Halpeh Extract-Derived Gold Nanoparticles: A Dual Force Against Microbes and for Phytochemical Enhancement

Halpeh Extract-Derived Gold Nanoparticles: A Dual Force Against Microbes and for Phytochemical Enhancement | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Halpeh Extract-Derived Gold Nanoparticles: A Dual Force Against Microbes and for Phytochemical Enhancement Ehsan Dahaz, Samer Asadi, Somayeh Lashgari This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4587822/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The increasing resistance of pathogenic microorganisms to conventional antibiotics has led to a growing interest in alternative antimicrobial agents. In this study, the antimicrobial activity (antibacterial and antifungal) of aqueous and methanolic extracts of Halpeh plant and gold nanoparticles synthesized using the extract of this plant was investigated. Additionally, the effect of different concentrations of synthesized gold nanoparticles on the phytochemical properties (antioxidant and reducing ability) of Halpeh plant extract was examined. The results of UV-Vis (Ultraviolet-Visible Spectroscopy), XRD (X-Ray Diffraction Analysis), TEM (Transmission Electron Microscopy), SEM (Scanning Electron Microscopy), and FTIR (Fourier Transform Infrared Spectroscopy) tests on gold nanoparticles confirmed their synthesis. SEM and TEM images showed that the gold nanoparticles were spherical in shape with an average size of 22.89 nm. According to the results, the methanolic extract exhibited significantly higher antibacterial and antifungal activity compared to the aqueous extract, with the maximum inhibition zone diameters observed for Escherichia coli (14 mm) and Aspergillus niger (15 mm). Moreover, the antimicrobial activity of gold nanoparticles showed that these nanoparticles have relatively good ability to inhibit gram-negative bacteria, with the largest inhibition zone diameters observed for Escherichia coli (18 mm) and Aspergillus niger (20 mm). The antioxidant and reducing power activity (phenolic flavonoids content) of Halpeh plant extract treated with different concentrations of synthesized gold nanoparticles increased with increasing nanoparticle concentrations up to 60 ppm (IC50 = 9.94 µg/ml and reducing power = 16.85 mMFe2+/mg sample), and decreased at higher concentrations. Gold nanoparticles Hellebore plant extract Antibacterial Antifungal Antioxidant activity Reducing power Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Introduction In recent years, the increasing prevalence of antibiotic-resistant bacteria has become a global health concern. These microorganisms, including bacteria, viruses, parasites, and fungi, have developed resistance to commonly used drugs. Their ability to adapt and evolve poses a significant challenge to healthcare systems, pharmaceutical companies, and regulatory agencies. Among various strategies to combat this issue, the proper use of antibiotics in pharmacies, hospitals, and community settings plays a crucial role in preventing the spread of resistance. Monitoring the availability of over-the-counter (OTC) drugs in pharmacies, adherence to prescription guidelines, and educating healthcare professionals are essential steps in curbing antibiotic resistance [ 1 ]. Increasing antibiotic stability in pathogenic microbes is one of the major problems facing medical science [ 2 – 4 ]. The value of plants is not only in providing food for humans and animals, but also healing and relieving a lot of human pain and needs that have been and will be found in plants since the beginning of creation. Research in the field of drugs of plant origin is in the realm of pharmacognosy and the origin of its earliest dates can be seen in the history of the customs of the ancient tribes and civilizations that provided healing solutions from plant components [ 5 ]. Medicinal herbs with antimicrobial effects and fewer side effects are a good choice than synthetic antimicrobial treatments [ 6 – 9 ]. The therapeutic properties of plant extracts against microbial and non-microbial diseases have been known since ancient times, and many studies were carried out on plant species and their essential oil effects on microorganisms. In fact, the antimicrobial, antiviral and anti-inflammatory effects of medicinal plants are due to the phytochemical properties of their extracts [ 10 ]. Therefore, the extract of medicinal plants can be used to fight pathogenic microorganisms. Nanotechnology is considered as one of the most active research areas in materials science. Nanoparticles have completely new or improved properties that are defined based on certain characteristics such as size, shape, crystalline structure, and morphology. When metals are defined at the nanoscale, they exhibit unique properties. In addition to the inhibitory effect of the particle, metal nanoparticles have a high antimicrobial effect due to their small size, large surface area to volume ratio and cause more contact with outer space [ 11 ]. One of these metals that has been considered in the field of nanotechnology in recent years is gold (Au). Gold at the nanoscale exhibits properties that make it an important metal in nanotechnology processes. 12–14 Due to the ease of supply, gold nanoparticles are more widely used than other nanoparticles. Due to the growing application of gold nanoparticles and the growing need to provide methods for providing nanoparticles that do not involve environmental pollution, the necessity of producing these nanoparticles is felt by biological systems [ 12 ]. Gold nanoparticles have good antimicrobial and antibacterial properties that resist human infectious agents [ 15 ]. The antimicrobial properties of gold nanoparticles include a wide range of bacteria. Gold nanoparticles are synthesized by extracellular and intracellular methods, and the results of experiments conducted so far show that extracellular nanoparticles have better antibacterial effect on a scale with Nanoparticles produced are in a cell manner and their ability to inhibit the growth of pathogenic bacteria is much greater [ 12 , 15 ]. Today, with the advancement of technology, it is possible to produce nanoparticles from gold with different biological methods that have good antimicrobial properties and show good resistance to Gram-positive and negative bacteria [ 16 ]. Conventional physical and chemical methods used to synthesize metal nanoparticles also bring toxic chemicals into the environment, in addition to being expensive. Biological methods such as the use of microorganisms and plant extracts can be a good alternative to conventional methods of synthesizing nanoparticles because they are simple, inexpensive, biocompatible and have the ability to produce nanoparticles on a large scale [ 17 , 18 ]. The synthesis of nanoparticles using different types of microorganisms is used in medicine because of their biocompatibility. However, the production of microorganisms is often more time-consuming, and it is more expensive than obtaining plant extracts. The use of plants in comparison to microorganisms eliminates the time required for cell culture and therefore has a high capacity for large-scale synthesis [ 19 , 20 ]. Considering that the extracts of medicinal herbs and metal nanoparticles, especially gold nanoparticles, have good antimicrobial effects. Therefore, the synthesis of gold nanoparticles using the extract of these plants can enhance their antimicrobial effects. Numerous studies have been done on the synthesis of metal nanoparticles with plant extracts, which have been found that nanoparticles are reduced by plant extracts [ 21 – 24 ]. The decreasing factor in plant extracts is due to phytochemical substances in them, especially phenolic compounds. By improving the phytochemical properties of plants, in addition to increasing their antimicrobial properties, nanoparticles with highly desirable purity and concentration can be synthesized. These phytochemicals have properties in plants Prevention of Diseases [ 10 , 25 ]. Plants produce phytochemicals to protect themselves. Recent research has shown that these substances are also useful for protecting humans from disease. In 2024, Rodolfo et al. conducted a study on the biosynthesis of gold nanoparticles using an extract of Anemopsis Californica . They were able to synthesize gold nanoparticles of 5 to 30 nm and concluded that these nanoparticles have good antibacterial properties, especially against Gram-positive bacteria [ 26 ]. In 2023, Tran et al. synthesized gold nanoparticles using an extract of Andrographis Paniculata leaves in an average size of 10 to 15 nm, which had desirable antibacterial properties [ 27 ]. In this study, gold nanoparticles were synthesized using the extract of the Halpeh plant for the first time. The antimicrobial activity (including antibacterial and antifungal properties) of both aqueous and methanolic extracts of the Halpeh plant, as well as the synthesized gold nanoparticles, was investigated and compared. Additionally, the phytochemical changes (antioxidant and reducing ability) of the extract of Halpeh plant is investigated by treatment with different concentrations of synthesized gold nanoparticles. Halpeh , a herbaceous plant reaching heights of 10 to 35 cm, belongs to the mint family and is characterized by its white, cotton-like appearance. It commonly grows in food-impoverished regions, rocky beaches and sand fields of different parts of Europe, the Mediterranean region, North Africa and southwest Asia, including Iran. This plant is widely distributed in south and central Iran. Its leaves are narrow, long, and covered with cotton cracks. Its flowers are also seen in white, yellowish white, yellow or even purple. It is a powerful reliever for gastrointestinal pains such as indigestion. The flowering branches of the plant have tonic, vigorous and anticonvulsant effects. In addition, the fever is high, and the boiled plant is used in cases of colds. More importantly, it is very useful in regulating kidney function, relieving asthma, increasing appetite and blood sugar. The use of the hemp plant in this study was collected from mountainous areas of Dashti County, Bushehr Province (Fig. 1). Experimental Chemicals All chemicals used in this study, which were of high purity and obtained from the reputable company Merck, included 2,2-diphenyl-1-picrylhydrazyl (DPPH), sodium hydroxide, hydrochloric acid, acetic acid, ascorbic acid (ASA), butylated hydroxytoluene (BHT), iron(III) chloride, iron(II) chloride, aluminum chloride, sodium carbonate, sodium acetate, gallic acid, quercetin, Folin’s reagent, iron sulfate, 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ), methanol, water, ethanol, and gold(III) chloride (HAuCl4). Devices The UV-Vis spectrophotometer model 2100 made by Unico company was used for recording absorptions. The pH measurement was also performed using the pH meter model pH lab 827. Additionally, for weight measurements, a digital scale with a precision of 0.0001 grams, model Kern, was used. For separation, the centrifuge model Sigma 2–7 was used; for dissolving extracts, the ultrasonic model SB-100D was used; for essential oil extraction, the Clevenger apparatus was used; and for the analysis of the essential oils, the gas chromatography system model GC-7890B connected to the mass spectrometer model MS5977A