Abstract-- Saffron, containing
carotenoids such as crocin, is a natural colorant with potential uses in food
and pharmaceutical products and health benefits. In this study, the
water-soluble crocin, the color substance of saffron was produced with
spray-drying microencapsulation by maltodextrin as a wall material. The saffron
before and after microencapsulation was researched in terms of physicochemical
properties (moisture content and color values), total phenolic and flavonoid
compounds, antioxidant capacity, and microencapsulation retention. Also, the
bioactive and sensory properties in a model beverage obtained by adding these
saffron microcapsules to apple juices in different proportions (30-180 ppm)
were investigated. The result of the study showed that coloring apple juice
with the lowest dose of 30 ppm saffron microcapsules improved its bioactive
compounds, antioxidant activity, color, and overall flavor. This is the first
study to use saffron microcapsules as a colorant and the results show that
saffron microcapsules can be used as colorants and enrichments in food
formulation.
Keywords-- Crocus Sativus, bioactivity, sensorial properties, microencapsulation retention, food
model systems.
I. INTRODUCTION
Today, consumers increasingly prefer food products with natural
colors due to the health effects of nutrition. For this reason, for the food
industry to use natural products, it is of great importance to perform studies
on these components (Pinela et al., 2019).
Saffron (Crocus sativus L.) is a spice that has
dried stigmas and contains components of crocin, picrocrocin, and safranal,
which give intense aroma, yellow color, and bitter taste. Crocin, a
water-soluble carotenoid, is the color substance of saffron. When saffron is
dissolved in water, it gives a color ranging from yellow to red depending on
the concentration. The high solubility of saffron colors in the water, being
soluble and easily dispersible in water-based foods provides great advantages
as a food color compared with other carotenoids (Karasu et al., 2019).
Thanks to its functional characteristics such as coloring, aromatic and
flavoring, saffron is used in different foods sectors such as cookery,
beverages, confectionery, and bakery (Sarfarazi et al., 2019).
Bioactive
compounds of saffron provide many medically important health benefits. These compounds
are providing of antiinflammatory, anticancer, antihypertensive, antidiabetic,
antioxidant, antidepressant, and cytotoxic relaxant effects (Srivastava et
al., 2010; Ozkan et al., 2021). In addition to its positive effects
on learning and memory enhancements, it raises blood flow in the retina,
reduces the physical symptoms of morphine withdrawal, stomach ailments,
premenstrual syndrome, cardiovascular disease, insomnia, and anxiety (Melnyk et
al., 2010; Alavizadeh and Hosseinzadeh, 2014; Bolhassani et al.,
2014).
Having very
sensitive bioactive components, saffron is easily affected by environmental
conditions such as heat, light and oxygen, and its quality decreases (Rajabi et al., 2015). The microencapsulation
techniques are used to increase the stability and resolution of the bioactive
compounds (Tupuna et al., 2018). The choice of coating agent is very
important, as it affects the stability and encapsulation efficiency of the
capsules. Maltodextrin is the most widely used encapsulating material because
of its high-water solubility, low viscosity, low price, and flavorless. Among
the various encapsulation methods, spray drying is the most effective and
commonly used method in the food industry, since it is flexible and inexpensive
(Šturm et al., 2019).
Apple is one of
the fruits with high antioxidant value, which is widely used in fruit juice
production and contains essential nutrients for human health (Khanali et
al., 2020). The quality and health functionality of these products can be
further improved by adding ingredients rich in bioactive ingredients to formulated
products (Armellini et al., 2018).
The natural
color components from saffron are affected by environmental factors. Thus, in
this study, the saffron extracts were microencapsulated by spray drying
technique with maltodextrin as a wall material. There are few studies (Rajabi et
al., 2015; Karasu et al., 2019) on the spray-dried microencapsulated
color pigments extract from saffron stigmas and its bioactive properties. These
studies are different from the content of our research. Despite the importance
of saffron as a potential color substance and having bioactive compounds, the potential and effect of
microencapsulated saffron stigmas extract added in apple juice is not known. At
the same time, no studies have been carried out so far to detect the antioxidant
capacity, total phenolic and flavonoid compounds of saffron microcapsules.
