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    Food Ghemisry 63 (2021) 130881 Contents Food Chemistry Journal homepage: veww.clseviercomiocatetfoodehem ts avallable at Sclencebirect, ELSEVIER iy Chemical and organoleptic changes of curd cheese stored in new and reused active packaging systems made of Ag-graphene-TiOz-PLA Anca Peter’, Leonard Mihaly Cozmuta“, Camelia Nicula", Anca Mihaly Cozmuta*, Catalina Mihaela Talasman °, Goran Drazic , Antonio Peiias ', Antonio Jestis Calahorro", Gianni Sagratini, Stefania Silvi’ Tcl dey of Np, Dagar of Ohi, View Bas, 7643008 Bat Me, Romania Car, Alvan an Ce 810019 Bal, Romania Neal bt of Chen, Heo 19 PO 65.100 tans Soe ‘ andoe om Cafe de aPC ier 22600 Mr Se, Spa “testo omer, Sk el arr Port al ie, a Mado le Care 9, 62982 Cami MG, Rey ‘try of Comer, Sek Boxcar «Mena Varnara, Va Cae da Varn, 2052 Cerin, MC aly ARTICLE INEO ABSTRACT Keprots “The end reports obtaining, characterization and evaluation ofthe preservation efcacy of an Ag-saphene Ave Pad ‘NO, polyacte aed (PLA) fll dung the storage ofthe curd chess. The eusblity of the active package was o ‘also evaluate. The mechanical resistance ofthe fm was improved by 80% following the compost ation. ‘Viner vapour permeability deceased by 11-27% when the eompsite was incorporated nto PLA In a mas percentage ranged berween 0.5 and 3%. The highest fat pemeabllty was obtained for PLAS at 4 °C. The ‘oxygen perenblty of PLA is 24.6% lowe han tht of eat PLA, PLAO.S0 and PLA presented the highest otibacterial activity. PLAS achieved the lowest solbity in food simulans at 4 °C empated to PLA and Feference. The best active packages for cur storage were PLAO.S% and PLAS. They showed also the lowest ‘depvelcon fron the Fst t the second se Teste roles kee 10% ad ASM respeeiey, during the ond, ean esas Miron i ot silat 1. Introduction ‘The active packaging system contains key component that exerts complex effect on the food, namely Co reduce mierobiologieal conta “The increasing ws of non degradable pass for packaging slid and liquid foods has ensed the accumulation of large amounts of waste inducing serious environmental problems. Alternative solutions vo these iajor environmental challenges are the biodegradable materials, “They are derived from renewable sources of bioniass sch as vege table fats and oils corn starch or sugarcane. Examples of such materials are polylactc aid, polyeaprolactone (PCL), polybutylene adipate tere phthalate (PBAT) and polyhydroxyburyrste (PHB), polybutylene ad Date terephthalate (PRAT), paper, cardboard and starch (Plsver el 2020). PLA and PHB have thermal and mechanieal properties comps rable fo those of the common polymers and have arouse considerable Interest. PHB is known to have poor processing properties and is bridle ‘at room) temperature. PLA has better mechaniesl strength than PHB, "hile PHB has better batrler properties, * conesponding autor ‘naton and minimize spilage/fermentation processes, thus prolonging the validity (Lukicet ol, 2020), Nanostructured composites based on Ag (an ec a, 2019), THO, (Apiok eal, 2019, Peter eta, 2019), graphene, Aluntnosilieates (Risyon et al, 2020), SiO, (Sepulveda etal, 2020), chitosan (Rips etal, 2016, Apjok et a, 2019), plant extracts (akie etal, 2020, Roy and him 2020, Suwanamornler etal, 2020) are sed as key components inthe active packages. Polylactieseld (PLA) Is produced from renewable soures, such as sugar beet and comm stare oF fom other products and waste from renewable biomass (lukic et sl, 2020). Currently, high moleculae weight PLA is obtained, with good physical-mechanical properties and low cost. However, biodegradable plastic aso has disadvantages such 9s fragility, degradation during processing and reduced capacity of reey «ling @avee ral, 2020). In order to overcome these aspects and to mail address anc. peter@ewiborsicnjio (A. Pete, sniono penasarndaiocoig (A. Pei), ghniisngitini@vnionn.t (Sagat, series (6 Sv, beps/Ao.og/10.1016/ foc, 202113041 Receive @Febriaty 2021; Receives in revised fom 6 May 2021; Ace 8 Jane 2021 ‘Available ale 10 June 2021 (0308 14676 2021 Hscier i All sights reserved Improve the preservation properties of fod, the enrichment of PLA with various active principles (TiO, Ag, SiO2, aluminosiliates, natural antioxidant extract) is intensively studied, whieh besides dhe anim ‘robial and. antioxidant properties, confer improved mechanical strength and barrier properties tothe new material Lic et al. (2020) have preduced polylacic ac ims with 586 (wt) fof poly(e-aprolactone) loaded with thymol, carvacrol and their ‘mixture. They demonstrated thatthe mitre of thymol snd earvacrol loaded into PLA film showed synergy and low percentage of migration into distilled water, revealing great potential for application a8 food packaging materia. Sepulveda el, (2020) have prepared by extrusion nanocomposite fms based on PLA with 1.096 (w/w) silee nanopariles and