Evaluation of the Antimicrobial and Anticancer Properties of Myrrh Resin Extract and Its Application in Cacao Beverages

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Goda, Eman G Ayad, Menna H. Amin, Mahmoud Youssef, Jianrong Shi, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4218698/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 37 You are reading this latest preprint version Abstract Due to the potential health risks of synthetic food preservatives, there has been a noticeable increase in interest in finding natural food preservatives during the past few decades. The goal of this study was to investigate the use of a natural extract of Commiphora Myrrh as an antimicrobial agent. The antioxidant properties of Myrrh resin extract (MRE) were analyzed using HPLC and GC-MS. The results showed that MRE contained potent antioxidant compounds, including 19 compounds, with the dominant compound being kaempferol, which had the highest value of 1896 µg/g. Quercetin was found to be the second most abundant compound, with a value of 520 µg/g. The efficacy of MRE as an antimicrobial agent against both Gram-positive and Gram-negative bacteria was tested, and its application in Cacao beverage was also studied. The results demonstrated that MRE was highly effective against all the tested bacteria both in vitro and in the total bacterial count of the produced cacao beverage. Additionally, the fungi in the cacao beverage were completely inhibited at all tested concentrations of MRE. The total soluble solids (TSS), pH value, and acidity of the produced untreated, treated cacao beverage with MRE and sodium benzoate were carried out, and all values mentioned were almost the same, with no differences noted. The sensory evaluation of Cacao beverage showed that the MRE had a minor impact on taste, odor, color, and texture of the produced cacao beverage in comparison with the control sample, which was very acceptable for judgments and recorded 95, 88, and 94 for the control and treated samples, respectively. Furthermore, the anti-cancer properties of MRE were evaluated, revealing significant cytotoxic effects against colon cancer (HCT) and liver cancer (HEPG2) cell lines. The IC50 values for HCT and HEPG2 cells were 55.69 µg/ml and 70.78 µg/ml, respectively, indicating the potential of MRE as an anti-cancer agent. Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Some preservatives are classed as carcinogens or cancer-causing chemicals because they are known to be hazardous to human health. Certain preservatives are considered carcinogenic or cancer-causing substances due to their known harmful effects on human health. According to Gupta and Yadav (2021) , artificial food additives interact with the cellular structure of the body, leading to a range of food-related disruptions, Mirza et al. ( 2017 ) also corroborate these findings. When used in excess, it might cause negative effects such skin rashes and itching, breathing problems, sneezing, or stomach issues (Silva and Lidon., 2016) . M. M. Silva and F. Lidon, 2016 . Certain food preservatives are classified as carcinogens or cancer-causing agents due to their hazardous effects on human health. 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 stomach issues when used excessively. Due to the possible health risks associated with synthetic food preservatives, consumers have become more cautious about consuming foods containing these additives, leading to an increase in demand for natural food preservatives over the past few decades. ( Atwaa et al., 2022 and Teshome et al 2022 ). Commiphora myrrh contains phytochemical components such as terpenoids (monoterpenoids, sesquiterpenoids, volatile/essential oil), diterpenoids, triterpenoids, and steroids. It has been utilized throughout history to treat various ailments such as wounds, mouth ulcers, pains, fractures, stomach issues, microbial infections, and inflammatory diseases, as reported by Mohammed et al., 2015 and Hanu LO et al ., 2005) . Akbar 2020 and Mohammed et al., 2015 have also documented the medicinal properties of Commiphora myrrh. Myrrh is known for its antiseptic, astringent, anthelmintic, and expectorant properties. Studies by Batiha et al., 2023 and Akbar., 2020 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 , 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. Al-Sabri et al., 2014 have also conducted research on myrrh's potential as an antibacterial agent. According to Khalil, et al., 2020 , Commiphora myrrh ether and ethanol extracts were tested for their antibacterial properties, these extracts showed antimicrobial activity against the Gram-negative organisms tested alongside Omer. , et al ., 2011 , and Alqahtani. , et al . 2021 . 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, Omer. , et al 2011 and Khalil., et al 2020. In addition to its antimicrobial properties, the potential anticancer effects of Commiphora myrrh have also garnered attention ( Suliman et al., 2022 ). 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. Material and methods 2.1. Materials Myrrh Resin Extract (MRE), Cacao powder, vegetable 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 B. subtilis ATCC6633, and S. aureus (MRSA) ATCC43300 as well as Gram-negative bacteria P. aeruginosa (ATCC27853), and E. coli (ATCC25922). 2.3. Chemicals and reagents All chemicals and reagents of the analytical methods used in the present study were analytical grades. Citric acid, sodium benzoate, CMC, xanthan gum, plate count agar (PCA), and potato dextrose agar (PDA) were purchased from El-Gamhouria Trading for Chemicals and Drugs Company, Cairo, Egypt. 2.4. Extraction of antioxidants from MER One hundred grams of Commiphora Myrrh resin added to excessive distilled water: ethanol (2:8 v/v) and incubated at room temperature for 24 h, then the slurry was filtered through filter paper. The water extract was concentrated using rotary evaporator under reduced pressure and the residues were dissolved in 50 mL of distilled water. 2.5. LC-ESI-MS/MS analysis of Myrrh resin extract profile 2.5.1. Instrument The analysis of the sample was performed by liquid chromatography–electrospray ionization–tandem mass spectrometry (LC-ESI-MS/MS) as described by El-Ssayad et al ., 2023.. 2.5.2 Positive ionization mode: Separation was performed with a Ascentis® Express 90 Å C18 Column (2.1×150 mm, 2.7 µm). The mobile phases comprised of two eluents A: 5 mM ammonium formate pH 3; B: acetonitrile (LC grade) El-Ssayad et al ., 2023. 2.6. Determination of antioxidants using HPLC HPLC analysis was carried out using an Agilent 1260 series.The separation was carried out using Eclipse C18 column (4.6 mm x250 mm i.d., 5 µm).Themobile phase consisted of water(A) and 0.05% trifluoroacetic acid in acetonitrile (B) at a flow rate 0.9 ml/min. The mobile phase was programmed consecutively 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 (82%A).The multi-wavelength detector was monitored at 280 nm. The injection volume was 5 µl for each of the sample solutions. The column temperature was maintained at 40°C. 2.7. Minimal inhibitory concentration (MIC) determination In accordance with the disk diffusion assay protocol, a volume of 1 mL from each bacterial suspension was evenly distributed onto a solid growth medium within a Petri dish. Six sterile paper disks measuring 6 mm in diameter were carefully positioned on the surface of each agar plate. These disks were then saturated with 15 µL of the diluted MER. The plates were incubated at 37°C for 24 hr. The antibacterial activity, measured as the minimum inhibitory concentration (MIC), was assessed by determining the lowest concentration of either diluted MER that resulted in the formation of an inhibition zone around a disk after a 24hr incubation period. Negative controls consisted of disks impregnated with sterile distilled water. Replicates were conducted at each concentration (Lima-Filho and de Aguira Corderio, 2014) . 2.8. Preparation of Cacao beverage In preparation for 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 the formula-2 and 0.1% of Sodium benzoate was incorporated into the formula-3. A quantity of tape water was added and the heated at 80 ± 2°C for 5 minutes (Faiqoh et al., 2021) , the formulations of cocoa beverages are presented in Table 1 . Table 1 formulations of cocoa beverages Cacao beverage % formula-1 formula-2 formula-3 Cacao powder 7.5% 7.5% 7.5% Sugar 15% 15% 15% Vegetable cream 6% 6% 6% Salt 0.03% 0.03% 0.03% Corn starch 1.5% 1.5% 1.5% Xanthan gum 0.1% 0.1% 0.1% Myrrh extract - 1% - Sodium benzoate - - 0.1% Water 69.87% 68.87% 69.77% 2.9. 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 to 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. According to the method described by the AOAC (2012). Total titratable acidity (TTA) was determined in 10mL of each mango juice and cacao beverage, which was homogenized with 90 ml of distilled water, and titrated with 0.1 mol/L NaOH, using phenolphthalein as an indicator (AOAC, 2012) . 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, $$\% acidity= \frac{N\times V1\times Eq. wt.}{V2\times 1000}\times 100$$ Where, N = normality of NaOH (mEq/mL); V 1 = volume of titrant (mL); Eq. wt. = equivalent weight of acid (; V 2 = volume of sample (mL); 1,000 = factor relating mg to grams (mg/g) (1/10 = 100/1000). 2.10. Microbial assay The total microbial count was determined using the method described by Herrera in 2001. A 5g sample of mango juice or 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 were 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 (Herrera, 2001) . 2.11. 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) CO 2 atmosphere at 37°C. For the SRB assay, aliquots of 100 µL cell suspension (5x10^3 cells) were seeded into 96-well plates and incubated in complete media for 24 hours. 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 hour. The TCA solution was then removed, and the cells were washed 5 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 minutes. 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) ( Skehan et al., 1990 ). 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 2.12. Sensory evaluation The sensory analysis was carried out using a composite score scale according to Lawless and Heymann (1993) . Ten panelists semi-trained (The age of participants was between 25–35 years old) from Food Science and Technology Department, Faculty of Agriculture, Al-Azhar University, Egypt, assessed the sensory quality of mango juice and cacao beverage. Three different treatments were scored for taste (30), odor (30), color (20), texture (20), acceptability (score 100). 2.13. Statistical analysis: The studies were reproduced at least three times, and the values are reported as the mean ± standard deviation (SD). A variance analysis (ANOVA) was performed to investigate the differences among the samples. Results and Discussions Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of Commiphora Myrrha extract (MRE) revealed the presence of more than 20 positive and negative substances including Phlorizin, Choline, alpha-D-Glucose-1,6-diphosphate, Sinapoyl malate, and N, N-Dimethylglycine. These findings are presented in Figures 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% (Arın et al ., 2021) . 