Uv vis uv1601 spectrophotometer

Freeze-dried and ground leaf samples were extracted with 100% acetone and subjected to an ultra-sound bath (VWR International, Germany) for 1 min to ensure complete disaggregation of the leaf material. Extraction occurred in the dark for 24 h at −20 °C, after which the samples were centrifuged at 4000× g at 4 °C for 15 min. Supernatants were scanned from 350 nm to 750 nm in 1 nm steps, using a dual-beam spectrophotometer (Shimadzu UV/VIS UV1601 Spectrophotometer, Country) and the absorbance data were analysed employing the Gauss-Peak Spectra (GPS) method [22 (link),24 (link)]. The De-Epoxidation State (DES) was calculated as

Ground freeze-dried leaf samples were extracted with 100% acetone added and subjected to an ultra-sound bath for 1 min to ensure complete disaggregation of the leaf material. Extraction occurred in the dark for 24 h at −20 °C, after which the samples were centrifuged at 4000× g at 4 °C for 15 min. Supernatants were scanned from 350 nm to 750 nm in 1 nm steps, using a dual-beam spectrophotometer (Shimadzu UV/VIS UV1601 Spectrophotometer). Finally, the detected pigment sample absorption spectra were analyzed and quantified employing Gauss-Peak Spectra (GPS) method [58 (link)]. The sample spectrum was analyzed, through the GPS fitting library, using SigmaPlot Software. This method is based on the sample spectrum fitting, by a linear combination, to the Gauss-peak spectra, that describes each pigment in the detected spectrum, identifying the samples pigment profile, chlorophyll a, chlorophyll b, auroxanthin, antheraxanthin, β-carotene, lutein, violaxanthin, and zeaxanthin.
For a better evaluation of the light-harvesting and photoprotection mechanisms, the De-Epoxidation State (DES) was calculated as: $$\mathrm{DES}=\frac{\left(\left[\mathrm{Antheraxanthin}\right] + \left[\mathrm{Zeaxanthin}\right]\right)}{\left(\left[\mathrm{Violaxanthin}\right] + \left[\mathrm{Antheraxanthin}\right] + \left[\mathrm{Zeaxanthin}\right]\right)}$$