## 2 Materials and Methods
### Sample Pre-Treatment  
Before analysis, the cheese samples were defrosted at 4 \({}^{\circ}\)C, cut into cubes, stored in beakers, and freeze-dried for 72 h. Containers were protected from light with aluminum foil. Later, the samples were finely crushed, homogenized, and stored at \(-20\)\({}^{\circ}\)C in dark plastic tubes until analyses. Tocopherl Extraction and Determination by High-Performance Liquid Chromatography Coupled to a Fluorescence Detector (HPLC-FD)  
The concentrations of \(\alpha\)-, \(\delta\)-, and \(\gamma\)-tocopherol (i.e., vitamin E) in cheese samples were determined by adapting an analytical method that was recently developed for milk sample analyses [28]. The outcomes were compared with those obtained according to the [PERSON] et al. [29] method, and the results showed that our procedure is reliable, quick, and has good recoveries. Briefly, 2 g of lyophilized cheese were poured into a dark screw-cup Pyrex glass test tube, added with \(n\)-hexane (2 mL), and homogenized (10 s at room temperature). Then 2 mL of potassium hydroxide methanolic solution (2M) were added, the oxygen removed with nitrogen flow, the vessel capped, and the resulting mixture mixed (10 s at room temperature). After that, the final mixture was sonicated at 20 \({}^{\circ}\)C for 10 min and subsequently centrifuged (10 min at 4000 rpm, 4 \({}^{\circ}\)C). The supernatant was isolated, filtered through a 0.2 \(\mu\)m OlimPeak syringe filter with nylon membrane (Teknokroma, Monza Brianza, Italy), transferred into an amber vial, and analyzed by HPLC/FD. Chromatographic analysis was performed using a Shimadzu (Milan, Italy) HPLC system LC-20 AD equipped with a fluorescence detector RF-20A, a CBM-20A controller, a CTO-20A column oven, one LC-20 AD pump, and a DGU-20A3 degasser. The analyses were performed using a _LiChrosorb(r) Si 60 (5 \(\mu\)m)_ column (4.6 mm I.D. \(\times\) 250 mm), protected by a guard column with the same stationary phase. Analyses were run at 40 \({}^{\circ}\)C in isocratic conditions using a mobile phase composed of \(n\)-hexane/ethyl acetate (90:10 \(\psi\)/\(\psi\)). The injection volume was 20 \(\mu\)L, and the flow rate was 0.8 mL/min. Fluorescence excitation and emission wavelengths were 295 nm and 330 nm respectively, and \(\alpha\)-, \(\delta\)-, and \(\gamma\)-tocopherol were identified using commercial standards. The quantitative analysis was carried out with the standard external method using suitable calibration curves and any quantification estimated as the mean value of three repeated measurements and referred to the cheese samples before the lyophilization process. The results are expressed as mean \(\pm\) standard deviation. Carotenoid Extraction and Analysis by High-Performance Liquid Chromatography Coupled to a Diode Array Detector with Atmospheric Pressure Chemical Ionization and Mass Spectrometry (HPLC-DAD-APCI-MS)  
Carotenoids (\(\alpha\)-carotene, \(\beta\)-carotene, and lutein) were extracted from cheese lyophilized samples adjusting an extracting method proposed by [PERSON] et al. [30] for bovine milk sam ples, and all treatments were performed under subdued light. In short, in a test tube, 3 g of lyophilized sample, added with ethanol (3 mL) and a saponification solution (3 mL of KOH 25% _v/v_ in deionized water), were fluxed with nitrogen, capped, vortexed (1 min) and subjected to ultrasound-assisted treatment (5 min at 20 \({}^{\circ}\)C). Then, the mixture was heated at 40 \({}^{\circ}\)C for 30 min for the saponification. Following, 6 mL of _n_-hexane/ethyl acetate (70:33, _v/v_) were added, and after another vortex-stirring (1 min) and sonication (5 min) treatment, the sample was centrifuged (10 min at 800 rpm, 25 \({}^{\circ}\)C) and the upper layer was collected. The ultrasound-assisted extraction was repeated with a fresh _n_-hexane/ethyl acetate mixture for three cycles; the upper organic phases were pooled and dried under nitrogen.