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 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. The remaining compounds, including Gallic acid, Chlorogenic acid, Catechin, Coffeic acid, Syringic acid, Rutin, Coumaric acid, Vanillin, Ferulic acid, Naringenin, Cinnamic acid, Apigenin, and Hesperetin, showed varying concentrations as presented in Table 3 and Figure 3. 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), Brasha et al ., (2023), Rahmani et al ., (2022), Fatani et al ., (2016), and Rasha et al ., (2023) 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. Table [2]: Active components of Commiphora Myrrha extract have been analyzed using LC-MS/MS Chemical structure Area Compounds C 18 H 30 O 2 4.63×10 8 gamma-Linolenic acid N,N-Dimethylglycine Choline Carnosine Uridine 5'-monophosphate Phlorizin Sinapoylmalate alpha-D-Glucose-1,6-diphosphate C 4 H 9 NO 2 5.9×10 8 C 5 H 14 NO 6.86×10 8 C 9 H 14 N 4 O 3 5.48×10 8 C 9 H 13 N 2 O 9 P 1.24×10 9 C 21 H 24 O 10 1.75×10 8 C 15 H 16 O 9 8.37×10 8 C 6 H 14 O 12 P 2 8.8×10 8 Table 3: Antioxidants compounds of Myrrh resin extract analyzed by HPLC. Bioactive compound Area Conc. (µg/ml) Gallic acid 17.11 1.48 Chlorogenic acid 6.12 0.84 Catechin 5.83 1.44 Methyl gallate 0.00 0.00 Caffeic acid 7.94 0.61 Syringic acid 15.04 1.02 Pyro catechol 19.24 2.77 Rutin 5.14 0.60 Ellagic acid 0.00 0.00 Coumaric acid 31.97 1.01 Vanillin 17.25 0.76 Ferulic acid 34.54 2.36 Naringenin 10.12 1.22 Daidzein 0.00 0.00 Querectin 64.25 8.85 Cinnamic acid 7.22 0.13 Apigenin 22.41 1.71 Kaempferol 415.80 32.24 Hesperetin 0.00 0.00 Table 4 Antibacterial activities and MIC of MRE against Gram-positive and negative bacteria Organisms Inhibitory activity against the tested organism (zone of inhibition in mm) Concentration 100% 50% 25% 12.5% 6.25% 3.13% 1.56% Gram-positive bacteria Staphylococcusaureus (MRSA) - ATCC43300 10 8.5 7.5 7 NA NA NA Bacillussubtilis ATCC6633 10 8 6.5 6 NA NA NA Gram-negative bacteria Escherichiacoli ) ATCC 25922 ( NA NA NA NA NA NA NA Pseudomonas aeruginosa (ATCC27853) 20 14 11.5 8.5 NA NA NA 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 subtilis ATCC6633 at concentrations of 6.25, 3.13, and 1.56% and gradually inhibited the same strains from 10-7, 10-6 mm for the tested microorganisms respetivly.Concerning the antibacterial activity of MER against Gram-negative bacteria, the results were better and more promising, and the MER completely inhibited Escherichia coli) ATCC 25922( at all tested concentrations, while gradually inhibiting Pseudomonas aeruginosa (ATCC27853) from 20–8.5 mm and completely inhibiting at 6.25, 3.13, and 1.56%. 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), and Klebsiella pneumonia ( K. pneumonia , sputum) were examined using the same method as MEO. Highest bactericidal activity was observed against Ps. aeruginosa while 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 Auda ., et al . 2017 . 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 P. auroginosa and E. coli at concentrations of 80 to 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 Alharbi, and Alhashmi., 2022 . 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 Al-Sabri AE; Moslem M 2016 . Table 5 Application of MER as antibacterial activity on total count of bacteria by Log cfu/mL in Cacao Beverage Storage at 4 °C Control MER (1%) Sodium benzoate (0.1%) Total bacteria count by Log cfu/Ml Zero 1.3±0.13 Nil Nil 7 2.2±0.08 1.5±0.1 1.1±0.06 14 2.9±0.3 1.7±0.05 1.1±0.09 Mold and yeast count by Log cfu/mL Zero Nil Nil Nil 7 Nil Nil Nil 14 Nil Nil Nil The MRE was applied at concentration 1% in cacao beverage as antibacterial agent against 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 Commiphora myrrh (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 Commiphora myrrh 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 Commiphora myrrh. ( Abbas et al ., 2020) . The best antibacterial activity was seen in a hydroalcohol extract that was extracted using an ethanol: phosphate buffer pH 7 (85:15) ratio, Almekhlafi S et al 2014 . 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 Boffa et al, 2023. 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 ( Alshibly et al .,2022). In this study, we investigated the anti-cancer properties of Commiphora myrrh extract against colon cancer (HCT) and liver cancer (HEPG2) cell lines. Our findings indicate that the extract has promising cytotoxic effects, Figure 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 Commiphora myrrh extract may have potential as a cytotoxic agent against these cancer cell types. 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 Commiphora Myrrh resin induced apoptosis of human hepatocellular carcinoma HepG2 cells with an IC50 of 3.6 μM. 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. This suggests a potential mechanism by which Commiphora myrrh 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 Commiphora myrrh extract supports its potential as an anti-cancer agent against colon and liver cancer cell lines. Table 6: total soluble solids, pH value and acidity of produced treated cacao beverage. Test Control Myrrhextract Sodium benzoate T.S.S 10.60±0.9 a 10.60±0.9 b 10.60±0.8 a pH value 6.80±1.6 a 6.80±0.8 b 6.70±0.8 a Acidity (as lactic acid) 0.10% 0.10% 0.13% The means within the same row having different superscripts are significantly varied (P ≤0.05). 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 samples. The pH range of the cocoa drinks was 6.44 to 7.10. The highest pH was 7.10 Rossi Indiarto et al ., 2022 . pH value of the sample was between 7.61-7.72, while total soluble solids of the chocolate beverages varied between 10.38-10.75 oBrix Salwa Al Aribah et al., 2020 . Table 7: The effect of the addition of extract on the sensory properties of cacao beverages Test Control Myrrh extract Sodium benzoate Taste (30) 29.0±0.9a 26.0±0.9b 28.0±0.8a Odor (30) 27.9±1.6a 25.0±0.8b 28.4±0.8a Color (20) 19.0±0.9a 18.0±1.9a 18.6±1.3a Texture (20) 19.2±0.8a 18.6±0.8a 18.8±0.8a Acceptability (score 100) 95.8±2.6a 88.4±1.1b 94.0±2.7a The means within the same row having different superscripts are significantly varied (P ≤0.05). 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. Conclusion The results of this study highlight the potential of Myrrh resin extract (MRE) as a natural antimicrobial agent for food preservation and as a potential anti-cancer agent. The study analyzed MRE's antioxidant properties using HPLC and LC-MS/MS and identified two potent antioxidant compounds, kaempferol and quercetin. These compounds not only contribute to MRE's effectiveness in inhibiting microbial growth but also exhibit anti-cancer properties, as they have a cytotoxic effect against colon cancer (HCT) and liver cancer (HEPG2) cell lines. The study also investigated the incorporation of MRE into cacao beverages and found no significant changes in their physical or chemical properties, as indicated by stable measurements of total soluble solids (TSS), pH value, and acidity. Furthermore, sensory evaluation revealed that treated samples did not have a significant impact on taste, odor, color, and texture and received very acceptable ratings comparable to untreated samples. Overall, the study suggests that Myrrh resin extract has multiple benefits, making it a promising natural alternative to synthetic food preservatives and a potential adjunct therapy in cancer prevention and treatment. With its demonstrated effectiveness in preserving food quality and safety, minimal sensory impact, and anti-cancer properties, it could be applied more broadly in food and beverage products, as well as in oncological interventions. Declarations Informed Consent Statement Every participant on the sensory panel has given written consent to participate in the research study. Before the study commenced, participants were notified of the purpose and methods of the research and assured of the strict confidentiality of their data. Participation in the study was voluntary, and subjects had the autonomy to withdraw their engagement at any time. The individuals demonstrated exceptional physical health and had no recorded allergies to the components. The research lacks institutional approval due to Egypt's rules regarding human research. The research protocol is in accordance with the ethical standards of our institution, Al-Azhar University, Faculty of Agriculture, and National Research Center, Egypt, and approval is not needed according to Egypt's rules regarding sensory evaluation. Data Availability Statement All relevant information has been included in the manuscript itself Acknowledgements We thank the Jiangsu Province Science and Technology Support Program (BZ2022001) China, Nanjing Microtest Biotechnology Co., Ltd, Nnjing, 210061, Jiangsu, China, and the National Research Centre, Cairo, Egypt. References Abbas, R.K., Al-Mushhin, A.A.M., Elsharbasy, F.S. and Ashiry, K.O. (2020). Nutritive Value, Polyphenol Constituents and Prevention of Pathogenic Microorganism by Different Resin Extract of Commiphora myrrh . J Pure Appl Microbiol ., 14(3):1871-1878. doi: 10.22207/JPAM.14.3.26 Akbar, S. (2020). Commiphora myrrh a (Nees) Engl. (Burseraceae). In: Akbar S (ed) Handbook of 200 medicinal plants: a comprehensive review of their traditional medical uses and scientific justifications. Springer International Publishing, Cham, pp 701–706. Alharbi, N.M. and Alhashmi, H.M. (2022). The use of Commiphora myrrh extract as disinfectant agent against pathogenic microbes isolated from beauty. African Journal of Biological Sciences, 108-121. https://ssrn.com/abstract=4023517. Almekhlafi, S., Thabit, A. A., Alwossabi, A. M., Awadth, N., Thabet, A. A., & Algaadari, Z. (2014). Antimicrobial activity of Yemeni myrrh mouthwash. J Chem Pharm Res, 6(5), 1006-1013. Alqahtani, A.S., Herqash, R.N., Noman, O.M., Tabish Rehman, M., Shahat, A.A., Alajmi, M.F., Nasr, F.A. (2021). Impact of Different Extraction Methods on Furanosesquiterpenoids Content and Antibacterial Activity of Commiphora myrrha Resin. J Anal Methods Chem. 10;2021:5525173. doi: 10.1155/2021/5525173. PMID: 34336357; PMCID: PMC8289610. Al-Sabri, A., Moslem, M.A., Hadi, S., Yassin, M.A. and Ameen F. (2014). Antifungal Activity of Commiphora myrrha L. Against Some Air Fungi. JOURNAL OF PURE AND APPLIED MICROBIOLOGY. Vol. 8(5), p. 3951-3955. Al-Sabri, A.E. and Moslem, M. (2016). Antimicrobial efficacy of Arabian shrub 'Myrrh' (Commiphora myrrha L.) Editore ‏ :‎ LAP LAMBERT Academic Publishing (1 marzo 2016) Copertinaflessibile ‏ : ‎ 116 pagine. ISBN-10 :‎ 365983372X, ISBN-13 ‏ : ‎ 978-3659833724 Alshibly, N. M., Mohamed, R. M., Al-Shaikh T. M. and Soliman, A.M. (2022). Study of Commiphora myrrha (Nees) Engl. var. molmolExtract and Its Antibiogram Against Some Microbial Pathogens,Egyptian Journal of Chemistry., 65(5), 13-22. doi10.21608/EJCHEM.2021.98426.4584 Atwaa, E.S.H., Shahein, M.R., Radwan, H.A., Mohammed, N.S., Aloraini, M.A., Albezrah, N.K.A., Alharbi, M.A., Sayed, H.H., Daoud, M.A. and Elmahallawy, E.K. (2022). Antimicrobial Activity of Some Plant Extracts and Their Applications in Homemade Tomato Paste and Pasteurized Cow Milk as Natural Preservatives. Fermentation .,, 8 , 428. https://doi.org/10.3390/fermentation8090428 Auda, S.H., Salem-Bekhit, M.M., Alanazi, F.K. et al. (2017). Antimicrobial evaluation of novel buccoadhesive films containing Myrrh extract. Polym. Bull., 74, 4041–4054. https://doi.org/10.1007/s00289-017-1946-x. Batiha, G.E., Wasef, L., Teibo, J.O., Shaheen, H.M., Zakariya, A.M., Akinfe, O.A., Teibo, T.K.A., Al-Kuraishy, H.M., Al-Garbee, A.I., Alexiou, A., Papadakis, M. (2023). Commiphora myrrh: a phytochemical and pharmacological update. NaunynSchmiedebergs Arch Pharmacol. 