TOTAL PHENOLIC AND FLAVONOID CONTENTS, ANTIMICROBIAL AND ANTIOXIDANT POTENTIALS OF CAMPANULA STRIGOSA BANKS & SOL

to quantify total phenolic content. An aluminium chloride assay determined the total flavonoid concentration. Studies found that the plant extract had a TAS value of 4.974±0.259 mmol/L, a TOS value of 12.437±0.150 µmol/L, and an OSI value of 0.251±0.014. The plant extract was shown to be effective against bacterial strains at concentrations between 100 and 400 µg/mL, and against fungal strains at concentrations between 50 and 100 µg/mL. Ethanol extract also had a greater total phenolic concentration and total flavonoid content. C. strigosa was shown to have antibacterial and antioxidant properties in this study.


INTRODUCTION
Natural remedies including mushrooms, plants, and animals are employed in the context of alternative medicine. (Korkmaz et al., 2018). Vitamins, minerals, and other nutrients are plentiful in plant foods. They are not only useful for human health because of their nutritional value, but also because of the secondary metabolites they generate. (Mohammed et al. 2021a). Plants have been shown to provide several health benefits, including those related to cancer prevention, infection prevention, DNA protection, allergy relief, cell growth inhibition, inflammation reduction, and liver protection (Miastkowska and  Determining the biological functions of plants is, thus, crucial. Anti-oxidant and antimicrobial properties of C. strigosa were investigated here. Total phenolic and flavonoid concentrations were also calculated for the plant. More than 500 species and subspecies of bellflowers are included in the genus Campanula (Campanulaceae). There are both annuals and perennials in this genus. It is the temperate zones of the northern hemisphere and the Mediterranean and Caucasus with the highest diversity. It is generally grown as an ornamental plant (Tsiftsoglou et al. 2022).

MATERIALS AND METHODS
Gaziantep was the site of the gathering of plant specimens. (Turkey). The aerial parts were dried in a cool, dry place with plenty of shade. Plant samples were dried, then pulverised into powder. Then, after soaking the plant sample at 50 °C for around 6 hours, 30 g of it was extracted with 250 mL of ethanol. The same method was applied for the methanol extract.. In a rotary evaporator, the solvents were evaporated off of the resulting extracts.

Antioxidant tests
The sample of plant material was analysed for its status of antioxidants and oxidants with Rel Assay Diagnostics' TAS and TOS kits. Calibrators for TAS and TOS testing included trolox and hydrogen peroxide. TAS values were expressed as mmol Trolox equiv./L and TOS values as μmol H2O2 equiv./L (Erel, 2004;Erel, 2005). The OSI was calculated by dividing the TAS by the TOS (Sevindik, 2019).

Antimicrobial activity tests
The agar dilution technique was used to evaluate the plant sample for its antimicrobial activity against a panel of reference bacteria and fungal strains. The plant extract was diluted to amounts ranging from 12.5 to 8.0 µg/mL. To dilute, we used distilled water.

Total Phenolic and Flavonoid Tests
The plant extract was dissolved in distilled water to make a 1 mL stock solution, and then 1 mL of Folin-Ciocalteu reagent (1:9, v/v) was added and the mixture was vortexed. The resultant solution was diluted to 0.75 mL with 1% Na2CO3 and let to incubation for 2 hours. Then, a reading was taken at 760 nm. Based on the gallic acid standard solution calibration curve (Yumrutaş et al. 2009), total phenolic content was calculated and reported as mg.GAE g -1 .
Aluminum chloride analysis was used to calculate the total flavonoid content of the plant extract. (Chang et al. 2002). The mixture included 0.1 mL of 10% Al(NO3)3, 0.1 mL of 1 M NH4CH3COO, 4.3 mL of methanol, 0.5 mL of Quercetin, and 0.5 mL of plant extract. It was then incubated for 40 minutes and absorbance was measured at 415 nm. The amount of flavonoids was expressed as mg.QE g -1 .
Humans find several uses for plants. Not only is it good for you from a nutritional standpoint, but it has also been used to cure a wide range of illnesses. In this research, we analysed Campanula strigosa Banks & Sol. for its antioxidant and antibacterial properties. Total phenolic and flavonoid concentrations were also calculated. For this purpose, a soxhlet apparatus was used to extract ethanol and methanol from the plant's above-ground portions. The effectiveness of antioxidants was measured using Rel Assay kits. The agar dilution technique was used to test for antimicrobial activity against various bacterial and fungus species. Folin-Ciocalteu reagent was used to quantify total phenolic content. An aluminium chloride assay determined the total flavonoid concentration. Studies found that the plant extract had a TAS value of 4.974±0.259 mmol/L, a TOS value of 12.437±0.150 µmol/L, and an OSI value of 0.251±0.014. The plant extract was shown to be effective against bacterial strains at concentrations between 100 and 400 µg/mL, and against fungal strains at concentrations between 50 and 100 µg/mL. Ethanol extract also had a greater total phenolic concentration and total flavonoid content. C. strigosa was shown to have antibacterial and antioxidant properties in this study.

