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Article

Impact of Functional Supplement Based on Cornelian Cherry (Cornus mas L.) Juice in Sourdough Bread Making: Evaluation of Nutritional and Quality Aspects

by
Ioanna Mantzourani
1,*,
Maria Daoutidou
1 and
Stavros Plessas
2,*
1
Laboratory of Food Processing, Department of Agricultural Development, Democritus University of Thrace, 68200 Orestiada, Greece
2
Laboratory of Microbiology, Biotechnology and Hygiene, Department of Agricultural Development, Democritus University of Thrace, 68200 Orestiada, Greece
*
Authors to whom correspondence should be addressed.
Appl. Sci. 2025, 15(8), 4283; https://doi.org/10.3390/app15084283
Submission received: 16 February 2025 / Revised: 14 March 2025 / Accepted: 9 April 2025 / Published: 13 April 2025
(This article belongs to the Section Food Science and Technology)
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Abstract

:
The production of functional bread has been of great interest lately to the Food Industry. Regarding this, the enrichment of bread with natural raw materials rich in phenolic antioxidants, such as fruits, has become a new trend. Likewise, the aim of the current study was to evaluate novel supplements based on freeze-dried Cornelian cherry juice, both unfermented and fermented by probiotic L. plantarum ATCC 14917, in sourdough bread production. The outcome showed that the fermented supplement led to sourdough bread with elevated nutritional features in terms of its total phenolic content (99.5 mg GAE/100 g) and antioxidant activity (213 mg TE/100 g for ABTS and 4.7 μmol TE/g for DPPH), as well as a reduction in phytic acid (93.3%) compared with all the other bread samples. In addition, the same sample contained higher amounts of lactic (2.91 g/Kg bread) and acetic acid (1.23 g/Kg), as well as formic (0.11 g/Kg), n-valeric (0.12 g/Kg) and caproic (0.05 g/Kg) acids compared with all the other samples, leading to a higher preservation time (13 days) regarding rope and mold spoilage. All breads exhibited the same sensorial characteristics, proving that the supplement did not affect bread quality. This outcome is very interesting since powdered supplements have recently been endorsed in the bread industry for enabling nutritional and technological improvements.

1. Introduction

The bread manufacturing industry has made remarkable progress regarding the application of novel biotechnology and other technologies, targeting the satisfaction of consumers’ requests for healthy and high-nutritional-value products [1,2]. Likewise, several research studies have focused on the enhancement of the nutritional value of bread [3]. In this vein, an upgraded protein content, enhanced antioxidant activity, a high load of bioactive compounds and longer preservation times, among other factors, are the main targets. Interestingly, vegetable- and fruit-derived materials have been studied in bread production, such as carrot [4], mango peel [5], chokeberry fruit [6], apple [7], banana [8], pomegranate peel [9], orange peel [10,11] and apple pomace, most of them in the form of dried powder. The outcomes were very promising, since fruits contain respectable amounts of fibers, minerals, vitamins, polyphenols, carotenoids and glucosinolates, comprising high nutritional value and simultaneously delivering healthy benefits. In addition, fruits do not contain gluten and can be easily applied in various formulations in gluten-free breads. However, drawbacks may appear in terms of the quality attributes of bread, such as the deterioration of sensorial and physicochemical properties [6,12]. Thus, the evaluation of proper formulations and an adequate concentration of all the ingredients in bread production is crucial in order for most of the quality parameters (nutritional value, physical properties and shelf life) to be in balance and finally be accepted by consumers.
Recently, Cornelian cherry (Cornus mas L.) was applied as a powder in the formulation of bread production, where the proper addition (0, 1, 2, 5 and 10% w/w) into wheat flour improved the physical properties of the produced bread and did not affect the sensorial characteristics of the bread [13]. It is a very good source of sugars, organic acids, anthocyanin and phenolic compounds; it has antibacterial and radioprotective properties against several pathogens and exhibits antihistamine, cytotoxic, anti-malarial and anti-inflammatory effects [14]. It was applied as a substrate in functional fruit juice production through fermentation by probiotic Lactobacillus plantarum ATCC 14917, leading to a product with high amounts of probiotic bacteria and significant nutritional properties [15]. The same outcome was found when the L. paracasei K5 strain, either free or immobilized on delignified wheat bran, was used for fermentation [16].
Likewise, the strategy of applying fermented fruit juices via LAB in freeze-dried powdered form seems quite interesting. Recently, a supplement containing freeze-dried fermented pomegranate juice via probiotic L. plantarum ATCC 14917 was successfully applied in sourdough bread making, leading to a final product with (i) improved nutritional quality in terms of total phenolic content (TPC), antioxidant activity (AC) and phytate reduction, (ii) longer preservation times and (iii) accepted sensory properties [17]. This specific work was a challenge in terms of examining the same strategy in the same conditions while replacing only the fruit juice (Cornelian cherry instead of pomegranate juice), not just for comparing these two fermentation systems but primarily investigating the consistencies of this form of Cornelian cherry supplement in sourdough bread’s properties. For instance, even though Cornelian cherry was applied as a powder in the formulation of bread production [13], in the current study, (i) Cornelian cherry was applied in its fermented form, while (ii) sourdough technology was used and not direct bread making technology (based on baker’s yeast).
The aim and perspectives of the current study are presented in Figure 1, which are dealing with the examination of, for the first time, (i) unfermented Cornelian cherry juice and (ii) fermented Cornelian cherry juice via L. plantarum ATCC 14917 as novel, nutritious supplements. The supplements were used in freeze-dried powdered form, and various physicochemical (pH, acidity and loaf volume), nutritional (TPC, AC and phytate reduction), sensory and microbiological properties (rope and mold spoilage) in the produced sourdough breads were determined and evaluated.