surface functionalized silica nanoparticles with lati acid, respectively. They Ihave proved thatthe permesbility to oxygen ofthe non modified PLA is ‘by 44% lower than ofthe modified PLA as well asthe plasticiing effet ‘of cynamualdelyde incorporated into PLA. matrix, Moreover, they ‘showed thatthe incorporation offnctionalied nanoparticles SiO with lactic acid decressed the difsion cooficient, and the filing effect ofthe silica nanoparticles was obtained, thus delaying the release ofthe active ‘compound, Fan el. 2019) have Incoeporated 1 ws, 5 wis, 10 wei, 15 W196, and 20 winano-Ag into PLA flim and showed that high pressure and migration time would make the films rougher and increase the crystal lization temperature of the PLA nanocomposite fs. {Chi etal, (2019) have showed chat PLA with Ag and TIO, could ‘extend the postharvest life of mangoes upto 15 days. in another study, the same research team (Li ets, 2017) showed that blend PLA with nano-Ag and nano-TiO, presents a good antimirobial activity against £. coll and Listeria monocytogenes and thatthe release of NPs from the ‘nano-blend films ves within the standard limit. api et al. (2019) have tibutyl O acetyl erate Improved the me ‘chanical and therm properties of PLA. Moreover, they established that PLA with chitosan is no toxie for germination of eueumber and radish seeds Every year, 448 milion tons of pastes are produced. They consn about 896 of raw material resoees. Ths, the plasis industry will use 2096 ofthe total amount oferade coil extracted until 2050, The ecological factor as well asthe inctease in the price of raw materials argued the use ‘of renewable raw materials aud finding solutions forthe packages reuse (@avee eta, 2020). Plavee eal. 2020) have modeled the reeyeabilty ‘of the biodegradable blend and showed that PLA/PHB type is sitable for multiple processing and material reeyeling. According to the best of ‘our knowledge, there are limited reports inthe terature related to the ‘reuse and reeycling of biodegradable polymeric materials. In this regard, this study ims to prepare and characterize polyactie ac (PLA) f enhanced with nanostructured composite based on silver, titania and ‘graphene and (0 establish ts preservation activity over curd cheese as ‘ell sits recycling potential “The packaging reise should be taken from two perspectives: (1) 10 test the reproducibility of the active package and (2) to be able to recover and reuse the packaging. This could be done both in the household by the consumer and at the store level. After cheese ‘consuming, the package ean be cleaned and disinfected and can be used. to store another cheese or food, ths avoiding throwing it. At the com mercial level, the strategy isto have in each supermarket a collection space fortis typeof packaging and he collected packages to be cleaned and disinfected and reused for the packaging of new products 2, Material and methods 2.1. Preparation of Ag graphene-TIO, composite ‘The composite was prepared by sol-gel method, followed aging, ‘ying and calcination, The procedure is detailed in Supplementary material S. Fao emt 969 (2021) 190941 2.2, Characterization ofthe Ag graphene T10> compose FTIR spectroscopy, UV-Vis analysis (Peter et aly 2021), Xray Aifration and Transinision eleetron microscopy (STEM) were per formed in order the characterize the composite and the procedures are detailed in Supplementary material 8.2 2.3. Obraining of Ae-araphene TO, PLA fis ‘The composite powder was incorporated into the PLA matrix and the composite PLA film was obtained by extrusion. The procedure is Aetaled in Supplementary material SS, 2A. Characterization of new prepared an reused Ag graphene TIO, PLA Fis Films characterization consisted fn SEM microscopy, mechani resistance, permenbility to water vapor, grease and oxygen, ove rigration and antibacterial aeivty. The procedures are detailed in Supplementary material $4 25. Storage of curd cheese imo the Ag graphene TiO, PLA fms ‘The eurd cheese was homemade prepared according to the tad tional recipe used inthe rural households from North Western Romania The reipe does not include additives and/or preservatives. The fresh ow mk was kep for natural fermentation ito ceane pots for several days and then the precipitated easoin was filtered into a white, clean, cotton cloth and the obtained cheese piece was allowed to dry for several hours, Four types of materials were tested as packages, as follows: virgin PLA (coded PLA), PLA modified with 0.5% (w) composite (coded PLAOS), PLA modified with 9% (wi) composite (coded PLAS) and reference commercial bag nade of polypropylene (coded PP). [Ananiount of 100g curd cheese was wrapped ina 40 em/40 cm sheet fom each type of material and the obtained packages were stored UV- ‘lumsinned reeigeator at 4 °C for 17 days. Periodically, sumples were extracted and analyzed in terms of organolepica changes, mass Tos, DH, titratable