Stigmas and Petals of Crocus sativus L. (Taliouine, Morocco): Comparative Evaluation of Their Phenolic Compounds, Antioxidant, and Antibacterial Activities

The dried stigmas of Crocus sativus L. produce saffron, a precious spice used for its culinary and medicinal properties since ancient times, while its petals are considered the main by-product of saffron production. The present study aimed to comparatively evaluate the phenolic content, antioxidant capacity, and antibacterial activity of methanolic extracts of stigmas and petals of Crocus sativus L. from Taliouine. The polyphenol content was measured using the Folin–Ciocalteu method, the antioxidant activity was determined using the DPPH free radical scavenging method, and the well-diffusion method was used to assess antibacterial activity against seven pathogenic bacterial strains (Bacillus subtilis, Escherichia coli, Listeria monocytogenes, Proteus vulgaris, Pseudomonas aeruginosa, Salmonella enterica, and Staphylococcus aureus). Furthermore, the minimum inhibitory concentration (MIC) of the extracts was determined using the microdilution broth test. Our findings revealed that stigmas and petals contained phenolic compounds at the rate of 56.11 ± 4.70 and 64.73 ± 3.42 mg GAE/g, as well as DPPH radical scavenging capacity with IC50 of 1700 µg/ml and 430 µg/ml, respectively. Petal extract showed more effective antibacterial activity, with inhibition diameters ranging from 10.66 ± 0.57 to 22.00 ± 1.00 mm and MIC values ranging from 2.81 to 5.62 mg/ml, compared to the stigma extract, which displayed inhibition diameters from 10.00 ± 0.00 to 18.67 ± 0.76 mm and MIC from 2.81 to 11.25 mg/ml, against five of the seven bacterial strains tested, including S. aureus, E. coli, P. vulgaris, P. aeruginosa, and S. enterica. Statistical analyses were performed to determine the significance of these results. Thus, stigmas and petals of Crocus sativus L. might serve as a suitable source of natural antioxidant and antimicrobial agents for application in the food and pharmaceutical industries.


Introduction
Aromatic and medicinal plants have always had a close relationship with humankind throughout the development of all ancient civilizations, serving various purposes in felds, such as cooking, perfumery, cosmetics, and medicine.Despite progress in the pharmaceutical feld, the therapeutic use of plants remains a common practice in all countries, including Morocco [1].According to the World Health Organization, almost 80% of the world's population in developing countries relies on plant extracts or their active ingredients to meet their primary healthcare needs [2].Several considerable socioeconomic and ecological benefts are described in the scientifc literature concerning the use of aromatic and medicinal plants, making their development a key objective in national agricultural development strategies.
In this context, research into the properties of aromatic and medicinal plants is more important than ever.Several studies have focused on medicinal plants as a valuable resource to identify new bioactive compounds with antioxidant properties, aiming to prevent complex diseases induced by oxidative stress, a primary contributor to most deaths, including cancers [3].Additionally, research has also focused on the antimicrobial properties of these plants, in an efort to discover new natural antimicrobial agents with fewer side efects [4][5][6] and combat the emergence of antibiotic resistance in bacteria, due to the expanding use of chemical drugs [7].Society itself is showing increasing interest in the benefts of alternative and complementary medicine based on aromatic and medicinal plants, as they are regarded as a more natural and less invasive means of treating and preventing disease than conventional medicine.Foremost among these plants is the Crocus sativus L. fower, which has been used for thousands of years to treat a variety of ailments and diseases [8].Tis herbaceous autumnblooming fower belongs to the Iridaceae family and the large Crocus genus, which includes over eighty species.Crocus sativus L. the only Crocus species that generates safron by drying its red stigmas is Crocus sativus L.
Te reproductive method of this particular species is through vegetative propagation using bulbs [12,13].It is cultivated in many places worldwide, such as Iran, India, Afghanistan, Morocco, and Euro-Mediterranean countries such as Greece, Spain, and Italy, with a world production of 418 t year 1 [14].Iran has long been the world's largest producer and exporter of safron, accounting for 85-90% of global production [15,16].
According to the National Agency for the Development of Oasis and Argan Zones (ANDZOA), national safron production is estimated at over 10 t in 2020, located in the Taliouine and Taznakht area, places Morocco fourth in terms of production after Iran, India, and Greece [12].Safron is the most expensive spice in the world, nicknamed "red gold."Its labor-intensive cultivation takes around 150,000-200,000 (78 kg) fresh fowers and 160,000 petals to produce 1 kg of safron [13], which explains its scarcity and high cost.In addition to their use in gastronomy, this spice has potential pharmaceutical and medicinal properties due to its richness in bioactive components [14], in particular, crocin, safranal, and picrocrocin, which contribute, respectively, to the color, aroma, and specifc bitterness of safron and have been identifed as being responsible for these targeted therapeutic properties in the treatment of several diseases [15].Antioxidant [16], anti-infammatory, antitumor, antimicrobial, and antidiabetic priorities have also been described [17].Clinical studies have indicated that safron, in various doses, dose not present serious adverse side efects [17,18].Furthermore, in recent times, research has been undertaken to extract bioactive components not only from the stigmas but also from the petals of Crocus sativus L. Many studies have shown that these often neglected petals also contain interesting bioactive components [19,20], mainly polyphenols, notably carotenoids, favonoids, and anthocyanins, known for their antioxidant and antibacterial properties, as well as their multiple protective roles [21].Tey have been associated with numerous pharmacological efects [22].Te results of previous studies have suggested that diferent parts of Crocus sativus L. can be used for their antioxidant activity [23,24] and antibacterial efects against specifc bacteria [12,25,26].
Our study aims to compare safron, a highly esteemed spice, with its primary by-product, the petals of the endemic Crocus sativus L. variety from Taliouine, Morocco.We aim to evaluate their respective phenolic and antioxidant properties while also assessing their antibacterial activity against a range of resistant strains of pathogenic bacteria.To achieve this, we employed the methanol extraction method, known for its efciency in maximizing the extraction of bioactive compounds.By undertaking this investigation, we seek to shed light on the potential of safron and its petals as valuable sources of bioactive compounds with promising applications in various industries.