2. Results

2.1. Viable Cell Counts in the Sourdoughs

The microbiological load of the produced sourdoughs is presented in Table 1. Interestingly, the levels of LAB are differentially developed in all the samples. In particular, it seems that Cornelian cherry juice may act as an enhancer agent in the growth of LAB, since both sourdoughs that contained Cornelian cherry juice achieved higher levels compared to the control sample. Sourdough prepared with fermented Cornelian cherry juice by L. plantarum ATCC 14917 exhibited the highest LAB viability levels (10.1 Log cfu/g) due to the presence of the strain, while the sourdough prepared with unfermented juice contained LAB in a level of 8.9 Log cfu/g. The high phenolic content of Cornelian cherry ameliorating prebiotic properties [18,19] seems to be the reason for this outcome, since these compounds increase the growth of LAB. Yeast levels of all sourdough samples varied in the same levels (7.7–8.0 Log cfu/g) without a statistical difference.

2.2. Bread Volume Acidity Levels and Organic Acid Composition

The physicochemical characteristics of the three sourdough bread samples are presented in Table 2. Except for the specific loaf volume, of which all of the samples had almost the same levels (2.53–2.57 mL/g), statistically significant differences (p < 0.05) were observed in the evaluated parameters.
Specifically, the sourdough bread produced with fermented Cornelian cherry juice (BFJ) was more acidic in terms of pH (4.41) and TTA (9.82 mL NaOH 0.1 M) values compared to the BUFJ and CB. The same motive was observed in the case of minor and major organic acid levels. Specifically, the BFJ contained higher amounts of lactic (2.91 g/Kg bread) and acetic acid (1.23 g/Kg) as well as formic (0.11 g/Kg), n-valeric (0.12 g/Kg) and caproic (0.05 g/Kg) acids compared to all the other samples. The BUFJ was second in terms of organic acid load, followed by the CB. It is obvious that the addition of fermented Cornelian cherry juice significantly enhanced the acidity and organic acid levels in the produced breads. It seems that the level of LAB viability (Table 1) determined the acidity levels as well as the values of organic acid in all the sourdough bread samples. In addition, the high content of prebiotic compounds contained in the Cornelian cherry juice, such as phenolics, provoked the growth of LAB and improved the lactic acid fermentation in the sourdough breads. Similar results were obtained recently, when another substrate rich in phenolics based on pomegranate juice was applied as a powdered supplement in sourdough bread making [17].

2.3. Microbial Spoilage

The examination of rope and mold spoilage was conducted through macroscopic observations during bread storage, and the results are presented in Table 3. The BFJ exhibited the highest resistance in both types of spoilage (p < 0.05) compared to all the other samples and was preserved until the 13th day. The BUFJ was preserved for 10–11 days, while the CB was spoiled on day 7. The high organic acid load of the BFJ positively influenced the resistance of this sample regarding spoilage, as was expected. Lactic and acetic acid are organic acids well known for their antibacterial and antifungal activities [20].
Minor organic acids such as formic, n-valeric and caproic acid were also detected in the BFJ sample, delivering additional protection to the bread.

2.4. Total Phenolic Content (TPC) and Antioxidant Activity (AC)

The nutritional features of all the sourdough bread samples were evaluated through the determination of TPC and AC (Table 4). The BFJ expressed the highest values of TPC (99.5 mg GAE/100 g), followed by the BUFJ (83.1 mg GAE/100 g) and finally the CB (58.4 mg GAE/100 g). The same pattern was observed in the case of AC. In particular, the highest ABTS and DPPH values were recorded in the BFJ, followed by the BUFJ and CB. It seems that L. plantarum ATCC 14917 was the main reason for the high phenolic transformation rate in the sourdough bread matrix due to its great variety of enzymes, which are capable of degrading the phenolic compounds present in a food matrix to other compounds, increasing the antioxidant activity and total phenolic content [21].

