Christ alpha 1 4 lsc

Each cultivar was represented by at least 10 P. domestica fruits randomly collected from a single tree. After pitting, the fruits were sliced in liquid nitrogen and lyophilized for 24 h (Christ Alpha 1–4 LSC; Martin Christ Gefriertrocknungsanlagen GmbH; Osterode am Harz, Germany). The homogeneous powder was obtained by grinding the P. domestica fruits in a closed laboratory mill (IKA A.11; Staufen, Germany). The powder was protected against unfavorable external factors to avoid hydration for 48 h without access to light.

Fresh bamboo shoots (500 g) of P. pubescens were harvested on April 2016 from Ravenna cultivar (Emilia Romagna, Italy) and provided by Consorzio Bamboo Italia. They were collected and kept at −20 °C until needed. Leaves were removed and bamboo shoots were sliced and freeze-dried (Christ Alpha 1-4 LSC). Dried bamboo shoots were then finely grinded with mortar and pestle and sifted through mesh strainers (200–500 μm, Cecchinato).

For alkaline hydrolysis the isolated AGPs from Echinacea and Zostera were dissolved in 0.44 M sodium hydroxide solution in a screw-cap tube (Pyrotube C vial; Associates of Cape Cod Inc., East Falmouth, MA, USA) resulting in a final concentration of 6.7 mg/mL. The solutions were hydrolyzed for 20 h at 105 °C in a heating block (Bioblock Scientific, Thermolyne Corp., Ramsey, MN, USA). Afterwards the solution was neutralized with diluted hydrochloric acid and added to the four-fold volume of absolute ethanol and cooled overnight at 4 °C. Subsequently the precipitate was separated by centrifugation at 19,000× g for 30 min (Hereus Multifuge X3, Thermo Fisher Scientific Corp., Waltham, MA, USA) and additional washing with 80% (v/v) ethanol two times. The washed residue was redissolved in double-distilled water and freeze-dried (Christ Alpha 1–4 LSC, Martin Christ GmbH, Osterode, Germany).
A total of 20 mg of the dried sample was dissolved in 2 mL of 12.5 mM oxalic acid in a screw-cap tube and hydrolyzed for 5 h at 100 °C in the heating block. Precipitation and further treatment were performed as stated above.

PTN/HP-β-CD inclusion complexes were prepared by the freeze-drying method [10 (link),34 (link)]. Briefly, PTN and HP-β-CD (at a molar rate of 1:1) were dissolved in ethanol and ultrapure water, respectively. PTN was added stepwise to the HP-β-CD aqueous solution placed on a magnetic stirrer at 200 rpm and 25 °C for 48 h (IKA RT 15, IKA-Werke, Staufen, Germany). Insoluble PTN was removed by centrifugation at 16,800× g for 15 min and filtration (0.45 µm nylon filter, Pall Corporation, New York, USA). The obtained solution was then lyophilized in a freeze-dryer (Christ Alpha 1-4 LSC, Martin Christ Gefriertrocknungsanlagen, Osterode am Harz, Germany). The physical mixture was prepared by mixing PTN and HP-β-CD (in a 1:1 molar ratio) in a porcelain mortar with grinding for 15 min and then stored in a desiccator until further analysis.

Freeze-thaw cycles were performed in accordance to the procedure suggested by Safaei et al., with some modifications. First, the algae suspension was lyophilised (Christ Alpha 1–4 LSC, Martin Christ Gefriertrocknungsanlagen GmbH, Germany). Subsequently, the dry biomass was resuspended in distilled water to the original volume of 75 mL and freeze thaw cycles were repeated three times. Freezing was performed at −70 °C for 24 h, followed by thawing the suspension at 4 °C for another 24 h. Without accounting for losses in the cooling device, the estimated energy input for all three cycles was 111,900 J in total.

Fruits—“Smoky” cultivar of Saskatoon berry and rye flour type 720—were used in this work. Saskatoon berry was collected in the year 2019 from BIOGRIM company (Wojciechow, Lublin, Poland), and the rye flour was from Good Mills Poland company (Stradunia, Poland). The freeze drying pure fruit powder (FP) as additives was prepared as described previously by Lachowicz et al. [18 (link)]. Meanwhile, the freeze drying encapsulated fruit powder with maltodextrin (FPM) and inulin (FPI) as additives was prepared as described previously by Lachowicz et al. [19 (link)]. The resulting Saskatoon berry fruit was ground in Thermomix (Wuppertal, Vorkwek, Germany) at 40 °C for 10 min. For encapsulation fruit, 70% of grounded fruit (w/w) and 30% of carrier (maltodextrin or inulin) were mixed. Furth, grounded fruit and encapsulated fruit were frozen at −80 °C. Next, the freeze drying method of FP and FPM as well as FPI (≈200 g of each sample) was carried out in a freeze dryer (Christ Alpha 1–4 LSC; Germany) for 24 h at a reduced pressure of 65 Pa. The temperature in the drying chamber was −60 °C, while the heating plate reached 30 °C.

The fruit and their fractions, such as peel, flesh, and seeds of seven selected Saskatoon berry genotypes were used in these studies. The research material contained “Martin”, “Pembina”, “Smoky”, and “Thiessen” cultivars (cvs.) and “no. 5/6”, “type N”, and “type S” clones [16 (link),17 (link)]. The fruit samples were collected from Skierniewice, Poland (51°55′24″ N, 020°5′58″ E). The raw material was directly frozen in liquid nitrogen and freeze-dried (24 h; Christ Alpha 1–4 LSC; Germany). The homogeneous dry material was obtained by crushing the dried tissues using a closed laboratory mill (IKA A.11, Germany). The powders were kept in a refrigerator (–80 °C) until extract preparation.