Antioxidant activity
The metabolic processes of all living creatures result in the production of reactive oxygen species (ROS), also known as free radicals. (Baba et al. 2020). At low concentrations, it has no ill effects, but at higher concentrations, it may cause serious damage to cells. The antioxidant defence system helps mitigate these negative consequences. (Bal et al. 2017). When antioxidant defence mechanisms are overwhelmed, oxidant chemicals become unchecked, leading to oxidative stress. Cancer, Parkinson's, cardiovascular problems, neurological illnesses, depression, and Multiple Sclerosis are only some of the major diseases that have been linked to oxidative stress in humans (Gürgen et al. 2020;Eraslan et al. 2021). Antioxidant supplements are useful in preventing or alleviating illnesses caused by oxidative stress. (Bal et al. 2019). Plants are significant here because they provide additional antioxidants from a natural source. Antioxidant status of C. strigosa was evaluated in this research. In addition, oxidant status were measured to calculate the oxidative stress index.  scutatus, G. glabra and G. aparine. The TAS value is an indicator of the whole of the antioxidant compounds found in natural products. A high TAS value indicates that the natural product has a high antioxidant potential (Krupodorova and Sevindik, 2020). In our study, it is seen that C. strigosa has an overall lower TAS value compared to the studies reported in the literature. However, it was determined that C. strigosa has antioxidant potential. TOS value is an indicator of all endogenous oxidant compounds produced in natural products (Krupodorova and Sevindik, 2020). The TOS value of C. strigosa used in our study was determined to be higher than M. longifolia ssp. longifolia, R. scutatus and A. kurdica, and lower than A. calocephalum, S. papposa, G. glabra and G. aparine. In this context, it was observed that the oxidant values of C. strigosa were at normal levels. OSI value shows how much oxidant compounds are suppressed by antioxidant compounds (Krupodorova and Sevindik, 2020). The OSI value of C. strigosa used in our study was determined by M. longifolia ssp. longifolia, R. scutatus and G. glabra, and lower than A. calocephalum, S. papposa, G. aparine and A. kurdica. In this context, it is thought that C. strigosa can be used as a natural antioxidant source.

Antimicrobial Activity
The Because of these characteristics, plants may be used as antibiotics. In this context, ethanol and methanol extracts of C. strigosa were used in our study and antimicrobial activity was determined against standard bacterial and fungal strains. The obtained results are shown in Table 3.  A study on the antimicrobial activity of C. strigosa was found in the literature. Several Campanula species have been discovered to possess antimicrobial properties.In these studies, it was reported that volatile oil and aqueous extracts of Campanula portenschlagiana were effective against Bacillus cereus, Enterococcus faecalis, Staphylococcus aureus, Clostridium perfringens, Listeria monocytogenes, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Candida albicans, Penicillium sp. and Rhizopus stolonifer at different concentrations (Politeo et al. 2013). Staphylococcus aureus, Bacillus cereus, Enterococcus faecium, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumanii, and Candida albicans were all shown to be inhibited by methanol and water extracts of Campanula retrorsa at varying doses, according to another research. (Alhage et al. 2018). Our research demonstrated that the activity of C. strigosa ethanol extracts was often greater than that of metanol extracts. Extract concentrations between 100 and 400 µg/mL were effective against several bacterial strains. The extract's efficacy against fungal strains was determined to be at its peak between 50 and 100 µg/mL. It exhibited the highest activity against fungal strains. E. coli and P. aeruginosa were the two bacterial species it was most effective against. These findings suggest that C. strigosa may have antibacterial properties.

Total phenolic and flavonoid contents
Phenolic and flavonoid chemicals may be found in plants. Their medicinal value is an essential function of these substances. (Iamkeng et al. 2022). C. strigosa was extracted in both ethanol and methanol, and their total phenolic and flavonoid contents were analysed. Table 4 displays the findings collected. Ethanol extract of C. strigosa employed in our research had a greater total phenolic and flavonoid content. The total phenolic content of C. lyrata subsp. lyrata methanol extracts has been reported in the literature to be 48.97 mg/g, while the total flavonoid content has been reported to be 67.02 mg/g. (Ayaz, 2021). Based on our findings, C. strigosa is a superior source of phenolic and flavonoid content. C. strigosa has been identified as a potential natural source due to its high phenolic and flavonoid content in this context.

CONCLUSION
The research here looked at the antimicrobial and antioxidant properties of aerial parts of C. strigosa plant.. Total phenolic and flavonoid concentrations were also calculated for the plant. Plant extracts were shown to have antioxidant and antimicrobial properties, as hypothesised by the study. The phenolic and flavonoid concentrations found in its structure further suggest it may be a naturally occurring source.