Biosynthesis of silver nanoparticles using aerial parts of Cardaria draba L Phytochemical analysis c

Abstract

Keywords: Cardaria Draba L.; Colorectal Adenocarcinoma Cancer; Enzyme İnhibition; Green Synthesis; Silver Nanoparticles

Introduction

The field of nanotechnology is the most dynamic research area in material sciences, and the synthesis of nanoparticles is increasing significantly worldwide. Nanotechnology is a field in which nanomaterials are produced using particles in the <100 nm size range . As a result of the research and discoveries, AgNPs (silver nanoparticles) have garnered interest due to their chemical, physical, and biological properties and their widespread use in both industrial and health fields [1]. In addition, the synthesis of metal nanoparticles, such as AgNPs, which exhibit antioxidant and antimicrobial activities has gained importance in biochemical applications [2]. Recently, many studies have proven that plants can act as effective precursors for metal nanoparticle synthesis. These studies have enabled people to realize new production models in an environmentally friendly way despite the pollution they experience in modern times [3]. The green synthesis method has become quite popular in recent years [4]. In the green synthesis method, biological resources such as plants, bacteria, fungi, and algae, among others, are used as reducing agents to form nanoparticles. Thus, high costs and waste generation are prevented by using materials that are suitable for large-scale commercial production, non-toxic, highly stable, environmentally friendly, and inexpensive. Since the plants involved in green synthesis form compounds containing functional groups such as phenolics and flavonoids, the nanoparticles formed by the reduction of these compounds have antioxidant, antibacterial and catalytic properties [5]. Thus, metal oxide nanoparticles produced via green synthesis are applied in many areas such as the enzyme industry, drug delivery systems, and biomedical and pharmaceutical applications [6, 7, 8]. Cardaria draba (Brassicaceae), known as watercress, is an annual herbaceous plant. Animal feed and treatment: the aboveground parts are fed to animals. The leaves are crushed and applied externally to wounds on the heads of animals infected with anthrax; these leaves are changed several times a day, and it is said that this method heals the wound. The aboveground parts are used as food [9, 10, 11], animal feed [12], a pain reliever [11], a sedative, a wound healer [13], a diuretic, and to remove kidney stones [14]. Although C. draba prefers alkaline conditions, it can be found in different soil types with adequate moisture, growing in a wide variety of habitats such as shady open spaces, meadows, fields, croplands, grasslands, and roadsides [15]. C. draba has been used against various ailments in the past [16]. The aerial parts of C. draba contain alkaloids, saponins, flavonoids, terpenoids, tannins, triterpenoids, and leukoanthocyanins. The essential oil contains o-xylene, p-xylene, nonane, styrene, hexane-3,3,4-trimethyl, octane-2,6-dimethyl, heptane, 3-methyl-2-ethyl, decane, and nonan-4methyl; it is also registered to have nonane-2-methyl, benzene-1-ethyl-2-methyl, cyclohexene-2-methylpropyl, mesitylene, dodecane, limonene, benzenacetaldehyde, tetradecane, and 5-(methylthio)-pentane compounds. While

