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226 Wild Edible Plants
the active components in carob remain stable during extraction, showcasing carob as
a product with signicant nutritive potential. To quantitatively analyze the carotenoids
and chlorophylls in the experimental carob samples, lipid extracts were prepared and
evaluated using UV/ Vis spectroscopy (Figure 8.2).
Distinct peaks observed in the 425 to 480 nm wavelength range conrm the presence
of biologically active substances in carob samples, specically carotenoids such as α-
carotene, β- carotene, lycopene, and zeaxanthin. Absorption maxima at 410 and 670 nm
indicate the presence of chlorophyll compounds. The UV- Vis spectra analysis of the
lipid carob extracts revealed a high concentration of carotenoids in the carob seeds and
signicant chlorophyll content in the pod pulp.
8.3.2 Essential Minerals Content
In terms of mineral content, Moldovan carob pods demonstrated signicantly higher
levels of Ca, Mg, and Fe (4506.7 mg/ kg DM, 1864.4 mg/ kg DM, and 78.19 mg/ kg DM,
respectively) compared to Spanish, Italian, or Algerian carob seeds (Table 8.1).
Several studies have investigated the mineral composition of carob seeds, reporting
that content of Ca, Mg, and Fe range from 2510 to 4207 mg/ kg, 630 to 894 mg/ kg, and
12.5 to 42.57 mg/ kg, respectively (Fidan et al., 2020; Musa Özcan et al., 2007). While
the bulk of the research has focused on Turkish carob, studies on European carob remain
limited, making the obtained data particularly valuable for the European agrifood sector.
According to the amount of accumulated mineral elements, the studied carob samples
are arranged as follows:
Calcium (Ca): Moldova > Italy > Spain > Algeria
Magnesium (Mg): Moldova > Algeria > Spain > Italy
Iron (Fe): Moldova > Italy > Spain > Algeria
The information on the mineral content of carob pods is limited, with the exception
of Ayaz et al. (2007), who reported that Anatolian carob pods contained, mg/ 100
g FW: calcium, 300; magnesium, 60.0, and iron 1.88. In a later study, the same author
documented that home- prepared carob pod our had Ca, Mg, and Fe content of 304.0
mg/ 100 g; 55.4 mg/ 100 g, and 1.51 mg/ 100 g, respectively (Ayaz et al., 2009). Notably,
TABLE 8.1 Mineral content in carob cultivated in European and North Africa agrifood
sectors, mg/ kg dry weight (DW)
SAMPLE CA MG FE
Spanish carob seeds 2522.5±11.23
Italian carob seeds 2702.2±9.87
Algerian carob seeds 2178.5±14.22
Carob pods pulp from Moldova 4506.7±22.57
Carob seeds from Moldova 3816.4±17.56
a– d
averages in columns, signed by the same letter, are not statistically different (p ≤ 0.05).
b
b
a
d
c
1235.5±6.45
1047.1±4.25
1348.1±8.32
1864.4±9.81
1487.7±6.59
ab
a
b
c
bc
42.57±0.98
45.42±0.37
39.65±0.26
78.19±0.89
61.28±0.58
ab
b
a
d
c
Edible Wild Carob as a Source of Nutrients in Food Production 227
carob pod pulp exhibited higher content of each mineral element, underscoring the
potential for valorizing these by- products.
8.3.3 Antioxidant Activity
8.3.3.1 Antiradical activity
Additionally, to mineral content, the antioxidant activity was measured using
both the DPPH (2,2- Diphenyl- 1- picrylhydrazyl) and ABTS (2,2′- azino- bis- (3ethylbenzothiazoline- 6- sulfonic acid) radical scavenging assays. The total phenolic
content, total avonoid content, and total tannin content were also determined to provide a thorough understanding of the phenolic compounds present within the extracts
(Table 8.2).
The results obtained indicate a signicant direct correlation between the total
avonoids, tannins, and phenolic compounds and the antioxidant capacity of carob seeds
and pod pulp (0.86 ≤ R2 ≤ 0.98). This correlation can be attributed to the antioxidant
potential of the phenolic compounds present in the studied products, as well as the fact
that it underwent similar processing treatments. Consequently, it can be concluded that
carob is a valuable agro- food source for obtaining functional compounds. Comparable
ndings were reported by Biernacka et al. (2017) in their study of the total phenol content
in carob ber. According to Singh et al. (2016), the observed correlation is due to
the electron- donating properties of phenolic compounds.
