1 Introduction

Preservatives are frequently classified as carcinogens or cancer-causing chemicals due to their hazardous impacts on human health [1], artificial food additives interact with the cellular structure of the body, resulting in a spectrum of food-related disruptions [2].

Excessive use of these might cause negative effects such as skin rashes and itching, breathing problems, sneezing, or stomach issues [3]. Synthetic food additives interact with the body's cellular structure, leading to various food disruptions and negative effects like skin rashes, itching, breathing problems, sneezing, and gastrointestinal disturbances when used excessively. Due to the possible health risks associated with synthetic food preservatives, consumers have become more vigilant about consuming foods containing these additives, leading to an increase in demand for natural food preservatives over recent decades [4, 5]. Myrrh resin is a resin known for its rich phytochemical profile and contains components such as terpenoids (monoterpenoids, sesquiterpenoids, volatile/essential oil), diterpenoids, triterpenoids, and steroids. It has been utilized throughout history to treat various ailments, including wounds, mouth ulcers, pain, fractures, digestive issues, microbial infections, and inflammatory diseases [6, 7]. The medicinal properties of Myrrh resin are well-documented, highlighting its antiseptic, astringent, anthelmintic, and expectorant characteristics [6, 8]. Recent studies by Batihaet al., 2023 and Akbar. 2020 [8, 9] have proven the traditional use of myrrh in treating bacterial infections and its antibacterial and antifungal properties. According to Mohammed et al. 2015 and, Khalil et al. 2020 [6, 10], myrrh extract is promoted as a source of chemicals that can be used to create safer and environmentally friendly antibacterial agents to fight various pathogenic fungi [11], they have also conducted research on myrrh's potential as an antibacterial agent.

Myrrh resin ether and ethanol extracts were tested for their antibacterial properties; these extracts showed antimicrobial activity against the Gram-negative organisms tested alongside [10, 12, 13]. However, the ether extract demonstrated antimicrobial action both against Candida albicans and the Gram-positive organisms that were tested, with the antifungal activity being stronger [12, 13]. In addition to its antimicrobial properties, the potential anticancer effects of Myrrh resin have also garnered attention [14]. Integrating its anticancer properties into the context of food preservation presents an intriguing avenue for research. Therefore, the primary objective of this study is to explore the antimicrobial and anticancer properties of Myrrh Resin Extract (MRE) and evaluate its application as a natural antimicrobial agent in cacao beverages.

2 Material and methods

2.1 Materials

Myrrh resin extract (MRE), Cacao powder, (protein 20.3/100 g, fat 9/100 g, and carbohydrate 56.3/100 g), non-dairy cream, salt, corn starch, and granulated sugar were purchased from a local market and herbal market in Cairo, Egypt.

2.2 Microbial strains

Gram-positive bacteria Bacillus subtilis ATCC6633, and Staphylococcus aureus (MRSA) ATCC43300, as well as Gram-negative bacteria Pseudomonas aeruginosa (ATCC27853), and Escherichia coli (ATCC25922).

2.3 Chemicals and reagents

The chemicals and reagents used in the analytical methods for this study were of analytical-grade quality. These included citric acid, sodium benzoate, CMC, xanthan gum, plate count agar, and potato dextrose agar, all purchased from the El-Gamhouria Trading for Chemicals and Drugs Company in Cairo, Egypt.

2.4 Extraction of antioxidants from MER

A sample of 100 g of Myrrh resin was combined with a mixture of distilled water and ethanol (2:8 v/v) and incubated at room temperature for 24 h. The resulting suspension was then filtered, and the aqueous extract was concentrated via a rotary evaporator at 35 °C under reduced pressure. The remaining solids were dissolved in 50 ml of distilled water.

2.5 HPLC standards

Standards of polyphenols such as Caffeic acid, P-Coumaric acid, ( +)-Catechin, and Chlorogenic acid were obtained from Tokyo Chemical Industry Co., Ltd. (Kita-Ku, Tokyo, Japan). Additional phenolic standards, including Rosmarinus acid, Gallic acid, Quercetin, Apigenin, kaempferol, and Rutin, were sourced from Sigma-Aldrich (St. Louis, MO, USA).

