Ceylon cinnamon ( Cinnamomum zeylanicum ): Unveiling the health potential

: Cinnamomum zeylanicum provides numerous health benefits linked to its phytochemical composition, which varies by maturity, storage conditions, and genotypes. As previous studies have not examined how these factors affect the antioxidant, antimicrobial, and anticancer properties of C. zeylanicum , this study aims to address this gap and explore its broader applications. Both leaves and bark of seed-propagated (N) and vegetatively propagated varieties, Sri Gemunu (G) and Sri Vijaya (V) were used in the study. Three maturity stages, i.e.: 1-3 years (M3), 3-5 years (M2), and >5 years (M1) were subjected to the analysis in each case. Antioxidant activity was assessed using DPPH and FRAP assays, considering storage conditions (room temperature, 4°C) and storage duration of 1 week to 12 months. The most promising samples were then evaluated for antimicrobial and anticancer activity against MCF-7 and Hep-G2 cells. The results showed that NM1 leaves displayed heightened RSA [IC 50 : 0.27 ± 0.04 mgmL -1 ] whereas NM1 bark gave varied radical scavenging activity (RSA) values exhibiting the highest RSA (0.60 ± 0.09 mgmL -1 ). Maturity-influenced antioxidant activity of GM1 leaves exhibited the highest FRAP values at 2358.8 µmol FeSO 4 /g dry weight (DW). Bark of GM1 recorded FRAP value of 2770.0 ± 3.19 µmol FeSO 4 /g DW. Enhanced methanolic extracts (MeOH-Ex) from NM1 leaves and the bark demonstrated potent antimicrobial effects, inhibiting Staphylococcus aureus (MIC: 2.5 mgmL -1 ) and displaying robust anti-fungal activity against Candida albicans (MIC: 0.078 µgmL -1 ). In addition, MeOH-Ex of leaves showed the most potent cytotoxicity against cancer cells, Hep-G2 (cell viability: 11.53 ± 0.63%) after 48 hours. The research underscores the remarkable antioxidant, antimicrobial, and anticancer properties in C. zeylanicum leaves and bark, in NM1. This highlights the health benefits of seed-propagated C. zeylanicum over G and V varieties, offering valuable insights for future research and product development.