Therefore, the aim of this study was to compare saffron, saffron microcapsules, and apple
juice colored with saffron microcapsules and their bioactive, physical
properties. As well as ıt was investigated sensorial properties of the food
model system. This study provides practical information on the physicochemical,
bioactive, and sensory properties of colored apple juices with different
proportions of saffron microcapsules.
II. METHODS
A. Material
Dried saffron stigmas were purchased from a local supplier in
Safranbolu, Turkey. The samples were stored at 4 oC in a dark place
for use in further analysis. All the chemicals were purchased from Merck
(Darmstadt, Germany).
B. Extraction of saffron with ultrasonic water bath
50 mg of powdered saffron was extracted with 50 ml distilled water
at 50 oC and 100% sonication amplitude for 5 minutes. The extracts
were centrifuged at 4500 rpm and filtered through 2.5 µm
filters (Whatman No. 42). These conditions were chosen
based on a study (Karasu et al., 2019). The extracts were kept at 4 °C
until analysis. Analysis was performed in triplicate.
C. Preparation of spray-dried saffron microcapsules
Before spray drying, the saffron extracts were prepared with an
ultrasonic bath (WiseClean, DH.WUC.D10H, Germany). 10 g of dry saffron stigmas
was extracted 500 ml distilled water, 100% amplitude at 50 oC. The
extracts filtered through 2.5 μm filters were concentrated in the evaporator
(BUCHI, Rotavapor R-210, Switzerland) device by adjusting the evaporation
conditions under the bath temperature 50 oC, boiling temperature 30 oC,
and pressure 85 mm bar vacuum and the °brix was brought to 4.3. Maltodextrin as
wall material 10% solid content was then added to the saffron extract to brix
15.7, and stirred (Ultra Turax, IKA, Germany) for 3 min at 10,000 rpm before
feeding to a spray dryer. The microencapsulation process was performed by a
spray dryer (BUCHI mini sprey dry B-290, Switzerland) with an airflow rate of
20%, a feed rate of 8 rpm, a rotary atomizer that an inlet air temperature of
175 °C and an outlet air temperature of 95 °C. The produced spray-dried saffron
microcapsules were packaged in polyethylene bags and kept at 4 °C for the
following studies (Karasu et al., 2019).
D. Preparation of juice colored with saffron microcapsules
Apple juice
used as a model system purchased from a market in Istanbul. Six different
formulations were prepared to achieve the best apple juice colored with saffron
microcapsules for our experiment. For this, 30-180 mg of saffron microcapsules
was added to 1L apple juice and homogenized (Fig. 1). The colored fruit juices
were stored at 4 °C until analysis.
E. Physicochemical properties
The pH of
the samples was measured using a pH meter (Seven compact pH/Ion S220,
Switzerland).
Color measurements were performed with HunterLab Color
Analyzer (USA) to obtain the CIELAB values as lightness (L*), red-green (a*),
yellow-blue (b*) (Wrolstad and Smith, 2017). The measurements were carried out
in triplicate.
F. Evaluation of bioactive properties
Total polyphenols and retention (%)
The total phenolic content (TPC) in the saffron, crocin
microcapsules, and colored apple juice with saffron microcapsules were analyzed
according to the Folin-Ciocalteu method reported by Singleton et al.
(1999). After mixing 2.5 ml Ciocalteu reagent (0.2 N) and 2 mL 7.5% (w / v) Na2CO3
into 0.5 mL extracts diluted 1:10 with distilled water, the mixture is kept at
room temperature for 30 min in the dark. The absorbance of the extracts was
detected spectrophotometrically at 760 nm (Shimadzu UV-1800, Japan) (Singleton et
al., 1999). The results were calculated as mg gallic acid equivalents (GAE)/g in dry stigmas.
The retention of TPC was determined with the following formula
(Ramakrishnan et al., 2018):
TPC %=
(TPC microcapsules/TPC extract) ×100 (1).