TABLE 8.2 Phenolic compounds’ content and antioxidant activities of biological
compounds in carob samples
TOTAL CONTENT OF ANTIOXIDANT ACTIVITY BY
Sample
Spanish carob
PHENOLICS,
MG GAE/ G
22.72±0.43a1.11±0.01a0.94±0.01ab77.36±0.22a19.87±0.23
FLAVONOIDS,
MG GAE/ G
TANNINS,
MG/ G DPPH, %
ABTS, MG
TEAC/ G
a
seeds
Italian carob
23.94±0.18a1.45±0.03b0.89±0.03a80.12±0.56b21.42±0.25
b
seeds
Algerian
25.15±0.12ab1.67±0.01c1.01±0.02b81.28±0.95b21.79±0.86
b
carob seeds
Carob pods
30.56±0.21c1.98±0.03d1.51±0.02d85.13±0.35c25.52±0.45
c
pulp from
Moldova
Carob seeds
27.75±0.27b1.74±0.05cd1.262±0.04c81.40±0.49b22.84±0.36
bc
from
Moldova
a– d
averages in columns, signed by the same letter, are not statistically different (p ≤ 0.05).
228 Wild Edible Plants
8.3.3.2 Antioxidant activity expressed via gastrointestinal
in vitro digestion
To assess the antioxidant activity of the carob samples, a time- course simulation of
gastrointestinal digestion was carried out. The results from the evaluation of antioxidant activity after induced gastric digestion (an acidic environment) (Figure 8.4a– e)
indicated lower values for the Spanish seeds (40.00– 64.25)±0.1%, Italian seeds (38.15–
60.25)±0.2%, and Algerian seeds (39.28– 62.61)±0.6% compared to the Moldovan
carob samples, which ranged between (47.00– 70.07)±0.05% for the pulp of carob pods
and (48.00– 74.24)±0.03% for carob seeds.
The gradual increase in antioxidant activity over a period of 3 hours can be
attributed to the progressive release of biologically active compounds during gastric
digestion. Another signicant factor was the impact of solution pH and enzymatic
interactions within the samples. It was observed that the content of biologically active
compounds with antioxidant properties, such as polyphenols, avonoids, tannins, and
other compounds like carotenoids and chlorophyll groups, can enhance the antioxidant capacity of the samples analyzed. Data revealed that gastric digestion did not substantially alter the qualitative and quantitative composition of these compounds, which
maintained high stability under acidic conditions. This acidic environment, combined
with digestive enzymes, facilitated the release of biologically active compounds, thereby
boosting the antioxidant capacity of the carob samples studied.
During the gastrointestinal digestion process, the next phase simulated involved
intestinal digestion, where samples were incubated in an alkaline environment (pH = 8.2)
and antioxidant activity was measured over a 3- hour period. Results showed that the
antioxidant activity of Moldovan carob samples (pod pulp and seeds) was superior to
that of carob seeds from Spain, Algeria, and Italy. The values ranged between (40.05–
19.89)±0.06% for Moldovan carob pod pulp; (35.25– 18.69)±0.12% for Moldovan
carob seeds; (31.22– 14.65)±0.08% for Italian carob seeds; (30.04– 17.47)±0.34% for
Algerian carob seeds; and (32.56– 16.78)±0.58% for Spanish carob seeds. Following
intestinal digestion, a gradual reduction in antioxidant activity was observed over the 3-
hour period for both Moldovan carob samples and those from Spain, Italy, and Algeria.
This reduction can be explained by the low stability of biologically active compounds
under alkaline conditions (pH = 8.2) and the formation of metabolites that inhibit their
antioxidant activity in the samples analyzed. This process is a normal physiological
occurrence, as the absorption of biologically active substances typically takes place
within 2– 3 hours during their peak antioxidant activity.
FIGURE 8.4 Antioxidant activity of carob powders expressed via gastric and intestinal
digestion: (a) Moldovan carob pulp, (b) Moldovan carob seeds, (c) Spanish carob seeds,
(d) Italian carob seeds, (e) Algerian carob seeds.