Plant Material.
Stigmas and petals of Crocus sativus L. fowers, utilized in this study, were collected in October and November 2022 from the plant's natural biotope in Taliouine, Taroudant province, Souss Massa region, Morocco.Fresh samples were then shade-dried at room temperature and stored in a dark place awaiting analysis.

Preparation of Methanolic Extracts of Stigmas and Petals.
Dried stigmas and petals were ground to powder and separately extracted by maceration with 80% methanol at an extraction ratio of 1 : 10 (v/w).To determine phenolic compounds and antioxidant activity, the macerate was shaken for 20 min at 350 rpm in the dark at room temperature and subsequently fltered through pleated flter paper (180-F, Ø � 150 mm, Dorsan, Barcelona).A second extraction was conducted under identical conditions, and the two fltrates obtained were combined and evaporated under vacuum using a rotary evaporator (IKA RV) at 45 °C.Te extract obtained was then reconstituted with 80% methanol and stored at 4 °C [27].
To assess antibacterial activity, the macerate was shaken at 350 rpm for 24 h in the dark at ambient temperature.Te resulting extract was then fltered and concentrated to dryness with the aid of a rotary evaporator.Te concentrates obtained were then dissolved and taken up in 10% dimethyl sulfoxide (DMSO) at various concentrations of 45, 180, and 360 mg/ml and stored at 4 °C until use.
Te extraction yield of each extract derived from stigmas and petals of Crocus sativus L. was calculated according to the following equation: where W dry extract is the weight of each dry extract obtained in grams and W sample is the weight of stigmas and petals' samples used in grams.[28].

Bacterial
A volume of 0.5 ml of each prepared extract was combined with 1 ml of the Folin-Ciocalteu reagent (diluted 10fold), and the mixture was thoroughly mixed and left to stand at room temperature for 5 min.Afterward, 0.8 ml of 7.5% Sodium Bicarbonate solution was added to the mixture, and it was left to incubate in the dark for 1 hour.Subsequently, the absorbance was measured at 760 nm using a UV-vis spectrophotometer (Agilent technologies).Te results were expressed as milligrams of gallic acid equivalents (GAE) per gram of dry extract.