was used. Other devices used included the XRD model PW1730 from PHILIPS, FT-IR model AVATAR from Thermo, SEM model VEGA3 from TESCAN, and TEM model CM120 made by Philips Holland. Water Extract Preparation The process of extracting the aqueous extract was carried out from the aerial parts of the plant known as “ Halpeh .” The plant extract is used for the reduction of gold ions. Therefore, precision in the extraction methods to obtain a pure extract is very important and noteworthy. For this purpose, approximately 10 grams of the powdered plant were carefully weighed using a digital scale and transferred into an Erlenmeyer flask containing about 100 milliliters of doubly ionized distilled water. Then, it was stirred for half an hour at a temperature of 30–40 ºC using an electric mixer and, after cooling, was filtered through Whatman filter paper No. 42, resulting in the aqueous extract of the mentioned plant. This extract contains a series of impurities that may originate from the filter paper, as it cannot separate all the components of the plant. For this reason, to remove the impurities, a centrifuge was used at 10,000 revolutions per minute for a duration of 10 minutes. Synthesis of Gold Nanoparticles by Aqueous Extract of Halpeh Plant For synthesis of gold nanoparticles, 1 ml of aqueous extract of the plant and 8 mL of 1 mM HAuCl4 solution were poured into 10 mL and placed on magnetic stirrer. After about an hour at room temperature, it was observed that the color of the solution had changed to purple, which itself indicated the formation of gold nanoparticles (Fig. 2). Aqueous extract of this plant has been used as a reducing agent and stabilizer for synthesis of gold nanoparticles. Optimization of pH of reaction solution In order to optimize the acidity of the reaction to obtain the most favorable conditions for the synthesis of gold nanoparticles, solutions including 1 ml of Halpeh extract and 8 ml of gold salt solution with a concentration of 1 mM with pH (2, 3, 4, 5, 6 and 8) were fabricated according to Table 1 and the absorption spectra of all of them were taken by UV-Vis and the optimum pH was selected. Their acidity was adjusted by the base solution and sodium hydroxide and hydrochloric acid with a concentration of 0.1 M. As shown in Table 1, the highest absorption rate of UV-Vis has been obtained at pH 5, which indicates the synthesis of nanoparticles with higher quality and optimum concentration. Table (1): Solutions made to optimize pH for synthesis of gold nanoparticles Soluble Series Extract Volume (ml) Volume of HAuCl 4 Solution (ml) Total volume (ml) pH Absorbance 1 1 8 10 2 0.297 2 1 8 10 3 0.832 3 1 8 10 4 0.859 4 1 8 10 5 1.194 5 1 8 10 6 1.069 6 1 8 10 8 0.190 Preparation of aqueous and methanolic extracts from plant for evaluation of antimicrobial activity In order to prepare aqueous and methanolic extracts of Halpeh plant for evaluation of their antimicrobial activity, 10 g of plant powder was added to 100 ml of water and methanol solvent and stirred on shaker for 24 hours and smoothed with 42 wattman filter paper. Then, to remove impurities, a centrifugal device was used at a speed of 10,000 minutes and 15 minutes. The extracts were stored at 4°C for later use. Antimicrobial activity of synthesized gold extracts and nanoparticles The antimicrobial activity of extracted gold extracts and nanoparticles was evaluated by disk diffusion method (NCCLS115), on gram-positive bacteria ( Staphylococcus aureus and Bacillus cereus ), gram-negative bacteria ( Escherichia coli and Salmonella typhimurium ) and fungi ( Aspergillus niger and Candida albicans ). First, each bacterium and fungi were cultured in Nutrient Broth medium (37°C, shaker at 200 rpm for 24 hours). After 24-hour culture, a suspension equivalent to half of McFarland concentration was prepared and each suspension was inoculated with a volume of 1 ml (108 cfu/ml) of Muller Hinton agar medium and cultured with swaps. Afterwards, paper discs (6 mm) impregnated with a solution of gold nanoparticles, aqueous and methanolic extracts were placed on the culture site of the relevant bacteria and fungi. Finally, the plates were placed in an incubator with a temperature of 37°C and the diameter of the inhibition zone around each disk was measured after 24 to 48 hours. In order to compare the antimicrobial effect of gold nanoparticles, aqueous and methanolic extracts against the tested bacteria and fungi, their sensitivity to antibiotics discs of ampicillin, gentamicin and clotrimazole was also investigated. Effect of gold nanoparticles on phytochemical properties of Halpeh plant Before flowering stage, the spray of gold nanoparticles on the aerial organs of the plant was performed. First, gold nanoparticles were synthesized under optimum conditions (by adding 1ml of aqueous extract of Halpeh plant with 8 ml of HAuCl4 (1 mM) salt solution at pH = 5) and their concentration (C) was determined using the Eq. ( 1 ). $$C={N_{total}}/N*V$$ 1 N total : The sum of gold atoms N: The average number of gold atoms per nanoparticle V: The final volume The Eq. (2) is used to obtain the average number of gold atoms in each nanoparticle (N). N A : Avogadro number \(\pi\) : The number pi D: Medium Diameter of Nanoparticles \(\rho\) : : Density of gold atom (g/cm 3 ) M: Atomic weight By placing \(\rho =19.3{\text{ g/c}}{{\text{m}}^3}\) ، \(M=196.96{\text{ g/gmole}}\) and the average diameter of the nanoparticles determined using the TEM image, the average number of gold atoms per nanoparticle (N) is obtained. Then, in Eq. ( 1 ), the concentration of nanoparticles was obtained about 315 ppm which was considered as the standard concentration, then concentrations of 0, 20, 40, 60 and 80 ppm were fabricated and sprayed on the aerial parts of the plant during three weeks. Results and Discussion UV-Vis Analysis Results During the synthesis process, Au + ions are exposed to reducing agents in the plant extract. This initiates the reduction of gold nitrate salts. Complete reduction of Au + ions to gold nanoparticles was achieved, as evidenced by the color change in the environment. This process occurred approximately five minutes after the reaction began. Figures (3) corresponds to the UV-Vis spectroscopic analysis of the solution containing the Halpeh plant extract and HAuCl4 solution. As observed in this figure, the spectrum at a wavelength of approximately 420 nm exhibits a peak (indicating maximum absorption), which confirms the synthesis of gold nanoparticles [ 28 ]. XRD Analysis Results The XRD pattern presented in Figure (4) is a testament to the crystalline nature of the gold nanoparticles synthesized through an environmentally benign process. The distinct peaks corresponding to the Miller indices (111), (200), (220), and (311) are indicative of a well-defined face-centered cubic (FCC) crystalline structure, which is characteristic of elemental gold [ 29 ]. The specific 2θ angles of 38.04°, 44.5°, 64°, and 77.7° align perfectly with the standard reference, confirming the high crystallinity of the nanoparticles. The absence of any superfluous diffraction peaks is a clear indicator of the high purity of the gold nanoparticles. This suggests that the synthesis process, aided by the Halpeh plant extract, is not only effective in reducing the gold ions to their nanoparticle form but also in preventing the formation of impurities or by-products. The calculated average size of 22.8 nm for the gold nanoparticle crystals, derived from the Debye-Scherrer equation, points to a uniform and controlled nanoparticle formation. This size is ideal for a multitude of applications, ranging from medical to electronic, due to its potential for high surface area and reactivity. Moreover, the calculated d-spacing of approximately 2.5 nm for the (111) plane further corroborates the FCC structure and is consistent with the expected values for gold nanoparticles. The ICDD (International Centre for Diffraction Data) reference number PDF (Powder Diffraction File) # 04-0784 serves as a benchmark for comparing and validating the crystal structure, ensuring that the synthesized nanoparticles conform to the recognized standards for gold. The successful synthesis of gold nanoparticles using the Halpeh plant extract highlights the precision of the green synthesis approach. These nanoparticles, with their confirmed structure and high purity, hold promise for diverse functional applications. FTIR Analysis Results The FTIR spectroscopy provides insightful details about the molecular interactions within the synthesized gold nanoparticles (Figure (5)). The observed shift in the O-H absorption band from 3438 to 3428 cm − 1 suggests a strong interaction between the phenolic compounds in the Halpeh plant extract and the surface of the gold nanoparticles [ 30 ]. This shift could indicate the formation of a new chemical environment or bond formation, which stabilizes the nanoparticles. The presence of distinct absorption bands at 2929, 1624, 1389, and 1278 cm − 1 in both the plant extract and nanoparticle spectra reveals the retention of functional groups such as C-H, C = O, C-N, and type I amide after the synthesis process. These functional groups are likely involved in the capping and stabilization of the gold nanoparticles, preventing agglomeration and ensuring uniformity in size. Moreover, the absorption bands at 1042 and 1058 cm − 1 associated with ether compounds (C-O-C) further confirm the attachment of plant-derived organic moieties on the nanoparticle surface. This attachment could play a crucial role in enhancing the biocompatibility and potential biomedical applications of the synthesized nanoparticles. The FTIR analysis thus not only confirms the successful synthesis of gold nanoparticles using the Halpeh plant extract but also highlights the critical role of plant-based compounds in imparting stability and functionality to the nanoparticles. SEM Analysis Results Figure (6) showcases the SEM analysis of the synthesized gold nanoparticles, providing a detailed view of their three-dimensional structure. The spherical morphology of the nanoparticles is clearly visible, which is a common and desirable shape in nanotechnology due to its large surface area to volume ratio. This spherical form can be advantageous for applications such as drug delivery, where uniformity in particle size and shape facilitates better control over the release rates of therapeutic agents. The observed agglomerations, typical in nanoparticle synthesis, indicate interactions that can be effectively dispersed via sonication. The SEM images also provide insights into the efficacy of the synthesis method. The relatively uniform size and shape of the nanoparticles indicate a controlled synthesis process, which is essential for ensuring reproducibility and reliability in subsequent uses. TEM Analysis Results Figure (7) illustrates the TEM analysis of the gold nanoparticles synthesized using the Halpeh plant extract, demonstrating their spherical shape with diameters less than 50 nanometers. Figures (8), which presents a histogram of the size distribution of the gold nanoparticles, indicates an average diameter of 22.89 nm. The TEM images provide valuable insights into the uniformity and quality of the nanoparticles. The spherical morphology is advantageous for many applications due to its symmetrical properties and even distribution