Edible Wild Carob as a Source of Nutrients in Food Production 229
8.3.4 Pigment and Bioactive Compound Composition
in Carob Extracts
The carob extracts were analyzed for their content of essential pigments and bioactive
compounds, such as β- carotene, lycopene, and zeaxanthin. The presence of chlorophyll
a and chlorophyll b was also quantied, providing a comprehensive overview of the
pigment prole in the carob samples (Table 8.3).
Carotenoids are among the most prevalent pigments found in nature and serve as an
essential source of dietary vitamin A (Delgado- Vargas et al., 2000). However, there is a
notable scarcity of data regarding the carotenoid content in carob. According to Khatib
and Vaya (2010), carob pods from the Mediterranean region contain 0.2 mg of total
carotenoids per 100 g DW, including α- and β- carotene (0.08 mg/ 100 g DW), lycopene
(0.03 mg/ 100 g DW), and lutein (0.02 mg/ 100 g DW).
In this study, the primary carotenoids identied in the carob samples included β-
carotene, zeaxanthin, and lycopene. Italian carob seeds were notably rich in lycopene
(1.558 mg/ 100 g DW), while the Spanish and Algerian samples had mean lycopene
contents of 0.647 mg/ 100 g DW and 0.467 mg/ 100 g DW, respectively. Italian carob
seeds demonstrated the highest overall carotenoid content, with β- carotene at 1.126 mg/
100 g DW, lycopene at 1.558 mg/ 100 g DW, and zeaxanthin at 1.453 mg/ 100 g DW.
In contrast, the Spanish and Algerian samples exhibited lower carotenoid levels, with
β- carotene at 0.542 mg/ 100 g DW and 0.267 mg/ 100 g DW, lycopene at 0.647 mg/ 100
g DW and 0.467 mg/ 100 g DW, and zeaxanthin at 0.715 mg/ 100 g DW and 0.427 mg/
100 g DW, respectively.
Table 8.3 illustrates that the difference in carotenoid content between Moldovan
samples and those from other regions was statistically signicant (P ≤ 0.05). The
Moldovan carob seeds exhibited carotenoid levels more than ten times higher than the
TABLE 8.3 Carotenoid and chlorophyll content in carob cultivated in European and
North Africa agrifood sectors, mg/ 100 g DW
SAMPLE β- CAROTENE LYCOPENE ZEAXANTHIN CHLOROPHYLL A CHLOROPHYLL B
Carob seeds,
0.542±0.021b0.647±0.011ab0.715±0.009ab0.152±0.008a0.234±0.009
Spain
Carob seeds,
1.126±0.016ab1.558±0.038b1.453±0.015b0.501±0.004b0.605±0.005
Italy
Carob seeds,
0.267±0.011a0.467±0.022a0.427±0.014a0.146±0.021a0.122±0.007
Algeria
Carob pods
2.749±0.027c3.879±0.108c3.804±0.123c0.749±0.043c0.482±0.002
pulp,
Moldova
Carob seeds,
13.610±0.142d19.882±0.209d20.709±0.213d0.278±0.011d0.352±0.001
Moldova
a– d
averages in columns, signed by the same letter, are not statistically different (p ≤ 0.05).
a
b
c
d
ad
230 Wild Edible Plants
industrially sourced samples, particularly in the case of zeaxanthin, which reached up
to 20.71±0.42 mg/ 100 g DW. This discrepancy can be attributed to the varying pre-
treatment conditions of the raw materials. Despite being considered industrial waste and
relatively unused in the food industry, Moldovan carob pod pulp contains substantial
amounts of β- carotene, lycopene, and zeaxanthin, highlighting its signicant biological
potential.