INTRODUCTION
Cinnamomum zeylanicum Blume, commonly known as true cinnamon or Ceylon cinnamon has gained global recognition for its therapeutic virtues and distinct spicy flavor attributed to its unique chemical composition.The bark oil contains cinnamaldehyde as the major volatile component, constituting 65.00 to 80.00%, while the leaf oil contains eugenol as the major component, comprising 70.00 to 95.00%.(Singh et al., 2007;Rao & Gan, 2014;Weisany et al., 2023).This botanical member of the family Lauraceae is believed to have originated in the central hilly terrain of Sri Lanka (Suriyagoda et al., 2021).While Sri Lanka hosts seven native cinnamon species, the commercial cultivation is predominantly focused on C. zeylanicum, setting it apart from the rest.In the competitive global market, Cinnamomum cassia stands as a primary rival to C. zeylanicum (Liyanage et al., 2017;Pathirana & Senaratne, 2020).However, C. zeylanicum distinguishes itself by exhibiting significantly lower levels of coumarin, a toxic compound.This aspect imparts C. zeylanicum with a heightened appeal over C. cassia, particularly in the realms of beverage, food, and pharmaceutical industries.The desirability of C. zeylanicum lies in its sought-after dried inner bark and volatile oils, derived from both leaves and bark, attributed to their reduced coumarin content (Blahová & Svobodová, 2012;Killday et al., 2015;Drobac et al., 2020).
Ayurvedic medicine has historically employed various C. zeylanicum extracts to support an array of health concerns, including toothaches, gastrointestinal issues, malignancies, arthritis, hypertension, and more.The modern medical landscape has equally embraced C. zeylanicum, exploring its functional attributes.Consequently, C. zeylanicum has been associated with a spectrum of potential medicinal effects encompassing anti-mutagenic, antioxidant, antimicrobial, anti-inflammatory, anticancer, antityrosinase, anti-diabetic, and neurological disorder alleviation, including Alzheimer's and Parkinson's diseases (Ranasinghe et al., 2012;Akilen et al., 2013;Yulug & Cankaya, 2019;Ahmadi et al., 2022).These diverse biological activities of C. zeylanicum can be attributed to its distinct phytochemical profile.Extracts from different parts of C. zeylanicum's inner bark, leaves, roots, fruits, and flowers have been reported to exhibit significantly varied phytochemical compositions, thereby suggesting unique sets of pharmacological activities for each plant component (Jayaprakasha & Rao, 2011;Narayanankutty et al., 2021).Nonetheless, C. zeylanicum's phytochemical makeup is subject to fluctuations based on factors such as plant maturity, soil composition, climatic variations, and physical growth stage, all of which necessitate an understanding of their influence on the plant's pharmacological properties (Madhushika & Bulugahapitiya, 2022).
Despite extensive documentation of C. zeylanicum's antioxidant activity across various extraction methods and volatile oils (Shahid et al., 2018), there remains a dearth of confirmed reports validating their origin from Ceylon cinnamon.Moreover, the potential degradation of antioxidant properties over prolonged storage periods and under varying environmental conditions poses further challenges.As dietary antioxidants have been linked to mitigating non-communicable diseases (NCDs) by reducing oxidative stress, determining optimal sample collection criteria and storage conditions for C. zeylanicum assumes paramount importance for both industrial and medicinal applications.While the literature has documented some data on the antioxidant, antimicrobial, and anticancer potential of seed-propagated C. zeylanicum extracts, there is a notable gap in comparative studies between seed and vegetative propagated C. zeylanicum varieties.To the best of our knowledge, no research has been published to date on the antioxidant, anticancer, and antimicrobial activities of different genotypes, maturity stages, storage conditions, and durations of seed and vegetative propagated C. zeylanicum varieties.This study aims to fill these research gaps by investigating the in-vitro antioxidant properties of C. zeylanicum leaf and bark extracts derived from both seed and vegetative propagated varieties, specifically Sri Gemunu and Sri Vijaya.The exploration will encompass diverse genotypes, stages of maturity, storage conditions, and duration.Beyond identifying the most potent antioxidant-containing C. zeylanicum extracts, the research will extend its focus to assess antimicrobial and anticancer properties.This comprehensive approach seeks to enhance our understanding of C. zeylanicum's potential and versatility, contributing valuable insights to the scientific community.

Cinnamon Leaf and Bark Sample Collection
Cinnamon leaves and bark of seed-propagated varieties were systematically collected from the premises of Mathurata Plantations (Pvt) Ltd, located in Matara, Sri Lanka (Table 1).Vegetatively propagated Sri Wijaya (V) and Sri Gemunu (G) accessions were received from the "Cinnamon Research and Training Center", Thihagoda, Palolpitiya, Matata, Sri Lanka.To ensure taxonomic accuracy, the botanical specimens were dispatched to a qualified botanist affiliated with the University of Ruhuna, Matara, for definitive species identification.Subsequent to species authentication, a voucher specimen was meticulously prepared and officially deposited within the herbarium of the Department of Botany at the University of Ruhuna, Sri Lanka.This specimen, bearing the unique identifier Cin/Sw/001, Cin/Sg/002, Cin/Cz/003, serves as a lasting record of the collected plant material.

Preparation of Extracts
The pulverized form of each sample of leaves and bark, weighing 100 g, underwent individual maceration for 3 days.This process occurred in 200 mL of solvents comprising methanol and water separately.Subsequently, the resultant mixture was meticulously filtered through Whatman® No.1 filter papers.The methanol solvent was subjected to evaporation using the rotary evaporator (HAHNVAPOR, HS-2005S) while maintaining a temperature of 40°C followed by freeze-drying using the SJIALAB freeze dryer (SJIA-10N-80C) to yield powdered crude samples.

Cancer Cell Lines
The human triple-positive adenocarcinoma cell line (MCF-7) and the human hepatocellular carcinoma cell line (Hep G2) were purchased from the European Collection of Authenticated Cell Cultures (ECACC), an entity under Public Health England situated in London, USA.(The Department of Biochemistry and Clinical Chemistry at the Faculty of Medicine, University of Kelaniya, provided laboratory facilities and guidance for the investigation into the anticancer properties of C. zeylanicum).