of surface area. The size distribution histogram further confirms the consistency in nanoparticle size, which is crucial for reproducibility in industrial and research applications. The sub-50 nm size range of these nanoparticles makes them suitable for high-precision tasks, including targeted drug delivery and sensing applications, where small size and large surface area are beneficial. Differences in sample preparation and imaging techniques contribute to discrepancies in nanoparticle size measurements between TEM and SEM [ 31 ]. The results of the antimicrobial activity of the aqueous and methanolic extracts of the Halpeh plant and the synthesized gold nanoparticles This research focused on the antibacterial and antifungal effects of extracts and synthesized gold nanoparticles, and their comparative efficacy against three antibiotics: ampicillin, gentamicin, and clotrimazole. The study found that these extracts and nanoparticles demonstrated a commendable ability to inhibit the growth of the tested pathogenic microorganisms, suggesting their potential as alternative antimicrobial agents. As per the data presented in Figure (9) and Table (2), the synthesized gold nanoparticles exhibited significantly superior antibacterial and antifungal activity compared to the aqueous and methanolic extracts. This was particularly evident in the case of the gram-negative bacteria Escherichia coli and the fungus Aspergillus niger, where the zones of inhibition were notably larger (18mm and 20mm respectively) [ 32 ]. This enhanced antimicrobial effect of the gold nanoparticles could potentially be attributed to their combination with the extract of the medicinal plant Halpeh . This suggests that the integration of traditional medicinal plant extracts with modern nanotechnology could lead to more potent antimicrobial treatments. Furthermore, the study inferred from the data that the synthesized nanoparticles were effective against all tested microbes. This is a significant finding as it highlights the broad-spectrum antimicrobial activity of the nanoparticles. In contrast, each of the antibiotics was found to be ineffective against some of the microbes. This underscores the potential of these nanoparticles as a more effective antimicrobial agent, capable of overcoming the limitations of traditional antibiotics. Table (2): Results of the antimicrobial activity of the aqueous and methanolic extracts of the Halpeh plant and synthesized gold nanoparticles compared with antibiotics. Microorganism The diameter of the growth inhibition zone (mm) Biosynthesized Au NPs Aqueous Extract Methanolic Extract Ampicillin Gentamicin Clotrimazole Escherichia coli 18 6 14 26 28 - Salmonella Typhimurium 16 6 11 27 29 - Staphylococcus aureus 14 8 13 25 26 - Bacillus cereus 15 7 12 27 27 - Aspergillus niger 20 10 15 - - 30 Candida albicans 19 9 13 - - 28 The Effects of Gold Nanoparticles on Phytochemical Compounds in Halpeh Plant Extract After extracting the essence from the Halpeh plant using a Cloninger apparatus, the results of analyzing the plant essence through gas chromatography-mass spectrometry (GC-MS) are presented in Table (3). Various concentrations of gold nanoparticles (0, 10, 20, 40, 60, and 80 ppm) have significantly impacted the yield and composition of the plant essence. Specifically, the changes in Halpeh plant essence yield were 1.10, 0.118, 0.125, 0.133, 0.151 and 0.113% for the respective concentrations. Figure (10) illustrates the comparison of the composition percentages in the Halpeh plant essence under the influence of different concentrations of gold nanoparticles. The major constituents of the essence, such as geranyl acetate, showed the following percentages: 8.131, 6.147, 8.280, 12.418, and 5.629% with the highest percentage observed in the 60-ppm treatment. Regarding Citral (E), its percentages were 29.650, 16.044, 25.122, 8.748, 33.075 and 18.96%, with the highest percentage found in the 60-ppm treatment (33.075%). Citral (Z) had percentages of 22.437, 19.457, 17.394, 20.336, 23.281, and 13.130%, with the highest observed in the 60-ppm treatment (23.281%), although there was no significant difference compared to the 0-ppm treatment. Additionally, Piperitone had percentages of 6.912, 8.030, 9.003, 3.758, 4.725, and 3.733% with the highest percentage observed in the 20-ppm treatment (9.003%). This value showed a noticeable difference from the 0-ppm treatment, and in all cases, the 80-ppm treatment had lower compound percentages. The percentage of Geraniol composition was 14.666, 9.818, 11.715, 8.747, 13.260, and 7.431% with the highest percentage in the 0-ppm treatment. As a result, the percentage of geraniol composition has relatively decreased under the influence of different concentrations of gold nanoparticles. Furthermore, the phytochemical compounds Phytol, Lianool, and cis-verbenol in the plant essence had percentages of 0.929, 2.310, and 1.919% respectively, in the 60-ppm treatment, which were higher than in other treatments. In summary, gold nanoparticles can act as a catalyst, influencing the extraction of secondary compounds in Halpeh plant essence. Depending on the specific compound type, they may either enhance or reduce the synthesis of these secondary constituents. Table (3): Composition Percentages of Halpeh Plant Essence Constituents Affected by Different Synthesized Gold Nanoparticle Concentrations Compound Name The Composition Percentage at the Concentration of 0 ppm The Composition Percentage at the Concentration of 10 ppm The Composition Percentage at the Concentration of 20 ppm The Composition Percentage at the Concentration of 40 ppm The Composition Percentage at the Concentration of 60 ppm The Composition Percentage at the Concentration of 80 ppm Octan 1.600 1.308 13.61 0.385 - 21.032 1-Octen-3-ol 0.77 0.396 0.302 0.695 0.592 - 6-Methyl-5-hepten-2-one 1.036 0.638 0.949 0.999 0.815 1.035 Decane 1.15 - 4.57 0.645 0.595 7.436 2-Cyclohexen-1-one, 3-methyl 0.713 0.322 0.344 0.537 0.467 - Linalool 0.783 0.524 0.443 0.529 0.929 - Spiro[4.5]decan-7-one, 1,8-dimethyl-8,9-epoxy-4-isopropyl 0.603 0.498 0.195 - - - Linalool formate 0.768 - - 0.592 0.670 - D(+)-10-Camphorsulfonyl chloride 0.404 - - 0.582 0.479 - Gerinic acid - - 0.986 3.133 - - Cis-Geranyl acetate - - - 2.201 0.616 - Caryophyllene - - 0.180 - - 0.548 Cubebene - - - - - 1.099 Results of Investigating Antioxidant Properties and Reducing Power of Halpeh Plant Extract under Different Synthesized Gold Nanoparticle Treatments The Half-Maximal Inhibitory Concentration (IC50) of Halpeh plant extract under various gold nanoparticle treatments with concentrations of 0, 10, 20, 40, 60, and 80 ppm is demonstrated in Figure (11) using the DPPH radical scavenging method. As observed in this figure, the IC50 values for the plant extract in different gold nanoparticle treatments 11.61, 10.48, 8.98, 9.94, and 12.67 µg/ml . Consequently, the antioxidant properties of the Halpeh plant extract increased in the 10, 20, 40, and 60 ppm treatments compared to the untreated sample (0 ppm). However, in the 80-ppm treatment, the antioxidant activity significantly decreased. Notably, in this test, the plant extract treated with gold nanoparticles at a concentration of 40- ppm exhibits the highest antioxidant properties with an IC50 value of 8.98 µg/ml relative to other treatments. The results indicate that the impact of gold nanoparticles on the antioxidant properties of Halpeh plant extract in the DPPH method is bidirectional. Up to a specific concentration, it enhances and stimulates the plant’s antioxidant properties. The reducing power of Halpeh plant extract under treatment with synthesized gold nanoparticles at concentrations of 0, 10, 20, 40, 60, and 80 ppm is shown in Figure (12) based on the calibration curve obtained from the standard FeSO 4 .7H 2 O (FRAP method). The reducing power of the plant extract has increased in the 10, 20, 40, and 60 ppm treatments compared to the 0-ppm treatment, while it has decreased in the 80-ppm treatment. Specifically, in the 40 and 60 ppm treatments, the plant extract demonstrates the highest reducing power with 16.71 and 16.85 mMFe2+ /mg sample, respectively. These results indicate that the impact of gold nanoparticles on the reducing properties of the plant extract in the FRAP method is bidirectional, and up to a specific concentration, it enhances and stimulates the extract’s reducing power, confirming their antioxidant properties. Conclusion Nanotechnology’s pivotal role in plant biology involves using nanomaterials to boost secondary metabolites and modify plant physiology. These materials can target genes linked to metabolite biosynthesis and plant growth. This research pioneered the green synthesis of gold nanoparticles with Halpeh plant extract as a reducing agent. Characterization confirmed their spherical, face-centered cubic structure, averaging 22.89 nm in size and peaking at 420 nm UV-Vis absorption. The study assessed the antimicrobial efficacy of these nanoparticles and Halpeh plant extracts against select bacteria and fungi, alongside phytochemical changes at varying nanoparticle concentrations (0–80 ppm) using DPPH and FRAP assays. The methanolic extract outperformed the aqueous extract in antimicrobial activity, with the gold nanoparticles showing even greater efficacy, notably against Escherichia coli and Aspergillus niger. Phytochemical analysis revealed that antioxidant and flavonoid levels increased with nanoparticle treatments up to 60 ppm but decreased at 80 ppm. The 40-ppm concentration yielded the highest reducing power in the plant extract. Abbreviations Declarations Authors' contributions S. A. and S. L. wrote the main manuscript text and E. D. prepared Figures and Tables. S. A. and S. L. reviewed the manuscript. Funding The authors did not receive support from any organization for the submitted word. Availability of data and materials All data generated or analyzed in this study are included in this article. Ethics Declaration Conflicts of interest/Competing interest The authors have no relevant financial or non-financial interests to disclose. There is no conflict of interest to report. 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Biosensors and Bioelectronics: X 17, 100457. https://doi.org/10.1016/j.biosx.2024.100457. Khan MAR, Al Mamun MS, Habib MA, Nazmul Islam ABM, Mahiuddin M, Karim KMR, Naime J, Saha P, Kumar Dey S, Ara MH (2022) A review on gold nanoparticles: Biological synthesis, characterizations, and analytical applications . Results in Chemistry 4, 100478. https://doi.org/10.1016/j.rechem.2022.100478. Gnanadesigan M, Anand M, Ravikumar S, Maruthupandy M, Vijayakumar V, Selvam S, Dhineshkumar M, Kumaraguru AK (2011) Biosynthesis of silver nanoparticles by using mangrove plant extract and their potential mosquito larvicidal property. Asian Pacific Journal of Tropical Medicine 4: 799- 803. https://doi.org/10.1016/S1995-7645(11)60197-1. Sivakama Valli J, Vaseeharan B (2012) Biosynthesis of silver nanoparticles by Cissus quadrangularis extracts. Materials Letters 82: 171- 173. https://doi.org/10.1016/j.matlet.2012.05.040. 