Determination of Antioxidant Activity.
Te antioxidant potential of methanolic extracts of Crocus sativus L. stigmas and petals was assessed through the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay.Tis widely recognized method evaluates the antifree radical potential of plant compounds or extracts by quantifying their capability to neutralize the chemical radical DPPH [29,30].DPPH is characterized by its dark violet hue.Te strength of the extracts' antioxidant properties can be determined by how much color fading or decrease occurs when they are present.
For each extract, a volume of 0.5 ml was combined with 1.8 ml of a DPPH solution (0.1 mM).Te mixture was thoroughly mixed and allowed in darkness at room temperature for 30 min.
Following the incubation period, the absorbance of each sample was measured at 517 nm against a blank utilizing a UV-vis spectrophotometer (Agilent Technologies) [31].Triplicate measurements were conducted for each sample, and antioxidant activity is expressed as a percentage of DPPH inhibition, and wish is calculated using the formula as follows: where Abs control is the absorbance of DPPH solution in methanol (Control).Abs sample is the absorbance of the DPPH solution in the presence of the sample extracts tested.Antioxidant activity is often expressed in terms of IC 50 (half concentration inhibition), representing the extract concentration necessary to reduce the DPPH radicals by 50%.Tis value was determined through a graphical linear regression analysis of inhibition percentages against the various extract concentrations used [29].

Measurement of the Antibacterial Activity.
Measurement of the antibacterial activity of Crocus sativus L. stigma and petal extracts at diferent concentrations was assessed using the agar well difusion method, as outlined by Tagg and McGiven [32].After checking the purity of the test organism, the bacteria were incubated at 37 °C for a duration of 24 h in tryptone soy broth (TSB) (Biokar, Beauvais, France).Te bacterial growth was adjusted to the 0.5 McFarland turbidity standard, corresponding to approximately 10 8 CFU/ml.Subsequently, 15 ml of Mueller-Hinton agar (MHA) (Biokar, Beauvais, France) was aseptically poured into each Petri dish.Using a sterilized stainless-steel cylinder, 8 mm-diameter wells were made in the agar inoculated with the bacterial culture to be evaluated, and then each well was flled with 100 μl of dilution of the prepared extracts.Chloramphenicol is employed as a positive control, while a 10% DMSO was included as a negative control.Te Petri dishes were then placed in a refrigerator at 4 °C for 1 h to facilitate the difusion of the extracts into the medium.Following this, they were incubated at 37 ± 1 °C for 16 to 24 h.Te assessment of antibacterial activity involved measuring the diameter of the inhibition zone by use of a transparent scale and expressing it in mm.Tis test was repeated three times, and the average of the results was subsequently reported.

Minimum Inhibitory Concentration.
Following confrmation of the antibacterial activity of methanolic petal and stigma extracts against tested bacterial strains, the MIC was determined using the microdilution broth sensitivity test, in line with the Clinical and Laboratory Standards Institute recommendations [33].Te extract was serially diluted in a 96-well microplate using the TSB medium.A standardized bacterial suspension adjusted to a concentration of 10 6 cfu/ ml was then added to each dilution; positive controls (culture medium with inoculum) and negative controls (culture medium without inoculum) were also included.After incubation at 37 °C for 24 h, the well exhibiting the lowest extract concentration displaying no turbidity (indicating the absence of bacterial growth) was identifed as the MIC, signifying that this specifc extract concentration efectively inhibits bacterial growth.Each experiment was repeated three times.

Extraction Yield. Te extracts of stigmas and petals of
Crocus sativus L. from Taliouine used in our study were obtained using the 80% methanol maceration method.Tis extraction method gave yields ranging from 49.4 to 66% and from 45 to 64% for stigmas and petals, respectively.Tese yields vary according to the processing conditions and extraction methods used.Numerous studies have claimed that methanol extraction achieves higher extraction yields compared to other solvents, for Crocus sativus L [34], as well as for other plants [35].