In-Vitro Antioxidant Activity
DPPH radical scavenging activity: A concentration series of the extracts were prepared by employing methanol as the solvent and encompassing concentrations within the range of 5000 to 100 µgmL -1 .In each instance, 100.0 µL of the extract was combined with 3.90 mL of a DPPH solution.Subsequently, the solutions were placed in a dark environment at room temperature for 30 minutes.
The blank solution for this process consisted of methanol, while the control involved DPPH solution devoid of antioxidants.The absorbance measurements of these solutions were conducted at a wavelength of 517 nm using a HITHACHI spectrophotometer (UH5300) connected to HITHACHI UH 5300 UV VIS SPECTROPHOTOMETRE software (Abeysuriya et al., 2021;Brand-Williams et al., 1995;Muthoni-Guchu et al., 2020).The resultant data were used to generate a scatter plot illustrating the relationship between the percentage of radical scavenging activity and the respective concentrations.This plot was subjected to a linear fit represented by the equation y = mx + c, where the x value corresponding to y = 50 was determined, denoting the IC 50 value.Ascorbic acid was employed as the standard solution to construct a standard curve, yielding an R² value of 0.8376.The equation representing the standard curve is expressed as y = 0.7125x + 23.051.

Ferric reducing antioxidant power (FRAP) assay:
A freshly prepared FRAP reagent (3.0 mL), consisting of a blend of 300 mM acetate buffer with a pH of 3.6, 10 mM TPTZ dissolved in 40 mM HCl, and 20 mM FeCl 3 •6H 2 O in a proportion of 10:1:1, was thoroughly combined with 100.0 µL of the plant extract, which had been diluted to a concentration of 1000 µgmL -1 from the initial stock solution.The reaction led to a vivid blue color formation as a consequence of reduction.After an incubation period of 30 minutes, the absorbance of this resultant solution was measured at a wavelength of 593 nm relative to a blank reagent composed of distilled water.To establish a Calibration was performed using an aqueous solution of FeSO 4 •7H 2 O ranging from 100 to 1200 mM.mg TE/g of crude extract (Abeysuriya et al., 2021;Benzie & Strain, 1999;Muthoni-Guchu et al., 2020).

Antimicrobial Activity
Disk diffusion assay: The plant extract (20.0 µL) was applied to sterile 6 mm blank disks, resulting in a final concentration of 400 µg per disk.As separate controls, the vehicle control (DMSO) (20.0 µL) and an appropriate positive control (20.0 µL) were applied to sterile blank disks.Subsequently, the disks were dried under aseptic conditions to facilitate the evaporation of excess solvent.A microorganism inoculate equivalent to the 0.5 McFarland turbidity standard (approximately 1.5 × 10 8 CFU/mL) was evenly dispersed on the surface of the suitable medium of agar plates using a sterilized cotton swab.Then the prepared discs were placed onto the respective agar plate and incubated at a temperature of 35 ± 2°C for 24 hours.The diameters of the inhibition zones (mm) were measured, including the diameter of the disk itself (Heatley, 1944;Balouiri et al., 2016).
Well diffusion assay: Using a sterile cork borer, wells with a diameter of six millimeters were created on the media plate that had been inoculated with a microorganism culture equivalent to the 0.5 McFarland turbidity standard.
The plant extract (20.0 µL) was introduced into these wells, achieving a final concentration of 400 µg per well.Wells containing 20.0 µL of DMSO and the appropriate antibiotic were used as the negative and positive controls, respectively.The plates were then incubated at a temperature of 35 ± 2°C for 24 hours.Following the incubation period, the diameters of the inhibition zones (mm) were assessed and recorded (Magaldi et al., 2004;Balouiri et al., 2016).