2.5. Phytic Acid Content

Application of the supplements on sourdough bread production (BFJ and BUFJ) led to reduced levels of phytic acid compared to the CB (Figure 2). More specifically, the reduction of phytic acid in the BFJ was 93.3%, while in the BUFJ was 88.9%, and it was 81.8% in the CB. This outcome is very important since the reduction in phytic acid in bread is desirable, making various minerals more bioavailable [22].

2.6. Sensorial Features

The results of the sensorial features (consumer preference) of the produced sourdough breads are illustrated in Table 5. Even though some optic differences were observed, such as slightly darker color and more strengthened gluten network (Figure 3), minor differences were recorded among the samples, showing that the employment of the supplements did not affect the sensorial features of the produced breads. Likewise, the consumers accepted the quality of the novel sourdoughs, scoring them almost the same as the CB, with no statistically significant difference. This result is quite encouraging, since a late report dealing the application of Cornelian cherry powder in bread production showed contradictory effects in the sensorial features [13].

3. Discussion

The development of functional bread has been considered a high priority by the Food Industry in recent years years, in addition to other food categories/products (dairy, fruit, etc.). Therefore, the bread manufacturing industry is facing various challenges, such as the production of products with reduced cost, longer preservation times without the addition of preservatives, improvements in nutritional value and the delivery of health benefits. In this vein, sourdough technology seems the most appropriate vehicle for the delivery of functional bread. The strategy that has been adopted lately is dealing with the application of novel wild strains and the use of various supplements for the nutritious fortification of products.
In the frame of this research survey, fermented Cornelian cherry juice by L. plantarum ATCC 14917 was applied for the first time in sourdough bread making. Even though Cornelian cherry has been previously applied as a supplement in bread production [13], the current research work has two basic differences: (i) the Cornelian cherry was applied in the form of fermented juice by LAB, and (ii) the sourdough technology was applied and not the direct yeast leavened method, as is usually performed. Initially, the respective produced sourdoughs showed an increased load of LAB compared to the control sourdough (Table 1). Specifically, the sourdough prepared with the addition of unfermented juice (UFJ) had higher levels of LAB (p < 005) compared to the control sourdough, leading handily to the conclusion that the composition of Cornelian cherry juice enhanced this effect. Indeed, the increased polyphenolic content (anthocyanins, cinnamic acids, flavonoids, benzoic acids, catechins and tannins) of Cornelian cherry acts as a prebiotic factor and stimulates the growth of probiotic strains such as L. plantarum ATCC 14917 [23]. Recently, polyphenols contained in fruits and vegetables have been officially included in the list of prebiotics [24].
The application of the sourdoughs to bread production revealed significant differences. The application of sourdough containing fermented Cornelian cherry juice by L. plantarum ATCC 14917 (BFJ) led to bread with the highest acidity and higher organic acid values, followed by the BUFJ and CB. This result implies that the high lactic acid fermentation rate by LAB in the sourdough matrix was mainly due (i) to the polyphenol content of the juice, which acts as enhancer, (ii) to the high acid tolerance that L. plantarum ATCC 14917 displays and (iii) to the greater LAB viability in the respective initially prepared sourdough [25,26].
The increased organic acid values of the sourdough breads led to predictable microbial resistance levels versus rope and mold spoilage. The BFJ was spoiled on the 13th day, which is considered a very satisfying result. Even though lactic acid is a well-established antimicrobial agent, the role of acetic acid has recently been found to be more enhanced, insinuating greater effectiveness. In the current study, the concentration of acetic acid reached a value of 1.23 g/Kg, which is the most elevated one compared to other relative studies that our research team performed in recent years [17,27,28,29,30,31]. More specifically, in the aforementioned previous research works, the sourdoughs were added in a concentration of 30% w/w for bread making, with employment of the following: (i) L. paracasei K5, which led to acetic acid with a concentration of 1.13 g/Kg of bread, (ii) L. plantarum ATCC 14917 with or without pomegranate juice, (iii) L. paracasei SP2, (iv) kefir grains, (v) L. paracasei SP5 free or immobilized and (vi) L. bulgaricus ATCC 11842, which led to acetic acid concentrations varying around lower than or almost 1 g/Kg. Likewise, the rope spoilage appeared on the 13th day, which is one of the most increased values recorded compared to previous studies. Only in two cases was the appearance of rope spoilage more extended, but there were