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Soni V, Raizada P, Singh P, Cuong HN, Rangabhashiyam S, Saini A, Saini RV, Van Le Q, Nadda AK, Le TT, Nguyen VH (2021) Sustainable and green trends in using plant extracts for the synthesis of biogenic metal nanoparticles toward environmental and pharmaceutical advances: A review . Environmental Research 202, 111622. https://doi.org/10.1016/j.envres.2021.111622. Nazri MKHM, Sapawe N (2020) A short review on green synthesis of iron metal nanoparticles via plants extracts . Materials Today: Proceedings 31: 48-53. https://doi.org/10.1016/j.matpr.2020.10.968. Dubey SP, Dwivedi AD, Lahtinen M, Lee C, Kwon YN, Sillanpaa M (2013) Protocol for development of various plants leaves extract in single-pot synthesis of metal nanoparticles . Spectrochimica Acta, Part A: Molecular and Biomolecular Spectroscopy 103: 134-142. https://doi.org/10.1016/j.saa.2012.11.021. Guo B, Yu Y, Wang M, Li R, He X, Tang S, Liu Q, Mao Y (2024) Targeting the JAK2/STAT3 signaling pathway with natural plants and phytochemical ingredients: A novel therapeutic method for combatting cardiovascular diseases . Biomedicine & Pharmacotherapy 172, 116313. https://doi.org/10.1016/j.biopha.2024.116313. Ávila-Avilés RD, Argueta-Figueroa L, García-Contreras R, Vilchis-Nestor AR (2024) Synthesis of biogenic silver and gold nanoparticles from Anemopsis californica extract with antibacterial and cytotoxic activities . Materials Today Communications 38, 108071. https://doi.org/10.1016/j.mtcomm.2024.108071. Dat TD, Cong CQ, Nhi TLH, Khang PT, Nam NTH, Tinh NT, Hue DT, Hieu NH (2023) Green synthesis of gold nanoparticles using Andrographis paniculata leave extract for lead ion detection, degradation of dyes, and bioactivities . Biochemical Engineering Journal 200, 109103. https://doi.org/10.1016/j.bej.2023.109103 . Lee KX, Shameli K, Yew YP, Teow SY, Jahangirian H, Rafiee-Moghaddam R, Webster TJ (2020) Recent Developments in the Facile Bio-Synthesis of Gold Nanoparticles (AuNPs) and Their Biomedical Applications. International Journal of Nanomedicine 15: 275-300. https://doi.org/10.2147/IJN.S233789. Hutchinson N, Wu Y, Wang Y, Kanungo M, DeBruine A, Kroll E, Gilmore D, Eckrose Z, Gaston S, Matel P (2022) Green Synthesis of Gold Nanoparticles Using Upland Cress and Their Biochemical Characterization and Assessment. Nanomaterials 12, 28. https://doi.org/10.3390/nano12010028. Susanna D, Balakrishnan RM, Ettiyappan JP (2023) Ultrasonication-assisted green synthesis and characterization of gold nanoparticles from Nothapodytes foetida: An assessment of their antioxidant, antibacterial, anticancer and wound healing potential. Journal of Drug Delivery Science and Technology 87, 104740. https://doi.org/10.1016/j.jddst.2023.104740. Safari-Talab A, Asadi S, Lashgari S (2024) From nature to nanoparticles: Synthesizing silver nanoparticles from Moortalkh plant leaves with potent antibacterial properties. Inorganic Chemistry Communications 165, 112458. https://doi.org/10.1016/j.inoche.2024.112458. Franzolin MR, Courrol DDS, Silva FRO, Courrol LC (2022) Antimicrobial Activity of Silver and Gold Nanoparticles Prepared by Photoreduction Process with Leaves and Fruit Extracts of Plinia cauliflora and Punica granatum . Molecules 27(20), 6860. https://doi.org/10.3390/molecules27206860. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4587822","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":320488707,"identity":"6f8b305b-e77f-4c5f-9c7e-2a920ddabda2","order_by":0,"name":"Ehsan Dahaz","email":"","orcid":"","institution":"Kherad Institute of Higher Education","correspondingAuthor":false,"prefix":"","firstName":"Ehsan","middleName":"","lastName":"Dahaz","suffix":""},{"id":320488708,"identity":"da1b8513-1d8b-4e52-9fb0-53051e93f52f","order_by":1,"name":"Samer Asadi","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA30lEQVRIiWNgGAWjYHCDxMYHDAwHSNPSbECqlgQ2CaK0yLefTnxcwWBnz9+e3FbNU3NHjp+B+eGjG3i0GJzJ3Wx4hiE5ccaZh223eY49M5ZsYDM2zsGnhSF3m2QDA3MCw41EoBa2w4kbDvCwSePTIt//dvvPBoZ6e3mglmKef0RoYbiRu42xgeEw4wagFmbeNiK0GNx4u1myweB44sYzD5sl5/YdNpZsJuAX+f7cjR8bKqrt5Y6nP/zw5tthOX725oeP8ToMYheEYuIBkcwElSMBxh+kqB4Fo2AUjIIRAwDtlFBsMcLYmAAAAABJRU5ErkJggg==","orcid":"","institution":"Kherad Institute of Higher Education","correspondingAuthor":true,"prefix":"","firstName":"Samer","middleName":"","lastName":"Asadi","suffix":""},{"id":320488709,"identity":"7ecb6cb8-1ab7-44e9-802e-d33d5448b32e","order_by":2,"name":"Somayeh Lashgari","email":"","orcid":"","institution":"Petrochemical Research \u0026 Technology Company","correspondingAuthor":false,"prefix":"","firstName":"Somayeh","middleName":"","lastName":"Lashgari","suffix":""}],"badges":[],"createdAt":"2024-06-15 21:23:22","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4587822/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4587822/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":60618144,"identity":"a347d895-498c-429a-b7c0-22e3101a883e","added_by":"auto","created_at":"2024-07-18 20:34:55","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":474289,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eThe \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eHalpeh\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e plant\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4587822/v1/c9d815e292b62b931d9a0950.png"},{"id":60617017,"identity":"984e8ed3-fbc4-43c5-88e9-fcd86462e46c","added_by":"auto","created_at":"2024-07-18 20:26:55","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":330029,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSolution of aqueous extract of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eHalpeh\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e (A) before and (B) after adding 1 mM HAuCl4 solution\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4587822/v1/7cbc1cb08372cfa865d71968.png"},{"id":60620168,"identity":"6d603d93-44ea-4ee4-955e-2847f014d7d4","added_by":"auto","created_at":"2024-07-18 20:50:55","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":260896,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eUV-Vis Spectroscopic Analysis Plot of Synthesized Gold Nanoparticles Using \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eHalpeh\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e Plant Extract\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4587822/v1/2d730d7507c2d15fbe99dff6.png"},{"id":60619220,"identity":"7f93bf8d-6ff6-443b-9e37-1e51670e6cfb","added_by":"auto","created_at":"2024-07-18 20:42:55","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":85110,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eXRD Analysis of Synthesized Gold Nanoparticles\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4587822/v1/393eb2aa404437af2251ca55.png"},{"id":60618147,"identity":"0d9afaad-0415-4349-a7b1-fea17401d815","added_by":"auto","created_at":"2024-07-18 20:34:55","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":130551,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFTIR spectroscopy related to (a) \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eHalpeh\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e plant extract and (b) synthesized gold nanoparticles.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-4587822/v1/b8007e647bc528a9f6a2fb49.png"},{"id":60617018,"identity":"d93d3036-70e7-4136-820e-c2c2b089b638","added_by":"auto","created_at":"2024-07-18 20:26:55","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":380586,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSEM micrograph of synthesized gold nanoparticles\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-4587822/v1/0675f321f572ccf8883eeb00.png"},{"id":60618151,"identity":"2cc9299b-7bd1-482d-bd2c-2c6d6329f21a","added_by":"auto","created_at":"2024-07-18 20:34:56","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":351752,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eTEM image of synthesized gold nanoparticles\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-4587822/v1/084c149a5aa3cf43eb97c406.png"},{"id":60617024,"identity":"67f2f44c-366c-47cb-96cb-87b64f67c8a6","added_by":"auto","created_at":"2024-07-18 20:26:55","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":123126,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ehistogram of the size distribution of gold nanoparticles synthesized using the extract of the \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eHalpeh\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e plant\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-4587822/v1/fa00cdc29414374176732a33.png"},{"id":60617026,"identity":"69dbf902-a89f-4a69-bd9a-c1cab45c9742","added_by":"auto","created_at":"2024-07-18 20:26:55","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":734779,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eImages obtained from antimicrobial tests of synthesized gold nanoparticles on (A) Escherichia coli, (B) Salmonella Typhimurium, (C) Staphylococcus aureus, (D) Bacillus cereus, (E) Aspergillus niger, and (F) Candida albicans.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-4587822/v1/8f41cd7bf656273a0fdd9b26.png"},{"id":60618148,"identity":"bcc198f6-8e80-4951-bfd0-f859ac65ab9b","added_by":"auto","created_at":"2024-07-18 20:34:55","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":118927,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eComposition Percentages of Halpeh Plant Essence Constituents under Treatment with Various Synthesized Gold Nanoparticle Concentrations\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-4587822/v1/f59e285e7ff720fad82fa5da.png"},{"id":60618152,"identity":"a0e6db18-a52b-48ac-8346-6ccbbb7f6ab0","added_by":"auto","created_at":"2024-07-18 20:34:56","extension":"png","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":71268,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eIC50 Values (Antioxidant Properties) of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eHalpeh\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e Plant Extract Treated with Various Concentrations of Synthesized Gold Nanoparticles\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"11.png","url":"https://assets-eu.researchsquare.com/files/rs-4587822/v1/3dcc1d70d956aa4b67c39338.png"},{"id":60618149,"identity":"f38c60df-f3f0-4e08-8312-d0982ebd1edd","added_by":"auto","created_at":"2024-07-18 20:34:56","extension":"png","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":63324,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eComparison of the Reducing Power of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eHalpeh\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e Plant Extract Treated with Various Concentrations of Synthesized Gold Nanoparticles\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"12.png","url":"https://assets-eu.researchsquare.com/files/rs-4587822/v1/267efad010edd46f9cc09b76.png"},{"id":70926183,"identity":"0c40b218-c2c1-4e69-8b9c-1fa6de2ccb68","added_by":"auto","created_at":"2024-12-09 09:09:46","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4225679,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4587822/v1/2cb526df-3763-4569-9f38-8f7d26b0eae1.