2.6 Analysis of Myrrh resin extract using LC–ESI–MS/MS

2.6.1 Instrument

The sample analysis was performed using liquid chromatography-electrospray ionization–tandem mass spectrometry (LC–ESI–MS/MS), using a 3200 QTRAP mass spectrometer (AB Sciex, Framingham, MA, USA) linked to a column oven and a binary gradient solvent pump, an autosampler, and a degasser, an Agilent 1200 Series HPLC system (Agilent Technologies, Santa Clara, CA, USA) was used [15].

2.6.2 Positive ionization mode

Separation was conducted using an Ascentis® Express 90 Å C18 Column (2.1 × 150 mm, 2.7 µm). The mobile phases included two eluents: (A) 5 mM ammonium formate, pH 3, and (B) acetonitrile (LC grade). Using a recently devised, straightforward, and quick approach that combined tandem mass spectrometry, electrospray ionization, and liquid chromatography, the concentrations of free flavonoid aglycones were ascertained. On a SB-C18 column, the compounds were separated at 25 °C. There was a 5 μL injection volume. Acetonitrile (B) and water with 0.1% HCOOH (A) were used for gradient elution. The 200 μL/min flow rate was used [15].

2.7 Determination of antioxidants using high performance liquid chromatography (HPLC)

HPLC analysis was performed using an Agilent Technologies 1260 infinity pump and a 1260 infinity II ultraviolet (UV) detector (Palo Alto, CA, USA). The chromatography system consisted of a 1260 infinity quaternion liquid. A microplate spectrophotometer (Epoch 2, Biotech Instruments, Winooski, VT, USA) was used in the assay of the scavenging activity on the DPPH radical. Separation was achieved with an Eclipse C18 column (4.6 × 250 mm i.d., 5 μm). The mobile phase comprised water (A) and 0.05% trifluoroacetic acid in acetonitrile (B), with a flow rate of 0.9 ml/min. The mobile phase was programmed in a linear gradient as follows: 0 min (82% A); 0–5 min (80% A); 5–8 min (60% A); 8–12 min (60% A); 12–15 min (82% A); 15–16 min (82% A); and 16–20 min (82% A). The multi-wavelength detector was set to 280 nm. A 5 μl injection volume was used for each sample solution, and the column temperature was maintained at 40 °C Amethanolic solution containing a standard stock solution (500 µg/ml) was prepared for each standard compound, and a calibration curve was established using 19 standards of polyphenolic compounds [16].

2.8 Minimal inhibitory concentration (MIC) determination

To determine the minimal inhibitory concentration (MIC), the disk diffusion assay protocol was followed. A volume of 1 mL from each bacterial suspension was evenly spread onto a solid Mueller–Hinton agar plate. Six sterile paper disks, each 6 mm in diameter, were placed on the surface of the agar. These disks were then saturated with 15 µL of diluted Myrrh extract (MRE) at concentrations of 100%, 50%, 25%, 12.5%, 6.25%, 3.13%, and 1.56%. The plates were incubated at 37 °C for 24 h. The MIC was determined by identifying the lowest concentration of MRE that produced an inhibition zone around the disk after incubation for a 24-h period. Disks of negative controls impregnated with sterile distilled water. Each concentration was tested in replicates, as described by Lima-Filho and de AguiraCorderio (2014) [17].

2.9 Preparation of Cacao beverage

In preparation for the cacao beverage, all materials utilized in this study underwent a sieving process using an 80-mesh sieve. A homogeneous mixture was prepared by combining white sugar (15%), cocoa powder (7.5%), and non-dairy creamer (6%) with 1.47% of corn starch, 0.03% of salt, and 0.1% of xanthan gum. The mixture was stirred continuously until it achieved homogeneity. 1% of Myrrh extract was incorporated into formula-2 and 0.1% of Sodium benzoate was incorporated into formula-3. A quantity of tap water was added and heated at 80 ± 2 °C for 5 min [18], the formulations of cocoa beverages are presented in Table 1.