Total flavonoid compounds and retention (%)
The total flavonoid compounds (TFC) of the samples were performed by
the aluminum chloride colorimetric method. 1 mL of sample extracts were diluted
with 4 mL of distilled water and 0.3 mL of NaNO2 (5%) solutions and
after 5 minutes 0.3 mL of AlCl3 (10%) solutions was added (Zhishen et
al., 1999). After 6 minutes of incubation at room temperature, 2 mL of NaOH
(1 M) was added and the mixture was completed to 10 mL with distilled water.
The absorbance of the reaction mixture was determined spectrophotometrically
(Shimadzu UV-1800 spectrophotometer, Japan) at 510 nm (Zhishen et al.,
1999). The total flavonoid compound was expressed as catechin equivalent (CAE),
mg CAE/g.
The retention of the TFC was determined with the following formula
(Ramakrishnan et al., 2018):
TFC % =
(TFC microcapsules/TFC extract) ×100 (2).
Antioxidant capacity and retention (%)
The antioxidant capacity (AC) of the samples was determined
according to the CUPRAC method described by Apak et al. (2004). The
method is based on the reaction to convert the copper (II)-neocuproine compound
resulting from the reaction of neokuproin-Nc with copper (II) to Cu (I)
-neocuproine chelate giving absorbance at 450 nm. After adding 1 ml of CuCl2
solution (0.01 M), 1 ml of 1 M ammonium acetate buffer (pH 7.0), and 1 ml of
neocuproin (7.5 mM) solutions to the test tube, 0.1 mL of sample was added and
the total volume was completed to 4.1 mL with distilled water. The mixture was incubated for 1 h at room temperature in the dark and
the absorbance was measured spectrophotometrically (Shimadzu
UV-1800, Japan) (Apak et al., 2004). The results as Trolox (mg TEAC/g)
were determined using Trolox standard. The retention of the AC (Antioxidant capacity) was determined with
the following formula (Ramakrishnan et al., 2018):
AC
retention%=(ACmicrocapsules/ACextract)×100 (3)
flavonoids
and the antioxidant capacity of the saffron extract showed that it has the
important potential of saffron as a natural source of antioxidants. The use of
high temperatures in the spray dryer in microcapsule production may cause the
phenolic compounds to degrade.
The total phenolic, flavonoid substance contents and
antioxidant capacity results applied to colored apple juices are shown in Table
2.
As shown in Table 2, the total amount of phenolic
substance increases as the amount of microcapsules added to the fruit juice
increases, whereas the total amount of flavonoid substance does not change. It
was found that the effect of added saffron microcapsule food coloring on the
total phenolic substance and antioxidant capacity in the samples were
statistically significant (p˂0.05), while total flavonoid compound was not
significant (p>0.05). In general, we found a depending on the increase in
the amount of phenolic substance, the antioxidant capacity also increases. Similar
to our study, Sarabandi et al. (2019) explained that the antioxidant
activity of eggplant peel powders was correlated with their total phenolic
compounds.
B. Effect
of encapsulation process on retention (%)
Successful microencapsulation occurs when the active substance is
retent at the maximum level in the microcapsule. The retention (%) of TPC, TFC,
and AC ranged between 27.10 and 47.53, respectively. Although the
microencapsulation process is performed under the same conditions for both the
bioactive compounds, the TFC retention (%) had a higher compared to TPC
retention (%). AC retention (%) was the lowest with 27.10%. This can be
explained by the lack of heat lability of antioxidant compounds. Our findings
confirmed with a study (Rajabi et al., 2015) who reported 41.86–91.03%.
The total solids contents were in their study 10-40% and in our study 15% of
the emulsions. From their studies, it appears that the solids content and wall
material composition significantly affect the retention of bioactive
components. The difference in retention of bioactive substances can be
explained by the difference in solids content. Microencapsulation is an
appropriate technique that protects sensitive materials from adverse environmental
conditions. However, there is no optimal condition for every component.
C. Physicochemical properties
The pH value of saffron extracts is 7.32 shown in Table 1. This
result was found to be comparable to the studies of Cardone et al.
(2020). In this study, in which the effects of the physical and chemical
properties of the soil on the pH value of saffron were examined, like the
current study, it was found in the range 6.41-8.11 (Cardone et al.,
2020).