Kaempferol, quercetin and isorhamnetin are the most abundant compounds in the ethanolic extract of the leaves, the most abundant compounds in the ethanolic extract of the seeds are ellagic acid, sinapic acid, p-coumaric acid and caffeic acid [17, 18]. it is also used against kidney stones due to its expectorant and diuretic effects. C. draba seeds and leaves have expectorant and laxative effects [19]. It has been used as a carminative and has antiscorbutic activity [20]. In the past, C. draba, as one of the existing traditional medicines, has been used against a wide variety of ailments. For example, it has been used in anti-inflammatory, anticancer, antimicrobial, antioxidant, hypoglycemic, and other areas. The selection of C. draba was made for a number of reasons. The antibacterial properties of medicinal plants are one of their beneficial aspects, and examining these properties is crucial for understanding their potential applications. Many bacteria have developed resistance to antibiotics as a result of the increased usage of antibiotics. Therefore, the search for novel sources of antibacterials is crucial. Using medicinal herbs is a top priority for this reason. Furthermore, compared to medicines, their potent antibacterial chemicals provide less risk to the user. Medicinal herbs have long been regarded as chemical alternatives because of their affordability, ease of use, low toxicity, and fewer adverse effects. Medicinal plants have made many contributions to the literature in the search for antibacterial sources and in the field of nanotechnology. Silver nanoparticles made with C.origanifolium extract were found to have antimicrobial properties that were superior to those of synthetic antimicrobial medications by inhibiting the growth of Pseudomonas aeruginosa bacteria, and Candida albicans pathogenic fungus [21]. Silver nanoparticles (AgNP) synthesized using extract of Amygdalus spinosissima were studied for their antibacterial, antioxidant, anticancer, and apoptotic properties. It was shown that AgNPs exhibited strong antibacterial activity against Gram-negative bacteria [22]. Cardaria draba (L.) was collected in July 2023 at approximately 2000 meters altitude and 39°54′0″N 41°15′0″E coordinates in Erzurum (Turkey), and silver nanoparticles were obtained from the aboveground parts. In our study, AgNPs were synthesized biologically using the aboveground parts of the C. draba plant. Characterization was performed with UV-Vis, XRD, FT-IR and SEM analyses. In addition to examining the antibacterial activities of AgNPs, enzyme inhibition on some enzymes of medical importance was examined. The cytotoxicity of the human colon cancer cell line (DLD-1) was investigated.

Materials And Methods

Preparation of Cardaria draba L. Extract and Synthesis of Silver Nanoparticles The aboveground parts of the plant were collected, washed thoroughly with pure water twice, dried, and ground thoroughly in a porcelain mortar. Thirty grams

Sigma J Eng Nat Sci, Vol. 43, No. 6, pp. 1943−1956, December, 2025

of ground plant parts were mixed in 100 mL of deionized water, with a magnetic stirrer for 30 minutes. Then, it was heated in a water bath at 60°C for 10 minutes. After cooling to room temperature, it was centrifuged at 3500 rpm for 10 minutes and filtered through filter paper to obtain the plant extract [23]. Synthesis of silver nanoparticles was carried out similarly to the working procedures of Manosalva et al (2019) [24]. To obtain silver nanoparticles, 20 mL of plant extract was added dropwise to 100 mL of 1 mmol (0.170 g) AgNO3 solution using a dropping funnel. The reaction continued for 2 hours at 50°C in a magnetic stirrer. The light yellow plant extract turned brown with the addition of silver nitrate solution. This brown colloidal solution was centrifuged at 10,000 rpm for 15 minutes to precipitate AgNPs. The AgNPs precipitated by centrifugation were washed several times with deionized water and then dried. Characterization of Silver Nanoparticles Ultra violet (UV-VIS) spectral analysis Whether pure Ag+ ions were reduced was determined by UV-Vis spectral analysis. UV-vis spectra of biosynthesized AgNPs were determined in the wavelength range of 300-700 nm using a spectrophotometer (Agilent CARY 60 UV-visible spectrophotometer). Scanning electron microscopy (SEM) analysis It was characterized by scanning electron microscopy (SEM) (Tescan Maıa3 XMU) to determine morphology and the particle size. X-ray diffraction (X-RD) analysis The morphology of the crystal structure of AgNPs was analyzed by XRD (RIGAKU ULTIMA IV XRD) in the range of 3° to 79° for 2θ. The crystal size of AgNPs was calculated by using the Debye-Scherrer equation (D=Kλ/β cosθ). Fourier transform infra red spectroscopy (FT-IR) analysis FT-IR (Perkin Elmer Spectrum 100 FTIR) spectroscopy analysis was performed to determine the functional groups present in the plant extract, as well as those playing a role in the reduction, in the range of 4000-400 cm-1 at the end of the reaction. GC/MS (Gas chromotography/mass spectrometry) analysis GC/MS analysis was performed according to Basar et al. (2024) [25]. Phenolic Component Analysis OF C. draba Extract by LC-MS/MS Method LC-MS/MS analysis of the phenolic compound was performed using an Agilent 6460 Triple Quadrupole model (Agilent Technologies, Wilmington, DE, USA). The column temperature was fixed at 40 °C. The elution gradient was formed from mobile phase A (water, 0.1% Formic acid) and mobile phase B (acetonitrile with 0.1% formic acid). The mobile phase gradient was programmed as follows: 98% A (v/v) from 0 to 4.0 min, 98–80% A (v/v) from 4.0–7.0

