Green Processing and Synthesis 2021; 10: 101–111 Research Article Rehman Ullah, Sumaira Shah*, Zahir Muhammad, Sajjad Ali Shah, Shah Faisal, Umbreen Khattak, Tauheed ul Haq, and Muhammad Taj Akbar In vitro and in vivo applications of Euphorbia wallichii shoot extract-mediated gold nanospheres https://doi.org/10.1515/gps-2021-0013 nanoparticles at 50, 100, and 200 mg/kg bw. Inhibition of received January 06, 2021; accepted January 26, 2021 gastrointestinal muscular contraction was 21.16%, 30.49%, Abstract: The current study was designed to investigate and 40.19% in mice feed with 50, 100, and 200 mg/kg bw, the potential of Euphorbia wallichii shoot extract for respectively. 3+ reducting Au and stabilizing gold nanoparticles. Keywords: Euphorbia wallichii, gold nanospheres, char- UV-visible spectra of gold nanoparticles showed obvious acterization, in vitro, in vivo applications surface plasmon resonance peak at 548 nm. Microscopy (SEM and TEM) showed spherical dimensions, and the energy dispersive X-ray spectra displayed the strongest optical absorption peak for gold (Au) at 2.1 keV. Dynamic 1 Introduction light scattering spectra represent polydispersed mixture ( ) - with particulate diameter of 2.5–103.2 nm. The IR spectra Centuries ago, gold nanoparticles AuNPs have been uti - confirm the potential functional groups of shoot extract lized by artists due to its interaction with light and cap responsible for the reduction of Au3+ to gold nanoparti- ability to produce attractive colors. Besides, biomedical cles which exhibit tremendous antibacterial potential of and electrical engineers are also taking advantages of the - 76.31%, 68.47%, 79.85%, 48.10%, and 65.53% against interesting and eye catching applications of AuNPs in - Escherichia coli, Staphylococcus aureus, Bacillus pumilus, nanomedicine and electronics. In this aspect, the impor Pseudomonas aeruginosa, and Klebsiella pneumoniae, tance of gold nanospheres cannot be neglected as it is [ ] respectively. Gold nanoparticles showed markedly elevated more potent and advanced than other metallic NPs 1 . fungicidal potency compared to the shoot extract alone Phytomedicines are moving from fringe to a major stream - against the tested fungal strains. IC for 2,2-diphenyl-1- and are used by greater number of people who seek reme 50 ff picrylhydrazyl scavenging was 31.52, 18.29, and 15.32 µg/mL dies for health, with no or low side e ects. In the recent at 30, 60, and 90 min of reaction time, respectively. era, a green route has emerged for NP fabrication, and Both shoot extract and nanoparticles revealed 71% mor- people from both developing and developed countries - tality at 100 µg/mL, with LD values of 310.56 µg/mL. have paid considerable attention to the eco friendly and 90 - - - Experimental mice acquired dose-dependent analgesia bio friendly plant based products in curing and pre [ ] of 54.21%, 82.60%, and 86.53% when treated with gold venting various human diseases 2 . Nano biotechnology is a new frontier for medicine with the application of nano-sized material for targeted cell or tissue-specific clinical intervention. Nanotechnology designed and * Corresponding author: Sumaira Shah, Department of Botany, employed techniques for the synthesis of systems that can Bacha Khan University Charsadda, Charsadda, KPK, Pakistan, ( ) fi- e-mail: [email protected] interact at molecular subcellular level with great speci Rehman Ullah, Zahir Muhammad, Tauheed ul Haq: Department of city in order to achieve maximum therapeutic potency with Botany, University of Peshawar, Peshawar, KPK, Pakistan minimal side effects. The toxic chemicals limit the biome- Sajjad Ali Shah, Shah Faisal: Department of Biotechnology, dical applications of chemically synthesized NPs [3].The Bacha Khan University Charsadda, Charsadda, KPK, Pakistan use of gold in nanostructure has become an attractive area Umbreen Khattak: Department of Botany, Islamia College University, University of Peshawar, Peshawar, KPK, Pakistan of research during last several years because of its diverse Muhammad Taj Akbar: Department of Microbiology, Abdul Wali applications in various fields of science and technology [4]. Khan University Mardan, Mardan, KPK, Pakistan A great deal of efforts has been employed for the biological Open Access. © 2021 Rehman Ullah et al., published by De Gruyter. This work is licensed under the Creative Commons Attribution 4.0 International License. 