396(3):405-420. doi: 10.1007/s00210-022-02325-0. Epub 2022 Nov 18. PMID: 36399185; PMCID: PMC9672555. Batiha, G.ES., Wasef, L., Teibo, J.O. et al. (2023). Commiphora myrrh : a phytochemical and pharmacological update. Naunyn-Schmiedeberg's Arch Pharmacol 396, 405–420. https://doi.org/10.1007/s00210-022-02325-0 Boffa, L., Binello, Ar., Boscaro, V.,Gallicchio, M., Amisano, G.,Fornasero, S. and Cravotto, G. (2023). Commiphora myrrha (Nees) Engl. extracts: Evaluation of antioxidant and antiproliferative activity and their ability to reduce microbial growth on fresh-cut salad. INTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY., 51(3) 625-632. DOI: 10.1111/ijfs.13018. http://doi.wiley.com/10.1111/ijfs.13018 Chen, Y., Zhou, C., Ge, Z., Liu, Y., Liu, Y., Feng, W., ... & Wei, T. (2013). Composition and potential anticancer activities of essential oils obtained from myrrh and frankincense. Oncology letters, 6(4), 1140-1146. Fatani, A.J., Alrojayee, F.S., Parmar, M.Y., Abuohashish, H.M., Ahmed, M.M., Al-Rejaie, S.S. (2016). Myrrh attenuates oxidative and inflammatory processes in acetic acid-induced ulcerative colitis. Exp Ther Med., 12(2):730-738. doi: 10.3892/etm.2016.3398. PMID: 27446268; PMCID: PMC4950638. Gupta, R. andYadav, R.K. (2021). Impact Of Chemical Food Preservatives On Human Health. PalArch’s Journal of Archaeology of Egypt / Egyptology , 18 (15), 811 - 818. https://archives.palarch.nl/index.php/jae/article/view/9009. Hamad, G. M., Taha, T. H., Alshehri, A., & El‐Deeb, N. M. (2017). Myrrh as a functional food with therapeutic properties against colon cancer in traditional meals. Journal of food processing and preservation , 41 (1), e12963. Hanuš, L.O., Řezanka, T., Dembitsky, V.M., et al. (2005). Myrrh—Commiphora chemistry. Biomed Papers 149:3–28 Khalil, N., Fikry, S., Salama, O. (2020). Bactericidal activity of Myrrh extracts and two dosage forms against standard bacterial strains and multidrug-resistant clinical isolates with GC/MS profiling. AMB Express 10:21. Khalil, N., Fikry, S., Salama, O. (2020). Bactericidal activity of Myrrh extracts and two dosage forms against standard bacterial strains and multidrug-resistant clinical isolates with GC/MS profiling. AMB Express. 28;10(1):21. doi: 10.1186/s13568-020-0958-3. PMID: 31993779; PMCID: PMC6987268. Mirza, S.K., Asema, U.K. And Kasim, S.S. (2017). To Study The Harmful Effects Of Food Preservatives On Human Health. Journal of Medicinal Chemistry and Drug Discovery 02, 02, pp. 610-616, 8 . www.jmcdd.org. Mohammed, S., Alhussaini, A.M., Saadabi, M. I., Alghonaim. and Khalid E. I. (2015). An Evaluation of the Antimicrobial Activity of Commiphora myrrha Nees (Engl.) Oleo-gum Resins from Saudi Arabia. Journal of Medical Sciences, 15: 198-203. DOI: 10.3923/jms.2015.198.203, URL: https://scialert.net/abstract/?doi=jms.2015.198.203 Obaid, Z.M. and Ahmed, M.A. (2020). Anti-Bacterial Effects of Commiphora Myrrha and Ziziphus Spina-Christ Leaves Extracts Against Streptococcus Mitis (Primary Colonizer of Dental Plaque) In vitro Study. J Res Med Dent Sci.,8(3): 57-64. Omer, S.A., Adam, S.E.I. and Mohammed O.B. (2011). Antimicrobial Activity of Commiphora myrrha Against Some Bacteria and Candida albicans Isolated from Gazelles at King Khalid Wildlife Research Centre. Research Journal of Medicinal Plants., 5: 65-71. https://scialert.net/abstract/?doi=rjmp.2011.65.71 Omer, S.A., Adam, S.E.I. and Mohammed, O.B. (2011). Antimicrobial Activity of Commiphora myrrha Against Some Bacteria and Candida albicans Isolated from Gazelles at King Khalid Wildlife Research Centre. Research Journal of Medicinal Plants., 5(1), 65-71. Rahmani, A.H., Anwar, S., Raut, R., Almatroudi, A., Babiker, A.Y., Khan, A.A., Alsahli, M.A., Almatroodi, S.A. (2022). Therapeutic Potential of Myrrh, a Natural Resin, in Health Management through Modulation of Oxidative Stress, Inflammation, and Advanced Glycation End Products Formation Using In Vitro and In Silico Analysis. Appl. Sci., 12 , 9175. https://doi.org/10.3390/app12189175 Rasha, A., Mansouri, A. A., Roushdy, M. M., Huda, F., Alshaibi., Mahmoud, R. (2023) "Pharmacological Studies on the Antidiabetic, Antioxidant, and Antimicrobial Efficacies of Commiphora myrrha Resin in Streptozotocin-Induced Diabetes in Rats: A Preclinical Study", Journal of Diabetes Research. , Article ID 5478267, 12 pages. https://doi.org/10.1155/2023/5478267 Rossi, I., Souvia, R., Edy, S., Nur A., Gardiantini P. and Aldila D. P. (2022). Antioxidant activity and characteristics of a cocoa drink formulated with encapsulated green coffee extract, International Journal of Food Properties., 25:1, 2477-2494. https://doi.org/10.1080/10942912.2022.2144883. Salwa, A., Adhitya, P.S., Dimas, R.A.M., Danar, P. (2020). Sensorial and physical properties of chocolate beverage prepared using low fat cocoa powder. AIP Conference ., 2219 (1): 070007. https://doi.org/10.1063/5.0003435 Silva, M.M., and Lidon, F. (2016). FOOD PRESERVATIVES – AN OVERVIEW ON APPLICATIONS AND SIDE EFFECTS. Emirates Journal of Food and Agricultur.e , vol. 28, no. 6, pp. 366-73. doi:10.9755/ejfa.2016-04-351. Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, et al. (1990). New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst 82(13): 1107-1112 Suliman, R. S., Alghamdi, S. S., Ali, R., Aljatli, D., Aljammaz, N. A., Huwaizi, S., & Rahman, I. (2022). The role of myrrh metabolites in cancer, inflammation, and wound healing: prospects for a multi-targeted drug therapy. Pharmaceuticals , 15 (8), 944. Teshome, E., Forsido, S.F., Rupasinghe, H.P.V., Olika Keyata, E. (2022). Potentials of Natural Preservatives to Enhance Food Safety and Shelf Life: A Review. Scientific World Journal., 9901018. doi: 10.1155/2022/9901018. PMID: 36193042; PMCID: PMC9525789. Additional Declarations No competing interests reported. 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As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4218698","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":300118779,"identity":"9ad4562e-647e-4e19-acc7-ecfa7c8c022b","order_by":0,"name":"Amira A. Goda","email":"","orcid":"","institution":"National Research Center","correspondingAuthor":false,"prefix":"","firstName":"Amira","middleName":"A.","lastName":"Goda","suffix":""},{"id":300118780,"identity":"06cdea6c-3cac-45bb-aaef-bed837aff427","order_by":1,"name":"Eman G Ayad","email":"","orcid":"","institution":"Helwan University","correspondingAuthor":false,"prefix":"","firstName":"Eman","middleName":"G","lastName":"Ayad","suffix":""},{"id":300118781,"identity":"9e63ce76-5ee8-4d51-9273-740f80fd9448","order_by":2,"name":"Menna H. 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Panels A and B depict the control and treated conditions for HCT cells, while panels C and D show the corresponding conditions for HEPG2 cells respectively. Panel E presents the IC50 values for Commiphora myrrh concerning its inhibitory effect on HCT and HEPG2 cells.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4218698/v1/1e100c63bb4f49fb96771635.png"},{"id":56158735,"identity":"25e85ccd-6b3b-4523-b5d1-641f1338a63f","added_by":"auto","created_at":"2024-05-09 08:52:57","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2515792,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4218698/v1/2bf6fa52-aa9c-4d5b-89b5-1dcb56cc2aed.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Evaluation of the Antimicrobial and Anticancer Properties of Myrrh Resin Extract and Its Application in Cacao Beverages","fulltext":[{"header":"Introduction","content":"\u003cp\u003eSome preservatives are classed as carcinogens or cancer-causing chemicals because they are known to be hazardous to human health. Certain preservatives are considered carcinogenic or cancer-causing substances due to their known harmful effects on human health. According to \u003cb\u003eGupta and Yadav (2021)\u003c/b\u003e, artificial food additives interact with the cellular structure of the body, leading to a range of food-related disruptions, Mirza et al. (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) also corroborate these findings.\u003c/p\u003e \u003cp\u003eWhen used in excess, it might cause negative effects such skin rashes and itching, breathing problems, sneezing, or stomach issues \u003cb\u003e(Silva and Lidon., 2016)\u003c/b\u003e. M. M. Silva and F. Lidon, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2016\u003c/span\u003e. Certain food preservatives are classified as carcinogens or cancer-causing agents due to their hazardous effects on human health. 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 stomach issues when used excessively. Due to the possible health risks associated with synthetic food preservatives, consumers have become more cautious about consuming foods containing these additives, leading to an increase in demand for natural food preservatives over the past few decades. \u003cb\u003e(\u003c/b\u003eAtwaa et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2022\u003c/span\u003e \u003cb\u003eand\u003c/b\u003e Teshome et al \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eCommiphora myrrh contains phytochemical components such as terpenoids (monoterpenoids, sesquiterpenoids, volatile/essential oil), diterpenoids, triterpenoids, and steroids. It has been utilized throughout history to treat various ailments such as wounds, mouth ulcers, pains, fractures, stomach issues, microbial infections, and inflammatory diseases, as reported by Mohammed et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2015\u003c/span\u003e \u003cb\u003eand Hanu LO\u003c/b\u003e \u003cb\u003eet al\u003c/b\u003e., \u003cb\u003e2005)\u003c/b\u003e. Akbar \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2020\u003c/span\u003e \u003cb\u003eand\u003c/b\u003e Mohammed et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2015\u003c/span\u003e have also documented the medicinal properties of Commiphora myrrh. Myrrh is known for its antiseptic, astringent, anthelmintic, and expectorant properties. Studies by Batiha et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2023\u003c/span\u003e \u003cb\u003eand Akbar., 2020\u003c/b\u003e have proven the traditional use of myrrh in treating bacterial infections and its antibacterial and antifungal properties. According to Mohammed et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2015\u003c/span\u003e \u003cb\u003eand Khalil.