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Halpeh Extract-Derived Gold Nanoparticles: A Dual Force Against Microbes and for Phytochemical Enhancement","fulltext":[{"header":"Introduction","content":"\u003cp\u003eIn recent years, the increasing prevalence of antibiotic-resistant bacteria has become a global health concern. These microorganisms, including bacteria, viruses, parasites, and fungi, have developed resistance to commonly used drugs. Their ability to adapt and evolve poses a significant challenge to healthcare systems, pharmaceutical companies, and regulatory agencies. Among various strategies to combat this issue, the proper use of antibiotics in pharmacies, hospitals, and community settings plays a crucial role in preventing the spread of resistance. Monitoring the availability of over-the-counter (OTC) drugs in pharmacies, adherence to prescription guidelines, and educating healthcare professionals are essential steps in curbing antibiotic resistance [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIncreasing antibiotic stability in pathogenic microbes is one of the major problems facing medical science [\u003cspan additionalcitationids=\"CR3\" citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. The value of plants is not only in providing food for humans and animals, but also healing and relieving a lot of human pain and needs that have been and will be found in plants since the beginning of creation. Research in the field of drugs of plant origin is in the realm of pharmacognosy and the origin of its earliest dates can be seen in the history of the customs of the ancient tribes and civilizations that provided healing solutions from plant components [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Medicinal herbs with antimicrobial effects and fewer side effects are a good choice than synthetic antimicrobial treatments [\u003cspan additionalcitationids=\"CR7 CR8\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. The therapeutic properties of plant extracts against microbial and non-microbial diseases have been known since ancient times, and many studies were carried out on plant species and their essential oil effects on microorganisms. In fact, the antimicrobial, antiviral and anti-inflammatory effects of medicinal plants are due to the phytochemical properties of their extracts [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Therefore, the extract of medicinal plants can be used to fight pathogenic microorganisms.\u003c/p\u003e \u003cp\u003eNanotechnology is considered as one of the most active research areas in materials science. Nanoparticles have completely new or improved properties that are defined based on certain characteristics such as size, shape, crystalline structure, and morphology. When metals are defined at the nanoscale, they exhibit unique properties. In addition to the inhibitory effect of the particle, metal nanoparticles have a high antimicrobial effect due to their small size, large surface area to volume ratio and cause more contact with outer space [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. One of these metals that has been considered in the field of nanotechnology in recent years is gold (Au). Gold at the nanoscale exhibits properties that make it an important metal in nanotechnology processes.\u003csup\u003e12\u0026ndash;14\u003c/sup\u003e Due to the ease of supply, gold nanoparticles are more widely used than other nanoparticles. Due to the growing application of gold nanoparticles and the growing need to provide methods for providing nanoparticles that do not involve environmental pollution, the necessity of producing these nanoparticles is felt by biological systems [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eGold nanoparticles have good antimicrobial and antibacterial properties that resist human infectious agents [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. The antimicrobial properties of gold nanoparticles include a wide range of bacteria. Gold nanoparticles are synthesized by extracellular and intracellular methods, and the results of experiments conducted so far show that extracellular nanoparticles have better antibacterial effect on a scale with Nanoparticles produced are in a cell manner and their ability to inhibit the growth of pathogenic bacteria is much greater [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Today, with the advancement of technology, it is possible to produce nanoparticles from gold with different biological methods that have good antimicrobial properties and show good resistance to Gram-positive and negative bacteria [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eConventional physical and chemical methods used to synthesize metal nanoparticles also bring toxic chemicals into the environment, in addition to being expensive. Biological methods such as the use of microorganisms and plant extracts can be a good alternative to conventional methods of synthesizing nanoparticles because they are simple, inexpensive, biocompatible and have the ability to produce nanoparticles on a large scale [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. The synthesis of nanoparticles using different types of microorganisms is used in medicine because of their biocompatibility. However, the production of microorganisms is often more time-consuming, and it is more expensive than obtaining plant extracts. The use of plants in comparison to microorganisms eliminates the time required for cell culture and therefore has a high capacity for large-scale synthesis [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eConsidering that the extracts of medicinal herbs and metal nanoparticles, especially gold nanoparticles, have good antimicrobial effects. Therefore, the synthesis of gold nanoparticles using the extract of these plants can enhance their antimicrobial effects. Numerous studies have been done on the synthesis of metal nanoparticles with plant extracts, which have been found that nanoparticles are reduced by plant extracts [\u003cspan additionalcitationids=\"CR22 CR23\" citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. The decreasing factor in plant extracts is due to phytochemical substances in them, especially phenolic compounds. By improving the phytochemical properties of plants, in addition to increasing their antimicrobial properties, nanoparticles with highly desirable purity and concentration can be synthesized. These phytochemicals have properties in plants Prevention of Diseases [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Plants produce phytochemicals to protect themselves. Recent research has shown that these substances are also useful for protecting humans from disease. In 2024, Rodolfo et al. conducted a study on the biosynthesis of gold nanoparticles using an extract of \u003cem\u003eAnemopsis Californica\u003c/em\u003e. They were able to synthesize gold nanoparticles of 5 to 30 nm and concluded that these nanoparticles have good antibacterial properties, especially against Gram-positive bacteria [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. In 2023, Tran et al. synthesized gold nanoparticles using an extract of \u003cem\u003eAndrographis Paniculata\u003c/em\u003e leaves in an average size of 10 to 15 nm, which had desirable antibacterial properties [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn this study, gold nanoparticles were synthesized using the extract of the \u003cem\u003eHalpeh\u003c/em\u003e plant for the first time. The antimicrobial activity (including antibacterial and antifungal properties) of both aqueous and methanolic extracts of the Halpeh plant, as well as the synthesized gold nanoparticles, was investigated and compared. Additionally, the phytochemical changes (antioxidant and reducing ability) of the extract of \u003cem\u003eHalpeh\u003c/em\u003e plant is investigated by treatment with different concentrations of synthesized gold nanoparticles.\u003c/p\u003e \u003cp\u003e \u003cem\u003eHalpeh\u003c/em\u003e, a herbaceous plant reaching heights of 10 to 35 cm, belongs to the mint family and is characterized by its white, cotton-like appearance. It commonly grows in food-impoverished regions, rocky beaches and sand fields of different parts of Europe, the Mediterranean region, North Africa and southwest Asia, including Iran. This plant is widely distributed in south and central Iran. Its leaves are narrow, long, and covered with cotton cracks. Its flowers are also seen in white, yellowish white, yellow or even purple. It is a powerful reliever for gastrointestinal pains such as indigestion. The flowering branches of the plant have tonic, vigorous and anticonvulsant effects. In addition, the fever is high, and the boiled plant is used in cases of colds. More importantly, it is very useful in regulating kidney function, relieving asthma, increasing appetite and blood sugar. The use of the hemp plant in this study was collected from mountainous areas of \u003cem\u003eDashti\u003c/em\u003e County, \u003cem\u003eBushehr\u003c/em\u003e Province (Fig.\u0026nbsp;1).\u003c/p\u003e"},{"header":"Experimental","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eChemicals\u003c/h2\u003e \u003cp\u003eAll chemicals used in this study, which were of high purity and obtained from the reputable company Merck, included 2,2-diphenyl-1-picrylhydrazyl (DPPH), sodium hydroxide, hydrochloric acid, acetic acid, ascorbic acid (ASA), butylated hydroxytoluene (BHT), iron(III) chloride, iron(II) chloride, aluminum chloride, sodium carbonate, sodium acetate, gallic acid, quercetin, Folin\u0026rsquo;s reagent, iron sulfate, 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ), methanol, water, ethanol, and gold(III) chloride (HAuCl4).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eDevices\u003c/h2\u003e \u003cp\u003eThe UV-Vis spectrophotometer model 2100 made by Unico company was used for recording absorptions. The pH measurement was also performed using the pH meter model pH lab 827. Additionally, for weight measurements, a digital scale with a precision of 0.0001 grams, model Kern, was used. For separation, the centrifuge model Sigma 2\u0026ndash;7 was used; for dissolving extracts, the ultrasonic model SB-100D was used; for essential oil extraction, the Clevenger apparatus was used; and for the analysis of the essential oils, the gas chromatography system model GC-7890B connected to the mass spectrometer model MS5977A was used. Other devices used included the XRD model PW1730 from PHILIPS, FT-IR model AVATAR from Thermo, SEM model VEGA3 from TESCAN, and TEM model CM120 made by Philips Holland.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eWater Extract Preparation\u003c/h2\u003e \u003cp\u003eThe process of extracting the aqueous extract was carried out from the aerial parts of the plant known as \u0026ldquo;\u003cem\u003eHalpeh\u003c/em\u003e.\u0026rdquo; The plant extract is used for the reduction of gold ions. Therefore, precision in the extraction methods to obtain a pure extract is very important and noteworthy. For this purpose, approximately 10 grams of the powdered plant were carefully weighed using a digital scale and transferred into an Erlenmeyer flask containing about 100 milliliters of doubly ionized distilled water. Then, it was stirred for half an hour at a temperature of 30\u0026ndash;40 \u0026ordm;C using an electric mixer and, after cooling, was filtered through Whatman filter paper No. 42, resulting in the aqueous extract of the mentioned plant. This extract contains a series of impurities that may originate from the filter paper, as it cannot separate all the components of the plant. For this reason, to remove the impurities, a centrifuge was used at 10,000 revolutions per minute for a duration of 10 minutes.