Iraffinity 1 spectrophotometer

Optical rotations were measured using an Anton Paar MCP-500 instrument (Anton Paar, Graz, Austria) and the circular dichroism (ECD) as well as the UV spectra were collected on a Jasco 820 spectropolarimeter (JASCO, Tokyo, Japan) in the 200–400 nm range (under N₂ protection). Infrared (IR) spectra were recorded on an IR Affinity-1 spectrophotometer (Shimadzu, Kyoto, Japan). All the 1D and 2D NMR data were recorded on a Avance-III 600 MHz HD spectrometer (Bruker, Bremen, Germany) with tetramethylsilane as an internal standard. HR-ESI-MS were collected on Bruker maXis high resolution mass spectrometer. A Shimadzu LC-20 AT equipped with an SPD-M20A PDA detector was used for HPLC analysis and preparative separations. An ACE 5 PFP-C₁₈ column (250 × 10.0 mm, 5 μm, 12 nm) was used for semipreparative HPLC separation, meanwhile, a CHIRAL-MD (2)-RH column (250 × 10.0 mm, 5 μm) was used for chiral-phase chromatography (Guangzhou FLM Scientific Instrument Co., Ltd., Guangzhou, China). Column chromatography material: commercial silica gel (200–300 mesh) was purchased from Qingdao Marine Chemical Plant (Qingdao, China); Sephadex LH-20 gel was purchased from Amersham Biosciences, Shanghai, China). All analytical grade solvents were purchased from Guangzhou Chemical Regents Company (Guangzhou, China). The natural sea salt was produced by Guangdong Yueyan saltern (Guangdong, China).

IR spectra were recorded on an IR Affinity1 spectrophotometer (Shimadzu, Kyoto, Japan) using attenuated total internal reflection (ATR) attachment with a ZnSe crystal as a measuring element. The spectral resolution was 4 cm⁻¹, and the number of accumulations was 512. The solutions of Fb and RG were successively passed through a flow microcell mounted on an ATR crystal for 1 h, alternating solutions of the studied substances and buffer 1. The spectrum for buffer 1 was registered first and used as background for all subsequent spectra, which allowed an automatic compensation for water and buffer salt contribution. Difference spectra were calculated subtracting the spectrum of pure adsorbed fibrinogen from the spectrum of fibrinogen with subsequently adsorbed RG. The spectrum of a pure RG solution was recorded separately, followed by washing, confirming a negligible adsorption of RG on the bare ATR surface. The cell was thermostated at 25 °C. The assignment of absorption bands in the IR spectra was based on the previous findings [36 (link),70 (link)]. All spectral manipulations were performed with OPUS 7.0 (Bruker) software.

All reactions were performed with the use of vacuum line and Schlenk techniques. Reagents were commercial grade and were used without further purification. ¹H and ¹³C{¹H} NMR spectra were obtained on a Bruker Avance dpx 400 spectrometer, and were recorded in CDCl₃, or CD₃CN solutions. ¹H, ¹³C{¹H} chemical shifts (δ) were determined relative to internal tetramethylsilane, Si(CH₃)₄ and are given in ppm. Low-resolution mass spectra were obtained by the staff at Cardiff University. High-resolution mass spectra were carried out by the staff at Cardiff University and the EPSRC National Mass Spectrometry Service at Swansea University, UK. Photophysical data was obtained on a Jobin Yvon-Horiba Fluorolog-3 spectrometer fitted with a JY TBX picosecond photodetection module in CHCl₃The pulsed source was a 355 nm output. Luminescence lifetime profiles were obtained using the Jobin Yvon – Horiba FluoroHub single photon counting module and the data fits yielded the lifetime values using the provided DAS6 deconvolution software. Quantum yields using [Ru(bipy)₃](PF₆)₂ as standard (1.6% in aerated MeCN)^{25 (link)} and the following equation:IR spectra were recorded on an ATR equipped Shimadzu IRAffinity-1 spectrophotometer. UV-vis data were recorded as solutions on a PerkinElmer Lamda 20 spectrophotometer.

Reagents and solvents used for experimentation were of laboratory grade (SD Fine- ChemLimited, Mumbai; Molychem, Mumbai, India). Reaction monitoring was per-formed using thin-layer chromatography (TLC) with pre-coated Silica gel-G plates. Purification was performed through recrystallization. Thiele’s melting point apparatus was used to determine the melting points of all synthesized compounds. FT-IR spectra of all derivatives were recorded with Shimadzu IR AFFINITY-1 spectrophotometer using the KBr pellet technique. ¹H NMR and ¹³C spectra data was obtained on Bruker Advance II 400 NMR Spectrometer using CDCl₃ or DMSO-d₆ as a solvent. Chemical shifts are expressed as δ (ppm) values.

After 7 days of incubation, culture broths of the four positive fungal strains was used to separate the produced SeNPs. Cultures were filtered through Whatman No0.1 filter paper and SeNPs were collected by the centrifugation for 20 min at 20,000 rpm. The collected SeNPs were washed in ethanol thrice then in deionized water and dried in a hot air (50 °C) oven. Fine powders of SeNPs obtained from each fungal strain were separately dissolved in ethanol, prior to characterization, and treated ultrasonically for their dispersion (El-Sayed et al. 2020a (link)).
SeNPs from each fungal strain was characterized by the following techniques; UV-Vis spectroscopy (3101PC Spectrophotometer, Shimadzu, Japan), X-ray diffraction (XRD) patterns recorded in the range 20°≤2ϴ≤80° (Cu-Kα radiation, wavelength of 1.5406°A, at 40 KV, and 40 mA) using a D8 DISCOVER BRUKER diffractometer DAVINCI design, USA), Dynamic light scattering (DLS) analyses (Zetasizer Nano ZS, Malvern Instruments, Worcestershire, UK), High resolution Transmission Electron Microscopy (JOEL Transmission Electron Microscope, model 2100, Japan), and Fourier Transform Infrared spectroscopy recorded at 400–4000 cm^− 1 (FT-IR, IRAffinity-1 spectrophotometer, Shimadzu, Japan).