Total Polyphenol Content.
Phenolic compounds have been widely studied for their bioactive properties, particularly their protective efects against pathologies.A comparative analysis of total polyphenols in stigma and petal extracts of Crocus sativus L. from Taliouine was conducted using the Folin-Ciocalteu method.To facilitate this analysis, a standard calibration curve was generated (y � 0.0102 x + 0.0212 and R 2 � 0.9951).
Te results revealed higher levels of phenolic compounds in petals than in stigmas.Te values in petals and stigmas were 64.73 ± 3.42 and 56.11 ± 4.75 mg GAE/g, respectively (Table 1).
Te polyphenol content recorded in our study appears to be higher for Taliouine safron stigma extracts than that described by Karimi et al. for Iranian safron (6.45 mg GAE/g) [36].However, it appears slightly close to that reported for the aqueous extract of Moroccan safron stigmas (72.47 mg GAE/g) [37].In our study, the phenolic compound levels found in the petal extracts of Crocus sativus L. were similar to those reported by Jadouali et al. and by Ouahhoud et al. for Taliouine safron [38,39].Tey reported concentrations of 65.34 mg GAE/g DM and 64.66 mg GAEeq/g DM, respectively.However, the values obtained appear higher than those reported by Asgarpanah et al. for methanolic extracts of Iranian safron petals (4.09 to 17.34 mg GAE/g MS) [40].Although some studies have shown that petals of Crocus sativus L. contain more polyphenols than stigmas [39,41], other studies have reported the opposite, with a concentration of 97.99 mg GAE/g for stigmas and 69.18 mg GAE/g for petals [42].Tus, there is no clear consensus as to which of these foral parts of Crocus sativus L. contain the highest polyphenol content.
Indeed, extraction methods and other intrinsic and extrinsic factors could vary the phenolic compound content of plants.Moreover, the diference in polyphenol content recorded between organs of the same plant has been reported in many plants [43].Tis variation refects known variations in other secondary plant metabolites.It could also be interspecifc and linked to the genotypic potential of plants in diferent countries, as has been described for wheat or according to the evolutionary stage or physiological state of the plants or according to the vegetative stage of the plants (vegetation or fowering period) [44].Indeed, total polyphenol levels can vary quantitatively between leaves (fresh or dry), roots, stems, and fowers.In addition, intrinsic factors such as ecological and/or climatic conditions (temperature, sun exposure, drought, and soil) are known to condition the biosynthesis of secondary plant metabolites [45].

Antioxidant Activity by the DPPH Test.
Te antioxidant potential of methanolic extracts from the stigmas and petals of Crocus sativus L. was determined by the DPPH free radical scavenging test, which results in a characteristic color change from dark purple to pale yellow, indicating DPPH scavenging activity.Tis test showed a higher antioxidant activity potential for methanolic extracts of Crocus sativus L. than for aqueous and ethanolic extracts [36,46].Te signifcant infuence of solvent extraction power on polyphenol extraction yield and antioxidant power has been reported by numerous studies.Methanol appears to be particularly efective [47].
Te DPPH scavenging activity of methanolic extracts of safron stigmas and petals increased proportionally with extract concentration.A concentration of 1 mg/ml of extract gave a percentage inhibition of 34.74% for stigmas and 89.17% for petals (Table 1).
Te regression equations were used to calculate the IC 50 indicating the free radical scavenging power of the extracts tested (Figure 1).Te IC 50 value obtained was 430 µg/ml for the petal extract and 1700 µg/ml for the stigma extract.A lower IC 50 value indicates greater antioxidant activity; therefore, petals had a higher antioxidant capacity than stigmas and contained more antioxidant compounds.
Tese results are in line with those of the study by Ouahhoud et al. who reported that the radical scavenging capacity of petals was higher than that of stigmas, with an IC 50 of 80.73 µg/ml for petals and 1554.37 µg/ml for stigmas [39].In other studies, the IC 50 for stigmas was estimated at 207.16 µg/ml and 304 µg/ml for ethanolic and aqueous extracts, respectively [48].For petal ethanolic extracts, an IC 50 of 504.26 μg/ml was obtained by Jadouali Mohamed [49], while in the study by Wali et al., this value was 86.63 µg/ml [50].

Antibacterial Activity. Te antibacterial activity of methanolic extracts of stigmas and petals of Crocus sativus
L. from Taliouine was assessed using the agar well difusion method.
It is worth noting that various methods are available for determining bacterial susceptibility to antimicrobial agents, including disk difusion, well difusion, and broth or agar dilution.In our study, we opted the agar well difusion method, as it is commonly recommended for highlighting the antibacterial efect of tested substances or extracts [51].
Te results showed diferent degrees of inhibition by Crocus sativus L. stigma and petal extracts, depending on the pathogenic bacterial strains tested (Table 2).
Antibacterial activity against the strains tested increased signifcantly (P ≤ 0.01) with increasing concentrations of Corcus sativus L. extracts (Figure 2).Concentrations (45,180, and 360 mg/ml) showed zones of inhibition ranging from 10.66 ± 0.57 to 22 ± 1.00 for petals and from 10 ± 0.00 to 18.67 ± 0.76 mm for stigmas against the following bacteria: Staphylococcus aureus, Escherichia coli, Proteus vulgaris, Pseudomonas aeruginosa, and Salmonella spp.Te lowest concentration 45 mg/ml of the two extracts showed a slight inhibition of the growth of these seven bacteria, with zones of inhibition not exceeding 12.50 mm; however, this concentration in the stigma extract had no inhibitory efect on the growth of Salmonella.
Te negative control (10% DMSO) showed no antimicrobial activity against any of the bacteria tested, while the positive control with chloramphenicol demonstrated efcacy against the tested bacteria.
According to these results, Staphylococcus aureus was the most sensitive, which concurs with the results obtained by Lachguer et al. [12].However, no antibacterial activity was observed against Listeria monocytogenes and Bacillus subtilis, indicating resistance of these strains.Te extracts demonstrated efective antibacterial activity against Gram-negative bacteria, E coli, P vulgaris, P aeruginosa, and Salmonella, than against Gram-positive bacteria, except for S aureus.
Results have been reported by Lahmass et al. that methanolic extracts of petals showed antibacterial activity against several strains, including S. aureus and E. coli [52].In addition, several other studies have shown that safron possesses antibacterial properties against several strains.Methanolic extracts were efective against Clostridium perfringens, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginos, Shigella fexneri, and Staphylococcus aureus [34], and petroleum ether and methanol extracts were efective against Klebsiella pneumonia, Proteus vulgaris, Escherichia.coli, Staphylococcus aureus and Pseudomonas aeruginosa [53].Other studies have reported the antibacterial activity of aqueous extracts from Moroccan and Italian stigmas against bacterial strains Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa, Klebsiella pneumonia, Staphylococcus aureus, and Listeria monocytogenes [37].