no statistical differences. Specifically, bread spoilage appeared by approximately the 14th day of bread storage when sourdoughs were prepared with L. paracasei SP2 and SP5 [29,31]. Indeed, acetic acid exhibits advanced antibacterial properties against bacteria involved in rope spoilage and is being added alone as preservative in bread production in the form of vinegar [32]. It is noteworthy that three other minor organic acids (formic, n-valeric and caproic acid) embrace elevated functionality against various bread spoilage microorganisms through synergistic effects [29]. It is well known that the basic ingredient of bread, wheat, loses its nutritional value during industrial bread making with regards to proteins, minerals, fiber and antioxidant activity, and this leads to deficiencies. Consequently, the fortification of bread with the addition of various micronutrients (vitamins and minerals) and protein-rich sources is necessary [33,34].
In this frame, the proposed supplement consisted of fermented Cornelian cherry juice by L. plantarum ATCC 14917, which led to statistically significant increases in TPC and AC levels in the respective bread (BFJ) compared to all the other samples produced.
L. plantarum strains have proved their effectiveness in various biotransformation food systems, including bread, by degrading various bound phenols to soluble compounds with increased AC and bioavailability [21,35]. Interestingly, respectable amounts of TPC and AC levels were also achieved in the case of the BUFJ (compared to the CB), where unfermented juice was applied, without the presence of any LAB, including L. plantarum ATCC 14917. This outcome is in agreement with other researchers who used powder of the whole fruit of Cornelian cherry in bread production. In the current study, the results are more impressive since the sourdough technology and the form of the fruit (juice) fermented or unfermented elevated the TPC and AC levels. The main reason for this outcome is that LAB contained in the wheat flour were activated and enhanced by the phenolic content of the Cornelian cherry juice and afterwards transformed the phenolic compounds present in wheat flour into various new compounds with high antioxidant activity. Generally, the antioxidant activity of fortified breads is mainly derived from the phenolic rather than other compounds present in bread [36]. Specifically, the increase in the antioxidant compounds’ content and activity after baking is commonly attributed to the weakening of the cell wall matrix, enabling the release of phenolic acids from bounded forms, and to the decomposition or conversion of complex molecules (tannins, quercetin derivatives and rutin) to simpler phenolic compounds (mostly phenolic acids and quercetin derivatives) [37]. Sourdough seems to enhance even more this effect, resulting in antioxidant protection [13]. In addition, the freeze-drying procedure followed for the preparation of the supplement (fermented and not fermented Cornelian cherry juice) is considered to be a method that can preserve biologically active compounds destined for bakery products, as other researchers have proposed and experimentally verified [38].
Phytic acid is considered an antinutrient in bread because it provokes decreased bioavailability and deficiencies of various minerals in the human diet [37]. In particular, the greatest reduction of phytic acid in the BFJ verifies the effect of LAB and L. plantarum ATCC 14917. More specifically, sourdough fermentation enhances phytases’ activity, which are present in wheat flour, by lowering the pH values and thereby increasing the acidity, which is important for their activation. In addition, there are some LAB such as L. plantarum that display extracellular phytase activity [39,40].
The sensorial features of all the samples showed no statistically significant differences, revealing that the proposed supplement had no negative effect in bread. This outcome is very important, since fruit powders often cause problems with the sensorial characteristics of bread. The best examples of this fact are chokeberry and Cornelian cherry fruit and their addition to bread. Their proper formulations are applied for the tackling of this drawback [6,13]. On the other hand, in the current research, all of the bread samples were produced by the application of the sourdough method and not by the yeast method, which may also contribute to this outcome. Sourdough technology is well established as a more effective method compared to yeasted bread due to organic acid production, the stimulation of the endogenous enzymes of the flour and the synthesis of microbial secondary metabolites, which exhibit a direct positive effect in the development of more attractive sensorial characteristics [1,2]. In addition, more advantages have been reported, such as the increase in the in vitro protein digestibility and amount of soluble fiber and the reduction in the glycemic index and other antinutritional factors [1,2,3]. These scientific outcomes have promoted the sourdough technology global manufacture market, which satisfies consumer expectations for natural, highly nutritious and sustainable foods [3].