102 Rehman Ullah et al. application of AuNPs in drug delivery, bio-diagnostics, bio- 30 min at 35 ± 3°C till the color of the reaction mixture technology, and medicine as metallic gold is biologically changes from pale yellowish to ruby red. After the com- nonreactive [5]. AuNPs have excellent antimicrobial activ- pletion of reaction, the AuNPs is centrifuged and washed ities against a wide of range of microorganisms such as with deionized water through centrifugation. The AuNPs bacteria and fungi [6]. These days AuNPs are used widely were dried through freeze-drying technique. The synthe- in chemical catalysis, nonlinear optics, surface-enhanced sized AuNPs were then stored for further use. Raman scattering, nanoelectronics, gene expression, dis- ease diagnosis, targeted drug delivery, and a controlled release of a drug at the desired site [7]. Similarly, nanoscale particles of gold also caught distinct position in biosensing, 2.3 Characterizations enzyme electrodes, catalysis electronics, super conductors, and cancer therapy [8]. The AuNPs prove overwhelming The reduction of Au ions was confirmed by analyzing photocatalytic and therapeutic properties and are more through UV-visible (UV-Vis) spectroscopic analysis of effective against microbes [9]. the synthesized mixture run against ethanol mixture Both Euphorbia wallichii shoot extract (SE) and and distilled water as a blank. The analysis is carrying AuNPs possess some novel physiological and physio- out using UV-Vis spectrophotometer at a resolution of chemical features such as electromagnetic and thermal 1 nm from 300 to 800 nm. For further characterizations, conductance, intense plasmon resonance, biochemical the selected sample is freeze-dried. stability, catalytic potential, antimicrobial activity, anti- Perkin Elmer spectrometer is used for FT-IR analysis HIV, antiangiogenic agents, antiarthritic, and antimalarial in the range of 4,000–0cm−1 at a resolution of 4 cm−1. activities [10]. The current study is designed to biosynthe- Thin pellets of KBr are prepared by pressing with the size AuNPs from E. wallichii SE and eluate its antimicrobial hydraulic pellet press and the sample was put on KBr and other in vivo and in vitro applications. pellet, which is then subjected to FTIR analysis. After FTIR analysis, the AuNP solution was dried at 90°C for XRD analysis using JEOL JDX-3532 X-ray diffractometer (JDX-3532, JEOL, Japan). The average particle size of 2 Materials and methods AuNPs is calculated from the XRD pattern according to the full width at half maximum (FWHM) of the maximum fl 2.1 Plant collection and preparation of intensity re ection peak. The size of NPs is calculated through the Scherrer’s equation (Eq. 1). extract Kλ D =×100 (1) βθ1 cos E. wallichii shoots is collected from various localities of 2 Khyber Pakhtunkhwa, Pakistan. The plant is identified at where D is the average crystal size, K = 0.89 is the the Department of Botany University of Peshawar, Pakistan. Scherrer coefficient, λ = 1.5406 Å is the X-ray wavelength, A specimen of the collected plant is submitted to herbarium 2θ is the Bragg’s angle, β (β = sample, β reference) is with voucher No. Bot. 20210 (UOP). The shoots are separated FWHM in radians. and washed with distilled water and dried for 1 week in Surface morphology of AuNPs is inspected by using shade. The dry shoots are powdered using an electric FE-SEM. A dried sample of AuNPs is coated on a carbon grinder. The shoot powder of 500 g is soaked in 1,500 mL tape and then with gold before subjecting it to Spi- ethanol for 2 weeks. The extract is filtered through Whatman module sputter coater for analysis. filter paper and then evaporated at 40°C through a rotatory The distribution of AuNPs is evident from the TEM evaporator. The solid product is preserved at 4°C in refrig- image. The synthesized AuNPs is used for the analysis of erator for NP synthesis. particle size. Specimens for TEM analysis are prepared by adding 1 or 2 drops of AuNPs solution onto the copper grids and allowing to dry at normal temperature. TEM (Philips Tacna G2 FEI F12, Brown University, Rhode 2.2 Biosynthesis of AuNPs Island, USA) is used to evaluate the surface morphology and size of NPs. The particles are washed and isolated by The synthesis of AuNPs involved the mixing of 1 mL SE centrifugation method by taking 30 mL of suspension in - solution with 1 mM aqueous solution of HAuCl4 in dif distilled water containing AuNPs for 30 min at 12,000 rpm. ferent ratios (1:1 to 1:5) by volume and stirred for The pellets are collected and dried at 50°C in the oven to Gold nanosphares and their in vitro and in vivo applications 103 remove the rest of the water. The collected sample is used For antispasmodic activity, forenight fasted groups for EDX analysis. AuNPs are dried and drop coated on to of mice were fed with normal saline (10 mL/kg bw), atro- the carbon film. EDX analysis is then carried out by using pine sulfate (10 mg/kg bw), 50 mg/kg bw of each SE and SEM equipped with a thermo EDX attachment. AuNPs (po), 100 mg/kg bw of SE and AuNPs (po),and 200 mg/kg bw of each SE and AuNPs (po). After 15 min, the animals were fed with 0.2 mL of activated charcoal, sacrificed (after 30 min), dissected, and the intestine was 2.4 In vitro bioassay cut down from pyloric to cecum. The distance traveled by the charcoal relative to the total intestinal length was mea- For bactericidal examination, Mueller–Hinton agar was sured as percentage of propulsion.