Minimum inhibitory concentration assay for bacteria:
The Minimum Inhibitory Concentration (MIC) of the extracts was ascertained through a modified micro dilution method based on the guidelines from the Clinical and Laboratory Standards Institute (CLSI, 2001).The concentrations assessed ranged from 10.00 to 0.07 mgmL - 1 , encompassing values of 10.00, 5.00, 2.50, 1.25, 0.62, 0.31, 0.15, and 0.07 mgmL -1 .To achieve the highest concentration (80.0 mgmL -1 ), crude cinnamon extracts (4.0 g) were dissolved first in DMSO (1% v/v, 25.0 mL) and subsequently in Mueller Hinton broth (MHB; 25.0 mL).Serial dilutions were conducted on the microplate, resulting in a final volume of 100 μL in each well.Freshly prepared bacterial inoculum underwent a dilution of a hundredfold (300.0 µL of bacterial inoculum was mixed with 30.0 mL of MHB medium), yielding an inoculum of 1×10 6 CFU/mL.This inoculum (100.0 μL) was introduced into the wells containing extracts, negative controls, and positive controls.
The microplate was subjected to an incubation period of 24 hours at 35 ± 2°C, and subsequent assessment of microbial growth was performed by quantifying the optical density at 630 nm.This measurement was conducted using a Bio-Tek Instruments, Inc. instrument (2014), connected to the Gen 5.0 Version 2.07.17 software.The MIC endpoint was defined as the lowest concentration that resulted in over 90% inhibition of growth (Balouiri et al., 2016;Gajic et al., 2022).

Minimum inhibitory concentration assay for fungi:
The Modified Agar Dilution method, as outlined by Scorzoni et al. (2007), was employed to determine the Minimum Inhibitory Concentration (MIC) for specific fungal strains.The series of extractions were prepared using the two-fold dilution technique with 10% DMSO as the solvent.
Sterile glass vials with a capacity of 10 mL were employed in an aseptic environment.Sabouraud Dextrose Agar (SDA) medium was cooled to 50°C in a water bath, and approximately 4 mL of the cooled medium was transferred into each vial.Just prior to solidifying, 1.0 mL of the extract was individually added to the SDA media in order to achieve the desired concentration range from 4000 to 7.56 µgmL -1 .The medium was allowed to solidify at an angle of 30 0 degrees.For Candida spp., 20.0 µL of inoculate suspension was spread evenly on the surface of the slant.
In the case of other selected fungal species, the inoculation was carried out on the surface of the slant in three distinct areas per vial.The tops of the vials were loosely sealed, and the entire setup was then placed in an incubator set at a temperature of 35 ± 2°C.Incubation was carried out for a duration of 2 to 14 days (Menon et al., 2001).(3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide) colorimetric assay: Cells were seeded at a density of 1×10 4 cells/well in two separate 96well microtiter plates.These plates were pre-incubated at 37°C for 24 hours.On the subsequent day, the media within the wells were meticulously aspirated, and 200.0 µL of plant extracts and corresponding control solutions, spanning various concentrations (200, 100, 50, and 25 µgmL -1 ), were introduced to the wells.One of the plates was subjected to an incubation period of 24 hours at 37°C, while the other was incubated for 48 hours under identical conditions.Post-incubation, the media containing the plant extracts were aspirated, and a gentle addition of 200.0 µL of Dulbecco's phosphate-buffered saline (DPBS) was performed to facilitate the removal and rinsing of residual media.After a brief interval, the DPBS was aspirated.To initiate the assay, 200.0 µL of MTT solution (prepared by diluting 5 mgmL -1 MTT in phosphate-buffered saline (PBS) solution at a 1:9 ratio in complete media) was dispensed into each well.Subsequently, the micro-titer plate was placed in a dark setting at room temperature for 2 hours.Following the MTT incubation, the MTT media was carefully withdrawn, and 100.0 µL of dimethylsulfoxide (DMSO) was introduced to each well to dissolve the resultant dye.To ensure uniform distribution, the plate was subjected to agitation using a 96-well plate shaker for a duration of 15 minutes.The measurement of absorbance at a wavelength of 570 nm was conducted using the microplate reader (Radosevich et al., 1993;Mazloum-Ardakani et al., 2019;van Tonder et al., 2015).

