T, Germany). Carotenoid standards (979 ) had been bought from CaroteNature (M singen, Switzerland
T, Germany). Carotenoid requirements (979 ) were bought from CaroteNature (M singen, Switzerland). Pure tocopherols (95 ) have been obtained from Calbiochem (Darmstadt, Germany). Pyrogallol (99 ), magnesium carbonate standard (40 as MgO), 2,6-di-tert-butyl-4-methylphenol (99 ), and (all-rac)–tocopheryl acetate (96 ) were Fmoc-Gly-Gly-OH Purity & Documentation purchased from SigmaAldrich (Taufkirchen, Germany). In addition, -Amylase (five units/mg strong), pepsin (250 units/mg strong), and pancreatin (eight USP) from porcine pancrease were obtained from Sigma-Aldrich. Porcine bile extract was purchased from Santa Cruz Biotechnology (Heidelberg, Germany). Peanut oil was obtained from a neighborhood grocery store. LucantinYellow (ethyl-8 -apo–caroten-8 -oat) was obtained from BASF SE (Lampertheim, Germany); 2,two -Azobisisobutyramidinium chloride (98 ) and sodium chloride (99.5 ) were purchased from Fisher Scientific (Nidderau, Germany). Hydrochloric acid (32 ), sodium bicarbonate (99.five ), sodium sulfate (99 ), and dimethyl sulfoxide (99.eight ) had been obtained fromAntioxidants 2021, ten,three ofCarl Roth GmbH Co. KG (Karlsruhe, Germany); 2,two -azino-di-[3-ethylbenzthiazoline sulfonate (6)] (98 ) and pH buffer options (pH 4, pH 7, pH ten) were purchased from Sigma-Aldrich. Fluorescein (Reag. Ph. Eur.) was obtained from Riedel-de Ha (Seelze, Germany). Potassium dihydrogen phosphate (99.5 ) and manganese dioxide (90 ) had been purchased from Merck KGaA (Darmstadt, Germany). Randomly methylated cyclodextrin (RMCD) was obtained from TCI Deutschland GmbH (Eschborn, Germany). two.2. Chlorophyll Isolation A mixture of 1 g of wheat grass powder and oat grass powder was extracted with MeOH/THF (50:50 = v/v) containing 0.1 butylated hydroxytoluene (BHT). Soon after solvent removal, working with a rotary evaporator, the residue was dissolved in ten mL of n-hexane. The isolation of chlorophyll reference requirements was achieved by way of semi-preparative HPLC (Merck Hitachi 7000 series) in normal-phase mode (ACE five Sil, 250 7.75 mm) employing a variable wavelength detector at 662 nm. Gradient elution at 25 C was applied with an n-hexane-i-propanol eluent (97/3; v/v) at 0 min, with a linear increase of i-propanol (9 ) more than 60 min, at a flow price of 4.0 mL/min and injection volume of as much as 200 sample. Fractions of chlorophyll a and chlorophyll b have been isolated at retention times of 22.7 min and 29.9 min, at the same time as pheophytin at 16.4 min. Identity and purity of isolated fractions was confirmed by reversed-phase HPLC methodology described in Section 2.5. Concentrations of chlorophyll standards had been determined via VIS spectrophotometry utilizing wavelengths of 662 nm (chlorophyll a) and 644 nm (chlorophyll b). 2.three. 2-Bromo-6-nitrophenol Purity & Documentation samples Description Fresh kale was bought throughout winter from neighborhood distributors (Jena, Germany). Kale leaves had been manually separated from stems. A knife mill (Retsch Grindomix GM 200) served to shred leaves to puree-like portions at 8000 rpm for 30 s. Kale samples, intended for High-Pressure processing, had been transferred into 2 mL polypropylene cryo tubes. Highpressure processing was carried out at stress levels of 200 MPa, 400 MPa, and 600 MPa, such as a set of 3 holding instances (5 min, 10 min, and 40 min) for each and every pressure regime. Following HPP therapy, all samples have been right away deep-frozen at -25 C. 2.4. High-Pressure Processing A high-pressure pilot plant (Dieckers, Willich, Germany) was utilized for the stress remedy of kale puree. A mixture of water/ethylene glycol (50:50, v/v) served as pressure transduction liquid. The unit was equipped wi.