min, 80–10% A (v/v) from 7.0–14.0 min, 10% A (v/v) from 14.0–15.0 min, 10–98% A (v/v) from 15.0–17.0 min. The total run time was 17 min, and the column equilibration time between each run was 4 min [26]. Antibacterial Activity Antibacterial activity was determined by working on Gram negative (Escherichia coli ATCC 25922, P. aeruginosa ATCC 27853, A. baumannii ATCC 17978) and Gram positive (Bacillus cereus DSSM 4312, S. aureus ATCC 25923, E. faecalis ATCC 29212) bacteria. The disk diffusion method was used to determine antibacterial activity (NCCLS 1997). Nutrient broth liquid medium, which was inoculated with each microorganism using loop inoculation, was incubated for 12-16 hours at 37 °C. 100 µl sample of the test microorganisms, prepared from an overnight culture, was taken and spread on nutrient agar solid medium. Then, 10 µl of different concentrations of all extracts were taken and absorbed onto sterile paper disks, which were placed in petri dishes, and the spread was made. Inhibition zone diameters were measured after 24 hours, of incubation at 37 °C for bacteria. The same procedure was repeated for positive (Chloramphenicol) and negative controls (ethanol and water). Each test was performed in three replicates at different times [27]. Enzyme Inhibition Antiurease and anticollagenase enzyme inhibition were performed according to Bakır and Kurbanoglu (2022) [28]. Lipoxygenase (LOX) enzyme inhibition was performed according to Bakır et al. (2022) [29].

Results And Discussion

SEM Analysis Figure 1 illustrates the scanning electron microscopy (SEM) images of silver nanoparticles (AgNPs) produced via the green synthesis process using C. draba. It is evident from the SEM images that the particles exhibit a range of sizes. Silver nanoparticles were discovered to have spherical shapes with an average diameter of 41 nm. From the SEM images, it can be observed that the silver nanoparticles are larger than expected as a result of a series of reactions triggered by the evaporation of the solvent due to the aggregation of the nanoparticles during sample preparation. This may have caused the differences in particle size [30]. The size of the particles was found to be 30-70 nm in the SEM study of AgNPs generated from Vitis vinifera (grape) fruit [31]. According to studies by Shah et al. (2018) on Daphne mucronata and Santhoshkumar et al. (2017) on Gnidia glauca, a species in the Thymelaeaceae family, silver nanoparticles have a spherical shape [32, 33]. The size and surface charge of the synthesized particles have an important effect on the interaction with the cells. Accordingly, when establishing the optimal conditions, the criteria exhibiting the smallest values for these parameters in the synthesized nanoparticles were selected.

Sigma J Eng Nat Sci, Vol. 43, No. 6, pp. 1943−1956, December, 2025

Figure 1. Images of SEM analyze of synthesized silver nanoparticles.

