Cary 50

The initial parameters in must were measured on the same day of harvest and before crushing the grapes in the winery. Approximated 100 berries were hand-crushed and then the juice was used for the chemical parameter’s determination. The concentration of sugars (°Brix) was measured in the juice with a Pen-Harvest digital refractometer (Atago Co., Ltd., Tokyo, Japan). Total acidity, expressed as g L⁻¹ of tartaric acid, was measured by titrating juice samples (10 mL) with 0.1 N NaOH to a final pH of 8.2. The standard chemical characteristics of must and wines were summarized in Table S7. Wine parameters including alcohol, total acidity, pH, volatile acidity and reducing sugars were analysed with the FTIR method using WineScan (FOSS, Hillerød, Denmark). The absorbance at 280, 420 and 520 nm were determined 1 month after bottling with a UV–VIS spectrophotometer Cary-50 (Varian Inc., Mulgrave, Australia) and quartz cuvettes of 1 mm pathlength. Wine colour intensity and hue were calculated by assessing the absorbance at 420 and 520 nm and calculating their ratio.

Analogous to a laser flash experiment, a commercial UV-Visible spectrometer (Varian, Cary 50) was used to cover the dynamics in the millisecond to seconds dynamics of DmCRY. The sample was excited orthogonally by a pulsed high-power LED at 455 nm (M455L3, Thorlabs) with a flash of 10 ms duration and 20 mJ pulse energy, which was collimated by an anti-reflection coated aspheric lens (Thorlabs) to yield a pump fluence of approximately 10 mJ cm^–2. The LED flash was synchronized with the spectrometer recording. A small sample volume of 120 μL with an OD of ∼0.1 over 10 mm at the excitation wavelength was used, allowing excitation of the entire sample volume to avert diffusion effects on the long timescales measured. The absorbance changes were recorded at single wavelengths ranging from 400 to 700 nm in 10 nm steps with a time resolution of 12.5 ms in a time window of 10 s.

N₂ sorption studies were conducted at −196 °C using a 3Flex v1.00 (Micromeritics, Norcross, GA, USA) automated gas adsorption system. Prior to the analyses, the samples were degassed under vacuum at 350 °C for 24 h. Fourier-Transform Infrared Spectroscopy (FT-IR) spectra were recorded using a Nicolet iS10 FT-IR spectrometer (Thermo Fisher Scientific, Waltham, MA USA) with an ATR accessory. UV-VIS spectra were recorded on a Varian Cary 50 (Palo Alto, CA, USA) UV-VIS spectrophotometer. Transmission Electron Microscopy (TEM) measurements were carried out by means of a FEI (Lausanne, Switzerland) Tecnai Osiris microscope with an X-FEG Schottky field emitter operated at 200 kV. Scanning Transmission Electron Microscopy (STEM) imaging was performed using a high-angle annular dark-field (HAADF) detector. Before analysis, the samples were ultrasonically dispersed in ethanol and dropped on a Lacey type copper grid (200 mesh) (Agar Scientific, Stansted, UK). Conductivity measurements were performed using a multifunction computer meter CX-741 (Elmetron, Zabrze, Poland). A 0.01 M KCl solution of known specific conductivity was used to determine the specific conductivities of the measured solutions. Centrifugation was carried out with an MPW-250 (MPW Med. Instruments, Warsaw, Poland). A Spin 150 wafer spinner (APT GmbH, Korbach, Germany) was used for spin-coating.

The changes in the absorption spectra were followed by UV-Vis (Varian, Cary-50, Victoria, Australia) during the spectrophotometric titration of TDAP (ca = 1 × 10⁻⁵ mol/L) at different (varying) concentrations of TCPP in ethanol, at room temperature, using a quartz cell.

The iron concentration c(Fe) of the nanoparticle samples was determined photospectroscopically using the phenanthroline protocol [70 (link)]. A total of 10 µL of nanoparticles was dissolved in 20 µL hydrochloric acid (37%). After complete dissolution, 470 µL of H₂O was added. A total of 100 µL hydroxylamine hydrochloride (10%) and 700 µL 1,10-phenanthrolinehydrochloride (0.1%) were added to 200 µL of this solution. After a reaction time of 15 min, the absorbance of the formed ferroin complexes was measured by UV-Vis spectrometer (Cary 50, Varian, Palo Alto, CA, USA) at a wavelength of 510 nm, and the iron concentrations were calculated using an iron standard calibration curve (Iron (II,III) Oxid (Sigma Aldrich, Darmstadt, Germany) as standard in the range c(Fe) = 1.25 to 40 mM).

The UV spectra of the reaction mixtures were scanned during the incubations of CYP440A18 with fatty acid hydroperoxides with Varian Cary 50 spectrophotometer. Alternatively, the UV spectra of products were recorded online during the HPLC separations using an SPD-M20A diode array detector (Shimadzu, Japan). Products (Me esters or Me/TMS derivatives) were analyzed by GC–MS as described previously [21 (link)]. GC–MS analyses were performed using a Shimadzu QP5050A mass spectrometer connected to a Shimadzu GC-17A gas chromatograph equipped with an MDN-5S (5% phenyl 95% methylpolysiloxane) fused capillary column (length, 30 m; ID 0.25 mm; film thickness, 0.25 µm). Helium at a flow rate of 30 cm/s was used as the carrier gas. Injections were made in the split mode using an initial column temperature of 120 °C, injector temperature 230 °C. The column temperature was raised at 10 °C/min until it reached 240 °C. Electron impact ionization (70 eV) was used.