Chlorophyll, known for its antioxidant properties, can neutralize the damaging
effects of free radicals similarly to vitamins A, C, and E (Mehdipoor Damiri et al.,
2021; Pérez- Gálvez et al., 2020; Queiroz Zepka et al., 2019). The two primary types
of chlorophyll (chlorophyll a and chlorophyll b), with their content in carob samples
are presented in Table 8.3. Although chlorophyll is not typically an essential parameter in carob pods or seeds, its presence has not been extensively studied in these
products. Table 8.3 data reveal that, in all seed samples except for the Algerian carob
seeds, chlorophyll b is more abundant than chlorophyll a, with contents ranging from
0.122 to 0.605 mg/ 100 g DW. The chlorophyll content in Moldovan seeds is comparable to that of other samples; however, higher levels were found in Moldovan carob
pod pulp, reaching 0.749±0.04 mg/ 100 g DW for chlorophyll a and 0.482±0.02 mg/
100 g DW for chlorophyll b. This indicates the signicant antioxidant potential of
Moldovan carob, including the pod pulp. Notably, in Moldovan carob pods, an antagonistic effect was observed, with chlorophyll a prevailing over chlorophyll b. The
chlorophyll a content in the pods (0.749 mg/ 100 g DW) surpassed that of all seed
samples examined.
8.4 QUALITY AND NUTRITIONAL VALUE OF
INNOVATIVE CAROB- BASED FUNCTIONAL
PASTRY SAUCES
Pastries encompass a wide array of products rich in fats and sugars (Jagarlamudi,
2022). Due to their high sugar and fat content, pastries are often viewed as indulgent, contributing to a sense of happiness (Guptill et al., 2023). However, this consumption is frequently linked to increased obesity rates among children and adults
(Basdeki et al., 2023; Solís- Guevara et al., 2022). At the point of serving, pastries
are commonly paired with sauces to enhance their avor. These sauces can either
complement the pastries or serve as a signicant addition. Basic pastry sauces typically consist of various ingredients, with sugar, chocolate, caramel, cream, fruits,
and berries being the most common (Alba- Martínez et al., 2022). These sauces are
popular not only in catering establishments but are also readily available in stores
for individual consumption. Consequently, pastry sauces improve the commercial
appeal of products in terms of avor, color, and appearance. However, their composition often increases the caloric content of pastries, categorizing them as obesogenic
foods. As a result, numerous studies focus on developing new natural additives for
food products (Capcanari et al., 2023b; Covaliov et al., 2023), including confectionery
Edible Wild Carob as a Source of Nutrients in Food Production 231
sauces, to prevent an increase in calorie content (Selvasekaran & Chidambaram,
2021; Stabnikova et al., 2021).
Vegetable- based raw materials are increasingly being employed in the development
of functional foods, including various types of sauces: emulsied sauces (MirzanajaZanjani et al., 2019), tomato sauces (Ferro et al., 2021), and confectionery sauces
(Abushal et al., 2021). These ingredients expand the range of available products,
enhancing their appeal to consumers. However, it should be noted that the inclusion of
ingredients like butter, oils, and sugar can elevate the energy content of these products.
Despite this increase in energy content, the biological value of such products often
remains low.
For the production of functional pastry sauces, a standard chocolate sauce recipe
was utilized. The conventional ingredients included cocoa powder, pasteurized milk
(3.5% fat), butter (82.5% fat), powdered sugar, vanilla extract, and processed drinking
water. To create the functional pastry sauce, carob powder derived from both carob
seeds and carob pods was incorporated.
Three sauce samples were prepared by using two different types of carob powder
and their mix (SCSP), each sourced from distinct morphological parts of the carob
plant: carob seeds (SCS) and carob pod pulp (SCP). The process involved moderate heat
treatment. During the preparation, it was observed that carob powder acts as a thickening
agent; therefore, in the carob pastry sauce formulation, sugar was replaced with water.
Initially, a mixture of pasteurized milk and vanilla essence was prepared. Butter
was melted at a temperature of 30°C and added to the prepared mixture. Subsequently,
the remaining ingredients, including powdered sugar (or water), and either carob pod
pulp or carob seed powder, were incorporated. A brief heat treatment (5 minutes) at a
temperature of up to 80°C with continuous stirring was conducted to achieve a homogenous mixture, which was then cooled to 20°C.
8.4.1 Physico- Chemical Characteristics
The development of innovative food products, such as functional sauces, calls for a
comprehensive understanding of their physico- chemical characteristics. These properties are pivotal in determining the quality, stability, and sensory appeal of the nal
product. In the quest to enhance the health benets and functional properties of pastry
sauces, the incorporation of carob powder presents a promising solution. The physicochemical characteristics of pastry sauces formulated with carob powder were evaluated,
examining key parameters such as protein, carbohydrate, lipid content, mineral composition, energy value, and antioxidant activity (Figure 8.5).