Statistical Analysis
The study utilized the IBS SPSS 20.0 trial version for Windows to compute the mean and standard deviation.Tukey's post-hoc tests (Student's unpaired t-test) assessed significance, means, and standard deviation.The research employed one-way ANOVAs followed by Dunnet's Multiple Comparisons Test to compare mean values of different extracts at the same concentration.
Cinnamon bark, meanwhile, displayed a diverse range of RSA values, spanning from 0.6 to 18.58 mgmL -1 .The seedpropagated varieties exhibited robust RSA in cinnamon bark, surpassing vegetatively propagated Sri Gemunu and Sri Wijaya accessions.The cinnamon bark derived from seed-propagated varieties aged beyond five years exhibited the most pronounced RSA, surpassing the bark from the 1-3 years (M3) and 3-5 years (M2) maturity categories, which demonstrated values of 18.58 ± 1.11 mgmL -1 and 9.45 ± 0.78 mgmL -1 , respectively.This indicates a twofold increase in RSA as the plant transitions from the M3 to M2 stage.The disparity in our DPPH values compared to those reported by Abeysekera et al. (2019), which recorded 33.96 ± 0.47 for mature cinnamon leaves, is noteworthy.
The differences are likely attributable to variations in maturity stages and genotypic factors, underscoring the intricate interplay of these variables in influencing antioxidant properties.A previous investigation has also reported DPPH values for immature, partially matured, and matured leaves of Ceylon cinnamon in Sri Lanka.However, this study diverges from previous research by emphasizing the plant's overall maturity rather than solely focusing on the leaves' maturation (Abeysekera et al., 2013).Consequently, our study represents the first exploration of maturity-dependent antioxidant activity within the C. zeylanicum plant.Regarding Ceylon cinnamon leaves, the FRAP values exhibited a wide range spanning from 86.71 to 2358.8 µmol FeSO 4 /g DW.Concerning maturitydependent antioxidant activity, leaves of the Sri Gemunu accession surpassing five years of maturity showcased the highest FRAP value, followed by the seed-propagation variety and the Sri Wijaya accession exceeding five years of maturation.Regarding the bark samples, the top FRAP value was detected in GM1, registering at 2770.0 ± 3.19 µmol FeSO 4 /g DW, followed by NM1 with 2518.9 ± 3.08 µmol FeSO 4 /g DW, and VM1 with 1461.2 ± 2.90 µmol FeSO 4 /g DW.Notably, maturity-dependent antioxidant capacity was evident in cinnamon bark.

C. zeylanicum leaves and bark:
Illustrating dynamic fluctuations in antioxidant potential, Table 2 shows variations in RSA for both Ceylon cinnamon leaves and bark.Out of the two storage conditions examined, maintaining the leaves and bark of NM1 at a temperature of 4°C was found to be the most optimal for preservation, resulting in IC 50 values of 0.26 ± 0.02 mgmL -1 and 0.62 ± 0.04 mgmL -1 , respectively, after one week.Impressively, cinnamon leaves exhibited sustained RSA for about three months when stored at 4°C, with only marginal reduction in RSA, with the IC 50 increasing at a rate of 0.06 mgmL -1 per month.However, significant changes in RSA were evident after six months, even when stored at 4°C.For instance, the IC 50 value for NM1 (4 0 C) 6m (seed-propagated, stored at 4°C for six months) was measured at 1.75 ± 0.03 mgmL -1 .
As for storage conditions and durations, FRAP values of leaves exhibited a gradual decline with prolonged storage in both assessed conditions (room temperature and 4°C).Among the samples stored at 4°C, the highest FRAP value was retained in the N (4 0 C) 1w (seed-propagated, stored at 4°C for one week) sample.Notably, no significant difference emerged between the FRAP values of fresh cinnamon leaves (2144.6 ± 26.2 µmol FeSO 4 /g DW) and those stored for up to one week at 4°C.Furthermore, a robust antioxidant capacity was discernible in the leaves after six months of storage at 4°C (2112.4± 3.05 µmol FeSO 4 /g DW), in contrast to their initial form.
Cinnamon sticks exhibited the capacity for six-month storage periods at both 4°C and room temperature, maintaining RSA with negligible decline, underscoring 4°C as the preferred storage condition.Although no significant difference was observed between the FRAP values of N (RT) 1w (1683.2± 11.02 µmol FeSO 4 /g DW) and N (4 0 C) 1w (2480.62 ± 3.58 µmol FeSO 4 / g DW) when compared to the fresh form (2518.9 ± 3.08 µmol FeSO 4 /g DW), a notable decrease in antioxidant capacity was noted with prolonged storage.The most significant decline in antioxidant capacity was evident in bark samples tested after a one-year storage period at room temperature (90.07 ± 3.49 µmol FeSO 4 / g DW).In comparison, N (4 0 C) 1y exhibited higher FRAP values than N (RT) 1y.Additionally, there were no significant differences in the FRAP values among N (4 0 C) 1w, N (4 0 C) 1m, N (4 0 C) 3m, and N (4 0 C) 6m cinnamon bark samples (Abeysekera et al., 2019).
The results from DPPH and FRAP assays reveal distinct antioxidant capacities in cinnamon leaves and bark, attributed to their unique chemical compositions.Notably, cinnamon bark consistently demonstrates higher antioxidant capacity than leaves across various maturity stages.This trend persists in both seed-propagation and vegetative propagation varieties.The variance in RSA increases with plant maturity, with plants aged >5 years exhibiting robust antioxidant capacity in both seed-propagation and vegetative accessions.Furthermore, the antioxidant activity is highest in leaves and bark of seed propagation compared to vegetative propagation accessions.While both leaves and bark experience a gradual reduction in antioxidant capacity over time, bark consistently outperforms leaves across all tested storage conditions and durations, underscoring its sustained superior antioxidant potential.Consequently, leaves and bark aged >5 years from the seed propagation variety in their fresh form were selected for further analysis.

