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Potential of Cinnamon (Cinnamomum Cassia)
as an Anti-Oxidative and Anti-Microbial Agent
in Sudanese Yoghurt (Zabadi)
Alia H. Suliman1, Khogali E Ahmed1, Babiker E Mohamed1 and Elfadil E Babiker
1Department of Food Science and Technology, Faculty of Agriculture, University of Khartoum, Shambat, Khartoum North, Sudan
2Department of Food Science and Nutrition, College of Food and Agricultural Sciences, King Saud University, Saudi Arabia
Submission: May 03, 2019; Published: May 31, 2019
*Corresponding author: Elfadil E Babiker, Department of Food Science and Technology, Faculty of Agriculture, University of Khartoum, Shambat, Khartoum North, Sudan
How to cite this article: Alia H. Suliman, Khogali E Ahmed, Babiker E Mohamed,Elfadil E Babiker. Potential of Cinnamon (Cinnamomum Cassia) as an Anti-Oxidative and Anti-Microbial Agent in Sudanese Yoghurt (Zabadi). Dairy and Vet Sci J. 2019; 12(2): 555833. DOI: 10.19080/JDVS.2019.12.555833
This study evaluated the biological efficacy of cinnamon powder (CP) from Cinnamomum cassia in cow milk yogurt. CP had a total flavonoid and phenolic content of 132.87 and 818.73mg/100 g, respectively. CP exhibited significant free radical scavenging properties with antioxidant activity clearly correlated with CP concentration. CP extracts, especially the methanolic extract, produced large inhibition zones against pathogenic bacteria, comparable to that of penicillin. Yogurt (0.5%, and 1.0% CP) was refrigerated for 7 and 14 days. CP increased total yogurt solids with no effect from storage. Titratable acidity, pH, and lactose content significantly decreased with storage and CP concentration. Total bacterial, coliform, and E. coli counts significantly decreased with CP concentration. Lactic acid bacteria significantly increased with both concentration and storage. Sensory attributes following CP addition were rated slightly lower, except taste. Results indicated that cinnamon is effective against pathogenic bacteria, highlighting the possibility of processing yogurt with cinnamon.
Herbs and spices have been used since ancient times, not only as antioxidants and flavoring agents but also for their antimicrobial activity against degradation caused by food-borne pathogens and food spoilage bacteria. Herbs and spices are rich in phytochemical antioxidants and among the top 50 foods with antioxidants, the top five antioxidants were dried spices . Cinnamon has been known as one of the most common spices and food flavoring additives since ancient times. For instance, it has been used to flavor sweets and chewing gum due to its pleasant and refreshing sensory attributes. It also shows beneficial effects on oral health and is used for toothaches, oral infections, and to remove bad breath . Cinnamon is mainly used in the aroma and essence industries owing to its fragrance, which can be incorporated into different varieties of foodstuffs, perfumes, and medicinal products. The most important constituents of cinnamon are cinnamaldehyde and trans-cinnamaldehyde, which are present in the essential oil, thus contributing to the fragrance and various biological activities observed with cinnamon .
Cinnamon significantly lower low-density lipo-protein (LDL) or “bad” cholesterol, triglycerides, and total cholesterol, and also reduces blood sugar levels in treating type 2 diabetes . The considerable increase in demand for fermented milks observed in recent years has resulted, to a great extent, from consumer awareness of their beneficial effects. However, fermented milks are also highly valued for their unique taste and aroma, which also contributed to their growing popularity. Yogurt, produced by lactic acid bacteria (LAB), is one of the most popular fermented dairy products consumed globally. Moreover, it is more nutritious than many other fermented milk products since it contains a high level of milk solids in addition to nutrients produced during the fermentation process. Various types of yogurt are now available in markets, including stirred, set, frozen, liquid, and flavored yogurt . Sudanese yogurt is mostly made from cow milk, with no added ingredients. The Processing of it is carried out at a local level and the procedure includes boiling and cooling of milk and inoculation with a 1-day-old previous batch of zabadi which serves as starter . The milk and the starter mixture are then incubated at room
temperature (~25 °C) for 12-15 h. The shelf life of the yogurt is
limited to 3-4 days after processing and based on the favorable
characteristics of cinnamon, this study aimed to evaluate the
potentiality of cinnamon powder as an anti-oxidative and
antimicrobial agent in extending the shelf life Sudanese yogurt.
Fresh milk was obtained from the University of Khartoum
Farm. The milk samples were transferred in an ice box in a
stainless-steel container and kept refrigerated until used to
manufacture yogurt. Cinnamomum cassia bark was purchased
from the local market, Khartoum North, Sudan. Cinnamomum
cassia bark was ground to pass a 0.4 mm screen. All chemicals
used in this study were of analytical grade.