Minimum Inhibitory Concentration (MIC).
Determining the MIC is a crucial step in assessing the effcacy of antimicrobial agents, and dilution methods are considered the most appropriate for determining MIC values.Tese methods allow for the precise estimation of the concentration of the tested antimicrobial agent within the culture medium [51].Te microdilution technique involves preparing a series of diluted solutions of Crocus sativus L. stigma and petal extract in each well of the 96-well plate.Te extract is tested at various concentrations, starting with a larger concentration in the frst well and progressively lowering it in succeeding wells.After inoculating each well with a standardized bacterial suspension (10 6 cfu/ml), the plates are incubated at 37 °C for 24 h.Te lowest concentration at which no bacterial growth is seen in a well is the MIC (Table 3).
Petals and stigmas extracts exhibited MIC values signifcantly higher (p < 0.01) than those of the positive control (chloramphenicol).Te MIC values for petal extract were 2.81 mg/mL for S aureus CECT 976 and P vulgaris CECT 484 and 5. 62    Te Scientifc World Journal    [40], while, according to Razavi and Jafari, the inhibitory efect of methanolic extracts of stigmas showed MIC values lower than the results of the present study, ranging from 0.33 to 1.23 mg/mL against E. coli, P. aeruginosa, and S. aureus [55].
Based on the results obtained, it was observed that extracts from the petal of Crocus sativus L. were more efective in inhibiting bacterial growth when compared to extracts from the stigma.Furthermore, these extracts were found to have a more marked efect on Gram-negative bacteria than on Gram-positive bacteria.Tis diference in sensitivity is due to diferences in bacterial cell structure and composition.
Let uss recall that Crocus sativus L. possesses various phytochemical properties that have been linked to its antibacterial activity; this activity has been attributed to the presence of compounds such as safranal and crocin [55][56][57], as well as favonoids such as kaempferol, quercetin, and an isorhamnetin derivative [58].Tese compounds are volatile or water-soluble, enabling them to easily reach contaminating microorganisms and contribute to their destruction [59].
Te variability in antimicrobial potential recorded by the authors is linked to several direct and indirect factors that accompany the sample from implantation to the fnal product.Tese factors include the age of the corms [55], altitude and climate [60], drying temperature [61], storage and packaging [62], and extraction and analysis methods [36,63].All these factors can afect the chemical composition of the sample and therefore the results obtained during analysis.Consequently, it is important to consider these factors and control them as much as possible to minimize the variability of results and obtain reliable and reproducible data.Stigma extracts showed a signifcant positive correlation between a zone of inhibition and MIC (r � 0.926).In addition, a strong positive correlation was observed between total polyphenol content and antioxidant activity, with a correlation coefcient (r � 0.962) (Table 4).Tese results were in line with those reported by Acar et al. [64].For the petals, a positive correlation was observed between the zone of inhibition and MIC (r � 0.723).Tere was no signifcant correlation found between total phenolic content and antioxidant activity (Table 5).Tis fnding is consistent with the results of previous studies for other medicinal plant  Te Scientifc World Journal extracts [65,66], indicating that antioxidant capacity and total phenolic content were not correlated.Tis may be explained by the fact that, in addition to phenolic compounds, other phytochemicals contained in Crocus sativus L. extracts, such as favonoids, carotenoids, and vitamins, may contribute synergistically to this overall antioxidant capacity.On the other hand, determination of the total phenolic compound content using the Folin-Ciocalteu method does not represent an absolute measure of the quantity of phenolic compounds.