EDX Analysis As illustrated in Figure 2, the elemental composition of the AgNPs produced at room temperature was determined to be 78.11 % Ag, 8.8 % C, 4.9 % N, 3.05 % Cl, 2.00 % Ca and 1.38 % Fe . The strong peaks of the formed AgNPs in an optical absorption band at 3 eV indicate the presence of silver within the particles. It indicates that AgNPs are dense. At the same time, it was determined that some elements other than silver also formed weak peaks. In another study, it was reported that nanoparticles obtained from Salvia officinalis (Sage) plant contained 60.97% silver [34]. AgNPs synthesized from Murraya koenigii (Curry) plant were found as 60% Ag, 19% C, 12% O, and 6% Cl [35]. Also, the EDX spectrum shows the presence of Cl, N, O, K, and C atoms in different percentages. Weak signals such as chlorine, carbon, and oxygen in the EDX profile are caused by phytochemicals found on the surface of the nanoparticles. Bacteria have the genetic ability to transmit and acquire drug resistance to therapeutic drugs. Resistant bacteria pose a challenge in the treatment of various well-known infections and require new substances with antimicrobial properties. The composition of the remaining substances in EDX and their effects on antimicrobial purposes may have implications. FT-IR Analysis FT-IR analyses show that AgNPs are surrounded by biomolecules, reducing the potential for metal nanoparticles to come together and form clusters, and increasing their stability in the solution environment. The peaks in the FT-IR spectrum of C. draba plant extract indicate the presence of 3675.83 -OH and -NH (polyphenols and amine) groups. The peaks at 2988.26 and 2901.60 cm−¹ indicate the presence of –CH and -CH2 aliphatic groups, and the peaks at 1056.84 and 1028.01 cm−¹ indicate the presence of -C-O groups, namely terpenoids, flavones, and carbohydrate structures (Fig. 3). It has been reported that FT-IR peaks in

plant extract support the presence of antioxidant flavonoids and phenolic acids, and the presence of phenolics in the extract may be responsible for the reduction of metal ions and the formation of nanocomposites [36]. It was observed that AgNPs showed absorption peaks at 3675.72, 2988.77, 1513.93, 1393.93, 1230.26, 1141.96, 1066.27 cm−¹ (Fig. 3). The first peak is seen as a band around 3675 cm−¹. The peak observed in this region generally represents -OH (hydroxyl) groups. It shows that the AgNPs sample contains various organic functional groups and compounds. The presence of hydroxyl (-OH) and carbonyl (C=O) groups, in particular, suggests that there are water molecules, alcohol, a phenol, or aldehyde, or ketone, or other related compounds on the surface of the sample. The peak located around 1513 cm−¹ represents the stretching vibrations of carbonyl (C=O) and C=C double-bonded groups. Carbonyl groups can be found in aldehyde, ketone, or amide groups and the peak in this wavenumber region suggests the presence of such functional groups in the sample [37]. A comparison of the extract and nanoparticle peaks in FTIR analysis, conducted using Cornelian cherry fruit for the synthesis of AgNP, revealed that the 3159 cm-1 O-H, 1717 cm-1 C=O, 1224 cm-1 C-OO groups were the primary contributors to Ag reduction [37]. According to the FTIR analysis results, the -OH group at 3332 cm-1, the -C≡N group at 2145 cm-1, and the -C=O group at 1635 cm-1 participate in the synthesis process of nanoparticles [38]. FT-IR analysis shows that AgNPs have a complex chemical structure; organic and inorganic components are present on their surface. May have a suitable profile for use in potential biomedical and chemical applications.

Sigma J Eng Nat Sci, Vol. 43, No. 6, pp. 1943−1956, December, 2025

XRD Analysis Peaks from the (111), (200), (220), and (311) crystal structures were seen as a result of the resulting difraction patterns. These peaks corresponded to the angle values of 2θ=38.38°, 44.61°, 64.97°, and 78.09°, for metallic silver (JCPDS 04-0783) (Fig. 4). These findings led to the understanding that, in this context, a face-centered cubic structure contained metallic silver. Furthermore, the metals from the factory were identified as contaminants based on the difraction patterns. Using the Scherrer equation and the XRD data, the particle sizes of AgNPs were determined in this section of the study. The

particle size of AgNP is 18.51 nm according to the DebyeScherrer equation. According to Agarwal et al. (2018), who synthesized AgNPs from Cymbopogon citratus, the aforementioned peaks correspond to those of silver [39]. It was found that in Nigellla sativa plant, there were diffraction bands of (111), (200), (220) and (311) with 2ϴ values​​ around 38.35º, 44.50º, 64.76º and 77.80º [40]. The sharpness of the peaks reveals that the synthesized nanoparticles have a high crystalline structure. In addition, the absence of peaks belonging to any other phase in the graph supports the high purity of the synthesized nanoparticles.