The data demonstrated that the inclusion of carob powder in the sauces signi-
cantly increased the levels of minerals such as calcium, iron, and potassium (K) when
compared to the control sauce made with cocoa and powdered sugar (Figure 8.6).
Specically, the calcium content more than doubled, iron content increased up to
nine times, and potassium content by 1.5 times in the carob- enhanced sauces. In terms
of protein content, there was very little change, while the levels of carbohydrates and
lipids signicantly decreased. Notably, the energy value of the carob pastry sauces was
232 Wild Edible Plants
FIGURE 8.5 Nutritional value of carob pastry sauces.
Control – classic chocolate sauce; SCP – sauce with carob pulp; SCS – sauce with carob
seeds; SCSP – sauce with mix of carob seeds and pulp.
FIGURE 8.6 Mineral content of carob pastry sauces.
Control – classic chocolate sauce; SCP – sauce with carob pulp; SCS – sauce with carob
seeds; SCSP – sauce with mix of carob seeds and pulp.
reduced from 230.7 kcal/ 100 g to 85.1– 88.9 kcal/ 100 g, representing a reduction of 2.7
times compared to the control sauce. This data indicates that the experimental sauces
possessed a higher biological value but a lower energy value, making them suitable as
functional sauces.
Studies on carob’s functional potential indicate it is a rich source of polyphenols,
known for their strong antioxidant activity (Goulas & Georgiou, 2019; Ioannou et al.,
Edible Wild Carob as a Source of Nutrients in Food Production 233
FIGURE 8.7 Total polyphenol content and antioxidant activity of carob pastry sauces.
Control – classic chocolate sauce; SCP – sauce with carob pulp; SCS – sauce with carob
seeds; SCSP – sauce with mix of carob seeds and pulp.
2023; Quiles- Carrillo et al., 2019). Total polyphenol content and DPPH antiradical
activity in the experimental sauces are presented in Figure 8.7.
There was a positive correlation between antioxidant activity and total phenol con-
tent in the studied sauces. Sauce, which was prepared with carob pod powder (SCP),
had a higher polyphenol content, 29.12 mg GAE/ g, compared to sauce made with carob
seed powder (CSP), leading to an increased antioxidant activity. According to Turhan
et al. (2006), the total polyphenol content in carob pods is 17.50 mg/ g. Mahtout et al.
(2016) reported that the total phenol content in carob pods and seeds are 10.53 mg/
g and 17.23 mg/ g, respectively. Cavallaro et al. (2021) suggest that the variation in
phenol content can be attributed to factors such as genotype, region of origin, soil type,
and precipitation levels. The control sauce made with cocoa powder had the lowest
total polyphenol content, 5.11 mg GAE/ g, and antioxidant activity of 60%. According
to Urbańska and Kowalska (2019), the total polyphenol content in fresh cocoa beans
ranges from 50– 60 mg/ g, but can decrease to 9.96– 37.81 mg/ g depending on the bean’s
origin and roasting parameters.
8.4.2 Rheological Properties
Understanding the rheological properties provides insights into the basic quality
indicators of foods through structural and mechanical characteristics (GonzalezGutierrez & Scanlon, 2018). Viscosity measurements were used to enhance the properties like structure, texture, and shape of the sauces. Rheological stability was assessed
by subjecting samples to increased shear stress and rates. It was found that the viscosity
of the emulsions signicantly decreases with increased shear stress and rate, indicating
the breakdown of structure (Figure 8.8).
Comparatively, sauces with carob powder exhibited greater stability and could
withstand higher shear rates (up to 4000– 5200 s–). The viscosity of the sauces was
234 Wild Edible Plants
FIGURE 8.8 Effective viscosity of the carob pastry sauces.