C. cassia bark powder (5g) was extracted with 50ml methanol
or water at room temperature (24 °C) for 12h using an orbital
shaker. The extract was then filtered and centrifuged (Hettich
Zentrifugen, Tuttlingen, Germany) at 4000rpm for 10 minutes,
and the supernatant was concentrated under reduced pressure at
40 °C for 3hr using a rotary evaporator (IKA-WERKE RV06ML).
The total phenolic content was quantified using the Folin-
Ciocalteu reagent (FCR) method as described by Madaan .
A gallic acid standard curve was prepared to evaluate the total
phenolics in terms of gallic acid equivalents (mg GAE/100g) and
the total phenolic content was expressed as mg GAE/100g.
The colorimetric method described by Kim  was used to
analyze the total flavonoid content of cinnamon using catechin as
a standard. A calibration curve was prepared using catechin, and
the results were expressed as mg catechin equivalents per gram of
sample (mg CE/g).
DPPH scavenging activity was determined using the method
described by Lee . A solution of DPPH in methanolic acid was
used to assess the antioxidant activity of samples. The antioxidant
activity was calculated as follows:
Antioxidant activity (%) = (1 − A1/A0) × 100,
Where A0 and A1 are the absorbance of the control and extract,
The TRPA of the sample was determined according to the
method described by Oyaizu . The TRPA of the powder at
different concentrations was determined using vitamin C as a
positive control, and the results were expressed as vitamin C
Raw milk was subjected to heat treatment (70 °C) for ~20 min,
and then left to cool at room temperature (25 °C). A starter culture
(1-dayold) from a previous batch of zabadi was added to the milk
(5g/kg milk), thereafter, mixed with different concentrations
of CP (0%, 0.5% or 1.0%) and then incubated overnight at the
room temperature (25 °C). Part of the fresh zabadi was subjected
to analysis and the rest was packed in plastic containers, closed
tightly and stored for 7 and 14 days at 4 °C. Samples were reanalyzed
at the end of each storage period.
Lactose (%) was determined according to the method by Teles
. A standard solution of dry lactose in distilled water (%) was
used to determine the lactose content in treated and untreated
yogurt using a colorimeter at 520nm.
The disc diffusion method by Boyanova  was used to
assess the antimicrobial activity of cinnamon powder (CP) extract.Briefly, pure cultures of indicator microorganisms (Escherichia
coli ATCC 10536, Salmonella typhimurium ATCC 14028, Bacillus
cereus ATCC 14579, and Staphylococcus aureus ATCC 29737) were
cultivated on nutrient agar plates. Paper discs saturated with
aqueous or methanolic CP extract (10μg/mL) were suitability
positioned on the surface of the cultures and incubated at 37 °C for
24h. Then, inhibition zones surrounding the discs were measured.
Penicillin (10μg/disc), a standard antibiotic, was used as a control.
The method of Harrigan  was applied to determine the
microbial characteristics of yogurt at different periods of storage
by plate count. Coliform bacteria count was carried out by using
the Most Probable Number (MPN) technique. For S. aureus bairdparker
medium was used. MRS medium was used for counting
lactobacillus bulgaricus. The count of streptococcus spp. was done
by plating the suitable dilution in M-17 agar. The plates were
incubated at 37 °C for 48 hours. Sample pH was determined with
a pH meter probe (Corning Scientific Products, New York, USA).
Panels of 20 semi-trained members (men and women between
20-35 years old) were asked to evaluate the sensory properties of
treated and untreated yogurt on a 7-point hedonic scale. Prior to
sample evaluation, three training sessions were carried to acquaint
panelists with the attributes expected to be measured. Panelists
were asked to evaluate the texture, flavor, color, taste, juiciness,
and overall acceptability of each sample. Scores were measured
from “like extremely” to “dislike extremely,” corresponding to
ratings from 7 to 1, and tests were carried out three times at
each evaluation interval (0, 7, and 14 days). Mean scores for each
sample and session were calculated.
All measurements were carried out in triplicate, and three
batches of yogurt were produced. The effect of CP on parameters
was analyzed statistically using SAS software (v 8.1, SAS Institute
Inc., Cary, NC). Data on the parameters measured based on various
treatments, storage periods, and their interactions were analyzed
using the general linear model (Two-way ANOVA), and statistical
differenvces were estimated using Duncan’s multiple range tests.
Mean separation was determined using least significant difference,
and data are reported as the mean ± standard deviation (SD). The
significance level was accepted at P ≤ 0.05.