The pH analysis of apple juices colored with different
amounts of saffron microcapsules was determined by pH meter. Results are shown
in Table 2. In this study, in which different amounts of microcapsule dyes were
used to color fruit juices, it was determined that the effect of food coloring
on the change in the pH value of the samples was not statistically significant
(p> 0.05).
Moisture contents of saffron and saffron microcapsules
were reported as 6.64% and 6.45%, respectively, in our previous study (Karasu et
al., 2019).
Color Analysis
Color is a very important quality parameter especially for products
used as colorants. This is extremely important to make food products attractive
and marketable. The color values of saffron before and after microencapsulation
in our study are given in Table 1. When the color values L*, a*, and b* of
saffron in powder form were compared with the saffron microcapsule, a
significant increase was observed in these values. As the L* value approaches
100, the whiteness increases. It is thought that the increase in the L* value
is due to maltodextrin, which is white in color, and the increase in the a* and
b* values is due to the increase in the concentration of the crocin color
substance in the microcapsules, which has a color range from yellow to red. A
high b* value for microcapsules saffron represents greater yellowness. In a
similar study, Armellini et al. (2018) reported that the color of the
pasta enriched with saffron gradually increased a∗ and a∗/b∗
values depending on the increase in saffron concentration (0.2% and 0.4%).
The effect of food coloring on the CIE L* value
(brightness (100 = white, 0 = black)) of the samples was found to be
statistically significant (p˂0.05) in this study, in which saffron
microcapsules containing different amounts of crocin color were used for
coloring apple juices. The ES-coded control group constituted the highest L*
value among the samples. The highest a* value was seen in the sample coded with
OS in fruit juices colored with crocin microcapsules. It is seen that there is
a statistically significant difference (p <0.05) between the a* values of
the colored samples. Among the colored apple juices, the highest b* value
(yellowness (positive value)) was found as 5.96 in the OES-coded sample. It was
determined that the effect of food coloring on the CIE b* value, the measure of
the samples was statistically significant (p˂0.05).
D. Sensory Analysis
In the scope of the study, sensory analyzes were conducted with 17
trained panelists using the hedonic scale method to determine the effects of
microcapsule saffron dye added to apple juice samples in different amounts on
sensory quality (Lim et al., 2009). Sensory analysis was evaluated in
terms of color, odor, taste, flavor, aroma, and general taste in the range of
1-5 points. Sensory analysis findings of apple juice samples colored with
saffron microcapsule dyes are shown in Table 3. Color is one of the most
important quality parameters determining consumer preference in purchasing
food. Apple juices colored with different amounts of saffron microcapsules Fig.
1 shows apple juices colored with different amounts of saffron microcapsules.
As can be seen from Fig. 1, as the concentration of saffron microcapsules dyes
increases, the color gets darker.
Among the samples with color scores between 3.41 and
4, the most preferred sample in terms of color was the colored apple juice
coded US (4), while the least liked sample was determined as OS (3.41). In this
study, in which different amounts of saffron microcapsule dyes were used in
coloring apple juices, it was found that the effect of food coloring for the
color value of all samples compared to the control group (ES) was statistically
significant (p˂0.1) in the sensory analysis on the color parameter of the
samples.
Smell scores range from 3.76-4.00. While BS was the
most liked example in terms of odor, the least liked example was the OES
sample. In this study, in which different amounts of saffron microcapsule dyes
were used in colored apple juices, it was determined that the effect of food
coloring on the odor value of the samples was not statistically significant
(p> 0.05).
Taste scores range from 3.58-4.12. While OES was the
most popular example in terms of taste, the least liked US and BS samples were.
It was determined that the effect of food coloring on the taste value of all
samples compared to the control group (ES) was not statistically significant
(p> 0.05).
It has been observed that the flavor scores range
between 3.88-4.12. Accordingly, while the most delicious base and OES samples
were found, it was observed that the least flavorful sample was BS. In this
study, in which different amounts of saffron microcapsule dyes were used in
colored apple juices, it was determined that the effect of food coloring on the
flavor value of the samples was not statistically significant (p> 0.05).