Control – classic chocolate sauce; SCP – sauce with carob pulp; SCS – sauce with carob
seeds; SCSP – sauce with mix of carob seeds and pulp.
directly related to their composition. For instance, the control sauce had a viscosity
change from 11.0 to 10.7 Pa·s, while the sauce with carob seed powder showed a change
from 16.5 to 16.0 Pa·s. After 4 months of storage, the changes in effective viscosity were
minimal, conrming the stability of the carob pastry sauces. These ndings suggest
that carob powders positively impact the viscosity and stability of the sauces under
increased shear stress and rates.
8.4.3 Sensory Test
A consumer acceptance test was conducted with over 100 participants, aged between
18 and 65 years. Participants were provided with a tasting sheet that outlined the primary quality parameters of the sauce and the grading scale. The parameters evaluated
included overall appearance, colour, texture, odour, and taste. The sauce’s quality
was assessed using a 9- point hedonic scale, ranging from “dislike extremely” to
“like extremely.” The scale was numbered from 1 to 9, where 1 represented “dislike
extremely,” 2 was “dislike very much,” 3 was “dislike,” 4 was “dislike slightly,” 5 was
“neither like nor dislike,” 6 was “like slightly,” 7 was “like,” 8 was “like very much,”
and 9 was “like extremely.”
The results indicated that all sauces had a pleasant avor and aroma, with consistent
textures for each type of confectionery sauce. Sauce with carob seeds (SCS), was noted
for its excellent appearance, consistency, and a ne, pleasant caramel avor, achieving
an average score of 8.80, while sauce with carob pod (SCP) had a pronounced dark
chocolate avor and was highly rated by tasters with an average score of 8.52. The
Edible Wild Carob as a Source of Nutrients in Food Production 235
TABLE 8.4 A check- all- that- apply (CATA) questionnaire terms for carob pastry sauces
PRODUCT ATTRIBUTE CATA TERMS
Appearance glossy, matte, smooth, lumpy, uniform, streaky
Colour too light, just the right colour, golden colour, too dark, reddish-
brown, blackish- brown
Texture thick, thin, creamy, gritty, smooth texture, lumpy, velvety
Odor rich chocolate, bitter odour, burnt odour, nutty odour, caramel
odour, pleasant odour
Taste intense chocolate, chocolate taste, sweet, too sweet, sugary
taste, caramel taste, intense caramel, bitter, burnt, nutty taste,
artificial
Enjoyment delicious, satisfying, indulgent, comforting
Emotions happy, excited, disappointed, indifferent, disgusted
sauce combining carob pod and seed powders (SCPS) also received high marks, with
an average score of 8.55. Based on these scores, all carob pastry sauces were deemed
acceptable.
A check- all- that- apply (CATA) questionnaire was administered following the
methodology of Biró et al. (2020). During the experiment, assessors were tasked with
identifying the sensory attributes present in the sample. Typically, a product’s attributes
are derived from its sensory characteristics but can also encompass hedonic terms,
emotions, and non- sensory qualities. This method allowed assessors to select an unlimited number of terms (Biró et al., 2020). A consensus among trained assessors produced
a list of terms, which is presented in Table 8.4.
The CATA questionnaire comprised 45 terms (Table 8.4). Among the 19 least
utilized terms were “matte,” “too light,” “golden color,” “reddish- brown,” “velvety,”
“burnt odor,” “sugary taste,” “intense caramel,” “burnt,” “articial,” and “indulgent.”
Cochran’s Q test was applied to lter out characteristics that do not statistically differentiate the samples to enhance the accuracy of the ndings. Henceforth, the analysis
incorporates only the remaining 26 key features (Figure 8.9).
According to the correspondence analysis (Figure 8.9), each sample was
associated with distinct attributes. Evaluators remarked that the control sample’s
taste and odor were bitter and overly sweet, and its color was excessively dark.
Concurrently, this sample elicited a sense of satisfaction and enjoyment. The sauce
with carob pod (SCP) demonstrated characteristics such as a lumpy and gritty texture,
a pleasant odor, and a sweet taste. However, some panellists expressed indifferent
emotions and disappointment. The most favored sample was the sauce with carob
seed (SCS), characterized by a chocolate and nutty taste, and a creamy and smooth
texture. This sauce evoked emotions such as deliciousness, comfort, and excitement.
The sauce combining carob pod and seed powders (SCPS) was positioned between
the SCB and SCS samples.
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