\u003c/b\u003e, \u003cb\u003eet al\u003c/b\u003e., \u003cb\u003e2020\u003c/b\u003e, 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. Al-Sabri et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2014\u003c/span\u003e have also conducted research on myrrh's potential as an antibacterial agent.\u003c/p\u003e \u003cp\u003eAccording to Khalil, et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2020\u003c/span\u003e, Commiphora myrrh ether and ethanol extracts were tested for their antibacterial properties, these extracts showed antimicrobial activity against the Gram-negative organisms tested alongside \u003cb\u003eOmer.\u003c/b\u003e, \u003cb\u003eet al\u003c/b\u003e., \u003cb\u003e2011\u003c/b\u003e, and \u003cb\u003eAlqahtani.\u003c/b\u003e, \u003cb\u003eet al\u003c/b\u003e. \u003cb\u003e2021\u003c/b\u003e. However, the ether extract demonstrated antimicrobial action both against \u003cem\u003eCandida Albicans\u003c/em\u003e and the Gram-positive organisms that were tested, with the antifungal activity being stronger, \u003cb\u003eOmer.\u003c/b\u003e, \u003cb\u003eet al\u003c/b\u003e \u003cb\u003e2011 and Khalil., et\u003c/b\u003e \u003cb\u003eal\u003c/b\u003e \u003cb\u003e2020.\u003c/b\u003e In addition to its antimicrobial properties, the potential anticancer effects of Commiphora myrrh have also garnered attention \u003cb\u003e(\u003c/b\u003eSuliman et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). 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.\u003c/p\u003e"},{"header":"Material and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Materials\u003c/h2\u003e \u003cp\u003eMyrrh Resin Extract (MRE), Cacao powder, vegetable cream, salt, corn starch and granulated sugar were purchased from a local market and herbal market in Cairo, Egypt.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Microbial strains\u003c/h2\u003e \u003cp\u003eGram-positive bacteria \u003cem\u003eB. subtilis\u003c/em\u003e ATCC6633, and \u003cem\u003eS. aureus\u003c/em\u003e (MRSA) ATCC43300 as well as Gram-negative bacteria \u003cem\u003eP. aeruginosa\u003c/em\u003e (ATCC27853), and \u003cem\u003eE. coli\u003c/em\u003e (ATCC25922).\u003c/p\u003e \u003cdiv id=\"Sec5\" class=\"Section3\"\u003e \u003ch2\u003e2.3. Chemicals and reagents\u003c/h2\u003e \u003cp\u003eAll chemicals and reagents of the analytical methods used in the present study were analytical grades. Citric acid, sodium benzoate, CMC, xanthan gum, plate count agar (PCA), and potato dextrose agar (PDA) were purchased from El-Gamhouria Trading for Chemicals and Drugs Company, Cairo, Egypt.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section3\"\u003e \u003ch2\u003e2.4. Extraction of antioxidants from MER\u003c/h2\u003e \u003cp\u003eOne hundred grams of Commiphora Myrrh resin added to excessive distilled water: ethanol (2:8 v/v) and incubated at room temperature for 24 h, then the slurry was filtered through filter paper. The water extract was concentrated using rotary evaporator under reduced pressure and the residues were dissolved in 50 mL of distilled water.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5. LC-ESI-MS/MS analysis of Myrrh resin extract profile\u003c/h2\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e2.5.1. Instrument\u003c/h2\u003e \u003cp\u003eThe analysis of the sample was performed by liquid chromatography–electrospray ionization–tandem mass spectrometry (LC-ESI-MS/MS) as described by \u003cb\u003eEl-Ssayad\u003c/b\u003e \u003cb\u003eet al\u003c/b\u003e., \u003cb\u003e2023..\u003c/b\u003e\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003e2.5.2 Positive ionization mode:\u003c/h2\u003e \u003cp\u003eSeparation was performed with a Ascentis® Express 90 Å C18 Column (2.1×150 mm, 2.7 µm). The mobile phases comprised of two eluents A: 5 mM ammonium formate pH 3; B: acetonitrile (LC grade) \u003cb\u003eEl-Ssayad\u003c/b\u003e \u003cb\u003eet al\u003c/b\u003e., \u003cb\u003e2023.\u003c/b\u003e\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e2.6. Determination of antioxidants using HPLC\u003c/h2\u003e \u003cp\u003eHPLC analysis was carried out using an Agilent 1260 series.The separation was carried out using Eclipse C18 column (4.6 mm x250 mm i.d., 5 µm).Themobile phase consisted of water(A) and 0.05% trifluoroacetic acid in acetonitrile (B) at a flow rate 0.9 ml/min. The mobile phase was programmed consecutively 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 (82%A).The multi-wavelength detector was monitored at 280 nm. The injection volume was 5 µl for each of the sample solutions. The column temperature was maintained at 40°C.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e2.7. Minimal inhibitory concentration (MIC) determination\u003c/h2\u003e \u003cp\u003eIn accordance with the disk diffusion assay protocol, a volume of 1 mL from each bacterial suspension was evenly distributed onto a solid growth medium within a Petri dish. Six sterile paper disks measuring 6 mm in diameter were carefully positioned on the surface of each agar plate. These disks were then saturated with 15 µL of the diluted MER. The plates were incubated at 37°C for 24 hr. The antibacterial activity, measured as the minimum inhibitory concentration (MIC), was assessed by determining the lowest concentration of either diluted MER that resulted in the formation of an inhibition zone around a disk after a 24hr incubation period. Negative controls consisted of disks impregnated with sterile distilled water. Replicates were conducted at each concentration \u003cb\u003e(Lima-Filho and de Aguira Corderio, 2014)\u003c/b\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e2.8. Preparation of Cacao beverage\u003c/h2\u003e \u003cp\u003eIn preparation for 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 the formula-2 and 0.1% of Sodium benzoate was incorporated into the formula-3. A quantity of tape water was added and the heated at 80 ± 2°C for 5 minutes \u003cb\u003e(Faiqoh et al., 2021)\u003c/b\u003e, the formulations of cocoa beverages are presented in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e\u003cdiv class=\"gridtable\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eformulations of cocoa beverages\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003c/colgroup\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colspan=\"4\" nameend=\"c4\" namest=\"c1\"\u003e \u003cp\u003eCacao beverage\u003c/p\u003e \u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eformula-1\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eformula-2\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eformula-3\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCacao powder\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.5%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7.5%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7.5%\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSugar\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15%\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVegetable cream\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6%\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSalt\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.03%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.03%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.03%\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCorn starch\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.5%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.5%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.5%\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eXanthan gum\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.1%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.1%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.1%\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMyrrh extract\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSodium benzoate\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.1%\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWater\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e69.87%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e68.87%\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e69.77%\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/table\u003e\u003c/div\u003e \u003cp\u003e\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e2.9. Physicochemical analysis (pH value, total soluble solids, and total titratable acidity)\u003c/h2\u003e \u003cp\u003eThe 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 to 85% Brix – Romania) at 25\u003csup\u003e°\u003c/sup\u003eC. 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. According to the method described by the \u003cb\u003eAOAC (2012).\u003c/b\u003e Total titratable acidity (TTA) was determined in 10mL of each mango juice and cacao beverage, which was homogenized with 90 ml of distilled water, and titrated with 0.1 mol/L NaOH, using phenolphthalein as an indicator \u003cb\u003e(AOAC, 2012)\u003c/b\u003e. 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,\u003c/p\u003e\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$\\% acidity= \\frac{N\\times V1\\times Eq. wt.}{V2\\times 1000}\\times 100$$\u003c/div\u003e\u003c/div\u003e\u003cp\u003e\u003c/p\u003e \u003cp\u003eWhere, N = normality of NaOH (mEq/mL); V\u003csub\u003e1\u003c/sub\u003e = volume of titrant (mL); Eq. wt. = equivalent weight of acid (; V\u003csub\u003e2\u003c/sub\u003e = volume of sample (mL); 1,000 = factor relating mg to grams (mg/g) (1/10 = 100/1000).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e2.10. Microbial assay\u003c/h2\u003e \u003cp\u003eThe total microbial count was determined using the method described by Herrera in 2001. A 5g sample of mango juice or 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\u003c/p\u003e \u003cp\u003eto 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 were 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 \u003cb\u003e(Herrera, 2001)\u003c/b\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e2.11. Cytotoxicity assay:\u003c/h2\u003e \u003cp\u003eCell 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) CO\u003csub\u003e2\u003c/sub\u003e atmosphere at 37°C. For the SRB assay, aliquots of 100 µL cell suspension (5x10^3 cells) were seeded into 96-well plates and incubated in complete media for 24 hours. 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 hour. The TCA solution was then removed, and the cells were washed 5 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 minutes. 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) \u003cb\u003e(\u003c/b\u003eSkehan et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e1990\u003c/span\u003e). 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\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003e2.12. Sensory evaluation\u003c/h2\u003e \u003cp\u003eThe sensory analysis was carried out using a composite score scale according to \u003cb\u003eLawless and Heymann (1993)\u003c/b\u003e. Ten panelists semi-trained (The age of participants was between 25–35 years old) from Food Science and Technology Department, Faculty of Agriculture, Al-Azhar University, Egypt, assessed the sensory quality of mango juice and cacao beverage. Three different treatments were scored for taste (30), odor (30), color (20), texture (20), acceptability (score 100).