Table 1 Formulations of cocoa beverages
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2.10 Physicochemical analysis (pH value, total soluble solids, and total titratable acidity).

The pH was monitored using a digital pH meter (model 3505-JENWAY-UK) calibrated with buffers at pH 4.0 and 7.0. Total soluble solids (TSS) were determined in the filtered samples using an MA871 digital refractometer (Milwaukee 0–85% Brix – Romania) at 25 °C. The equipment was calibrated with distilled water before samples were measured. The sample was poured onto a prism of the refractometer, and T.S.S was directly measured. Total titratable acidity (TTA) was determined in 10 mL of cacao beverage, which was homogenized with 90 ml of distilled water, and titrated with 0.1 mol/L NaOH, using phenolphthalein as an indicator [19]. The results expressed as % lactic (molecular weight = 90) for cacao beverage and % Citric (molecular weight = 64). The titratable acidity was calculated using the following equation,

$$\begin{gathered} \,\,\,\,\,\,\,\,\,\,\,{\text{N}}\,X\,{\text{V1}}\,X\,\,\,{\text{Eqwt}} \hfill \\ \% {\text{ of acidity}}\, = \,\,X\,{1}00 \hfill \\ \,\,\,\,\,\,\,\,\,\,\,\,{\text{V2}}\,X\,{1}000 \hfill \\ \end{gathered}$$

where, N = normality of NaOH (mEq/mL); V1 = volume of titrant (mL); Eq. wt. = equivalent weight of acid (V2 = volume of sample (mL); 1000 = factor relating mg to grams (mg/g) (1/10 = 100/1000).

2.11 Microbial assay

The total microbial count was determined using the method described by Herrera in 2001. A 5 g sample of cacao beverage was obtained and subsequently diluted with 90 ml of water peptone solution at a ratio of 1:10. Following the homogenization process, 1 mL of the sample was combined with 10 mL of total plate count agar medium in a sterilized Petri plate. This mixture was then thoroughly mixed at a temperature of 45 °C. In addition, the samples were incubated at a temperature of 37 °C to observe and quantify the number of colonies that developed on each Petri dish. The recorded observations were documented as the number of colonies detected within a 1 mL sample, expressed as log cfu/mL. The enumeration of yeasts and molds was conducted as the experimental procedures for the total microbial count were identical, with the only variation being the substitution of the total plate count agar with a potato dextrose agar medium [20].

2.12 Cytotoxicity assay

Cell viability was assessed using the SRB (Sulforhodamine B) assay. Myrrh resin extract (MRE) was prepared as an ethanolic extract dissolved in DMSO to create a final 25 mM stock solution. Proper media were utilized to prepare MRE at their final tested doses. The DMSO vehicle control was prepared by adding the maximum volume of DMSO used in preparing the tested MRE to appropriate media types, ensuring that the final DMSO concentration did not exceed 0.2%. Colon cancer (HCT) and liver cancer (HEPG2) cell lines obtained from Nawah Scientific Inc., (Mokatam, Cairo, Egypt) were utilized in the assays. Cells were maintained in DMEM media supplemented with 100 mg/mL of streptomycin, 100 units/mL of penicillin, and 10% heat-inactivated fetal bovine serum in a humidified, 5% (v/v) CO2 atmosphere at 37 °C. For the SRB assay, aliquots of 100 μL cell suspension (5 × 10^3 cells) were seeded into 96-well plates and incubated in complete media for 24 h. Subsequently, cells were treated with another aliquot of 100 μL media containing MRE extract at various concentrations. After exposure, cells were fixed by replacing the media with 150 μL of 10% TCA and incubated at 4 °C for 1 h. The TCA solution was then removed, and the cells were washed five times with distilled water. Aliquots of 70 μL SRB solution (0.4% w/v) were added and incubated in a dark place at room temperature for 10 min. Plates were washed 3 times with 1% acetic acid and allowed to air-dry overnight. Then, 150 μL of TRIS (10 mM) was added to dissolve protein-bound SRB stain, and the absorbance was measured at 540 nm using an Infinite F50 microplate reader (TECAN, Switzerland) [21]. The IC50 value for each tested sample was calculated by nonlinear regression of log concentration versus the percentage survival, implemented in Graph Pad PRISM version 8.0, GraphPad Software, Inc., CA.