The total phenolic content, flavonoid content, and antioxidant
activity of CP are presented in Table 1. The total phenolic content
of CP was found to be 818.73 mg GAE/100 g. This result was
higher than that reported by Kumar and Prakash (2012) for Cassia
fistula and C. cassia. Yang and Chuang (2012) reported that the
highest phenolic content in ethanolic extracts of cinnamon was
observed in the bark, followed by the leaves and buds. Cinnamon
contained 132.87 CE mg/100 g of flavonoid compounds, higher
than the values reported by Kumar & Prakash  for C. fistula
and C. cassia. Prasad  reported 98.11mg/100 g of flavonoids
in 50% ethanol extracts of Chinese C. cassia leaves, while Yang
& Chuang  reported 2.030 g/100 g of the flavonoid content
of bark in 95% ethanolic extracts. The capacity of flavonoids to
act as antioxidants depends upon their molecular structure. The
position of hydroxyl groups and other features in the chemical
structure of flavonoids are important for their antioxidant and free
radical scavenging activities . Flavonoids are potent watersoluble
antioxidants and free radical scavengers which prevent
oxidative cell damage, and they have strong anticancer activity; as
antioxidants, flavonoids from spices provide an anti-inflammatory
As shown in Table 1, the DPPH scavenging activity of C. cassia
was 80.45%. Kumar & Prakash , who studied DPPH scavenging
activity in four medicinal plants, reported that the highest
antioxidant activity (91.66 ± 4.33) was observed in C. fistula
followed by C. cassia. This finding is consistent with the previous
literature reported by Lin. The H2O2 scavenging activity of C. cassia
at various concentrations differed significantly (P ≤ 0.05). H2O2
scavenging was found to be 59.27%, 44.30%, 33.07%, and 24.07%
for 1000, 500, 250, and 125μl of extract, respectively. Scavenging
activity decreased as CP concentration decreased. Similarly,
previous study  showed a highly positive linear relationship
between antioxidant activity and total phenolic content in some
spices. Many studies have reported that phenolic compounds in
spices and herbs significantly contribute to their antioxidant and
pharmaceutical properties . Hydrogen peroxide itself is not
very reactive, yet it is sometimes toxic to cells as it may produce
hydroxyl radicals. H2O2 scavenging activity by extracts may be
attributed to their phenolic compounds, which can neutralize
H2O2 to water by electron donation.
Values are mean (± SD) of triplicate samples. †diphenyl-2-picrylhydrazyl.
The total reducing power of C. cassia extract increased
along with CP concentration, indicating that it is concentration
dependent. This result was comparable to that of Kamleshiya
, who studied the reducing power of aqueous and methanolic
C. cassia extracts at different concentrations. The results of the
present study indicate that C. cassia extract exhibited high H2O2
and DPPH scavenging activities and reducing power ability. The
overall antioxidant activity of C. cassia might be attributed to its
phenolic and flavonoid contents as well as other phytochemical
The antimicrobial activity of aqueous and methanolic CP
extracts against two gram-negative (Escherichia coli ATCC 10536
and Salmonella typhimurium ATCC 14028) and two gram-positive
(Staphylococcus aureus ATCC 29737 and Bacillus cereus ATCC
14579) pathogenic bacteria is shown in Table 2. Aqueous C. cassia
extract exhibited antibacterial activity against all bacterial strains.
The inhibition zone of the aqueous extract was found to be 14.67
and 12.78 mm for E. coli and S. typhimurium, respectively, and that
of the methanolic extract were 18.67 and 17.88 mm, respectively.
The penicillin inhibition zone was significantly (P ≤ 0.05) larger
than that of aqueous and methanolic extracts against gramnegative
bacteria, but comparable to that of the methanolic extract
against gram-positive bacteria. The inhibition zone of the aqueous
extract was found to be 16.67 and 12.67 mm for S. aureus and B.
cereus, respectively, and that of the methanolic extract were 20.33
and 14.13 mm, respectively. Again, penicillin exhibited larger
zones. The inhibition zone of the methanolic extract, comparable
to that of penicillin, was found to be significantly (P ≤ 0.05) larger
than that of the aqueous extract for all pathogenic bacteria. This
may be due to the extraction of oil with an organic solvent as
reported by Huang, who stated that the essential oil from C. cassia
bark possessed antibacterial activities against four food spoilage
bacteria. These results agree with that reported by Anees ,
who found that methanolic C. cassia extract exhibited antibacterial
activity against isolated bacteria.
Values are means ± SD of triplicate samples. Means sharing a different superscript in a row are significantly different at P ≤ 0.05.