4. Materials and Methods

4.1. Chemicals

The chemicals used in this study for the production and treatment of breads and the methods of analysis were as follows: Std 0.1 M sodium hydroxide (NaOH) solution, 0.25 M phosphate buffer (KHPO4) (Sigma-Aldrich, St. Louis, MO, USA), phthalic acid 2.5 mM solution (Sigma-Aldrich, St. Louis, MO, USA), Tris(hydroxymethyl)aminomethane (C4H11NO3) 2.4 mM solution (Sigma-Aldrich, St. Louis, MO, USA), phosphate buffered saline (PBS) (Merck, Darmstadt, Germany), yeast extract and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) (Duchefa Biochemie, Haarlem, The Netherlands), gallic acid (Sigma-Aldrich, St. Louis, MO, USA), Folin–Ciocalteu reagent (Scharlab S.L., Barcelona, Spain), anhydrous sodium carbonate (Na2CO3) (Penta, Prague, Czech Republic), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) 7.4 mM solution (Sigma-Aldrich, St. Louis, MO, USA) and potassium persulfate (K2S2O8) 2.6 mM solution (Merck, Darmstadt, Germany).

4.2. Raw Materials, Microorganism and Media

The probiotic L. plantarum ATCC 14917 was cultured at 37 °C for 48 h in MRS broth. After 48 h, it was harvested by centrifugation (Sigma 3K12, Bioblock Scientific, Strasbourg France) at 5000 rpm for 10 min at 25 °C. Commercial white flour was used for bread making (Hellenic Biscuit CO S.A., Volos, Greece). Commercial baker’s yeast (S.I. Lesaffre, France) was also applied (70% w/w moisture).

4.3. Preparation and Fermentation of Cornelian Cherry Juice

Cornelian cherry fruits were delivered from a local market of Orestiada (Greece) and were prepared as previously described with initial sugar concentration and pH values of 55 g/L and 3.5, respectively [16]. In brief, after proper selection and washing, the fresh juice was obtained through blending, dilution with water, cloth filtration and finally pasteurization (80 °C for 5 min). Two grams (dry weight) of L. plantarum ATCC 14917 was added in the prepared juice (100 mL), and it was left to be fermented for 24 h at 30 °C.

4.4. Freeze-Drying

Freeze-drying of the cornelian cherry juice unfermented and fermented by L. plantarum ATCC 14917 was conducted by freezing at −44 °C (5 °C/min) and drying for 48 h (at 5–15 mbar and −45 °C) with a FreeZone 4.5 Freeze-Drying System (Labconco, Kansas City, MO, USA).

4.5. Sourdough Bread Making

Sourdough bread was produced by mixing all the ingredients and kneading manually. The dough was molded in 1.5 L baking pans. Initially, 2 mother sponges were prepared by mixing 300 g wheat flour and 160 mL tap water (i) with 1% w/w (on flour basis) of freeze-dried fermented juice and (ii) with 1% w/w (on flour basis) of freeze-dried unfermented juice for 15 min. Subsequently, 2 sourdough breads were produced containing 30% w/w (on flour basis) of the aforementioned mother sponges. Specifically, the composition of the sourdough breads was as follows: 150 g of each sourdough, 500 g wheat flour, approximately 270 mL tap water, 4 g salt and 1% w/w (on flour basis) pressed baker’s yeast. All doughs were fermented at 30 °C for 2 h, proofed at 40 °C for 60 min and baked at 230 °C for ~40 min. Therefore, 2 types of sourdough breads were produced containing sourdough with (i) 30% freeze-dried fermented juice (BFJ) and (ii) 30% freeze-dried unfermented juice (BUFJ).
Traditional sourdough (wild microflora) provided by a local bakery was used for the production of the control sourdough bread (CB) in an amount of 30% (on flour basis). All the conditions regarding the recipe and procedure for bread making were the same as the 2 sourdough breads described before. All trials were carried out in triplicate.

4.6. Analytical Methods

4.6.1. Microbial Cell Counts and Monitoring of Bread Spoilage

An amount of 1 g of sample was added in 9 mL of phosphate buffer (0.25 M solution of KH2PO4 diluted as 1.25 mL/L of distilled water) in order to determine the viable cell counts in the freeze-dried L. plantarum and the fermented pomegranate juice powders [28]. After decimal dilutions, the suspensions were plated on MRS agar (Fluka, Buchs, Switzerland) and incubated at 37 °C for 48–72 h. Similarly, viable cell counts of LAB and yeasts were determined.
The bread loaves were stored in sterile plastic bags at room temperature, and macroscopical observation for mold and rope spoilage followed [17]. The day when mold growth became visible was recorded as the mold spoilage time.