Sigma J Eng Nat Sci, Vol. 43, No. 6, pp. 1943−1956, December, 2025

UV-Visible Spectral Analysis UV-Vis spectra of AgNPs synthesized from C. draba plants were taken. As seen in Figure 5, absorbance values are in the range of 400 to 500 nm. According to previous studies, the absorbance ranges of AgNPs generally occur at 400-500 nm. It has been reported that the transformation from a light yellow to brown colloidal solution during the reaction allows the qualitative detection of AgNPs. The color change is due to the plasmon resonances (SPR) on their surfaces and indicates an increase in the concentration of AgNPs over time. When literature findings are evaluated, they show that the synthesis was carried out successfully in the research. This analysis confirmed the literature indicating that silver nanoparticles typically exhibit maximum absorption

values at approximately 400 nm wavelength. AgNPs were synthesized with extracts obtained from Rubus ellipticus, and it was determined that the maximum absorbance value was obtained around 430 nm [41]. AgNPs were synthesized using Punica granatum extract. The maximum UV-vis absorbance value of the plant extract was determined to be 367 nm. The maximum UV-vis absorbance value of AgNPs was determined as 462 nm [42]. The greatest absorbance peak was observed in the range of 420–450 nm when AgNPs were produced from the Salvia limbata (sage) plant [43]. This finding substantiates the hypothesis that when exposed to the bioactive elements of the plant extract, the silver ion undergoes a transformation into silver nanoparticles [44].

Sigma J Eng Nat Sci, Vol. 43, No. 6, pp. 1943−1956, December, 2025

Results

Gas chromatography-mass spectrometry (GC-MS) was used to analyze the essential oils obtained from the aerial part of the C. draba plant. The analysis revealed that the allyl isothiocyanate compound was responsible for 74.07 percent of the total weight of the plant extract. After the allyl isothiocyanate compound, thiocyanic acid, 2-propenyl ester compound, comprising 8.45 percent of the weight, constituted a significant part of the plant extract . Other components in the extract were less than 2 percent of the total extract weight (Table 1). Chemical analysis of C. draba flower, leaf, stem, and root sections gathered from Iran revealed that 6-10, 14-Trimethyl-2-pentadecanone (16.4%) and 3-butenyl isothiocyanate (26.4%) were the most prevalent components [45]. Chemical analysis of C. draba aerial organs and leaves obtained from France revealed that 6,10,14-trimethyl-2-pentadecanone (20.61%) and (E)-phytol (11.38%) were the most significant components [46]. C. draba roots, stems, leaves, flowers, and fruits gathered from Tunisia were examined for their primary fragrance volatiles, which included hexadecanoic acid (34.6%), 6-methyl-5-hepten-2-one (18.3%), decanal (15.0%), 6,10,14-trimethyl-2-pentadecanone (13.2%), and n-pentacosane (13%) [47]. Liquid Chromatography–Mass Spectrometry (LC-MS/ MS) Results In the LC-MS/MS results of the C. draba, Vitexin compound is the phenolic compound found in the highest amount with 8561.6281 ng/mL. Luteolin is the second most abundant phenolic compound after Vitexin. Rosmarinic acid, which has a large amount of reactive functional groups, was found at a concentration of 1.0837 ng/mL. Further data on other reactive phenolic compounds can be reviewed in Table 2. Since the focus was on the most

