Knowitall software

Fourier-transform infrared spectroscopy (FTIR) spectra of garlic extract in aqueous methanol (8:2) and DMSO were recorded at room temperature using a Perkin Elmer Frontier spectrum spectrophotometer, scanning over the frequency range of 4000 to 400 cm⁻¹. The spectrum of each sample was acquired using spectrum software version 5.3.1 and a diamond ATR (Perkin Elmer, MA, USA) and each spectrum was an average of 64 scans, with a resolution of 4 cm⁻¹. The aqueous methanol aliquots and DMSO aliquots of 10 µL were uniformly spread directly onto the ATR crystal before each spectrum was collected. Functional group analysis was carried out using Know-it-all software (Bio-Rad, Life Science, Hercules, CA, USA).

The average particle size of microplastics was measured using PSA (ELS-Z2Plus, Otsuka, Japan), and the actual shape and particle size were confirmed by SEM (JSM-6701F, JEOL Inc., Akishima, Tokyo, Japan) and 3D profile (Confocal Microscopy, Keyence, IL, USA). The chemical properties of the particles were analyzed using Raman spectroscopy (RAMANtouch, Nanophoton, Osaka, Japan). Briefly, after exploring the morphology of particles with a 20× objective lens, Raman spectra were collected in the range of 160–3000 cm⁻¹ using 300 lines per millimeter grid with a 50 µm slit width. The spectrum of particles was measured over a 16-bit dynamic range with Peliter-cooled charge-coupled element detectors. The acquisition time and accumulation number were adjusted for each scan to obtain enough signals for conducting a library search. The spectrometer was calibrated with silicon at a line of 520.7 cm prior to obtaining the spectrum. Raw Raman spectra underwent noise reduction by polynomial baseline correction and vector normalization to enhance spectral quality (LabSpec 6 software, Horiba Scientific, Kyoto, Japan). The Raman spectra were compared with those of the SLoPP Library of Microplastics and the spectral library KnowItAll software (Bio-Rad Laboratories, Inc., Hercules, CA, USA). Similarities above the hit quality index of 80 were thought to be satisfactory.

A375-v and A375RIV1 cells were incubated for 24 h in DMEM medium containing [U-¹³C]-glucose (10 mM). Metabolites were extracted³⁵ and redissolved in 550 µl of phosphate buffer (0.2 M; D₂O) at pH 7.4. 1D ¹³C NMR experiments were acquired with power-gated proton decoupling (3.1 kHz, Waltz-16) at 298 K on a Bruker Avance 500 equipped with a 5-mm cryoprobe. Acquisition parameters were as follow: relaxation delay 2 s, acquisition time 0.93 s, spectral width 35.2 kHz (280 ppm), and number of scans 20 K. ¹³C NMR spectra were processed using the Bruker TopSpin software 3.1, integrated with the KnowItAll® software (BIO-RAD) and normalized using the external standard and the amount of proteins in the sample as previously described³⁵ .

FTIR studies were carried out in a FT/IR-670 plus (Jasco, Tokyo, Japan) to detect interaction of each component before and after nanoparticulation. Sb, chitosan, PLGA, pluronic and the nanoparticles were pelletized individually with IR grade KBr in the ratio of 1∶100 in a hydraulic press at a pressure of 150 bar for 30 sec. The pellets were scanned over a range of 4000 to 400 cm⁻¹ at a resolution of 4 cm⁻¹ and the data stacked in Biorad KnowItAll software for analysis and overlap regions.

The FTIR spectra of standard fatty acid (FAME Mix C4-C24, Sigma-Aldrich, St Louis, MO, USA) and extracted lipids from A. caespitosus ASEF14 grown in SWW for ten days were recorded using ATR-FTIR spectroscopy (JASCO FT/IR-6300, Japan), with a diamond-enabled ATR sample holder and a DLaTGS detector in the spectral range of 400 to 4,000 cm^–1. The functional components in the samples were identified qualitatively by matching the maximum peak hit (> 99.5 peak region matching) with the IR Spectral Library using the KnowItAll software (BioRad Laboratories, Munchen, Germany). The analysis of the second derivative spectra (2800–3050 cm^–1 and 1350–1500 cm^–1) was done by the Means-Movement method (smoothing with a convolution width of 25) using the Jasco Spectra Manager software. All analyses were done thrice to confirm the reproducibility of the data.