\u003c/p\u003e \u003cp\u003e \u003cb\u003eSynthesis of Gold Nanoparticles by Aqueous Extract of\u003c/b\u003e \u003cb\u003eHalpeh\u003c/b\u003e \u003cb\u003ePlant\u003c/b\u003e\u003c/p\u003e \u003cp\u003eFor synthesis of gold nanoparticles, 1 ml of aqueous extract of the plant and 8 mL of 1 mM HAuCl4 solution were poured into 10 mL and placed on magnetic stirrer. After about an hour at room temperature, it was observed that the color of the solution had changed to purple, which itself indicated the formation of gold nanoparticles (Fig.\u0026nbsp;2). Aqueous extract of this plant has been used as a reducing agent and stabilizer for synthesis of gold nanoparticles.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eOptimization of pH of reaction solution\u003c/h2\u003e \u003cp\u003eIn order to optimize the acidity of the reaction to obtain the most favorable conditions for the synthesis of gold nanoparticles, solutions including 1 ml of \u003cem\u003eHalpeh\u003c/em\u003e extract and 8 ml of gold salt solution with a concentration of 1 mM with pH (2, 3, 4, 5, 6 and 8) were fabricated according to Table\u0026nbsp;1 and the absorption spectra of all of them were taken by UV-Vis and the optimum pH was selected. Their acidity was adjusted by the base solution and sodium hydroxide and hydrochloric acid with a concentration of 0.1 M. As shown in Table\u0026nbsp;1, the highest absorption rate of UV-Vis has been obtained at pH 5, which indicates the synthesis of nanoparticles with higher quality and optimum concentration.\u003c/p\u003e \u003cp\u003e \u003cb\u003eTable\u0026nbsp;(1): Solutions made to optimize pH for synthesis of gold nanoparticles\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Taba\" border=\"1\"\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSoluble Series\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eExtract Volume (ml)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eVolume of HAuCl\u003csub\u003e4\u003c/sub\u003e Solution (ml)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTotal volume (ml)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003epH\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAbsorbance\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.297\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.832\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.859\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.194\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.069\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.190\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003ePreparation of aqueous and methanolic extracts from plant for evaluation of antimicrobial activity\u003c/h2\u003e \u003cp\u003eIn order to prepare aqueous and methanolic extracts of \u003cem\u003eHalpeh\u003c/em\u003e plant for evaluation of their antimicrobial activity, 10 g of plant powder was added to 100 ml of water and methanol solvent and stirred on shaker for 24 hours and smoothed with 42 wattman filter paper. Then, to remove impurities, a centrifugal device was used at a speed of 10,000 minutes and 15 minutes. The extracts were stored at 4\u0026deg;C for later use.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eAntimicrobial activity of synthesized gold extracts and nanoparticles\u003c/h2\u003e \u003cp\u003eThe antimicrobial activity of extracted gold extracts and nanoparticles was evaluated by disk diffusion method (NCCLS115), on gram-positive bacteria (\u003cem\u003eStaphylococcus aureus\u003c/em\u003e and \u003cem\u003eBacillus cereus\u003c/em\u003e), gram-negative bacteria (\u003cem\u003eEscherichia coli\u003c/em\u003e and \u003cem\u003eSalmonella typhimurium\u003c/em\u003e) and fungi (\u003cem\u003eAspergillus niger\u003c/em\u003e and \u003cem\u003eCandida albicans\u003c/em\u003e). First, each bacterium and fungi were cultured in Nutrient Broth medium (37\u0026deg;C, shaker at 200 rpm for 24 hours). After 24-hour culture, a suspension equivalent to half of McFarland concentration was prepared and each suspension was inoculated with a volume of 1 ml (108 cfu/ml) of Muller Hinton agar medium and cultured with swaps. Afterwards, paper discs (6 mm) impregnated with a solution of gold nanoparticles, aqueous and methanolic extracts were placed on the culture site of the relevant bacteria and fungi. Finally, the plates were placed in an incubator with a temperature of 37\u0026deg;C and the diameter of the inhibition zone around each disk was measured after 24 to 48 hours. In order to compare the antimicrobial effect of gold nanoparticles, aqueous and methanolic extracts against the tested bacteria and fungi, their sensitivity to antibiotics discs of ampicillin, gentamicin and clotrimazole was also investigated.\u003c/p\u003e \u003cp\u003e \u003cb\u003eEffect of gold nanoparticles on phytochemical properties of\u003c/b\u003e \u003cb\u003eHalpeh\u003c/b\u003e \u003cb\u003eplant\u003c/b\u003e\u003c/p\u003e \u003cp\u003eBefore flowering stage, the spray of gold nanoparticles on the aerial organs of the plant was performed. First, gold nanoparticles were synthesized under optimum conditions (by adding 1ml of aqueous extract of \u003cem\u003eHalpeh\u003c/em\u003e plant with 8 ml of HAuCl4 (1 mM) salt solution at pH\u0026thinsp;=\u0026thinsp;5) and their concentration (C) was determined using the Eq.\u0026nbsp;(\u003cspan refid=\"Equ1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003cdiv id=\"Equ1\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equ1\" name=\"EquationSource\"\u003e\n$$C={N_{total}}/N*V$$\u003c/div\u003e\u003cdiv class=\"EquationNumber\"\u003e1\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003eN\u003csub\u003etotal\u003c/sub\u003e: The sum of gold atoms\u003c/p\u003e \u003cp\u003eN: The average number of gold atoms per nanoparticle\u003c/p\u003e \u003cp\u003eV: The final volume\u003c/p\u003e \u003cp\u003eThe Eq.\u0026nbsp;(2) is used to obtain the average number of gold atoms in each nanoparticle (N).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Tabb\" border=\"1\"\u003e \u003ccolgroup cols=\"1\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cimg src=\"data:image/png;base64,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\" width=\"632\" height=\"51\"\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"1\"\u003eN\u003csub\u003eA\u003c/sub\u003e: Avogadro number\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"1\"\u003e\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\pi\\)\u003c/span\u003e\u003c/span\u003e: The number pi\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"1\"\u003eD: Medium Diameter of Nanoparticles\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"1\"\u003e\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\rho\\)\u003c/span\u003e\u003c/span\u003e: : Density of gold atom (g/cm\u003csup\u003e3\u003c/sup\u003e)\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"1\"\u003eM: Atomic weight\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eBy placing \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\rho =19.3{\\text{ g/c}}{{\\text{m}}^3}\\)\u003c/span\u003e\u003c/span\u003e،\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(M=196.96{\\text{ g/gmole}}\\)\u003c/span\u003e\u003c/span\u003e and the average diameter of the nanoparticles determined using the TEM image, the average number of gold atoms per nanoparticle (N) is obtained. Then, in Eq.\u0026nbsp;(\u003cspan refid=\"Equ1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), the concentration of nanoparticles was obtained about 315 ppm which was considered as the standard concentration, then concentrations of 0, 20, 40, 60 and 80 ppm were fabricated and sprayed on the aerial parts of the plant during three weeks.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results and Discussion","content":"\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eUV-Vis Analysis Results\u003c/h2\u003e \u003cp\u003eDuring the synthesis process, Au\u003csup\u003e+\u003c/sup\u003e ions are exposed to reducing agents in the plant extract. This initiates the reduction of gold nitrate salts. Complete reduction of Au\u003csup\u003e+\u003c/sup\u003e ions to gold nanoparticles was achieved, as evidenced by the color change in the environment. This process occurred approximately five minutes after the reaction began. Figures\u0026nbsp;(3) corresponds to the UV-Vis spectroscopic analysis of the solution containing the \u003cem\u003eHalpeh\u003c/em\u003e plant extract and HAuCl4 solution. As observed in this figure, the spectrum at a wavelength of approximately 420 nm exhibits a peak (indicating maximum absorption), which confirms the synthesis of gold nanoparticles [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eXRD Analysis Results\u003c/h2\u003e \u003cp\u003eThe XRD pattern presented in Figure (4) is a testament to the crystalline nature of the gold nanoparticles synthesized through an environmentally benign process. The distinct peaks corresponding to the Miller indices (111), (200), (220), and (311) are indicative of a well-defined face-centered cubic (FCC) crystalline structure, which is characteristic of elemental gold [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. The specific 2θ angles of 38.04\u0026deg;, 44.5\u0026deg;, 64\u0026deg;, and 77.7\u0026deg; align perfectly with the standard reference, confirming the high crystallinity of the nanoparticles.\u003c/p\u003e \u003cp\u003eThe absence of any superfluous diffraction peaks is a clear indicator of the high purity of the gold nanoparticles. This suggests that the synthesis process, aided by the \u003cem\u003eHalpeh\u003c/em\u003e plant extract, is not only effective in reducing the gold ions to their nanoparticle form but also in preventing the formation of impurities or by-products.\u003c/p\u003e \u003cp\u003eThe calculated average size of 22.8 nm for the gold nanoparticle crystals, derived from the Debye-Scherrer equation, points to a uniform and controlled nanoparticle formation. This size is ideal for a multitude of applications, ranging from medical to electronic, due to its potential for high surface area and reactivity.\u003c/p\u003e \u003cp\u003eMoreover, the calculated d-spacing of approximately 2.5 nm for the (111) plane further corroborates the FCC structure and is consistent with the expected values for gold nanoparticles. The ICDD (International Centre for Diffraction Data) reference number PDF (Powder Diffraction File) # 04-0784 serves as a benchmark for comparing and validating the crystal structure, ensuring that the synthesized nanoparticles conform to the recognized standards for gold.\u003c/p\u003e \u003cp\u003eThe successful synthesis of gold nanoparticles using the \u003cem\u003eHalpeh\u003c/em\u003e plant extract highlights the precision of the green synthesis approach. These nanoparticles, with their confirmed structure and high purity, hold promise for diverse functional applications.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eFTIR Analysis Results\u003c/h2\u003e \u003cp\u003eThe FTIR spectroscopy provides insightful details about the molecular interactions within the synthesized gold nanoparticles (Figure (5)). The observed shift in the O-H absorption band from 3438 to 3428 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e suggests a strong interaction between the phenolic compounds in the \u003cem\u003eHalpeh\u003c/em\u003e plant extract and the surface of the gold nanoparticles [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. This shift could indicate the formation of a new chemical environment or bond formation, which stabilizes the nanoparticles.