The results of the present study indicated that CP had a great
effect on pathogenic bacteria and other food-borne microorganisms
and this could be due to antimicrobial mechanisms exhibited by
C. cassia bark extract. Similarly, Chaudhry & Tariq  reported
that the essential oil extracted from C. cassia bark exhibited
antibacterial effects against food borne pathogens such as S.
aureus, Listeria monocytogenes, Streptococcus oralis, Streptococcus
anginosus, and E. coli, and claimed that these antimicrobial
properties may come from cinnamaldehyde as well as a variety of
other active components.
Values are mean ± SD of triplicate samples. Means sharing a different superscript in a row (p, q, r, s.) or column (a, b, c, ..) are significantly different
at P ≤ 0.05.
The effects of C. cassia powder concentration and storage
period on the physicochemical properties of yogurt (total solids,
pH, titratable acidity, and lactose content) are shown in Table 3.
Total solids decreased slightly over longer storage periods but
increased significantly (P ≤ 0.05) with CP concentration, reaching
12.35% prior to storage with little reduction over the total period.
The results indicated that the addition of cinnamon increased
total yogurt solids alongside cinnamon concentration, although
this increase was lower than that reported by El-bakri & El-zubeir
. Sample pH decreased significantly over the storage period,
but cinnamon concentration had a significant (P ≤ 0.05) effect on
pH, especially at high concentrations, indicating that the addition
of CP at different concentrations may help stabilize yogurt pH.
The titratable acidity of yogurt was significantly (P ≤ 0.05)
affected by the storage period and by the concentration of C. cassia;
it was observed to decrease slightly as cinnamon concentration
increased due to increase in pH. Prior to storage, the titratable
acidity ranged from 0.83 to 0.85 and increased significantly (P ≤
0.05) to a range of 0.93-1.53 after 14 days due to decrease in pH.
The results agree with those reported by Eissa , who attributed
the increase in titratable acidity and decrease in pH to lactose
metabolism and reported that these effects occurred over time
during fermentation by LAB. Plain yogurt contained low levels of
lactose (3.47%) compared to that extended with 0.5% and 1% C.
cassia (4.73% and 4.87%, respectively). Lactose content increased
significantly (P ≤ 0.05) with cinnamon concentration. However,
the decline in lactose level was insignificant (P ≥ 0.05) until
day 7 of storage, after which it lowered significantly (P ≤ 0.05),
especially in yogurt containing 0.5% and 1% C. cassia. The increase
in LAB during storage may be responsible for the increase in acid
production due to lactose fermentation .
The microbial load of treated and untreated yogurt during
storage is shown in Table 4. The results showed that plain yogurt
contained the highest bacterial count (p ≤ 0.05) for all storage
periods, ranging between 4.97 and 7.85. Treatment of yogurt with
0.5% and 1% C. cassia powder significantly (p ≤ 0.05) decreased
the total viable bacterial count; the highest reduction was
observed in the latter sample. The storage period also significantly
(p ≤ 0.05) affected the total viable bacterial count in all samples
studied. The highest count was found on day 7 of storage, after
which it decreased significantly through day 14 for all samples.
This could be attributed to increased lactic acid levels produced
during the storage period .
Values are mean ± SD of triplicate samples. Means sharing a different superscript in a row (p, q, r, s, ..) or column (a, b, c, ..) are significantly different at P ≤ 0.05. †Cfu, colony forming units; ‡MPN, most probable number; §ND, not detected.
As shown in Table 4, the S. aureus count in untreated yogurt
was 2.78 cfu/g. Addition of 0.5 and 1% C. cassia significantly
decreased the count to 1.56 and 1.48, respectively. The results
showed that the S. aureus count decreased with increased C. cassia
concentration. The storage period was also found to affect the
S. aureus count; it dropped to 2.53 after 7 days and disappeared
completely on day 14 in the control sample. The disappearance
of S. aureus could be due to the reduction in pH as well as
competitive growth of other micro-organisms such as lactic acid
bacteria which had inhibitory effect on S. aureus growth . The
presence of S. aureus indicated that milk could be contaminated
during handling. Samples treated with either amount of C. cassia
showed a negative S. aureus count in the two storage periods,
possibly due to the antimicrobial action of C. cassia powder.
The coliform count in untreated yogurt was 22.00 MPN/g.
