Zein or gelatin nanofibers loaded with au nanospheres, SnO2 or black elderberry extract used as active and smart packaging layers for various fish fillets

Abstract

Fresh fish products spoil in a shorter time than other meat products. Consumers would prefer the freshest fish products that have higher initial quality. Therefore, it is important to investigate alternative methods to preserve the quality of fresh fish products and increase their shelf life. Furthermore, estimating the shelf life of fresh fish products in fast and easy methods has started to gain importance in recent years. These developments increase researchers' interest in active and smart packaging layers that are produced by nanotechnological methods. In this dissertation preparation, characterization, and practical application of biopolymer-based electrospun nanofibers as active and smart packaging layers are presented. First, the purpose and objectives are explained and recent studies are presented in Chapter l. Then, preliminary microbiological analyzes were applied to smoked fish products after their packages being opened (Chapter 2), suggesting that the smoking process inhibit microbial growth only if the package is not being opened. Then, the quality of three different fish meat samples (local salmon: YS, bream: Ç, sea bass: L) was determined in terms of the electrical conductivity value, surface tension values, and the ε″. Sensory evaluation results were also evaluated by photos and point scales. The electrical conductivity value of the YS, Ç, and L samples on the initial day (0.21, 0.24, and 0.233 mS cm-1) increased on the 4th day of storage (0.35, 0.39, and 0.47 mS cm-1), respectively (~65%, 63%, and 101% change). Dielectric loss factor values calculated at a frequency of 30 MHz were also increased with the highest change of L samples (29.68%) on the 4th day. On the contrary, surface tension values decreased with storage time. The surface tension of YS, Ç, L, declined from 38.22, 42.56, 37.57 mN m-1 to 16.2, 29.45, 25.94 mN m-1 on the 2nd day of storage. Results revealed these analyzes can be accepted as an important and rapid quality techniques in fish meat samples. Chapter 2 results revealed that active coating layers could be used to preserve shelf life and inhibit microbial growth. Therefore, in Chapter 3, skinless fish fillets nanocoated with fabricated AuZ-Nm (530 ± 377 nm) and TMAB growth at 4 ± 1 °C compared with the uncoated group during the 8-days storage. Microbiological results indicated that the use of zein nanofibers with gold nanospheres delayed the TMAB growth up to ~1 log CFU/g (p<0.05) and sensory deterioration. The monitoring dielectric properties of fresh fish fillets for evaluating their quality (Chapter 2), was also used in Chapter 3. In this sense, dielectric properties (ε′ and ε′′) of the fish fillets were treated with AuZ-Nm and were investigated. ε′ values of the uncoated fish samples were more variable (34.53%) compared to the nanocoated group (<30%) on the 7th day of storage. Similarly, a 40.55% decrease was recorded for the uncoated group according to the initial ε′′ value, while this decrease was 22.75% for nanocoated samples (p<0.05 between groups). It has been concluded that zein based nanofiber swith Au nanospheres have the potential as an active food packaging layer to extend the shelf life of fish fillets. In Chapter 4, the effect of electrical conductivity, ε′, ε′′, and loss tangent (ε′′/ε′) values (at 300 and 3000 MHz), on feed solution electospinnability was investigated. For the first time, the aggregation behavior of nanofibers was determined by ZetaSizer equipment. Electrospun samples dispersed in the ethanol had lower translational diffusion coefficient (water:2.03 μm2/s; ethanol: 1.85 μm2/s) and higher hydrodynamic radius (water: 242 nm; ethanol: 221 nm). Zein-gold nanofiber stability was also studied by zeta potential measurements (ethanol: +41.73 mV; water: +5.1 mV). In addition, the antimicrobial effects of AuZ-Nm were investigated and physicochemical characteristics were compared without Au. Specific shoulders in Au zein nanofiber spectrum indicated C=O carbonyl stretch vibrations in the amide I and amide II region, which does not appear for pure zein, but is observed also in the pure Au spectrum. After addition of Au, bands in the pure zein spectrum converted to stretching peaks, indicating the vibration frequency of Au–O ionic bond groups. These molecular observations and other signs (narrowing band, shifting wavenumber, transmittance changes) confirmed the successful integration of Au molecules. All these results indicated that although the process of smoking decreased the initial TMAB load, after opening the package, the TMAB values of the smoked fish increased more (Chapter 2). Using Au in nanofiber coating layers inhibited microbial growth more than the smoking process as explained in Chapter 3 and Chapter 4. However, since disruption is inevitable, it is also important to use these nanofiber materials as indicator layers to predict shelf life. In this context, gelatin-based nanofibers were developed to evaluate their color changing functions (Chapter 5). At first, the production of liquid gold NPs and dried gold nanopowders from gold salt (HAuCl4) was explained as stated in supplementary material. Then liquid gold added to feed solutions. SEM results of gelatin nanofibers the average diameter of pure gelatin nanofibers (GL) was 81.4 nm without any beads. The addition of 10% BE extract to the solution increased diameter to 277 nm (GLE). EDS elemental mapping peaks and Figures indicated that after 5% SnO2 incorporation into the feed solution, SnO2 NPs both encapsulated inside the nanofiber and attached to the surfaces of gelatin nanofibers (GLES). Small spots on nanofibers were observed with the addition of produced Au nanopowders at 2%, and the average diameter increased to 554 nm (GLESA). Nanofibers were deeply characterized by FTIR before and after exposed to fish meat for 30 hours (no direct contact). New methods were proposed for the first time to evaluate nanofiber stability by transmittance ratio values and produced degradation metabolites confirmed with the band differences in spectrums. Specific spectral changes indicated absorption/attachment of volatile amines to the nanofibers during the deterioration of fresh fish samples. Thermal stability between samples was compared by DSC and TGA. Thermal characteristic results of gelatin nanofiber samples also proved the new ionic bond complexes (initial) and thermal decomposition of absorbed volatiles (30th hour). Furthermore, L a b values of the nanofiber showed that interaction between flavonoids/phenolic acids and metal ions modified the color produced by the anthocyanins. Therefore, the lowest brightness values (L=64.23) with the highest redness (a=10.37) for the gelatin nanofiber that contains gold nanopowders (p<0.05) were obtained in the first measurement before the spoilage (GLESA). During 30 hour storage period the absorption of volatile amines influenced L a b in different directions. On the 24th hour, the color of the gold nanopowder added gelatin sample (GLESA) became more intense and turned to dark purple with the lowest b value (-3.11) and the lowest L value (24.14) (p<0.05; between nanofiber samples and between initial, 3rd, 6th hours). When all chapters are evaluated together, the results of this study enable a better insight into a) understanding of the determination of fish quality by rarely studied parameters such as electrical conductivity, surface tension, and dielectric loss factor, b) evaluation of various properties of prepared feed solutions for electospinnability, c) molecular, morphological, elemental characterization of nanofibers and NPs, d) searching aggregation behavior and stability of nanofibers by novel techniques such as DLS data calculations e) practical application of fabricated nanofibers containing Au nanospheres, SnO2, and BE extract on fish samples for gas sensing performance. These chapters guide possible nanoapplications of the nanofibers incorporated with antimicrobial agents, plant extracts, and metal-based NPs for active and intelligent packaging functions on meat products.