\u003c/p\u003e \u003cp\u003eThe presence of distinct absorption bands at 2929, 1624, 1389, and 1278 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e in both the plant extract and nanoparticle spectra reveals the retention of functional groups such as C-H, C\u0026thinsp;=\u0026thinsp;O, C-N, and type I amide after the synthesis process. These functional groups are likely involved in the capping and stabilization of the gold nanoparticles, preventing agglomeration and ensuring uniformity in size.\u003c/p\u003e \u003cp\u003eMoreover, the absorption bands at 1042 and 1058 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e associated with ether compounds (C-O-C) further confirm the attachment of plant-derived organic moieties on the nanoparticle surface. This attachment could play a crucial role in enhancing the biocompatibility and potential biomedical applications of the synthesized nanoparticles.\u003c/p\u003e \u003cp\u003eThe FTIR analysis thus not only confirms the successful synthesis of gold nanoparticles using the \u003cem\u003eHalpeh\u003c/em\u003e plant extract but also highlights the critical role of plant-based compounds in imparting stability and functionality to the nanoparticles.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eSEM Analysis Results\u003c/h2\u003e \u003cp\u003eFigure (6) showcases the SEM analysis of the synthesized gold nanoparticles, providing a detailed view of their three-dimensional structure. The spherical morphology of the nanoparticles is clearly visible, which is a common and desirable shape in nanotechnology due to its large surface area to volume ratio. This spherical form can be advantageous for applications such as drug delivery, where uniformity in particle size and shape facilitates better control over the release rates of therapeutic agents.\u003c/p\u003e \u003cp\u003eThe observed agglomerations, typical in nanoparticle synthesis, indicate interactions that can be effectively dispersed via sonication.\u003c/p\u003e \u003cp\u003eThe SEM images also provide insights into the efficacy of the synthesis method. The relatively uniform size and shape of the nanoparticles indicate a controlled synthesis process, which is essential for ensuring reproducibility and reliability in subsequent uses.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eTEM Analysis Results\u003c/h2\u003e \u003cp\u003eFigure (7) illustrates the TEM analysis of the gold nanoparticles synthesized using the \u003cem\u003eHalpeh\u003c/em\u003e plant extract, demonstrating their spherical shape with diameters less than 50 nanometers. Figures\u0026nbsp;(8), which presents a histogram of the size distribution of the gold nanoparticles, indicates an average diameter of 22.89 nm.\u003c/p\u003e \u003cp\u003eThe TEM images provide valuable insights into the uniformity and quality of the nanoparticles. The spherical morphology is advantageous for many applications due to its symmetrical properties and even distribution of surface area. The size distribution histogram further confirms the consistency in nanoparticle size, which is crucial for reproducibility in industrial and research applications. The sub-50 nm size range of these nanoparticles makes them suitable for high-precision tasks, including targeted drug delivery and sensing applications, where small size and large surface area are beneficial. Differences in sample preparation and imaging techniques contribute to discrepancies in nanoparticle size measurements between TEM and SEM [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e \u003cb\u003eThe results of the antimicrobial activity of the aqueous and methanolic extracts of the\u003c/b\u003e \u003cb\u003eHalpeh\u003c/b\u003e \u003cb\u003eplant and the synthesized gold nanoparticles\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThis research focused on the antibacterial and antifungal effects of extracts and synthesized gold nanoparticles, and their comparative efficacy against three antibiotics: ampicillin, gentamicin, and clotrimazole. The study found that these extracts and nanoparticles demonstrated a commendable ability to inhibit the growth of the tested pathogenic microorganisms, suggesting their potential as alternative antimicrobial agents.\u003c/p\u003e \u003cp\u003eAs per the data presented in Figure (9) and Table\u0026nbsp;(2), the synthesized gold nanoparticles exhibited significantly superior antibacterial and antifungal activity compared to the aqueous and methanolic extracts. This was particularly evident in the case of the gram-negative bacteria Escherichia coli and the fungus Aspergillus niger, where the zones of inhibition were notably larger (18mm and 20mm respectively) [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThis enhanced antimicrobial effect of the gold nanoparticles could potentially be attributed to their combination with the extract of the medicinal plant \u003cem\u003eHalpeh\u003c/em\u003e. This suggests that the integration of traditional medicinal plant extracts with modern nanotechnology could lead to more potent antimicrobial treatments.\u003c/p\u003e \u003cp\u003eFurthermore, the study inferred from the data that the synthesized nanoparticles were effective against all tested microbes. This is a significant finding as it highlights the broad-spectrum antimicrobial activity of the nanoparticles. In contrast, each of the antibiotics was found to be ineffective against some of the microbes. This underscores the potential of these nanoparticles as a more effective antimicrobial agent, capable of overcoming the limitations of traditional antibiotics.\u003c/p\u003e \u003cp\u003e \u003cb\u003eTable\u0026nbsp;(2): Results of the antimicrobial activity of the aqueous and methanolic extracts of the\u003c/b\u003e \u003cb\u003eHalpeh\u003c/b\u003e \u003cb\u003eplant and synthesized gold nanoparticles compared with antibiotics.\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Tabc\" border=\"1\"\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eMicroorganism\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"7\" nameend=\"c8\" namest=\"c2\"\u003e \u003cp\u003eThe diameter of the growth inhibition zone (mm)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBiosynthesized Au NPs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAqueous Extract\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMethanolic Extract\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAmpicillin\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eGentamicin\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eClotrimazole\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eEscherichia coli\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSalmonella Typhimurium\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eStaphylococcus aureus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eBacillus cereus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAspergillus niger\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCandida albicans\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eThe Effects of Gold Nanoparticles on Phytochemical Compounds in\u003c/b\u003e \u003cb\u003eHalpeh\u003c/b\u003e \u003cb\u003ePlant Extract\u003c/b\u003e\u003c/p\u003e \u003cp\u003eAfter extracting the essence from the \u003cem\u003eHalpeh\u003c/em\u003e plant using a Cloninger apparatus, the results of analyzing the plant essence through gas chromatography-mass spectrometry (GC-MS) are presented in Table\u0026nbsp;(3). Various concentrations of gold nanoparticles (0, 10, 20, 40, 60, and 80 ppm) have significantly impacted the yield and composition of the plant essence. Specifically, the changes in \u003cem\u003eHalpeh\u003c/em\u003e plant essence yield were 1.10, 0.118, 0.125, 0.133, 0.151 and 0.113% for the respective concentrations.\u003c/p\u003e \u003cp\u003eFigure (10) illustrates the comparison of the composition percentages in the \u003cem\u003eHalpeh\u003c/em\u003e plant essence under the influence of different concentrations of gold nanoparticles. The major constituents of the essence, such as geranyl acetate, showed the following percentages: 8.131, 6.147, 8.280, 12.418, and 5.629% with the highest percentage observed in the 60-ppm treatment. Regarding Citral (E), its percentages were 29.650, 16.044, 25.122, 8.748, 33.075 and 18.96%, with the highest percentage found in the 60-ppm treatment (33.075%). Citral (Z) had percentages of 22.437, 19.457, 17.394, 20.336, 23.281, and 13.130%, with the highest observed in the 60-ppm treatment (23.281%), although there was no significant difference compared to the 0-ppm treatment. Additionally, Piperitone had percentages of 6.912, 8.030, 9.003, 3.758, 4.725, and 3.733% with the highest percentage observed in the 20-ppm treatment (9.003%). This value showed a noticeable difference from the 0-ppm treatment, and in all cases, the 80-ppm treatment had lower compound percentages.\u003c/p\u003e \u003cp\u003eThe percentage of Geraniol composition was 14.666, 9.818, 11.715, 8.747, 13.260, and 7.431% with the highest percentage in the 0-ppm treatment. As a result, the percentage of geraniol composition has relatively decreased under the influence of different concentrations of gold nanoparticles. Furthermore, the phytochemical compounds Phytol, Lianool, and cis-verbenol in the plant essence had percentages of 0.929, 2.310, and 1.919% respectively, in the 60-ppm treatment, which were higher than in other treatments.\u003c/p\u003e \u003cp\u003eIn summary, gold nanoparticles can act as a catalyst, influencing the extraction of secondary compounds in \u003cem\u003eHalpeh\u003c/em\u003e plant essence. Depending on the specific compound type, they may either enhance or reduce the synthesis of these secondary constituents.\u003c/p\u003e \u003cp\u003e \u003cb\u003eTable\u0026nbsp;(3): Composition Percentages of\u003c/b\u003e \u003cb\u003eHalpeh\u003c/b\u003e \u003cb\u003ePlant Essence Constituents Affected by Different Synthesized Gold Nanoparticle Concentrations\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Tabd\" border=\"1\"\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCompound Name\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eThe Composition Percentage at the Concentration of 0 ppm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eThe Composition Percentage at the Concentration of 10 ppm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eThe Composition Percentage at the Concentration of 20 ppm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eThe Composition Percentage at the Concentration of 40 ppm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eThe Composition Percentage at the Concentration of 60 ppm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eThe Composition Percentage at the Concentration of 80 ppm\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eOctan\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.600\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.308\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e13.