Christ alpha 1 2 ld plus

All hydrogel samples were freeze-dried using a lyophilizer (Christ Alpha 1–2LD plus, Germany) and ground into a powder before characterization. The IR spectra of the Gel/SA/nano-ATP composite hydrogels were carried out by FTIR (Thermo Scientific Nicolet iS50, USA) at room temperature. Phase analysis of the composite hydrogel was performed by XRD (D/max-2500, Rigaku, Japan) with 2θ values of 5–50° in a step-scan mode of 5° per minute.
Thermogravimetry of the composite hydrogel was tested using a TGA analyzer instrument (TG 209 F3, Tarsus, Netzsch, Germany) with 10±0.1 mg samples weighed into a platinum pan for thermal analysis and heated at 10°C/min.
Evaluation of the surface wettability of the composite hydrogel was conducted by a contact angle test instrument (FM4000, Krüss, Germany) at specific time intervals. The contact angle was recorded as the average of three measurements.
A Laser Zeta meter machine (Malvern Instruments, Zetasizer Nano ZSE) was utilized to determine the value of the zeta potential of the samples in water solution (pH = 7.4).

Nucleotide exchange as preparation step for FTIR measurements was performed in the presence of alkaline phosphatase as described (8 (link)). Exchange rate to caged GTP was analyzed via RP-HPLC (LC-2010, Shimadzu, Kyoto, Japan) (mobile phase: 50 mM P_i (pH 6.5), 5 mM tetrabutylammoniumbromide, 7.5% acetonitrile; stationary phase: ODS-Hypersil C18 column) and was always >95% cgGTP. Samples were flash-frozen in liquid nitrogen, lyophilized-light-protected for 3 h at –55°C and 0.05 mbar in a Christ Alpha-1-2 LDPlus lyophilizer (Martin Christ GmbH, Osterode am Harz, Germany), and stored packed in parafilm and aluminum foil at –20°C until utilization.

Nanocrystals were prepared by combination of anti-solvent precipitation process followed by probe sonication method.31 (link) Briefly, organic feeds were prepared by dissolving 80 mg of ATR and Labrasol^® (0.5% w/w) in methanol. Then the organic feeds were injected instanta neously into polymer solution containing chitosan and were pre-homogenized at 12,000 rpm for 2 minutes (Ultra-turrax T-25 1KA, Germany). The dispersion was further processed for particle size reduction using probe sonication (Sonics Vibra cell, VCX 750; Sonics & Materials, Inc., Newtown, CT, USA) at 20–23 kHz for 4 minutes to prepare 1% w/w ATR nanocrystals. The process was established and repeated to fabricate ATR nanocrystals using Labrasol^® alone and in combination with three different molecular weight chitosan (CS_L, CS_M, CS_H). Further, the final colloidal dispersions of all batches were lyophilized (Christ alpha 1-2-LD plus, Germany) to obtain powder chitosan based ATR nanocrystals.

Encapsulation of truffle aroma in β‐CD using the paste method similar to the procedures described by Shrestha et al. (2017 (link)), with some modification, were prepared at two different ratios and their formulation strategies are shown in Table 1. For truffle‐β‐CD complexes 1:1, 200 ml of deionized water was mixed with 200 g of food‐grade β‐CD (CAVAMAX® W7 Food) (Wacker Chemie AG, Victoria, Australia) to form a paste before adding 200 g of frozen truffles. For truffle‐β‐CD complexes 1:2, 400 ml of deionized water was mixed with 400 g of food‐grade β‐CD (CAVAMAX® W7 Food) (Wacker Chemie AG) to form a paste before adding 200 g of frozen truffles. Each mixture was homogenized using a shear blender (Kambrook KSB7, Breville Group Ltd, NSW, Australia) for 2 min at the minimum speed of the blender (setting 1). The complexed paste was freeze‐dried (Christ Alpha 1–2 LD plus, Martin Christ Gefriertrocknungsanlagen GmbH) at −30°C, 0.37 mbar until constant weight. Next, the dried complexes were powdered using a mortar and pestle. A set of negative control corresponding to each method was prepared in the same manner, except no truffles were added into the process. Each control set and encapsulated products were separately vacuum packed and stored at 4 ± 2°C until required for volatile and sensory analyses. Each experiment was performed in triplicate.