4.6.2. Organic Acids

HPLC was applied for the determination of the organic acids (lactic, acetic, formic, propionic, n-valeric and caproic) in the sourdough breads, as described before [27]. In brief, a Shimadzu HPLC system was used, which contained a Shim-pack ICA1 column, an LC-10AD pump, a CTO-10A oven (40 °C) and a CDD-6A conductivity detector. Phthalic acid (2.5 mM) and tris(hydroxymethyl) aminomethane (2.4 mM; pH 4.0) were applied as the mobile phase (1.2 mL/min). The concentration of the diluted sample was 5% v/v, and the injection volume was 60 μL. All organic acid concentrations were determined by means of standard curves of lactic and acetic acids (Fluka) and valeric, isovaleric, and hexanoic acids (Sigma-Aldrich).

4.6.3. pH and Total Titratable Acidity (TTA)

For the pH measurement of the sourdough bread samples, a Sentron Argus pH meter (Sentron Europe B.V., Roden, The Netherlands) was used as described before [27]. Additionally, for the determination of total titratable acidity (TTA), 10 g of breadcrumbs was blended with 90 mL of deionized water, and titration of the suspension with 0.1 N NaOH to a final pH of 8.5 followed with phenolophthalein as the indicator. The TTA was expressed as the volume (mL) of NaOH consumed.

4.6.4. Specific Loaf Volume

The specific loaf volume (mL/g) of the sourdough breads was measured by the rapeseed displacement method [41]. In brief, after cooling, each loaf was placed in a container, and rapeseed was used to totally fill the container. After the removal of the loaf, the volume of the rapeseed was recorded. The loaf volumes were the result of the deduction of the rapeseed volume from the container volume. The specific loaf volume was expressed as mL/g.

4.6.5. Total Phenolic Content (TPC)

After the baking of the breads, cool down at room temperature for 3 h followed. Then, crumb samples were freeze-dried for 48 h (FreeZone 4.5, Labconco, Kansas City, MO, USA), and 1 g of each freeze-dried sample was added to 20 mL of phosphate buffered saline (PBS, pH 7.4) for 1 h under shaking at 37 °C. The residues were extracted once again with 20 mL of PBS. The collected extracts were combined and stored at −20 °C until analysis of TPC and AC [17].
More specifically, 200 μL of sample extract was mixed with 800 μL of Folin–Ciocalteu reagent and left to react for 2 min in the dark. Afterwards, 2 mL of sodium carbonate (7.5% v/v) and distilled water was added up to final volume of 10 mL. The mixture was left to react for 60 min at room temperature in the dark. The absorbance was measured at 765 nm. Gallic acid (GA) solutions and a blank were also prepared for standardization. The TPC was expressed as GA equivalents (mg GAE/100 g dried sample) [17].

4.6.6. Antioxidant Capacity (AC)

The antioxidant activity (AC) was evaluated through the (i) ABTS [2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)] assay as Trolox equivalents (mg TE/100 g dried sample) and (ii) the DPPH radical scavenging activity assay (mol TE/g dw) as described before [17].
In brief, an ABTS radical cation (ABTS_+) stock solution was prepared by mixing 7.4 mM ABTS (in water) and 2.6 mM potassium persulfate (in water) in equal volumes. The mixture was left for incubation at room temperature for 12 h. The above solution was diluted with water to an absorbance of 0.70 at 734 nm. Then, 100 μL of bread extract was mixed with 3.9 mL of the diluted ABTS_+ solution, and after 4 min, the absorbance was measured against a blank at 734 nm. A standard Trolox curve was also prepared.
As for the DPPH assay, 3 mL of a 137.6 μM methanolic DPPH solution and various amounts of bread extract (in the range of 0.05–1 mL) were added in several test tubes, and the volume was fixed with methanol to 4 mL. The samples were left for 60 min in the dark, and the absorbance was measured at 515 nm against an aqueous methanol solution as the blank. The antioxidant capacity was calculated from the Trolox calibration curve as well.

4.6.7. Phytic Acid

Phytic acid (phytate; myo-inositol-1,2,3,4,5,6-hexakisphosphate) was determined employing a Megazyme test-kit K-PHYT (Megazyme, Bray, Ireland) according to the manufacturer’s recommendations.

4.6.8. Sensory Evaluation

A blind sensory evaluation assessment took place for all the sourdough breads right after their baking, as described previously [17]. In brief, the sourdough breads were evaluated by 20 random, untrained testers (consumer-oriented testing), as far as their flavor, taste and overall appearance are concerned, based on a 1, dislike extremely, to a 9, like extremely, preference scale. The results were recorded as average scores plus standard deviations.

4.6.9. Statistical Analysis

Analysis of Variance (ANOVA) followed by Duncan’s post hoc multiple range test at the 5% level of significance was used to produce differences between the various treatments, i.e., the effects of the different sourdoughs on the physicochemical and sensory properties of the produced breads. The analysis was performed using SPSS Statistics 20.0 (IBM Corp., Armonk, NY, USA) software at an alpha level of 5%.