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003e2.13. Statistical analysis:\u003c/h2\u003e \u003cp\u003eThe studies were reproduced at least three times, and the values are reported as the mean ± standard deviation (SD). A variance analysis (ANOVA) was performed to investigate the differences among the samples.\u003c/p\u003e \u003c/div\u003e "},{"header":"Results and Discussions","content":"\u003cp\u003e\u0026nbsp;Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of Commiphora Myrrha extract (MRE) revealed the presence of more than 20 positive and negative substances including Phlorizin, Choline, alpha-D-Glucose-1,6-diphosphate, Sinapoyl malate, and N, N-Dimethylglycine. These findings are presented in Figures 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% \u003cstrong\u003e(Arın \u0026nbsp;\u003cem\u003eet al\u003c/em\u003e., 2021)\u003c/strong\u003e.\u003c/p\u003e\n\u003cp\u003eThe antimicrobial properties of Myrrh resin extract (MRE) are attributed to a group of antioxidant compounds, as determined by high-performance liquid chromatography (HPLC). The analysis revealed 19 active antioxidant compounds in MRE, with kaempferol being the most abundant at 1896 \u0026micro;g/g, followed by quercetin at 520 \u0026micro;g/g. Notably, Pyrocatechol exhibited the highest value among the compounds analyzed, at 162 \u0026micro;g/g. The remaining compounds, including Gallic acid, Chlorogenic acid, Catechin, Coffeic acid, Syringic acid, Rutin, Coumaric acid, Vanillin, Ferulic acid, Naringenin, Cinnamic acid, Apigenin, and Hesperetin, showed varying concentrations as presented in Table 3 and Figure 3.\u003c/p\u003e\n\u003cp\u003eFurther 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 \u003cstrong\u003eAbbas \u003cem\u003eet al\u003c/em\u003e. (2020), Brasha \u003cem\u003eet al\u003c/em\u003e., (2023), Rahmani \u0026nbsp;\u003cem\u003eet al\u003c/em\u003e., (2022), Fatani \u003cem\u003eet al\u003c/em\u003e., (2016), and Rasha \u0026nbsp;\u003cem\u003eet al\u003c/em\u003e., (2023)\u003c/strong\u003e identified significant polyphenolic compounds such as Chlorogenic acid, Gallic acid, Catechin, Caffeine, Syringic acid, Coumaric acid, Ferulic acid, Naringenin, 4\u0026apos;,7-Dihydroxyisoflavone, Propyl Gallate, Vanillin, Quercetin, and Ellagic acid in various concentrations, underscoring the potential therapeutic applications of myrrh extracts.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable [2]: Active components of Commiphora Myrrha extract have been analyzed using LC-MS/MS\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable dir=\"\" border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"37.967914438502675%\" valign=\"top\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cstrong\u003eChemical structure\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"22.994652406417114%\" valign=\"top\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cstrong\u003eArea\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"39.037433155080215%\" valign=\"top\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cstrong\u003eCompounds\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"37.967914438502675%\" valign=\"top\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"22.994652406417114%\" valign=\"top\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003e4.63\u0026times;10\u003csup\u003e8\u003c/sup\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"39.037433155080215%\" rowspan=\"8\" valign=\"top\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003egamma-Linolenic acid\u003c/span\u003e\u003cspan dir=\"LTR\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003cp dir=\"LTR\"\u003eN,N-Dimethylglycine\u003c/p\u003e\n \u003cp dir=\"LTR\"\u003eCholine\u003c/p\u003e\n \u003cp dir=\"LTR\"\u003eCarnosine \u0026nbsp;\u003c/p\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/span\u003e\u003cspan dir=\"LTR\"\u003eUridine 5\u0026apos;-monophosphate\u003c/span\u003e\u003c/p\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003ePhlorizin\u003c/span\u003e\u003cspan dir=\"LTR\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eSinapoylmalate\u003c/span\u003e\u003cspan dir=\"LTR\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003ealpha-D-Glucose-1,6-diphosphate\u003c/span\u003e\u003cspan dir=\"LTR\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\u0026nbsp;\u0026nbsp;\u003cp dir=\"LTR\"\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"62.280701754385966%\" valign=\"top\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eC\u003csub\u003e4\u003c/sub\u003eH\u003csub\u003e9\u003c/sub\u003eNO\u003csub\u003e2\u003c/sub\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"37.719298245614034%\" valign=\"top\"\u003e\n \u003cp dir=\"LTR\"\u003e5.9\u0026times;10\u003csup\u003e8\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"62.280701754385966%\" valign=\"top\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eC\u003csub\u003e5\u003c/sub\u003eH\u003csub\u003e14\u003c/sub\u003eNO\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"37.719298245614034%\" valign=\"top\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003e6.86\u0026times;10\u003csup\u003e8\u003c/sup\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"62.280701754385966%\" valign=\"top\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eC\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e14\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"37.719298245614034%\" valign=\"top\"\u003e\n \u003cp dir=\"LTR\"\u003e5.48\u0026times;10\u003csup\u003e8\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"62.280701754385966%\" valign=\"top\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eC\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e13\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003eP\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"37.719298245614034%\" valign=\"top\"\u003e\n \u003cp dir=\"LTR\"\u003e1.24\u0026times;10\u003csup\u003e9\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"62.280701754385966%\" valign=\"top\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eC\u003csub\u003e21\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eO\u003csub\u003e10\u003c/sub\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"37.719298245614034%\" valign=\"top\"\u003e\n \u003cp dir=\"LTR\"\u003e1.75\u0026times;10\u003csup\u003e8\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"62.280701754385966%\" valign=\"top\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"37.719298245614034%\" valign=\"top\"\u003e\n \u003cp dir=\"LTR\"\u003e8.37\u0026times;10\u003csup\u003e8\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"62.280701754385966%\" valign=\"top\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eC\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e14\u003c/sub\u003eO\u003csub\u003e12\u003c/sub\u003eP\u003csub\u003e2\u003c/sub\u003e\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"37.719298245614034%\" valign=\"top\"\u003e\n \u003cp dir=\"LTR\"\u003e8.8\u0026times;10\u003csup\u003e8\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3: Antioxidants compounds of Myrrh resin extract analyzed by HPLC.\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"515\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003e\u003cstrong\u003eBioactive\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003ecompound\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e\u003cstrong\u003eArea\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e\u003cstrong\u003eConc. (\u0026micro;g/ml)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eGallic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e17.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e1.48\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eChlorogenic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e6.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e0.84\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eCatechin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e5.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e1.44\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eMethyl gallate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eCaffeic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e7.94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e0.61\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eSyringic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e15.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e1.02\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003ePyro catechol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e19.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e2.77\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eRutin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e5.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e0.60\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eEllagic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eCoumaric acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e31.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e1.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eVanillin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e17.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003e0.76\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eFerulic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e34.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e2.36\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eNaringenin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e10.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e1.22\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eDaidzein\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eQuerectin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e64.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e8.85\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eCinnamic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e7.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003e0.13\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eApigenin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e22.41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003e1.71\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eKaempferol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e415.