The addition of 0.5% and 1% C. cassia significantly (p ≤ 0.05)
decreased the coliform count to 14.00 and 12.33 MPN/g,
respectively. Coliform presence could be attributed to milk
contamination during handling. The results showed that coliform
count decreased significantly (p ≤ 0.05) as C. cassia concentration
increased. This may indicate the antimicrobial effect of C. cassia
powder in yogurt. The storage period was found to affect coliform
count, which dropped significantly (p ≤ 0.05) to 14.33 by day 7
of storage and disappeared completely on day 14 in the control
sample due to increase in lactic acid. Samples treated with 0.5%
and 1% C. cassia powder showed null coliform counts during the
two storage periods (7 and 14 days) investigated. The E. coli count
in untreated yogurt was 3.33 MPN/g and dropped significantly (p
≤ 0.05) to 1 and 0 for samples treated with 0.5% and 1% C. cassia,
respectively, clearly indicating an inhibitory effect on yogurt E.
coli by cinnamon. The storage period significantly (P ≤ 0.05)
affected the E. coli count in yogurt. All samples studied showed
no E. coli growth during the storage periods investigated (7 and
14 days). Again, the absence of coliform bacteria and E. coli in
treated samples could be due to the microbial action of C. cassia
resulting from its phytochemicals compounds such as flavonoids
In general, LAB (Streptococcus and Lactobacillus) increased
significantly (p ≤ 0.05) with both C. cassia concentration and
storage period, as shown in Table 4. Generally, Streptococcus
was dominant compared to Lactobacillus in plain and treated
yogurt, even during storage. Treatment of yogurt with 0.5% and
1% C. cassia significantly (P ≤ 0.05) increased the Streptococcus
count, indicating that the addition of cinnamon improves LAB
availability to reach acceptable levels. The storage period also
significantly (P ≤ 0.05) affected the Streptococcus count in all
samples studied. The count was highest on day 7, after which it
decreased significantly (p ≤ 0.05) in all samples. This could be
attributed to the reduction of nutrients in yogurt or the increase
of bacterial lactic acid production. The significant drop by day
14 in viable S. thermophilus cell counts in cow milk yogurt may
be attributed to the accumulation of organic acids. Treatment
of yogurt with 0.5% and 1% C. cassia significantly (P ≤ 0.05)
increased the Lactobacillus count, with the highest observed in
the latter samples. These results were similar to previous findings
by Eissa , who reported that fermentation improved proteolytic
activity and enhanced LAB growth in cow and goat milk. The
increase in Lactobacillus count lies within the acceptable level (106). The storage period also significantly (P ≤ 0.05) affected the
Lactobacillus count in all samples studied. The peak count was
found on day 7, after which it decreased significantly (P ≤ 0.05)
in all samples. This result agreed with a previous study that found
refrigerated storage significantly decreased the viable counts of
Lactobacillus spp. by day 14 of refrigerated storage. Additionally,
the reduction in Lactobacillus spp. counts could be associated with
the post-acidification of yogurt, which further reduces pH .
The effect of C. cassia powder on the final characteristics of
the yoghurt was evaluated by sensory analysis on samples at day
0 and during refrigerated storage until day 14 (Table 5). Twenty
panelists were asked to score color, flavor, texture, taste and overall
acceptability. During storage, the levels of the sensory attributes
of yoghurt were higher when CP was added. All panelists evaluated
the plain yoghurt as less consistent and more acid compared to
the yoghurt containing CP. This result could be attributed to the
low microbiological quality of the raw milk; in fact, although
pasteurization decreased the levels of TMM, the metabolites
of the indigenous microorganisms that grew before thermal
treatment could have impacted this aspect. On the contrary, CP
odors might have limited perception of such traits. Microbial
hydrolysis of yoogurt component during storage was found to be
the key deteriorating factor with regard to taste, color, flavor and
texture . The most notable differences between the control
and CP yoghurt were observed for color and flavor intensity. In
general, the data obtained suggested that color, flavor and texture
were the three most important parameters reflecting the overall
acceptability of yoghurt containing CP. Regarding texture, the
control yoghurt showed a decrease of the values with storage,
while CP yoghurt showed almost increasing levels. Similar results
were observed in yoghurt manufactured from goat’s milk .
Values are mean ± SD of triplicate samples. Means sharing a different superscript in a row (p, q, r, s, ..) or column (a, b, c, ..) are significantly different at P ≤ 0.05.
C. cexhibits significant antioxidant and antibacterial
activities and maintains LAB at acceptable levels. The use of C.
cassia bark powder in yogurt resulted in significantly decreased
acidity with an increase in concentration, improved product
taste, delayed yogurt acidity, and reduced total coliform count
and total viable counts of E. coli and S. aureus. C. cpowder
can be utilized as a functional additive to preserve food against
pathogens and extend shelf life. The results of this investigation
highlighted the possibility of processing yogurt with C. cassia.