2.13 Sensory evaluation

The sensory analysis was carried out using a composite score scale according to Lawless and Heymann (1993) [22]. Ten panelists semi-trained (the age of participants was between 25 and 35 years old) from the Food Science and Technology Department, Faculty of Agriculture, Al-Azhar University, Egypt, assessed the sensory quality of mango juice and cacao beverages. Three different treatments were scored for taste (30), odor (30), color (20), texture (20), and acceptability (score 100).

2.14 Statistical analysis

The studies were reproduced at least three times, and the values are reported as the mean ± standard deviation (SD). A variance analysis (one way-ANOVA) was performed to investigate the differences among the samples.

3 Results and discussions

The chemical structure and constituents of MER were determined using liquid chromatography-tandem mass spectrometry (LC–MS/MS). Myrrh resin extract (MRE) revealed the presence of more than 20 using positive and negative ionization modes including Phlorizin, Choline, alpha-D-Glucose-1,6-diphosphate, Sinapoyl malate, and N, N-Dimethylglycine. These findings are presented in Figs. 1, 2, and Table 2. Additionally, 106 major components were identified in myrrh essential oil, with other components detected at concentrations ranging from 0.01 to 1.88%. Terpenoids (monoterpenoids, sesquiterpenoids, volatile/essential oil), diterpenoids, triterpenoids, and steroids are among the constituents of myrrh resin, which is renowned for its abundant phytochemical profile. It has been used historically to treat a wide range of conditions, such as microbial infections, inflammatory illnesses, wounds, mouth ulcers, pain, fractures, and digestive problems [6, 7, 23].

Fig. 1
figure 1

LC–MS/MS analytical scan of Myrrh resin extract (MER) in positive ion mode. The chromatogram displays the detected peaks corresponding to various bioactive compounds present in the extract. The identified compounds are highlighted with their respective m/z (mass-to-charge) ratios and retention times, demonstrating the extract’s complex profile and the presence of key bioactive constituents

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Fig. 2
figure 2

LC–MS/MS analytical scan of Myrrh Resin Extract (MER) in negative ion mode. The chromatogram illustrates the mass-to-charge (m/z) ratios of various compounds detected in the extract. Key peaks corresponding to specific bioactive components are labeled, highlighting the compound profile in the negative ion mode

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Table 2 Active components of Myrrh resin extract analyzed using LC–MS/MS
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The antimicrobial properties of Myrrh resin extract (MRE) are attributed to a group of antioxidant compounds, as determined by high-performance liquid chromatography (HPLC), the results are in agreement with [6, 10]. The analysis revealed 19 active antioxidant compounds in MRE, with kaempferol being the most abundant at 1896 µg/g, followed by quercetin at 520 µg/g. Notably, Pyrocatechol exhibited the highest value among the compounds analyzed, at 162 µg/g.

Also, the extract contains other active compounds at low concentrations, which include Gallic acid, Chlorogenic acid, Catechin, Coffeic acid, Syringic acid, Rutin, Coumaric acid, Vanillin, Ferulic acid, Naringenin, Cinnamic acid, Apigenin, and Hesperetin at concentrations of 1.48, 0.84, 1.44, 0.60, 1.02, 60, 1.01, 0.76, 2.36, 1.22, 0.13, 1.71, and.00.00 µg/ml, respectively, as presented in Table 3 and Fig. 3.

Table 3 Antioxidant compounds of Myrrh resin extract analyzed by HPLC
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Fig. 3
figure 3

HPLC chromatogram of antioxidant compounds in Myrrh Resin Extract (MRE). The figure displays the chromatographic profile of the extract, showing the distinct peaks corresponding to various antioxidant compounds. Each peak represents a specific compound identified in the extract, highlighting their relative abundance and contributing to the overall antioxidant activity of MRE

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Further studies utilizing HPLC to evaluate the polyphenol constituents of various myrrh resin extracts (ethanol, ethyl acetate, petroleum ether, and chloroform) demonstrated their efficacy against respiratory infections such as COVID-19. These extracts exhibited a range of biological activities, including anti-inflammatory, antioxidant, antimicrobial, neuroprotective, anti-diabetic, and anticancer properties. Studies by Abbas et al. (2020), Rashaet al., (2023), Rahmani et al., (2022), Fatani et al., (2016), identified significant polyphenolic compounds such as Chlorogenic acid, Gallic acid, Catechin, Caffeine, Syringic acid, Coumaric acid, Ferulic acid, Naringenin, 4',7-Dihydroxyisoflavone, Propyl Gallate, Vanillin, Quercetin, and Ellagic acid in various concentrations, underscoring the potential therapeutic applications of myrrh extracts [24,25,26,27].

Table 4 illustrates the antimicrobial activity of MRE at different concentrations against Gram-positive (Staphylococcus aureus (MRSA)-ATCC43300; Bacillus subtilis-ATCC6633) and Gram-negative bacteria (Escherichia coli -ATCC25922; Pseudomonas aeruginosa-ATCC27853)). The MRE had potent antimicrobial activity against Gram-positive bacteria, and copmpelety inhibited Staphylococcus aureus (MRSA) (ATCC43300) ATCC43300-Bacillus subtilisATCC6633 at concentrations of 6.25, 3.13, and 1.56% and gradually inhibited the same strains from 10 to 7, 10 to 6 mm for the tested microorganisms respectively. Concerning the antibacterial activity of MER against Gram-negative bacteria, the results were better and more promising, and the MER completely inhibited Escherichia coli (ATCC25922) at all tested concentrations, while gradually inhibiting Pseudomonas aeruginosa (ATCC27853) from 20 to 8.5 mm and completely inhibiting at 6.25, 3.13, and 1.56%.

Table 4 Antibacterial activities and MIC of Myrrh Resin Extract (MRE) against Gram-positive and Gram-negative bacteria
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Using the viable count approach, the bactericidal effects of myrrh hexane extract and myrrh essential oil (MEO) against Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (Ps. aeruginosa) were studied. After a two-hour contact period, MEO demonstrated superior efficacy, killing both tested strains in > 99.999% of the cases. Four multidrug resistant isolates of S. aureus (MRSA, sputum), Escherichia coli (E. coli, urine), Ps. aeruginosa (wound) [28], and Klebsiella pneumonia (K. pneumonia, sputum) were examined using the same method as MEO. The highest bactericidal activity was observed against Ps. Aeruginosa, while the lowest was against K. pneumonia (99.59 and 54.04% killing, respectively. MEO has antibacterial properties against various bacterial strains and antifungal properties against Candida albicans [29]. The most effective efficacy was against S. aureus, which displayed 100% total growth suppression. While myrrh extract exhibited no effect against any of the pathogens under study at concentrations (12, 6, and 3 mg/mL), it completely inhibited the gram-negative bacteria Ps. auroginosa and E. coli at concentrations of 80–60 mg/mL. The result indicates that myrrh is an antibacterial agent that can be used in the future by making appropriate doses. The antibacterial activity of the C. myrrh extract was superior to that of the fungal isolates [30]. Myrrh's in-vitro effectiveness in combating bacteria and some airborne fungus. Twenty airborne fungi and ten harmful bacteria were examined in-vitro using extracts of myrrh in methanol, ethanol, hot water, and normal. Compared to extracts in boiling and plain water, methanol and ethanol demonstrated stronger activity against fungus [31, 32].

The MRE was applied at a concentration 1% in cacao beverage as an antibacterial agent against the total bacterial count, and the produced beverage was stored for 14 days. Results in Table 5 cleared the efficacy of MRE as an antibacterial agent since it could inhibit the microorganisms in the produced cacao beverage at zero time as typical of sodium benzoate, although the control cacao beverage had 1.3 cfu/mL, and the total bacterial count slightly appeared after 7 and 14 days to reach 1.5 and 1.7, cfu/mL, while the control and sodium benzoate samples had 2.2, 2.9, and 1.1, respectively. The results demonstrate the efficiency of MRE against microorganisms contaminated cacao beverage. Also, MRE was tested in cacao beverage against mold and yeast counts and gave strong efficiency against these counts, and all tested beverage samples were free from mold and yeast at zero time and during storage. The effect of Myrrh resin (ethanol, ethyl acetate, petroleum ether, and chloroform) resin extract against four different pathogenic bacteria, Salmonella typhimurium, Pseudomonas aeruginosa, Escherichia coli, and Bacillus cereus, was examined by measuring inhibition zone (diameter mm). The results revealed that there were significant differences between the bacteria and different extraction methods. Aqueous, ethyl acetate, and petroleum ether extracts of the Myrrh resin seed have excellent activity against the Candida albicans fungus. The purpose of the study was to determine the nutritional value, polyphenol content, and antibacterial and antifungal properties of Myrrh resin [24]. The best antibacterial activity was seen in a hydroalcohol extract that was extracted using an ethanol: phosphate buffer pH 7 (85:15) ratio, [33]. The highest antibacterial and antifungal activity was found in the ethanol extract. Fresh-cut salads treated with these two myrrh extracts showed noticeably less bacterial growth than untreated salads, Luisa [34]. Escherichia coli was shown to have a zone of inhibition caused by C. myrrha extract at 29 mm, Staphylococcus epidermidis at 27 mm, Candida albicans at 27 mm, and Aspergillus brasiliensis at 16 mm [35].

Table 5 Application of Myrrh Extract Resin (MER) as an antibacterial agent in cacao beverages
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Myrrh resin exhibits significant therapeutic benefits due to the activity of its diverse metabolites. The extract has shown promising effects against cancer and inflammatory conditions, with evidence of its safety and lack of harmful impact on primary fibroblasts [14].

In this study, we investigated the anti-cancer properties of Myrrh resin extract against colon cancer (HCT) and liver cancer (HEPG2) cell lines. Our findings indicate that the extract has promising cytotoxic effects, Fig. 4 illustrates the IC50 values of the extract against HCT and HEPG2 cell lines, indicating significant cytotoxic effects with values of 55.69 μg/ml for HCT cells and 70.78 μg/ml for HEPG2 cells. These IC50 values are noteworthy as they are close to the recommended threshold for significance set by the American Cancer Institute, which is IC50 < 30 µg/mL for crude extracts. This suggests that Myrrh resin extract may have potential as a cytotoxic agent against these cancer cell types.

Fig. 4
figure 4

The cytotoxic impact of Myrrh resin on the growth of colon cancer (HCT) and liver cancer (HEPG2) cell lines, both in control conditions and after treatment. A, B depict the control and treatment conditions for HCT cells, respectively. Notice the loss of cellular integrity and increased cell fragmentation in Panel B, indicating the cytotoxic effects of Commiphora myrrh. Similarly, C, D show HEPG2 liver cancer cells under control and treated conditions, with significant morphological alterations such as cell shrinkage and detachment observed in Panel D. Panel E presents the IC50 values for Myrrh resin concerning its inhibitory effect on HCT and HEPG2 cells. The IC50 value for each tested sample was calculated by nonlinear regression of log concentration versus the percentage survival

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The anticancer effects observed in our study with Commiphora myrrh resin extract may be closely linked to its antioxidant properties. Antioxidants are crucial in safeguarding cells from the detrimental effects of oxidative stress, which is a significant contributor to cancer development and progression. Reactive oxygen species, produced as byproducts of normal cellular metabolism, can induce DNA damage and drive the mutations that initiate and propel cancer. Antioxidants mitigate this risk by neutralizing these reactive species, thereby preventing or diminishing the oxidative damage that can lead to cancer.

Our results are consistent with previous research; Chen et al. (2013) investigated the effects of myrrh extract on liver cancer and found that furano-sesquiterpenoids isolated from Arabic Myrrh resin induced apoptosis of human hepatocellular carcinoma HepG2 cells with an IC50 of 3.6 μm [37]. Furthermore, Hamad et al. (2017) reported that myrrh, in combination with protocatechuic acid, induces apoptotic cell death in colon cancer cells by suppressing the Bcl-2 gene [38]. This suggests a potential mechanism by which Myrrh resin extract exerts its anti-cancer effects on colon cancer cells. Our phytochemical and antioxidant analyses revealed significant levels of bioactive compounds in the extract. Among them, kaempferol, quercetin, and ferulic acid stood out for their well-documented anti-cancer properties. Kaempferol, a flavonoid, has been extensively studied for its ability to induce apoptosis, inhibit cancer cell proliferation, and suppress tumor growth through various molecular mechanisms. Quercetin, another flavonoid, exhibits anti-proliferative effects by interfering with cellular processes involved in cancer development and possesses antioxidant and anti-inflammatory properties. Ferulic acid, a polyphenolic compound, exerts anti-cancer effects by inhibiting cancer cell proliferation, inducing apoptosis, and suppressing inflammation and angiogenesis. The presence of these dominant compounds in the Myrrh resin extract supports its potential as an anti-cancer agent against colon and liver cancer cell lines.

The total soluble solids (TSS), PH value, and acidity of the produced untreated and treated cacao beverage with MRE and sodium benzoate were carried out as shown in Table 6, and the results show that the TSS of all tasted samples had the same value of 10.60, there was no difference between untreated or treated samples, and the addition of MRE or sodium benzoate hadn’t any effect. The same results were found for the pH value, which recorded almost 6.8 for all tested samples, as well as the acidity, which recorded almost 0.10% for untreated and treated samples, and all tested values were almost similar to control. The pH range of the cocoa drinks was 6.44 to 7.10. The highest pH was 7.10 [39]. The PH value of the sample was between 7.61 and 7.72, while the total soluble solids of the chocolate beverages varied between 10.38 and 10.75 Brix [40].

Table 6 Total soluble solids, pH value, and acidity of the produced treated cacao beverage
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The effect of the addition of extract on the sensory properties of cacao beverages was measured as shown in Table 7. Results cleared that the MRE had an insignificant effect on taste, odor, color, and texture of the produced cacao beverage in comparison with the control sample, and the score of taste decreased from 29 to 26 and 28 for the control and treated samples, respectively. The same trend has been shown with odor, color, and texture, which have had a slight decrease from 27, 19, and 19 of control to 18, 18, and 18, respectively. In general, the general acceptability of treated beverages was very acceptable for judgments, which recorded 95, 88, and 94 for control and treated samples, respectively.

Table 7 The effect of the addition of Myrrh Extract Resin (MER) on the sensory properties of cacao beverages
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4 Conclusion

In conclusion, Myrrh resin extract (MRE) emerges as a versatile natural agent with significant implications for both food preservation and oncological interventions. This study has demonstrated MRE’s dual functionality as an effective antimicrobial agent for food safety and its potential as an anti-cancer therapeutic. The analysis using HPLC and LC–MS/MS highlighted MRE’s antioxidant properties, identifying kaempferol and quercetin as key compounds that not only inhibit microbial growth but also exhibit cytotoxic effects against colon cancer (HCT) and liver cancer (HEPG2) cell lines. Moreover, the incorporation of MRE into cacao beverages showed no adverse effects on their physical or chemical properties, maintaining stable measurements of total soluble solids (TSS), pH, and acidity. Sensory evaluation confirmed minimal impact on taste, odor, color, and texture, with treated samples receiving favorable ratings comparable to untreated samples. These findings underscore MRE’s potential as a natural alternative to synthetic food preservatives and its promise in oncological interventions, particularly in cacao products. The ability of MRE to preserve food quality without compromising sensory attributes positions it favorably for broader application in the food and beverage industry. Furthermore, its anti-cancer properties suggest potential therapeutic benefits that could be explored further in oncological research and product development. Considering these results, future studies could delve deeper into optimizing MRE formulations for specific food applications and refining its efficacy in cancer prevention and treatment strategies. Overall, MRE represents a promising avenue for innovation in both food science and oncology, paving the way for safer, healthier food products and potential therapeutic advancements.