5. Conclusions

The fortification of bread has recently become common in the Food Industry in order to overcome various nutrient deficiencies caused in bread manufacturing and to even deliver health benefits. In this vein, novel supplements consisting of fermented Cornelian cherry juice unfermented or fermented by L. plantarum ATCC 14917 are proposed in this study. The results are quite encouraging, since the produced bread, especially in the case of the fermented supplement application, embraced high nutritive value, a high organic acid load and a high preservation time. In addition, the proposed supplement did not affect the physicochemical features and sensorial properties of the product (e.g., loaf volume) even though in other cases proper formulations were required. The application of the proposed supplement in sourdough bread technology seems feasible and could be adopted by the Food Industry. However, more research is needed for the corroboration of possible health benefits in order for this technological proposal to be more attractive and have more possibilities for commercialization. In addition, future work is planned targeting the determination of the qualitative and quantitative profiles of phenolic compounds both in the raw materials and final products in order for more accurate outcomes to surface and be addressed.

Author Contributions

Conceptualization, I.M. and S.P.; methodology, M.D.; software, S.P.; validation, S.P. and I.M.; investigation, M.D.; resources, S.P. and I.M.; data curation, S.P. and I.M.; writing—original draft preparation, I.M. and S.P.; writing—review and editing, S.P.; supervision, S.P. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Data are contained within the article.

Conflicts of Interest

The authors declare no conflicts of interest.