abundant compounds in the plant extract, seven compounds are included in Table 2. Antibacterial Activity In the study, the paper disc method was used to examine the antibacterial activity of Cardaria draba extract and AgNPs biologically generated from Cardaria draba extract. The antibacterial impact of the extract on both Gram-positive and Gram-negative bacteria, including B. subtilis, S. aureus, and E. faecalis was investigated The antimicrobial activity tests yielded an inhibitory zone of 18–30 mm generated by chloramphenicol antibiotic discs in Petri dishes, which served as positive controls. The aqueous extract of Cardaria draba demonstrated a significant bactericidal effect on both Gram-negative and Grampositive bacteria, with zones of inhibition ranging from 6 mm to 9 mm. The most effective antibacterial dose against P. aeruginosa was 9 (Fig. 6). For S. aureus ATCC 25923 and E. faecalis ATCC 29212, AgNPs showed an inhibitory zone for the bacteria E. coli ATCC 25922, P. aeruginosa ATCC 27853, B. subtilis DSMZ 1971, and S. enteritidis ATCC 13075 (Table 3). When the antibacterial activity of silver nanoparticles synthesized from Trema orientalis (L.) was examined, it was found that silver nanoparticles formed a 9 mm zone at a concentration of 25 µg/mL, for S. aureus [48]. The antibacterial activity of silver nanoparticles synthesized using Ferula gummosa essential oil against Gram-positive and Gram-negative bacteria was shown with inhibition zones of 10 mm and 8 mm in diameter for E. coli and S. aureus, respectively [49]. In the antibacterial activity of silver nanoparticles made from Mirabilis jalapa leaf extract, Staphylococcus aureus showed a 10 ± 1.58 mm inhibition zone, Pseudomonas aeruginosa showed a 6 ± 1.15 mm inhibition zone, and Staphylococcus aureus AgNPs showed a 10 ± 1.58 mm inhibition zone [50]. Inhibition was observed in

Table 1. Biologically active chemical compounds of Cardaria draba aqueous extract Name of compounds (molecular formula)

Ethanone, 1-(1,3a,4,5,6,7-hexahydro-4-hydroxy-3,8-dimethyl-5-azulenyl)-

Propanoic acid, 2-methyl-, 3-hydroxy-2,4,4-trimethylpentyl ester

Sigma J Eng Nat Sci, Vol. 43, No. 6, pp. 1943−1956, December, 2025

Table 2. Most abundant phenolic compounds in quantitative analysis results Molecule name

Sigma J Eng Nat Sci, Vol. 43, No. 6, pp. 1943−1956, December, 2025

Table 3. Zone diameters (in mm) and mean ± SE values of AgNPs’ effects against bacterial strains, as determined by the disk diffusion method, are given in the table

the antibacterial effect of silver nanoparticles formed with the aqueous extract of Aquilegia pubiflora against E. coli (10.9 ± 0.29), B. subtilis (10.2 ± 0.31), Klebsiella pneumoniae (9.8 ± 0.33), S. epidermidis (9.3 ± 0.37), and P. aeruginosa (9.1 ± 0.34) [51]. The antibacterial impact of C. draba may be linked to the presence of phenolic chemicals, which have demonstrated antimicrobial action. The antibacterial activity of Brassicaceae members against clinical bacterial isolates was examined by Panghal [52]. The generally higher antibacterial activity of AgNPs may be due to the large surface area of nanoparticles, which provides better contact with microorganisms. According to the literature, the precise mechanism of AgNPs remains unclear. AgNPs have antibacterial properties because they create pits in bacterial cell walls [53].

Figure 6. a) Zone diameters of AgNPs against Gram-negative bacteria, b) Zone diameters of AgNPs against Gram-positive bacteria.