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp\u003e32.24\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"49.320388349514566%\"\u003e\n \u003cp\u003eHesperetin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.04854368932039%\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.631067961165048%\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003e0.00\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003cstrong\u003eTable 4 Antibacterial activities and MIC of MRE against Gram-positive and negative bacteria \u0026nbsp;\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" align=\"\" width=\"690\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"31.884057971014492%\" rowspan=\"3\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eOrganisms\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.971014492753623%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"60.14492753623188%\" colspan=\"6\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eInhibitory activity against the tested organism (zone of inhibition in mm)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.702127659574469%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"88.29787234042553%\" colspan=\"6\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentration\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"11.48936170212766%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e100%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.595744680851062%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e50%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.319148936170214%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e25%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.957446808510639%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e12.5%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.957446808510639%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e6.25%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.340425531914894%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.13%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.340425531914894%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.56%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"9\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGram-positive bacteria\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"31.884057971014492%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eStaphylococcusaureus\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e(MRSA) - ATCC43300\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.826086956521739%\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.304347826086957%\" colspan=\"2\"\u003e\n \u003cp\u003e8.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.434782608695652%\"\u003e\n \u003cp\u003e7.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.869565217391305%\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.869565217391305%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.405797101449275%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.405797101449275%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"31.884057971014492%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eBacillussubtilis\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eATCC6633\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.826086956521739%\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.304347826086957%\" colspan=\"2\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.434782608695652%\"\u003e\n \u003cp\u003e6.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.869565217391305%\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.869565217391305%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.405797101449275%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.405797101449275%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"9\"\u003e\n \u003cp\u003e\u003cstrong\u003eGram-negative bacteria\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"31.884057971014492%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eEscherichiacoli\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003e\u003cspan dir=\"RTL\"\u003e)\u0026nbsp;\u003c/span\u003e\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eATCC\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e25922\u003c/strong\u003e\u003cstrong\u003e\u003cspan dir=\"RTL\"\u003e(\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.826086956521739%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.304347826086957%\" colspan=\"2\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.434782608695652%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.869565217391305%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.869565217391305%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.405797101449275%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.405797101449275%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"31.884057971014492%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;(ATCC27853)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.826086956521739%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.304347826086957%\" colspan=\"2\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.434782608695652%\"\u003e\n \u003cp\u003e11.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.869565217391305%\"\u003e\n \u003cp\u003e8.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.869565217391305%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.405797101449275%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.405797101449275%\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eTable 4 illustrates the antimicrobial activity of MRE at different concentrations against Gram-positive (\u003cem\u003eStaphylococcus aureus\u0026nbsp;\u003c/em\u003e(MRSA) - ATCC43300;\u003cem\u003e\u0026nbsp;Bacillus subtilis - ATCC6633\u003c/em\u003e) and Gram- negative bacteria (\u003cem\u003eEscherichia coli\u0026nbsp;- ATCC25922; Pseudomonas\u0026nbsp;aeruginosa\u003c/em\u003e - ATCC27853)). The MRE had potent antimicrobial activity against Gram-positive bacteria and copmpelety inhibited \u003cem\u003eStaphylococcus aureus\u0026nbsp;\u003c/em\u003e(MRSA) (ATCC43300) ATCC43300-\u003cem\u003e\u0026nbsp;Bacillus subtilis\u003c/em\u003eATCC6633 at concentrations of 6.25, 3.13, and 1.56% and gradually inhibited the same strains from 10-7, 10-6 mm for the tested microorganisms respetivly.Concerning the antibacterial activity of MER against Gram-negative bacteria, the results were better and more promising, and the MER completely inhibited \u003cem\u003eEscherichia coli) ATCC\u003c/em\u003e25922( at all tested concentrations, while gradually inhibiting \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e (ATCC27853) from 20\u0026ndash;8.5 mm and completely inhibiting at 6.25, 3.13, and 1.56%.\u003c/p\u003e\n\u003cp\u003eUsing the viable count approach, the bactericidal effects of myrrh hexane extract and myrrh essential oil (MEO) against \u003cem\u003eStaphylococcus aureus\u003c/em\u003e (\u003cem\u003eS. aureus\u003c/em\u003e) and \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e (\u003cem\u003ePs. aeruginosa\u003c/em\u003e) were studied. After a two-hour contact period, (MEO) demonstrated superior efficacy, killing both tested strains in \u0026gt; 99.999% of the cases. Four multidrug resistant isolates of \u003cem\u003eS. aureus\u003c/em\u003e (MRSA, sputum), Escherichia coli (E. coli, urine), \u003cem\u003ePs. aeruginosa\u003c/em\u003e (wound), and \u003cem\u003eKlebsiella pneumonia\u003c/em\u003e (\u003cem\u003eK. pneumonia\u003c/em\u003e, sputum) were examined using the same method as MEO. Highest bactericidal activity was observed against \u003cem\u003ePs. aeruginosa\u003c/em\u003e while lowest was against K. pneumonia (99.59 and 54.04% killing, respectively. MEO has antibacterial properties against various bacterial strains and antifungal properties against \u003cem\u003eCandida albicans\u003c/em\u003e \u003cstrong\u003eAuda ., \u003cem\u003eet al\u003c/em\u003e. 2017\u003c/strong\u003e. \u0026nbsp;The most effective efficacy was against \u003cem\u003eS. aureus\u003c/em\u003e, 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 P. auroginosa and E. coli at concentrations of 80 to 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 \u003cstrong\u003eAlharbi, and Alhashmi., 2022\u003c/strong\u003e. \u0026nbsp;Myrrh\u0026apos;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 \u0026nbsp;\u003cstrong\u003eAl-Sabri AE; Moslem M 2016\u003c/strong\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 5 Application of MER as antibacterial activity on total count of bacteria by Log cfu/mL in Cacao Beverage\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"602\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.865671641791046%\"\u003e\n \u003cp\u003eStorage at 4 \u0026deg;C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.081260364842453%\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.03150912106136%\"\u003e\n \u003cp\u003eMER (1%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.021558872305143%\"\u003e\n \u003cp\u003eSodium benzoate (0.1%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"4\"\u003e\n \u003cp\u003eTotal bacteria count by Log cfu/Ml\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.865671641791046%\"\u003e\n \u003cp\u003eZero\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.081260364842453%\"\u003e\n \u003cp\u003e1.3\u0026plusmn;0.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.03150912106136%\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.021558872305143%\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.865671641791046%\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.081260364842453%\"\u003e\n \u003cp\u003e2.2\u0026plusmn;0.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.03150912106136%\"\u003e\n \u003cp\u003e1.5\u0026plusmn;0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.021558872305143%\"\u003e\n \u003cp\u003e1.1\u0026plusmn;0.06\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.865671641791046%\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.081260364842453%\" valign=\"top\"\u003e\n \u003cp\u003e2.9\u0026plusmn;0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.03150912106136%\" valign=\"top\"\u003e\n \u003cp\u003e1.7\u0026plusmn;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.021558872305143%\" valign=\"top\"\u003e\n \u003cp\u003e1.1\u0026plusmn;0.09\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"4\"\u003e\n \u003cp\u003eMold and yeast count by Log cfu/mL\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.865671641791046%\"\u003e\n \u003cp\u003eZero\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.081260364842453%\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.03150912106136%\" valign=\"top\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.021558872305143%\" valign=\"top\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.865671641791046%\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.081260364842453%\" valign=\"top\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.03150912106136%\" valign=\"top\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.021558872305143%\" valign=\"top\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.865671641791046%\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.081260364842453%\" valign=\"top\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.03150912106136%\" valign=\"top\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.021558872305143%\" valign=\"top\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eThe MRE was applied at concentration 1% in cacao beverage as antibacterial agent against 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 Commiphora myrrh (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 Commiphora myrrh 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 Commiphora myrrh. (\u003cstrong\u003eAbbas \u003cem\u003eet al\u003c/em\u003e., 2020)\u003c/strong\u003e. The best antibacterial activity was seen in a hydroalcohol extract that was extracted using an ethanol: phosphate buffer pH 7 (85:15) ratio, \u003cstrong\u003eAlmekhlafi S \u003cem\u003eet al\u003c/em\u003e 2014\u003c/strong\u003e. 