13.30%
60%
16.70%
3.30%
0%
10%
20%
30%
40%
50%
60%
70%
10⁰ Brix 20⁰ Brix 30⁰ Brix 40⁰ Brix 50⁰ Brix
Panelistscore
Standard Sugar Solutions
Fig n: Comparison chart of sweetness of extract (2°Bx) with
standard sugar solution by sensory method from 32 panelist

17. 6.6
6.9
7.2
7.5
7.8
8.1
8.4
8.7
9
Color Texture Taste Flavor Appearance Overall
Acceptance
a
a
a
a
a a
b
b
b
b b b
a b
b
c c b
c c
c c
d c
d
d
d
c
e a
Sensoryscore
Sensory Parameters
A
B
C
D
E
The order of superiority of Sample A, B, C, D and E representing 0%, 0.15%, 0.25%, 0.3% and
0.35% Stevia extract in yogurt respectively can be summarized as;
Color: Sample B>Sample C>Sample A>Sample D>Sample E
Texture: Sample B=Sample C>Sample D>Sample E>Sample A
Taste: Sample B=Sample C>Sample D>Sample E>Sample A
Flavor: Sample B>Sample C=Sample D>Sample E>Sample A
Appearance: Sample B>Sample C=Sample D=Sample E> Sample A
Overall acceptance: Sample B>Sample C>Sample D>Sample E>Sample A
Fig o: Mean sensory score of 30 panelist
Superscript with different alphabets in the column differs significantly at (p<0.05)

18. Parameters Best Formulation (Sample B)
Moisture (%) 85±0.0
Total Solid 15.03±0.05
SNF 12.07±0.05
Fat 3.97±0.03
Acidity 1.0±0.01
Protein 5.33±0.05
The values are Mean ±S. Deviation for triplicate data.
Table 3. Chemical analysis of best yoghurt formulation

19. Conclusion
• Normal table sugar (sucrose) can be effectively
substituted by incorporating 15% steviol glycosides of
15°Bx in yoghurt
• Steviol glycosides i.e. water extract of stevia leaf powder
was at least thirty times sweeter than sucrose
• Highly nutritious, sweet and full of health benefits
aromatic yoghurt was developed which could easily
replace mostly-avoided, sucrose incorporated sweet
yoghurt

20. Recommendations
• Extract of stevia can be further purified to pure white
form by nano filtration process
• The stevioside and rebaudioside content can be
determined
• Chemical analysis as antioxidant activity, phenolic
content, tannin content can be carried out
• Stevia can be used instead of sucrose in many other
bakery products
• Bitter taste or plant like raw taste can be eliminated by
adding essence other than cardamom

21. Acknowledgement
Sincere thanks goes to my supervisor, Mr.
Pravin Ojha
Special thanks goes to my research
partner Miss. Alisha Karmacharya
Thanks to my college, National College of
Food Science and Technology
Mr. Pravin Ojha
Miss. Alisha Karmacharya