Principal Component Analysis (PCA).
Te efect of concentrations (45, 180, and 360 mg/ml) of petal and stigma extracts on the antibacterial activity of the diferent strains assessed was assessed using the statistical technique of principal component analysis (PCA) (Figure 3).Principal component 1 (PC1) contributed 92.80%, while principal component 2 (PC2) accounted for 5.81% of the total variability of the components represented.According to the results, similarities were highlighted by the small distance between the concentrations of stigma and petal extracts, suggesting that these extracts have signifcant efects in terms of zone of inhibition against bacteria (Staphylococcus aureus, Escherichia coli and Proteus vulgaris, Pseudomonas aeruginosa, and salmonella).In addition, the 360 mg/ml concentration was associated with the highest antimicrobial response, and the activity increased with the increasing concentration.Staphylococcus aureus proved the most sensitive one.However, a slight dissimilarity was observed with Salmonella, which proved less sensitive to extracts than the other bacterial strains evaluated.

Conclusion
Te extracts obtained from the stigmas and petals of Crocus sativus L., originating from Taliouine in the Taroudant Province of Morocco, exhibit a signifcant abundance of bioactive compounds, particularly polyphenols.Tese foral components demonstrate remarkable antioxidant and antibacterial activities, thereby ofering protective efects against cellular oxidation processes and pathogenic microorganisms.Te presence of these valuable properties in Crocus sativus L. from Taliouine highlights the    Te Scientifc World Journal underappreciated potential of safron and its petals.Te outcomes of this study open new avenues for the extensive utilization of Crocus sativus L. in diverse felds, including the pharmaceutical, food, and cosmetics industries.Te incorporation of Crocus sativus L. and its derived products holds promise for the development of innovative and effective solutions in these domains, capitalizing on their inherent bioactive properties.

Figure 1 :
Figure 1: Te percentage DPPH inhibition (%) in mg/ml of methanolic extracts: stigmas (a) and petals (b) of Crocus sativus L. from Taliouine, as a function of concentration.
bold indicate the highest inhibition zones, these values can be left in their usual format.

3. 6 .
Statistical Analysis 3.6.1.Correlation between TPC, Antioxidant Capacity, Antibacterial Activity, and MIC.Te results of the statistical measurements analyzed of Crocus sativus L. extracts indicated signifcant positive correlations according to Pearson's correlation analysis at a signifcance level p < 0.01 between the diferent parameters analyzed.

Figure 3 :
Figure 3: PCA efects of diferent concentrations of Crocus sativus L stigma and petal extracts against bacterial strains.

8
Analysis.Statistical analysis was conducted using IBM SPSS (Statistical Package for the Social Sciences) version 25.Means and standard deviations were calculated, and statistical correlation between the various parameters tested for Crocus sativus L. extracts was assessed using Pearson's bivariate correlation test option.Results were deemed statistically signifcant if the P value was less than 0.01.In addition, principal component analysis (PCA) was carried out using ORIGIN Lab Pro 2017 software to further analyze and interpret the data.

Table 1 :
Total polyphenol content and antioxidant activity of methanolic extracts of stigmas and petals of C. sativus L. Taliouine (n � 3).

Table 2 :
Antibacterial activities of methanolic extracts of stigmas and petals of Crocus sativus L from Taliouine.
In support of this, studies have reported that methanol and ethanol extracts of Crocus sativus L. demonstrated an MIC value of 6.5 mg/mL for Staphylococcus aureus according to Okmen et al. [54].Te MIC values in the study by Asgarpanah et al. were 31.2mg/mL for Staphylococcus aureus, 62.5 mg/mL for Salmonella, and 125 mg/mL for B. cereus in Crocus sativus L. petal extract

Table 3 :
MIC of methanolic extracts of stigmas and petals of Crocus sativus L. from Taliouine.

Table 4 :
Linearity among diferent analyzed parameters of Crocus sativus L. stigmas.Correlation is signifcant at the 0.01 level (two-tailed).Values in bold in the table indicate signifcant correlations.

Table 5 :
Linearity among diferent analyzed parameters of Crocus sativus L. petals.Correlation is signifcant at the 0.01 level (two-tailed).Values in bold in the table to indicate signifcant correlations.