Enzyme Inhibition To observe the inhibitory effects of C.draba extract and AgNPs synthesized from it, tests were performed on three different enzymes: urease, collagenase, and lipoxygenase. Percentage inhibition and IC50 values ​​were determined. Effect on Urease The urease inhibition test demonstrated significant inhibitory effects of C. draba extract and AgNPs. C. draba extract showed 34.44% (IC50: 2.60± 1.32 µg/mL) inhibition at 2 µg/mL concentration. AgNPs showed 44.64% (IC50: 2.23± 1.65 µg/mL) inhibition at the same concentration. AgNPs produced with C. draba extract showed strong inhibitory activity against urease. Thiourea, when used as a positive control, showed strong inhibition of 57.77% (Table 4). Ovais et al. evaluated the capacity of the green synthesis of AgNPs to inhibit urease enzyme activity using Olax nana Wall. ex Benth. AgNPs showed significant inhibitory activity with a urease inhibition of 39.23 ± 0.42% at a concentration of 0.2 mg/mL [54]. In the study conducted by Kaya et al., when Thymbra spicata L subsp. spicata L was tested at a concentration of 100 µg/mL, urease enzyme inhibition was observed, reducing its activity by 34.62±2.33%, [55]. AgNPs and AuNPs were synthesized with Crataegus oxyacantha extract. Among the synthesized nanoparticles, AgNPs showed inhibitory activity with an IC50 value of 1.38 ± 0.3 µM. The −NH, −OH, and C=O groups of the stabilizing phytochemicals, which interact with Ni in the urease enzyme, may be the cause of the notable urease inhibitory actions of NPs [56]. Effect on Collagenase Collagenase inhibition test showed significant inhibitory effects of C. draba extract and AgNPs. C. draba extract showed 12.59% inhibition at a 2 µg/mL concentration. AgNPs showed 35.55% inhibition at the same concentration. AgNPs produced with C. draba extract showed strong inhibitory activity against collagenase. EGCG (Epigallocatechin gallate), used as a positive control, showed 48.14% inhibition (Table 4). In the study by Kaya et al., Thymbra spicata L. subsp. spicata L. was determined to exhibit a low collagenase enzyme inhibitory activity (10.57±0.30%) [57]. Radwan et al.[ 58] stated that

Sigma J Eng Nat Sci, Vol. 43, No. 6, pp. 1943−1956, December, 2025

the nanoparticles they obtained inhibited the collagenase enzyme by an average of 75%. AgNPs obtained from F. vesiculosus have been shown to have stronger anti-collagenase activity than F. vesiculosus extract. Terpenoids and flavonoids, two naturally occurring antioxidants, work in concert to block the collagenase enzyme [59]. It may be claimed that various plant extracts are the source of bioactive compounds that can block the activity of the same enzymes, which is consistent with the findings of Andrade et al. (2021) [60] and Gomes et al. (2022) [61]. Effect on Lipoxygenase (LOX) The lipoxygenase inhibition test showed significant inhibitory effects of C. draba extract and AgNPs. C . draba extract showed 27.40% inhibition at 2 µg/mL concentration. AgNPs showed 38.88% inhibition at the same concentration. AgNPs produced with C .draba extract showed strong inhibitory activity against lipoxygenase. Nordihydroguaiaretic acid (NDGA), used as a positive control, showed strong inhibition of 54.44% (Table 4). This study did not show any LOX enzyme inhibition activity. This study highlights the possible variation in enzyme

inhibitory activity based on elements including the plant’s particular composition, the plant material’s place of origin, and the experimental setup. Cytotoxicity Analysis The cytotoxicity of AgNPs was investigated. Figure 7 illustrates the cell viability rates for the DLD-1 colorectal adenocarcinoma cell line when AgNPs was administered at varying concentrations (2500, 1250, 625, and 312.5 ppm). The lowest concentration of AgNPs applied in the study, when compared to the control group, was 312.5 ppm. The remaining applied concentrations were observed to exert a diminished cytotoxic effect. The lowest concentration in the AgNPs, that was found to be less cytotoxic than the control group, was determined to be 312.5 ppm. Furthermore, concentrations of 625 and 1250 ppm were demonstrated to be less cytotoxic than the specified threshold value. The human body’s digestive system is capable of transporting nanoparticles into the bloodstream, where they can subsequently be delivered the liver, brain, and other organs. Nanoparticles have the potential to deliver therapeutic medications to the brain and other organs [62]. The decrease

Table 4. Enzyme inhibitory activity of extracts and AgNPs from C. draba Enzyme Urease

Conflict Of Interest

The author declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Ethics

There are no ethical issues with the publication of this manuscript.

Statement On The Use Of Artificial Intelligence

Artificial intelligence was not used in the preparation of the article.

Share and Cite