It was observed that the aroma scores ranged from 4.00
to 4.24. While OS and OES coded samples were found with the most prominent
aroma, the samples with the least aroma were US and BS. It is estimated that a
more pronounced aroma is released as the amount of saffron microcapsule dye
dissolved in apple juices increases. When all samples were compared with the
control group (ES) in terms of aroma value, it was determined that the effect
of food coloring was not statistically significant (p> 0.05).
Fig 1. Apple juices colored with different
amounts of saffron microcapsules.
* ES: Control sample without microcapsules, US: 30 mg/L
microcapsule added apple juice, BS: 50 mg/L microcapsule added apple juice, OES:
120 mg/L microcapsule added apple juice, OS: 180 mg/L microcapsule added apple
juice.
It was observed that the general appreciation scores
varied between 4.10-3.82. It was observed that the samples coded the US (4.10)
received the most general appreciation among the samples stained with saffron
microcapsule dye. It was determined that the general taste
did not increase in parallel with the increase in food coloring, and
the effect of food coloring on the general taste scores was not statistically
significant (p> 0.05).
The most admired example in terms of color and the
general taste was the US sample. US (30 ppm) is the example in which the least
amount of food coloring is used. In this case, it has been concluded that the
use of a small amount of microcapsule saffron dye is sufficient in terms of
making the product more attractive and getting more appreciation.
Armellini et al. (2018) analyzed the sensory
analysis of different properties of pasta samples enriched with saffron, and
evaluated in terms of acceptability, color, aroma and chewiness; while the
color assessment is in line with our study, they found significant differences
between the samples for other parameters, unlike our study. Cam et al. (2014),
in the study in which the sensory properties of control samples and ice cream
samples containing microencapsulated pomegranate peel phenolics (0.5% and 1.0%) were determined, they were
observed that the sensory properties of enriched ice creams got higher scores
similar to our study, and there was no statistically significant difference
between the liking scores of the samples. Turgut and Diler (2020) found that
adding coriander to cheese in different proportions significantly affected
sensory characteristics such as appearance, texture, odor, flavor and general
acceptability, unlike our study.
According to the results of the sensory analysis, it
was determined that the color, odor, and flavor parameters of the fruit juice prepared
with the addition of 30 ppm microcapsule saffron dyes were positively affected.
It was concluded that the use of saffron microcapsule dyes in the coloring of
apple juices not only made the color more attractive but also increased the
total phenolic compounds and antioxidant capacity, so it was concluded that the
new product also gained functional properties.
V. CONCLUSIONS
Saffron is a natural pigment containing
crocin that has the potential to be used in food and pharmaceutical ingredients
with health benefits. This study provides scientific data on the physical and
bioactive properties of saffron, saffron microcapsules, and apple juices that
color with saffron microcapsule dyes. Compared to saffron stigmas and saffron
microcapsule extract, microcapsules have advantages such as being practical to
use, being easily soluble in water, and providing homogeneous distribution. Thus,
time and pre-treatment costs are saved. In addition, it is predicted that
operations can be made easier by determining the usage dosage while producing
with standard features. It has been observed that saffron microcapsule dyes,
which are used in minimum amounts (30 ppm) for coloring, not only increase the
appeal of apple juice but also enrich the product. The fact that crocin, the
color substance of saffron, is well soluble in water, shows that saffron
microcapsules can be easily used in water-based food formulations and to
develop functional products.
Colors are one
of the most important sensory perceptions in foods and affect the consumer's
choice. This is the first study to highlight the potential for use in model
food of saffron microcapsules, which are easy to apply as a natural color, have
new organoleptic and technological innovations with bioactive ingredients. By
adding natural food dyes rich in such bioactive compounds to formulated food
products, the quality and health functionality of these products can be further
improved.
Conflict
of Interest Statement: The authors declare that
there is no conflict of interest.
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Received: November 29, 2021
Sent to Subject Editor: December 15, 2021
Accepted: May 4, 2022
Recommended by Subject Editor Diego Lomonaco