Anticancer Activity of C. zeylanicum
The cytotoxic activity of Ceylon cinnamon extracts derived from leaves and bark of seed propagation accession against Hep G2 and MCF-7 cells was assessed using the MTT colorimetric assay at various extraction concentrations after 24 and 48 hours of incubation (Table 4).reported as 9 mgmL -1 and 4.8 mgmL -1 for 24 and 48 hours, respectively (Abd-Wahab & Adzmi, 2017).

CONCLUSION
In conclusion, this study is credited as the first detailed study on anti-oxidant properties based on different conditions of Ceylon cinnamon.The antioxidant capacity reveals a nuanced correlation with maturity stages and underscores a significant improvement in antioxidant properties with C. zeylanicum leaves that mature beyond five years.Notably, leaves from seed-propagated varieties, particularly those surpassing five years, demonstrate superior RSA compared to their vegetatively propagated counterparts (Sri Vijaya and Sri Gemunu).This emphasizes the relevance of maturity stages in evaluating the antioxidant potential of C. zeylanicum leaves.Similarly, the RSA of cinnamon bark displays a diverse spectrum of values, with seed-propagated varieties consistently exhibiting robust antioxidant activity, outperforming vegetatively propagated counterparts.
The most pronounced RSA is evident in cinnamon bark from seed-propagated varieties aged beyond five years, indicating a substantial increase in antioxidant capacity during later maturity stages.Emphasizing the importance of considering maturity stages in evaluating antioxidant potential, the study found the highest antioxidant activity in >5 years of matured samples from seed propagation varieties during the fresh stage, with better preservation at 4°C than at room temperature.
Furthermore, C. zeylanicum demonstrates potent antimicrobial activity against various bacteria and fungi, with susceptibility variations attributed to composition differences.Methanol extracts of leaves and bark exhibit antibacterial effects against S. aureus and moderate activity against E. coli and P. aeruginosa.The study also highlights potential anticancer properties, with methanol extracts inhibiting human carcinoma cells like human breast cancer cells (MCF-7) and hepatocellular carcinoma cells (Hep G2), suggesting potential applications in nutraceuticals and anticancer drugs.In conclusion, these findings significantly contribute to understanding C. zeylanicum 's diverse bioactive properties and its promising applications in various health-related fields, with a crucial emphasis on the impact of maturity stages.