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Figure 1. Perspectives of the current study dealing with the production of functional sourdough bread.
Figure 1. Perspectives of the current study dealing with the production of functional sourdough bread.
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Figure 2. Phytic acid reduction in doughs after baking. BUFJ: bread containing sourdough produced with 30% unfermented Cornelian cherry juice; BFJ: bread containing sourdough produced with 30% fermented Cornelian cherry juice; CB: control bread made with 30% wild microflora sourdough. Different letters in a column indicate statistically significant differences (ANOVA, Duncan’s multiple range test, p < 0.05).
Figure 2. Phytic acid reduction in doughs after baking. BUFJ: bread containing sourdough produced with 30% unfermented Cornelian cherry juice; BFJ: bread containing sourdough produced with 30% fermented Cornelian cherry juice; CB: control bread made with 30% wild microflora sourdough. Different letters in a column indicate statistically significant differences (ANOVA, Duncan’s multiple range test, p < 0.05).
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Figure 3. Experimental bread slices of (A) BUFJ, (B) BFJ and (C) CB.
Figure 3. Experimental bread slices of (A) BUFJ, (B) BFJ and (C) CB.
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Table 1. Yeast and LAB levels in the produced sourdoughs.
Table 1. Yeast and LAB levels in the produced sourdoughs.
SourdoughLABYeasts
Log cfu/g
UFJ8.9 ± 0.2 b7.9 ± 0.1 a
FJ10.1 ± 0.1 a8.0 ± 0.2 a
C8.5 ± 0.1 c7.7 ± 0.2 a
UFJ: sourdough prepared with the addition of freeze-dried unfermented juice, FJ: sourdough prepared with the addition of freeze-dried fermented juice, C: control sourdough, LAB: lactic acid bacteria, cfu: colony forming units. Different superscript letters in a column indicate statistically significant differences (p < 0.05).
Table 2. Physicochemical characteristics of breads made with sourdoughs prepared with freeze-dried, free and immobilized L. plantarum ATCC 14917 and of the control sourdough.
Table 2. Physicochemical characteristics of breads made with sourdoughs prepared with freeze-dried, free and immobilized L. plantarum ATCC 14917 and of the control sourdough.
Bread SamplepHTTA
(mL 0.1 M NaOH)		SLV
(mL/g)		Organic Acids (g/kg Bread)
LacticAceticFormicn-ValericCaproic
BUFJ4.55 ± 0.03 b7.25 ± 0.06 b2.55 ± 0.05 a2.49 ± 0.08 b0.87 ± 0.02 b0.05 ± 0.01 b0.05 ± 0.01 btr
BFJ4.41 ± 0.04 c9.82 ± 0.05 a2.53 ± 0.04 a2.91 ± 0.05 a1.23 ± 0.02 a0.11 ± 0.01 a0.12 ± 0.01 a0.05 ± 0.01
CB4.76 ± 0.03 a6.32 ± 0.05 c2.57 ± 0.05 a2.22 ± 0.06 c0.68 ± 0.02 ctrtrtr
BUFJ: bread containing sourdough produced with 30% unfermented Cornelian cherry juice; BFJ: bread containing sourdough produced with 30% fermented Cornelian cherry juice; CB: control bread made with 30% wild microflora sourdough; TTA: total titratable acidity; SLV: specific loaf volume; tr: traces (<0.01 g/Kg). Different superscript letters in a column indicate statistically significant differences (ANOVA, Duncan’s multiple range test, p < 0.05).
Table 3. Evaluation of mold and rope spoilage in the sourdough bread samples.
Table 3. Evaluation of mold and rope spoilage in the sourdough bread samples.
Bread SampleMold SpoilageRope Spoilage
Days
BUFJ11 ± 0.5 b10 ± 0.5 b
BFJ13 ± 0.5 a13 ± 0.5 a
CB7 ± 0.5 c7 ± 0.5 c
BUFJ: bread containing sourdough produced with 30% unfermented Cornelian cherry juice; BFJ: bread containing sourdough produced with 30% fermented Cornelian cherry juice; CB: control bread made with 30% wild microflora sourdough. Different superscript letters in a column indicate statistically significant differences (ANOVA, Duncan’s multiple range test, p < 0.05).
Table 4. Total phenolic content (TPC) and antioxidant activity (AC) (on dry weight basis) of the produced sourdough breads.
Table 4. Total phenolic content (TPC) and antioxidant activity (AC) (on dry weight basis) of the produced sourdough breads.
Bread SampleTPC AC
ABTSDPPH
(mg GAE/100 g)(mg TE/100 g)(μmol TE/g)
BUFJ83.1 ± 3.2 b188.0 ± 3.7 b3.6 ± 0.2 b
BFJ99.5 ± 2.5 a213.8 ± 5.1 a4.7 ± 0.2 a
CB58.4 ± 2.7 c179.1 ± 4.0 c3.2 ± 0.1 c
BUFJ: bread containing sourdough produced with 30% unfermented Cornelian cherry juice; BFJ: bread containing sourdough produced with 30% fermented Cornelian cherry juice; CB: control bread made with 30% wild microflora sourdough; TPC: total phenolic content; AC: antioxidant capacity; ABTS: 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid; DPPH: 1,1-diphenyl-2-picryl-hydrazyl. Different superscript letters in a column indicate statistically significant differences (ANOVA, Duncan’s multiple range test, p < 0.05).
Table 5. Sensory evaluation test (consumer preference) of sourdough breads.
Table 5. Sensory evaluation test (consumer preference) of sourdough breads.
Bread SampleFlavorTasteAppearanceOverall Quality
BUFJ8.5 ± 0.2 a8.8 ± 0.1 a8.5 ± 0.1 a8.4 ± 0.1 a
BFJ8.8 ± 0.1 a9.0 ± 0.2 a8.5 ± 0.1 a8.6 ± 0.1 a
CB8.7 ± 0.1 a8.7 ± 0.1 a8.6 ± 0.1 a8.5 ± 0.1 a
BUFJ: bread containing sourdough produced with 30% unfermented Cornelian cherry juice; BFJ: bread containing sourdough produced with 30% fermented Cornelian cherry juice; CB: control bread made with 30% wild microflora sourdough; TTA: total titratable acidity; SLV: specific loaf volume; tr: traces (<0.01 g/Kg). Different superscript letters in a column indicate statistically significant differences (ANOVA, Duncan’s multiple range test, p < 0.05).
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Mantzourani, I.; Daoutidou, M.; Plessas, S. Impact of Functional Supplement Based on Cornelian Cherry (Cornus mas L.) Juice in Sourdough Bread Making: Evaluation of Nutritional and Quality Aspects. Appl. Sci. 2025, 15, 4283. https://doi.org/10.3390/app15084283

AMA Style

Mantzourani I, Daoutidou M, Plessas S. Impact of Functional Supplement Based on Cornelian Cherry (Cornus mas L.) Juice in Sourdough Bread Making: Evaluation of Nutritional and Quality Aspects. Applied Sciences. 2025; 15(8):4283. https://doi.org/10.3390/app15084283

Chicago/Turabian Style

Mantzourani, Ioanna, Maria Daoutidou, and Stavros Plessas. 2025. "Impact of Functional Supplement Based on Cornelian Cherry (Cornus mas L.) Juice in Sourdough Bread Making: Evaluation of Nutritional and Quality Aspects" Applied Sciences 15, no. 8: 4283. https://doi.org/10.3390/app15084283

APA Style

Mantzourani, I., Daoutidou, M., & Plessas, S. (2025). Impact of Functional Supplement Based on Cornelian Cherry (Cornus mas L.) Juice in Sourdough Bread Making: Evaluation of Nutritional and Quality Aspects. Applied Sciences, 15(8), 4283. https://doi.org/10.3390/app15084283

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