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 Boffa et al, 2023. 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 (\u003cstrong\u003eAlshibly \u003cem\u003eet al\u003c/em\u003e .,2022).\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn this study, we investigated the anti-cancer properties of Commiphora myrrh extract against colon cancer (HCT) and liver cancer (HEPG2) cell lines. Our findings indicate that the extract has promising cytotoxic effects, Figure 4 illustrates the IC50 values of the extract against HCT and HEPG2 cell lines, indicating significant cytotoxic effects with values of 55.69 \u0026mu;g/ml for HCT cells and 70.78 \u0026mu;g/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 \u0026lt; 30 \u0026micro;g/mL for crude extracts. This suggests that Commiphora myrrh extract may have potential as a cytotoxic agent against these cancer cell types. Our results are consistent with previous research. \u003cstrong\u003eChen \u003cem\u003eet al\u003c/em\u003e. (2013)\u003c/strong\u003e investigated the effects of myrrh extract on liver cancer and found that furano-sesquiterpenoids isolated from Arabic Commiphora Myrrh resin induced apoptosis of human hepatocellular carcinoma HepG2 cells with an IC50 of 3.6 \u0026mu;M. Furthermore, \u003cstrong\u003eHamad \u003cem\u003eet al\u003c/em\u003e. (2017)\u003c/strong\u003e reported that myrrh, in combination with protocatechuic acid, induces apoptotic cell death in colon cancer cells by suppressing the Bcl-2 gene. This suggests a potential mechanism by which Commiphora myrrh 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 Commiphora myrrh extract supports its potential as an anti-cancer agent against colon and liver cancer cell lines.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 6: total soluble solids, pH value and acidity of produced treated cacao beverage.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"602\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.865671641791046%\"\u003e\n \u003cp\u003eTest\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.081260364842453%\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.03150912106136%\"\u003e\n \u003cp\u003eMyrrhextract\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.021558872305143%\"\u003e\n \u003cp\u003eSodium benzoate\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.865671641791046%\"\u003e\n \u003cp\u003eT.S.S\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.081260364842453%\"\u003e\n \u003cp\u003e10.60\u0026plusmn;0.9\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.03150912106136%\"\u003e\n \u003cp\u003e10.60\u0026plusmn;0.9\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.021558872305143%\"\u003e\n \u003cp\u003e10.60\u0026plusmn;0.8\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.865671641791046%\"\u003e\n \u003cp\u003epH value\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.081260364842453%\"\u003e\n \u003cp\u003e6.80\u0026plusmn;1.6\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.03150912106136%\"\u003e\n \u003cp\u003e6.80\u0026plusmn;0.8\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.021558872305143%\"\u003e\n \u003cp\u003e6.70\u0026plusmn;0.8\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.865671641791046%\"\u003e\n \u003cp\u003eAcidity (as lactic acid)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.081260364842453%\"\u003e\n \u003cp\u003e0.10%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.03150912106136%\"\u003e\n \u003cp\u003e0.10%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.021558872305143%\"\u003e\n \u003cp\u003e0.13%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"4\" valign=\"bottom\"\u003e\n \u003cp\u003eThe means within the same row having different superscripts are significantly varied (P \u0026le;0.05).\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe 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\u0026rsquo;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 samples. The pH range of the cocoa drinks was 6.44 to 7.10. The highest pH was 7.10 \u003cstrong\u003eRossi Indiarto \u003cem\u003eet al\u003c/em\u003e., 2022\u003c/strong\u003e. pH value of the sample was between 7.61-7.72, while total soluble solids of the chocolate beverages varied between 10.38-10.75 oBrix \u003cstrong\u003eSalwa Al Aribah \u003cem\u003eet al.,\u003c/em\u003e 2020\u003c/strong\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 7: The effect of the addition of extract on the sensory properties of cacao beverages\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"602\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"21.92691029900332%\"\u003e\n \u003cp\u003eTest\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"21.92691029900332%\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.07641196013289%\"\u003e\n \u003cp\u003eMyrrh extract\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.069767441860463%\"\u003e\n \u003cp\u003eSodium benzoate\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"21.92691029900332%\"\u003e\n \u003cp\u003eTaste (30)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"21.92691029900332%\"\u003e\n \u003cp\u003e29.0\u0026plusmn;0.9a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.07641196013289%\"\u003e\n \u003cp\u003e26.0\u0026plusmn;0.9b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.069767441860463%\"\u003e\n \u003cp\u003e28.0\u0026plusmn;0.8a\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"21.92691029900332%\"\u003e\n \u003cp\u003eOdor (30)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"21.92691029900332%\"\u003e\n \u003cp\u003e27.9\u0026plusmn;1.6a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.07641196013289%\"\u003e\n \u003cp\u003e25.0\u0026plusmn;0.8b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.069767441860463%\"\u003e\n \u003cp\u003e28.4\u0026plusmn;0.8a\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"21.92691029900332%\"\u003e\n \u003cp\u003eColor (20)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"21.92691029900332%\"\u003e\n \u003cp\u003e19.0\u0026plusmn;0.9a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.07641196013289%\"\u003e\n \u003cp\u003e18.0\u0026plusmn;1.9a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.069767441860463%\"\u003e\n \u003cp\u003e18.6\u0026plusmn;1.3a\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"21.92691029900332%\"\u003e\n \u003cp\u003eTexture (20)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"21.92691029900332%\"\u003e\n \u003cp\u003e19.2\u0026plusmn;0.8a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.07641196013289%\"\u003e\n \u003cp\u003e18.6\u0026plusmn;0.8a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.069767441860463%\"\u003e\n \u003cp\u003e18.8\u0026plusmn;0.8a\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"21.92691029900332%\"\u003e\n \u003cp\u003eAcceptability (score 100)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"21.92691029900332%\"\u003e\n \u003cp\u003e95.8\u0026plusmn;2.6a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.07641196013289%\"\u003e\n \u003cp\u003e88.4\u0026plusmn;1.1b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.069767441860463%\"\u003e\n \u003cp\u003e94.0\u0026plusmn;2.7a\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"4\" valign=\"bottom\"\u003e\n \u003cp\u003eThe means within the same row having different superscripts are significantly varied (P \u0026le;0.05).\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eThe 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.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe results of this study highlight the potential of Myrrh resin extract (MRE) as a natural antimicrobial agent for food preservation and as a potential anti-cancer agent. The study analyzed MRE's antioxidant properties using HPLC and LC-MS/MS and identified two potent antioxidant compounds, kaempferol and quercetin. These compounds not only contribute to MRE's effectiveness in inhibiting microbial growth but also exhibit anti-cancer properties, as they have a cytotoxic effect against colon cancer (HCT) and liver cancer (HEPG2) cell lines. The study also investigated the incorporation of MRE into cacao beverages and found no significant changes in their physical or chemical properties, as indicated by stable measurements of total soluble solids (TSS), pH value, and acidity. Furthermore, sensory evaluation revealed that treated samples did not have a significant impact on taste, odor, color, and texture and received very acceptable ratings comparable to untreated samples. Overall, the study suggests that Myrrh resin extract has multiple benefits, making it a promising natural alternative to synthetic food preservatives and a potential adjunct therapy in cancer prevention and treatment. With its demonstrated effectiveness in preserving food quality and safety, minimal sensory impact, and anti-cancer properties, it could be applied more broadly in food and beverage products, as well as in oncological interventions.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eInformed Consent Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eEvery participant on the sensory panel has given written consent to participate in the research study. Before the study commenced, participants were notified of the purpose and methods of the research and assured of the strict confidentiality of their data. Participation in the study was voluntary, and subjects had the autonomy to withdraw their engagement at any time. The individuals demonstrated exceptional physical health and had no recorded allergies to the components. The research lacks institutional approval due to Egypt\u0026apos;s rules regarding human research. The research protocol is in accordance with the ethical standards of our institution, Al-Azhar University, Faculty of Agriculture, and National Research Center, Egypt, and approval is not needed according to Egypt\u0026apos;s rules regarding sensory evaluation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll relevant information has been included in the manuscript itself\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank the Jiangsu Province Science and Technology Support Program (BZ2022001) China, Nanjing Microtest Biotechnology Co., Ltd, Nnjing, 210061, Jiangsu, China, and the National Research Centre, Cairo, Egypt.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAbbas, R.K., Al-Mushhin, A.A.M., Elsharbasy, F.S. and Ashiry, K.O. (2020). Nutritive Value, Polyphenol Constituents and Prevention of Pathogenic Microorganism by Different Resin Extract of \u003cem\u003eCommiphora myrrh\u003c/em\u003e. \u003cem\u003eJ Pure Appl Microbiol\u003c/em\u003e., 14(3):1871-1878. doi: 10.22207/JPAM.14.3.26\u003c/li\u003e\n\u003cli\u003eAkbar, S. (2020). \u003cem\u003eCommiphora myrrh\u003c/em\u003ea (Nees) Engl. (Burseraceae). In: Akbar S (ed) Handbook of 200 medicinal plants: a comprehensive review of their traditional medical uses and scientific justifications. Springer International Publishing, Cham, pp 701\u0026ndash;706.\u003c/li\u003e\n\u003cli\u003eAlharbi, N.M. and Alhashmi, H.M. (2022). The use of Commiphora myrrh extract as disinfectant agent against pathogenic microbes isolated from beauty. African Journal of Biological Sciences, 108-121. https://ssrn.com/abstract=4023517.\u003c/li\u003e\n\u003cli\u003eAlmekhlafi, S., Thabit, A. A., Alwossabi, A. M., Awadth, N., Thabet, A. A., \u0026amp; Algaadari, Z. (2014). 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Editore\u003cspan dir=\"RTL\"\u003e\u0026rlm;\u003c/span\u003e :\u0026lrm; LAP LAMBERT Academic Publishing (1 marzo 2016) Copertinaflessibile\u003cspan dir=\"RTL\"\u003e\u0026rlm;\u003c/span\u003e : \u0026lrm; 116 pagine. ISBN-10 :\u0026lrm; 365983372X, ISBN-13 \u003cspan dir=\"RTL\"\u003e\u0026rlm;\u003c/span\u003e : \u0026lrm; 978-3659833724\u003c/li\u003e\n\u003cli\u003eAlshibly, N. M., Mohamed, R. M., Al-Shaikh T. M. and Soliman, A.M. (2022). Study of Commiphora myrrha (Nees) Engl. var. molmolExtract and Its Antibiogram Against Some Microbial Pathogens,Egyptian Journal of Chemistry., 65(5), 13-22. doi10.21608/EJCHEM.2021.98426.4584\u003c/li\u003e\n\u003cli\u003eAtwaa, E.S.H., Shahein, M.R., Radwan, H.A., Mohammed, N.S., Aloraini, M.A., Albezrah, N.K.A., Alharbi, M.A., Sayed, H.H., Daoud, M.A. and Elmahallawy, E.K. (2022). 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Composition and potential anticancer activities of essential oils obtained from myrrh and frankincense. Oncology letters, 6(4), 1140-1146.\u003c/li\u003e\n\u003cli\u003eFatani, A.J., Alrojayee, F.S., Parmar, M.Y., Abuohashish, H.M., Ahmed, M.M., Al-Rejaie, S.S. (2016). \u003cem\u003eMyrrh\u003c/em\u003e attenuates oxidative and inflammatory processes in acetic acid-induced ulcerative colitis. Exp Ther Med., 12(2):730-738. doi: 10.3892/etm.2016.3398. PMID: 27446268; PMCID: PMC4950638.\u003c/li\u003e\n\u003cli\u003eGupta, R. andYadav, R.K. (2021). Impact Of Chemical Food Preservatives On Human Health. \u003cem\u003ePalArch\u0026rsquo;s Journal of Archaeology of Egypt / Egyptology\u003c/em\u003e, \u003cem\u003e18\u003c/em\u003e(15), 811 - 818. https://archives.palarch.nl/index.php/jae/article/view/9009.\u003c/li\u003e\n\u003cli\u003eHamad, G. M., Taha, T. H., Alshehri, A., \u0026amp; El‐Deeb, N. M. (2017). Myrrh as a functional food with therapeutic properties against colon cancer in traditional meals. \u003cem\u003eJournal of food processing and preservation\u003c/em\u003e, \u003cem\u003e41\u003c/em\u003e(1), e12963.\u003c/li\u003e\n\u003cli\u003eHanu\u0026scaron;, L.O., Řezanka, T., Dembitsky, V.M., et al. (2005). Myrrh\u0026mdash;Commiphora chemistry. Biomed Papers 149:3\u0026ndash;28\u003c/li\u003e\n\u003cli\u003eKhalil, N., Fikry, S., Salama, O. (2020). Bactericidal activity of Myrrh extracts and two dosage forms against standard bacterial strains and multidrug-resistant clinical isolates with GC/MS profiling. AMB Express 10:21.\u003c/li\u003e\n\u003cli\u003eKhalil, N., Fikry, S., Salama, O. (2020). Bactericidal activity of Myrrh extracts and two dosage forms against standard bacterial strains and multidrug-resistant clinical isolates with GC/MS profiling. AMB Express. 28;10(1):21. doi: 10.1186/s13568-020-0958-3. PMID: 31993779; PMCID: PMC6987268.\u003c/li\u003e\n\u003cli\u003eMirza, S.K., Asema, U.K. And Kasim, S.S. (2017). To Study The Harmful Effects Of Food Preservatives On Human Health. Journal of Medicinal Chemistry and Drug Discovery 02, 02, pp. 610-616, 8 . www.jmcdd.org.\u003c/li\u003e\n\u003cli\u003eMohammed, S., Alhussaini, A.M., Saadabi, M. I., Alghonaim. and Khalid E. I. (2015). An Evaluation of the Antimicrobial Activity of \u003cem\u003eCommiphora myrrha \u003c/em\u003eNees (Engl.) Oleo-gum Resins from Saudi Arabia. \u003cem\u003eJournal of Medical Sciences, 15: 198-203.\u003c/em\u003e\u003cbr\u003e\u003cstrong\u003eDOI:\u003c/strong\u003e 10.3923/jms.2015.198.203,\u003cstrong\u003eURL:\u003c/strong\u003e https://scialert.net/abstract/?doi=jms.2015.198.203\u003c/li\u003e\n\u003cli\u003eObaid, Z.M. and Ahmed, M.A. (2020). Anti-Bacterial Effects of Commiphora Myrrha and Ziziphus Spina-Christ Leaves Extracts Against Streptococcus Mitis (Primary Colonizer of Dental Plaque) In vitro Study. J Res Med Dent Sci.,8(3): 57-64.\u003c/li\u003e\n\u003cli\u003eOmer, S.A., Adam, S.E.I. and Mohammed O.B. (2011). Antimicrobial Activity of \u003cem\u003eCommiphora myrrha\u003c/em\u003e Against Some Bacteria and \u003cem\u003eCandida albicans\u003c/em\u003e Isolated from Gazelles at King Khalid Wildlife Research Centre. \u003cem\u003eResearch Journal of Medicinal Plants., 5: 65-71. \u003c/em\u003e https://scialert.net/abstract/?doi=rjmp.2011.65.71\u003c/li\u003e\n\u003cli\u003e Omer, S.A., Adam, S.E.I. and Mohammed, O.B. (2011). Antimicrobial Activity of \u003cem\u003eCommiphora myrrha\u003c/em\u003e Against Some Bacteria and \u003cem\u003eCandida albicans\u003c/em\u003e Isolated from Gazelles at King Khalid Wildlife Research Centre. \u003cem\u003eResearch Journal of Medicinal Plants., 5(1), 65-71.\u003c/em\u003e\u003c/li\u003e\n\u003cli\u003eRahmani, A.H., Anwar, S., Raut, R., Almatroudi, A., Babiker, A.Y., Khan, A.A., Alsahli, M.A., Almatroodi, S.A. (2022). Therapeutic Potential of Myrrh, a Natural Resin, in Health Management through Modulation of Oxidative Stress, Inflammation, and Advanced Glycation End Products Formation Using \u003cem\u003eIn Vitro\u003c/em\u003e and \u003cem\u003eIn Silico \u003c/em\u003e Analysis. \u003cem\u003eAppl. Sci.,\u003c/em\u003e \u003cem\u003e12\u003c/em\u003e, 9175. https://doi.org/10.3390/app12189175\u003c/li\u003e\n\u003cli\u003eRasha, A., Mansouri, A. A., Roushdy, M. M., Huda, F., Alshaibi., Mahmoud, R. (2023) \u0026quot;Pharmacological Studies on the Antidiabetic, Antioxidant, and Antimicrobial Efficacies of \u003cem\u003eCommiphora myrrha\u003c/em\u003e Resin in Streptozotocin-Induced Diabetes in Rats: A Preclinical Study\u0026quot;, \u003cem\u003eJournal of Diabetes Research.\u003c/em\u003e, Article ID 5478267, 12 pages. https://doi.org/10.1155/2023/5478267\u003c/li\u003e\n\u003cli\u003eRossi, I., Souvia, R., Edy, S., Nur A., Gardiantini P. and Aldila D. P. (2022). Antioxidant activity and characteristics of a cocoa drink formulated with encapsulated green coffee extract, International Journal of Food Properties., 25:1, 2477-2494. https://doi.org/10.1080/10942912.2022.2144883.\u003c/li\u003e\n\u003cli\u003eSalwa, A., Adhitya, P.S., Dimas, R.A.M., Danar, P. (2020). Sensorial and physical properties of chocolate beverage prepared using low fat cocoa powder. \u003cem\u003eAIP Conference .,\u003c/em\u003e 2219 (1): 070007. https://doi.org/10.1063/5.0003435\u003c/li\u003e\n\u003cli\u003eSilva, M.M., and Lidon, F. (2016). FOOD PRESERVATIVES \u0026ndash; AN OVERVIEW ON APPLICATIONS AND SIDE EFFECTS. \u003cem\u003eEmirates Journal of Food and Agricultur.e\u003c/em\u003e, vol. 28, no. 6, pp. 366-73. doi:10.9755/ejfa.2016-04-351.\u003c/li\u003e\n\u003cli\u003eSkehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, et al. (1990). New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst 82(13): 1107-1112\u003c/li\u003e\n\u003cli\u003eSuliman, R. S., Alghamdi, S. S., Ali, R., Aljatli, D., Aljammaz, N. A., Huwaizi, S., \u0026amp; Rahman, I. (2022). The role of myrrh metabolites in cancer, inflammation, and wound healing: prospects for a multi-targeted drug therapy. \u003cem\u003ePharmaceuticals\u003c/em\u003e, \u003cem\u003e15\u003c/em\u003e(8), 944.\u003c/li\u003e\n\u003cli\u003eTeshome, E., Forsido, S.F., Rupasinghe, H.P.V., Olika Keyata, E. (2022). Potentials of Natural Preservatives to Enhance Food Safety and Shelf Life: A Review. Scientific World Journal., 9901018. doi: 10.1155/2022/9901018. PMID: 36193042; PMCID: PMC9525789.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"discover-food","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"discoverfood","sideBox":"Learn more about [Discover Food](https://www.springer.com/44187)","snPcode":"","submissionUrl":"","title":"Discover Food","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-4218698/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4218698/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eDue to the potential health risks of synthetic food preservatives, there has been a noticeable increase in interest in finding natural food preservatives during the past few decades. The goal of this study was to investigate the use of a natural extract of Commiphora Myrrh as an antimicrobial agent. The antioxidant properties of Myrrh resin extract (MRE) were analyzed using HPLC and GC-MS. The results showed that MRE contained potent antioxidant compounds, including 19 compounds, with the dominant compound being kaempferol, which had the highest value of 1896 \u0026micro;g/g. Quercetin was found to be the second most abundant compound, with a value of 520 \u0026micro;g/g. The efficacy of MRE as an antimicrobial agent against both Gram-positive and Gram-negative bacteria was tested, and its application in Cacao beverage was also studied. The results demonstrated that MRE was highly effective against all the tested bacteria both in vitro and in the total bacterial count of the produced cacao beverage. Additionally, the fungi in the cacao beverage were completely inhibited at all tested concentrations of MRE. The total soluble solids (TSS), pH value, and acidity of the produced untreated, treated cacao beverage with MRE and sodium benzoate were carried out, and all values mentioned were almost the same, with no differences noted. The sensory evaluation of Cacao beverage showed that the MRE had a minor impact on taste, odor, color, and texture of the produced cacao beverage in comparison with the control sample, which was very acceptable for judgments and recorded 95, 88, and 94 for the control and treated samples, respectively. Furthermore, the anti-cancer properties of MRE were evaluated, revealing significant cytotoxic effects against colon cancer (HCT) and liver cancer (HEPG2) cell lines. The IC50 values for HCT and HEPG2 cells were 55.69 \u0026micro;g/ml and 70.78 \u0026micro;g/ml, respectively, indicating the potential of MRE as an anti-cancer agent.\u003c/p\u003e","manuscriptTitle":"Evaluation of the Antimicrobial and Anticancer Properties of Myrrh Resin Extract and Its Application in Cacao Beverages","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-05-09 08:28:39","doi":"10.21203/rs.3.rs-4218698/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision 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