Table 1 :
Geographic and environmental data of Cinnamon Sample Collection Sites.
(Prabuseenivasan et al., 2006)utty et al., 2021)af extracts of cinnamon are limited, the current study's results align with the antimicrobial properties of essential oils present in cinnamon leaves, which have shown susceptibility against E. coli, S. aureus, and P. aeruginosa(Teles et al., 2019;Narayanankutty et al., 2021).Among the bacterial isolates, S. aureus exhibited the lowest Minimum Inhibitory Concentration (MIC) with 2.5 mgmL-1 for both MeOH and water extracts of C. zeylanicum leaf and bark.Additionally, MeOH extracts of C. zeylanicum bark displayed less inhibitory activity against P. aeruginosa.E. coli was tested only against MeOH leaf extracts, indicating an intermediate MIC level.In line with previous research, comparable results have been documented for MICs of bark MeOH extract against S. aureus and E. coli(Singh et al., 2023).Nonetheless, earlier studies reported minimum bactericidal concentration (MBC) values for cinnamon extracts against S. aureus and E. coli as 5% and 10%, respectively.These values are higher than those observed in the current study, as MBCs generally exceed MICs under similar conditions(Parisa et al., 2019).Patel et al. (2022), reported identical MIC results for S. aureus against alcoholic extracts of cinnamon bark.C. zeylanicum bark ethanolic extract MICs ranged from 64 μgmL -1 to 256 μgmL - 1 in detailed research on S. aureus sensitivity(Mandal, 2011).Moreover, previous authors have reported MICs of essential oils from C. zeylanicum bark.Compared to prior essential oil MIC values (0.8 -3.3 mgmL -1 ), the current study's results align reasonably with the investigated MIC range(Prabuseenivasan et al., 2006).
than bark MeOH extracts.Notably, E. coli, P. aeruginosa, and S. aureus displayed inhibitory effects with leaf MeOH extracts, while S. pyogenes remained resistant.Among these bacterial strains, S. aureus demonstrated the highest sensitivity to MeOH leaf extracts, while E. coli and P. aeruginosa exhibited moderate sensitivity.Intriguingly, all selected bacterial strains were resistant to aqueous leaf extracts.

Table 3 : Antibacterial and Antifungal Activity of Cinnamon Extracts Against various strains.
This table presents the diameter of inhibition zones (in mm) and Minimum Inhibitory Concentration (MIC) values for methanol (MeOH) and aqueous extracts of cinnamon bark and leaves against bacterial strains (E.coli, P. aeruginosa, S. aureus, S. pyogenes) and fungal strains(C.albicans,C. tropicalis, C. parapsilosis, C. krusei, M. gypseum, T. mentagraphytes, T. rubrum, F. dimerum).Positive controls are included for comparison.
Paranagama et al., 2001bition zone; DD= agar disk diffusion assay; WD= agar well diffusion assay; MIC= minimum inhibitory concentration; Zone of inhibition of mean ± standard deviations (SD) of triplicate assays are provided (p <0.05).(-)Nozone of inhibition.The diameter of the disk was included in the inhibition zone measurements.suggesttheantibacterialeffects of C. zeylanicum essential oils on Fusarium species, the present study found limited inhibitory effects on F. dimerum.Cinnamon bark's richness in cinnamaldehyde, with potent antimicrobial properties, as demonstrated byParanagama et al., 2001might be responsible for the antimicrobial actions observed against the tested strains. Abbreviations:

Table 4 : Cytotoxic Activity of Ceylon Cinnamon Extracts on MCF-7 and Hep G2 Cancer Cells.
This table displays the cytotoxic effects of cinnamon extracts (Lw, Bw, Lm, Bm, C) at various concentrations (25, 50, 100, 200 µg/mL) on MCF-7 and Hep G2 cancer cell lines after incubation periods of 24 and 48 hours.The table includes the mean cell viability percentages (± standard deviation) for each concentration and time point."Nt" indicates no test performed or not tested.Soxhlet extraction) and aqueous (maceration) extracts of C. zeylanicum bark against MCF-7 cells, yielding IC 50 values of 58 µgmL -1 and 140 µgmL - 1 for 48 and 24 hours of post-incubation, respectively.Additionally, IC 50 s of aqueous cinnamon bark extract were

Table 5 : IC 50 Values of Cinnamon Extracts on MCF-7 and Hep G2 Cancer Cell Lines.
This table presents the IC 50 values (µg/mL) of cinnamon extracts(Lw, Bw, Lm, Bm, C)for MCF-7 and Hep G2 cancer cell lines, measured at 24 and 48 hours.Values are expressed as mean ± standard deviation, with different letters indicating statistically significant differences between the samples."NA" indicates that the data was not available or not applicable.Lw-Aqueous extracts of cinnamon leaves; Bw-Aqueous extracts of cinnamon bark; Lm-Methanolic extracts of cinnamon leaves; Bm-Methanolic extracts of cinnamon bark; C-Control (Tamoxifen); NA-Not applicable.Mean ± Standard deviation (n = 3).Different letters (a-e) for each column donate significant differences (ρ>0.05) between the tested concentrations. Abbreviations: