J. Env. Bio-Sci., 2016: Vol.

30 (1):71-75
(71) ISSN 0973-6913 (Print), ISSN 0976-3384 (On Line)

PHYSICO - CHEMICAL AND NUTRITIONAL QUALITIES OF YOGHURT

ENRICHED WITH SPIRULINA AND NATURAL HONEY
N.Karthikeyan
Department of Livestock Products Technology (Dairy Science), Veterinary College and Research Institute
Namakkal, Pin code no:637 002, TamilNadu,India.
[Corresponding author E-mail:karthitamil_2007@rediffmail.com]

Received: 11-01-2016 Accepted: 13-03-2016

Spirulina is the most nutrient - dense food currently known with a protein content of more than 60% with high availability of
essential amino acid. It is an excellent source of nutrients viz., essential amino acids, essential fatty acids, carotenoids, minerals
and vitamins and thus can potentially represent an important staple in the human diet. The current research work focuses on
protein enriched yoghurt using Spirulinaas a source of protein and replacement of canesugar with natural honey. Different levels
of Spirulina (0, 0.2%, 0.4%, 0.6% and 0.8%) was added in the yoghurt to assess the optimum level of inclusion based on the sensory
evaluation and accordingly, Spirulina at the rate of 0.6 per cent was found to be superior in terms of sensory qualities like colour
and appearance, body and texture, whey separation, flavour and overall acceptability with that of control. Production of Spirulina
enriched and honey fortified yoghurt with 0.6 per cent Spirulina and up to 50 per cent replacement of cane sugar with 3% level of
honey was found to be more beneficial at the rate of 4.65 rupees per 50 ml with numerous therapeutic properties.

Yoghurt is widely consumed throughout the world for its
sensory and nutritional benefits and is made from milk with
high solid content, a lactic culture and sugar and can be
enriched with milk powder, proteins, vitamins, minerals and
fruits. Yoghurt is an excellent source of calcium, phosphorus
and other nutritional properties but as it is typical of all dairy
products, contains very little protein. Therefore, dairy products
are logical vehicles for protein fortification because they have
high nutritive values, reach target population and are widely
consumed1-2.
Spirulina is the most nutrient - dense food currently known
with a protein content of more than 60% with high availability
of essential amino acid. It is an excellent source of vitamins
including beta - carotene, thiamine, riboflavin, niacin, vitamin
C, vitamin E and biotin. The pigments include chlorophylls,
xanthophylls, carotenoids, and phytocyanin. It is highly
digestive 85-95% due to thin wall and low nucleic acid
contents(4%).The other valuable constituent is gamma-
linolenic acid which is a polyunsaturated fatty acid. This along
with superoxide dismutase acts as antioxidants and hence
useful in chemoprevention. They have antiviral, anti-
inflammatory, and antitumor effects and reduce blood lipid
profile, blood sugar, body weight, and wound healing time 3.
Use of sucrose in the yoghurt has a negative impact on the
diabetes patients. Bee honey, a natural food product, is the
NAAS Rating (2016)-4.20
oldest sweetening substance. It is a good source of many
valuable nutritious compounds for human body. Honey can be
used as sucrose replacer and/or supplemented in yoghurt
formulation. Honey has broad consumer appeal as natural
sweeteners and flavoring4. Considering the above facts, the
present study has been made to prepare a protein enriched
yoghurt using honey and Spirulina with good sensory,
therapeutic and nutritional properties.
MATERIAL AND METHODS
Different treatments were designed as PY- Control-without
addition of Spirulina and honey, SY- Spirulina 0.6% without
replacement of cane sugar, THSY1- Spirulina 0.6% and 1%
replacement of cane sugar with honey, THSY2- Spirulina 0.6%
and 2% replacement of cane sugar with honey, THSY3-
Spirulina 0.6% and 3% replacement of cane sugar with honey,
THSY4- Spirulina 0.6% and 4% replacement of cane sugar
with honey, THSY5- Spirulina 0.6% and 5% replacement of
cane sugar with honey, THSY6- Spirulina 0.6% and 6%
replacement of cane sugar with honey.
Preparation of Spirulina: Desired amount of Spirulina powder
was dissolved in 15 ml distilled water at 25°C and mixed with
the help of magnetic stirrer for 20 min. This solution was kept
for hydration for overnight at refrigeration temperature for
 NUTRITIONAL QUALITIES OF YOGHURT ENRICHED WITH SPIRULINA AND NATURAL HONEY
hydration. Just before using for product preparation, this was
remixed for 15 minutes using a magnetic stirrer5.
Preparation of Spirulina solution for incorporation into
yoghurt : Spirulina solution was dispersed in milk at 0.2, 0.4,
0.6, and 0.8 per cent levels for yoghurt preparation before
homogenization. Different levels of Spirulina (0, 0.2%, 0.4%,
0.6% and 0.8%) was added in the yoghurt treatment samples
to assess the optimum level of inclusion based on the sensory
evaluation5.
Procedure for the preparation of yoghurt: Plain yoghurt
was prepared using fresh milk. Skim milk powder at the rate
of 4 per cent (w/v) and sugar at the rate of 6 per cent (w/v)
were added to it and homogenized at 2500 psi. The contents
were mixed well and pasteurized at 850C for 30 minutes, cooled
to room temperature and inoculated with 2 per cent of yoghurt
cultures containing Lactobacillus delbrueckii ssp. bulgaricus,
and Streptococcus salivarius ssp. thermophilus. It was then
mixed well and incubated at 420C for 4 to 5 hours and finally
stored at 50C.
Physico chemical analysis of yoghurt : pH, viscosity and
acidity were estimated as per the procedure described in
IS:SP:18 (part XI)-19816. Syneresis (release of whey) was
measured by placing a 300 g yoghurt sample on a cheese
cloth placed on top of a funnel. After 2 h of drainage, the volume
of whey collected in a graduated cylinder was measured and
used as an index of syneresis7.
Nutritional analysis of yoghurt: Total solids content was
determined according to AOAC (1990), 15th edition8, Fat was
estimated as per the procedure described in IS:SP:18 (Part
XI) - 19816 and Protein was estimated as per the procedure
described in AOAC (1995), Vol.II, 16th edition9 .
Enumeration of yoghurt bacteria in yoghurt: Lactic acid
bacteria were determined from the colony counts on specific
lactic agar: MRS agar (pH 5.4) for Lactobacillus delbrueckii
subsp.bulgaricus and M17 agar for Streptococcus alivarius
spp. thermophilus. One gram yoghurt samples stored for 0, 7,
14 and 21 days were diluted with 9 ml of sterile peptone and
water diluent. Subsequent serial dilutions of each sample was
performed. From the suitable dilution 1 ml was transferred
into sterile petri plates in duplicates. Pre-melted MRS and
M17 agar (20-30 ml) were poured into the plate and mixed
(72)
well with the contents. After solidification the plates were
incubated at 41°C for 48 h10.
Organoleptic evaluation: Yoghurt samples were evaluated
for colour and appearance, taste and flavor, body and texture
and overall acceptability by a panel using a 9 point Hedonic
scale. All the samples were appropriately coded before
subjected for sensory evaluation11.
RESULTS AND DISCUSSION
Sensory evaluation of different inclusion level of spirulina
enriched and honey fortified yoghurt: Among the different level
of inclusion of Spirulina in yoghurt 0.6 per cent of inclusion
level scored high overall acceptability points. This inclusion
level was fixed as standard for different treatments with different
level of cane sugar replacement with honey yoghurt.
Physico-chemical properties of spirulina enriched and
honey fortified yoghurt
Titratable acidity (percent lactic acid): The titratable acidity
showed high significant difference (P>0.05) between control
and treatment yoghurt during different periods of storage. The
result of this study showed that with increase in cane sugar
replacement with honey from 1 to 6 percent resulted in increase
in acidity from 0.95 ±0.04 to 1.096 ±0.09per cent of lactic
acid (LA). Earlier researchers reported that, the acidity (lactic
acid content) goes on increasing as we increase the percentage
of honey11. This is because honey contains organic acids which
increases the acidity, also the lactose gets converted into
lactic acid
pH: The pH showed high significant difference (P>0.05)
between control and treatment yoghurt during different periods
of storage further increase in can sugar replacement with honey
from 1 to 6 percent resulted in decrease in pH from 4.28 ±0.01
to 4.10 ±0.1.5 per cent at 0 day of storage. The results are in
accordance with the results of earlier workers11. The pH value
goes on decreasing as we increase the percentage of honey.
This might be due to increase in acidity, as acidity and pH are
inversely proportional to each other.
Viscosity: There was no significant (P>0.05) difference
observed in viscosity of control (PY) and treatment yoghurt
samples viz., THSY1, THSY2 and THSY3 but significant
(P<0.05) difference was noticed between control (PY) and
(73) KARTHIKEYAN

Table-1.Nutritional properties of spirulina enriched and honey fortified yoghurt.

Different superscripts in a column differ significantly

Table-2.Viability of Lactobacillus delbrueckii spp. bulgaricus in treatment yoghurt (log10 cfu/ml).

(Different superscripts in uppercase in a row differ significantly)

( Different superscripts in lowercase in a column differ significantly )

Table-3.Viability of Streptococcus salivarius ssp. thermophilus treatment in yoghurt (log10 cfu/ml).

(Different superscripts in uppercase in a row differ significantly)

(Different superscripts in lowercase in a column differ significantly )
 NUTRITIONAL QUALITIES OF YOGHURT ENRICHED WITH SPIRULINA AND NATURAL HONEY
treatment yoghurt samples viz., THSY4, THSY5 and THSY6.
Some researchers opined that the apparent viscosity of
yoghurts fortified with protein and minerals (iron, magnesium,
zinc, manganese, molybdenum, chromium and selenium) did
not vary significantly (P >0.05) when compared to the control7.
Nutritional properties spirulina enriched and honey
fortified yoghurt: Nutritional properties viz., fat, total protein,
ash and total solids content of control and spirulina enriched
honey fortified yoghurt were showed in Table-1.
Total solids: The results showed that there was a significant
(P<0.05) difference in the total solids content between control
(PY) and different treatment of yoghurt. The increase in total
solids from 17.58 ±0.02 to 18.96 ±0.07. These results were
in accordance with the findings of some workers who defined
that the effect of enrichment of spirulina on gross composition
of yoghurt indicated that with increase in enrichment level
from 0 to 0.5 per cent, there was significant increase in total
solids from 17.85 to 18.96 per cent and while others reported
that, the total solid goes on increasing as we increase the
percentage of honey5,11. This is because honey contains fewer
amounts of water and more total solids.
Total protein: The increase in total protein from control to
treatment yoghurt is from 5.021 ±0.05 to 5.609 ±0.04. The
results revealed that there was a significant (P<0.05) difference
in the total protein content between control (PY) and different
treatment of yoghurt but no significant difference were observed
among the different treatments. As reported in earlier studies
that the increase in the level of spirulina from 2 to 8% caused
significant increased in the protein content of frozen yoghurt
containing 8% further enrichment of yoghurt with spirulina on
gross composition of yoghurt indicated that with increase in
enrichment level from 0 to 0.5 per cent, there was significant
increase in protein from 3.80 to 4.17 per cent5.
Fat content: No significant (P>0.05) difference was observed
in fat content between control and different treatment yoghurt.
The fat content ranged from 4.27 ± 0.01 to 4.79 ± 0.06 between
control and different treatment yoghurt. The results are in
accordance with findings of earlier workers who reported that
the increase in the level of spirulina from 2 to 8% caused no
significant increased in the fat content in frozen yoghurt7.
Ash content: The results revealed that there was no significant
(74)
(P<0.05) difference in the ash content between control (PY)
and different treatments of yoghurt. The non significant increase
in ash content from 0.84 to 0.88 per cent. The increase in the
level of spirulina in yoghurt caused non significant increment
of ash content in frozen yoghurt. Some workers defined that,
the effect of enrichment of yoghurt with spirulina on gross
composition of yoghurt indicated that increase in carbohydrate
and ash content in enriched spirulina yoghurt was found to be
non significant5,11.
Effect of enrichment of spirulina and honey fortification
on viability of yoghurt bacteria: The viability of Lactobacillus
delbrueckii ssp. bulgaricus and Streptococcus salivarius ssp.
thermophilus in developed yoghurt is presented in Table-2&3.
The results showed that there was no significant (P>0.05)
difference observed in count of Lactobacillus delbrueckii ssp.
bulgaricus and Streptococcus salivarius spp. thermophilus
between control and different treatments of yoghurt on 0, 7,
14 and 21 days. Whereas, the viable count significantly
(P<0.05) decreased in control and treatment yoghurt during
storage period and thus addition of spirulina and honey did not
affect the viability of Lactobacillus delbrueckii ssp. bulgaricus
and Streptococcus salivarius spp. thermophilus in yoghurt.
Earlier workers reported that S. platensis powder may
represent a unique source of nutrients for yoghurt bacteria
since it contains significant concentrations of amino acids,
precursors of nucleic acids, vitamins, mineral and etc., among
them also derivates of vitamin B which is a well known promoter
for the probiotic bacteria and reported that the viability of the
bacteria in all yoghurts increased when S. platensis powder
was added during the storage period7.
Sensory evaluation of spirulina enriched and honey
fortified yoghurt: Statistical analysis showed that a significant
(P<0.05) difference were observed between treatments with
regard to all sensory parameters. The results revealed that
enrichment of spirulina at 0.6% and fortification of honey at
upto 3 per cent level and 3 per cent replacement of can sugar
scored higher in sensory evaluation.
Cost of production for spirulina enriched and honey
fortified yoghurt: The cost of production for treatment yoghurt
was moderately higher than that of control, which can be
attributed to the cost of spirulina and natural honey. Though, it
is costlier a little bit, the resultant yoghurt is a healthier, as it
(75) KARTHIKEYAN
possesses numerous therapeutic/functional attributes which 2.
will definitely cater to the demands of the present day highly
health conscious consumers. Hence, production of enriched 3.
spirulina honey fortified yoghurt with 0.6 per cent spirulina with
4.
upto 50 per cent replacement of cane sugar with 3% level of
honey was found to be more beneficial at the rate of 4.65
rupees per 50 ml with numerous therapeutic properties.
5.
CONCLUSION
6.
The present study concluded that production of spirulina
enriched and honey fortified yoghurt with 0.6 per cent spirulina
and up to 50 per cent replacement of cane sugar with 3% level 7.
of honey was found to be more beneficial at the rate of 4.65
rupees per 50 ml with numerous therapeutic properties. Hence 8.
the developed yoghurt will not only provide nutrients qualities
but also posses the minimum required therapeutic levels of
10.
yoghurt probiotic cultures at above the level of 107cfu/g. which
will definitely meet the demand of the health conscious
11.
consumers at an reasonable cost.
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