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.385\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e21.032\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e1-Octen-3-ol\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.396\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.302\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.695\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.592\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e6-Methyl-5-hepten-2-one\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.036\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.638\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.949\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.999\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.815\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.035\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eDecane\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.645\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.595\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e7.436\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e2-Cyclohexen-1-one, 3-methyl\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.713\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.322\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.344\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.537\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.467\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLinalool\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.783\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.524\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.443\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.529\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.929\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSpiro[4.5]decan-7-one, 1,8-dimethyl-8,9-epoxy-4-isopropyl\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.603\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.498\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.195\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLinalool formate\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.768\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.592\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.670\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eD(+)-10-Camphorsulfonyl chloride\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.404\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.582\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.479\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eGerinic acid\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.986\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.133\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCis-Geranyl acetate\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.201\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.616\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCaryophyllene\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.180\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.548\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCubebene\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.099\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eResults of Investigating Antioxidant Properties and Reducing Power of\u003c/b\u003e \u003cb\u003eHalpeh\u003c/b\u003e \u003cb\u003ePlant Extract under Different Synthesized Gold Nanoparticle Treatments\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe Half-Maximal Inhibitory Concentration (IC50) of \u003cem\u003eHalpeh\u003c/em\u003e plant extract under various gold nanoparticle treatments with concentrations of 0, 10, 20, 40, 60, and 80 ppm is demonstrated in Figure (11) using the DPPH radical scavenging method. As observed in this figure, the IC50 values for the plant extract in different gold nanoparticle treatments 11.61, 10.48, 8.98, 9.94, and 12.67 \u003cem\u003e\u0026micro;g/ml\u003c/em\u003e. Consequently, the antioxidant properties of the \u003cem\u003eHalpeh\u003c/em\u003e plant extract increased in the 10, 20, 40, and 60 ppm treatments compared to the untreated sample (0 ppm). However, in the 80-ppm treatment, the antioxidant activity significantly decreased. Notably, in this test, the plant extract treated with gold nanoparticles at a concentration of 40- ppm exhibits the highest antioxidant properties with an IC50 value of 8.98 \u003cem\u003e\u0026micro;g/ml\u003c/em\u003e relative to other treatments. The results indicate that the impact of gold nanoparticles on the antioxidant properties of \u003cem\u003eHalpeh\u003c/em\u003e plant extract in the DPPH method is bidirectional. Up to a specific concentration, it enhances and stimulates the plant\u0026rsquo;s antioxidant properties.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e\u003cp\u003eThe reducing power of \u003cem\u003eHalpeh\u003c/em\u003e plant extract under treatment with synthesized gold nanoparticles at concentrations of 0, 10, 20, 40, 60, and 80 ppm is shown in Figure (12) based on the calibration curve obtained from the standard FeSO\u003csub\u003e4\u003c/sub\u003e.7H\u003csub\u003e2\u003c/sub\u003eO (FRAP method). The reducing power of the plant extract has increased in the 10, 20, 40, and 60 ppm treatments compared to the 0-ppm treatment, while it has decreased in the 80-ppm treatment. Specifically, in the 40 and 60 ppm treatments, the plant extract demonstrates the highest reducing power with 16.71 and 16.85 mMFe2+ /mg sample, respectively. These results indicate that the impact of gold nanoparticles on the reducing properties of the plant extract in the FRAP method is bidirectional, and up to a specific concentration, it enhances and stimulates the extract\u0026rsquo;s reducing power, confirming their antioxidant properties.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eNanotechnology\u0026rsquo;s pivotal role in plant biology involves using nanomaterials to boost secondary metabolites and modify plant physiology. These materials can target genes linked to metabolite biosynthesis and plant growth.\u003c/p\u003e \u003cp\u003eThis research pioneered the green synthesis of gold nanoparticles with \u003cem\u003eHalpeh\u003c/em\u003e plant extract as a reducing agent. Characterization confirmed their spherical, face-centered cubic structure, averaging 22.89 nm in size and peaking at 420 nm UV-Vis absorption. The study assessed the antimicrobial efficacy of these nanoparticles and \u003cem\u003eHalpeh\u003c/em\u003e plant extracts against select bacteria and fungi, alongside phytochemical changes at varying nanoparticle concentrations (0\u0026ndash;80 ppm) using DPPH and FRAP assays.\u003c/p\u003e \u003cp\u003eThe methanolic extract outperformed the aqueous extract in antimicrobial activity, with the gold nanoparticles showing even greater efficacy, notably against Escherichia coli and Aspergillus niger. Phytochemical analysis revealed that antioxidant and flavonoid levels increased with nanoparticle treatments up to 60 ppm but decreased at 80 ppm. The 40-ppm concentration yielded the highest reducing power in the plant extract.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003e\u003cimg 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\" width=\"377\" height=\"418\"\u003e\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthors' contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eS. A. and S. L. wrote the main manuscript text and E. D. prepared Figures and Tables. \u0026nbsp;S. A. and S. L. reviewed the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors did not receive support from any organization for the submitted word.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analyzed in this study are included in this article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics Declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of interest/Competing interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors have no relevant financial or non-financial interests to disclose. There is no conflict of interest to report.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003ePenny AA, Christine MS, Heleen HD (2024) Antibiotic resistance of bacterial pathogens isolated from the conjunctiva in the Antibiotic Resistance Monitoring in Ocular micRoorganisms (ARMOR) surveillance study (2009\u0026ndash;2021)\u003cem\u003e. 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Molecules 27(20), 6860. https://doi.org/10.3390/molecules27206860.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Gold nanoparticles, Hellebore plant extract, Antibacterial, Antifungal, Antioxidant activity, Reducing power","lastPublishedDoi":"10.21203/rs.3.rs-4587822/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4587822/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe increasing resistance of pathogenic microorganisms to conventional antibiotics has led to a growing interest in alternative antimicrobial agents. In this study, the antimicrobial activity (antibacterial and antifungal) of aqueous and methanolic extracts of \u003cem\u003eHalpeh\u003c/em\u003e plant and gold nanoparticles synthesized using the extract of this plant was investigated. Additionally, the effect of different concentrations of synthesized gold nanoparticles on the phytochemical properties (antioxidant and reducing ability) of \u003cem\u003eHalpeh\u003c/em\u003e plant extract was examined. The results of UV-Vis (Ultraviolet-Visible Spectroscopy), XRD (X-Ray Diffraction Analysis), TEM (Transmission Electron Microscopy), SEM (Scanning Electron Microscopy), and FTIR (Fourier Transform Infrared Spectroscopy) tests on gold nanoparticles confirmed their synthesis. SEM and TEM images showed that the gold nanoparticles were spherical in shape with an average size of 22.89 nm. According to the results, the methanolic extract exhibited significantly higher antibacterial and antifungal activity compared to the aqueous extract, with the maximum inhibition zone diameters observed for Escherichia coli (14 mm) and Aspergillus niger (15 mm). Moreover, the antimicrobial activity of gold nanoparticles showed that these nanoparticles have relatively good ability to inhibit gram-negative bacteria, with the largest inhibition zone diameters observed for Escherichia coli (18 mm) and Aspergillus niger (20 mm). The antioxidant and reducing power activity (phenolic flavonoids content) of \u003cem\u003eHalpeh\u003c/em\u003e plant extract treated with different concentrations of synthesized gold nanoparticles increased with increasing nanoparticle concentrations up to 60 ppm (IC50\u0026thinsp;=\u0026thinsp;9.94 \u0026micro;g/ml and reducing power\u0026thinsp;=\u0026thinsp;16.85 mMFe2+/mg sample), and decreased at higher concentrations.\u003c/p\u003e","manuscriptTitle":"Halpeh Extract-Derived Gold Nanoparticles: A Dual Force Against Microbes and for Phytochemical Enhancement","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-07-18 20:26:51","doi":"10.21203/rs.3.rs-4587822/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"4b8b7063-38b7-4523-8d41-f67a15c8dc0d","owner":[],"postedDate":"July 18th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-12-14T09:53:32+00:00","versionOfRecord":[],"versionCreatedAt":"2024-07-18 20:26:51","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4587822","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4587822","identity":"rs-4587822","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}