acidity, fat and protein content and peroxide index. ‘The organoleptic changes were monitored according to the proced ture deseribed by Necula (2010). The sensorial study was eonducted under conditions of room temperature and daylight, using a group of 10 trained panelists (6 wonen and 4 men, with ages fron 18 to 60 years) who were regular cheese consumers. The aspect, the consistency, the taste and the odor of the cheese were evalnted using a five-point he done seale (1-dstke extremely to 5 like extremely). The dtaled pro ‘este is deserbed in Suppleniensary misters S#. The correspondence between the evaluation unit and the characterstis ofthe urd cheese are displayed in Table S.. fom Supplementary mate Mass loss (ML) wes determined by weighting the packages atthe begining ane a the end of the experiment. The derailed procedure is deserbed in ipplerertary material St ‘pH and titratable acidity. "The acidity was determined by titration and the pi was recorded by using @ WTW pH meter. The detailed pro cedure is described in Supplementary material. Fat content. The fat content was determined by using the solvent extractor VELP Scientific. The detailed procedure is described in Si plementary material $4, Peroxide index. The peroxide index was determined as the content of peroxide o other oxiizing substances that oxidize potassium iodide, releasing iodine. The detailed procedure is deseribed in Supplementary ater 8. Protein content, The protein content was determined by Kjeldahl seth by sing the Velp Seientifen system The detiled procedure is deseribed in Supplementary material 8. 2.5.1. FTIR bvestigaions ofthe cur cheese after storage new and reused packages FTIR spectroscopy was used to establish the stracural changes ofthe ‘curd cheese during storage as well as the posible migration of the ‘package components into food, The detniled procedire is described in ‘Supplementary material S4 2.6, Recovery and reuse of the Ag gmphene TO, PLA fms “The used packaging resulted from the packaging of slice of fresh ‘cheese in exactly the same conditions a those wsed in Section 2.5. They ‘were cleaned from the remained cheese resides, washed for tee tines with aistiled water and with 959% ethanol aad died i ar. The as ob tained recovered packaging materials were used forthe wrapping of the ‘curd elieese from the same batch and in the same conditions as those ‘mentioned in Section 25. The reused packaging materials were also ‘characterized necording tthe procedures described in Section 2.4. 22. Seaisieal analysis ach experiment was conducted at Teast tte thmes and the results were expressed as mean, Statistica 7.0 software (StatSof, ne, Tus, USA) using One-Way ANOVA CTukey's tes was used, 3. Results 3.1. Characterzaion of Ag-graphene-T10, composite FTIR spectroscopy: The signals low frequency (600-900 em) ‘were artibuted to the wibration of TOT and C-O-Ti, which confirmed the formation of 710, and C-O~Ti bonds inthe composite @ivang e€ al, 2016) (Pigure 8.2. fom Supplementary material) The trsmission bans at 1234 em”? and 1382 em" are assigned to the -O—C—0- stretching (Hints etal, 2014, Atchudan et al, 2017). The spectrum also shows signals at 1620-1939 cm attributed to skeletal C=C vibration from graphene, ‘hich correspond to the sp? character of the carbons and the C—O stretching (Zhang et sl, 2016, Song et sl, 2018). The transmission bands at 1631-1647 ear * correspond 10 OH groups and the band at 1707-1728 ens assigned 0. C=O stretching vibration from carbonyl ‘and eacboxylic groups, indicating a possible oxidation of graphene at the ‘surface, The signals at 2908 en and 2960 cm are also the result of the OMH stretching (Tsai & Cheng, 1997). The signal in the region ‘3624-9448 emis assigned to the anatase pase ofthe titanium dioxide lattice (Tssi & Cheng, 1997 UV-Vis Spectroscopy: Figure S.. from Supplementary material Iustrates the UV-Vis absorption spectra of Ag-graphene-TIO, and com mercial TiO (reference). Low absorbance values were observed for ‘comiercial TiO2 (reference), which demonstrates low radiation sb somption near the Vis region, The highest absorbance value (0,629) was ‘obtained forthe Ag grapiene TIO, composite at 398 nm, which means thar ic absorbs radiation inthis wavelength range toa larger proportion "han TIO Base on this result, the gap energy (Eg) ofthe composite was calculated by using the Kubelka-Munk equation, The absorption coef ficent was replaced with F factor determined from the reflectance rests: ey oe “Thus, the calculated Ey for commercial 103 was 3.21 eV, while for ‘Agegraphene-TiO. was 3.04 eV. This shows thatthe compute f mil ‘asi activated by a radiation with a lower energy than that needed 10 excite TiO, XRD diffraction: According to Figure S4. frm Supplementary fey the XRD profile shows dhe presence of both anatase and rutile ery lied phases of titania. The XRD resis are confirmed by those exhibited FR) o Fao emt 969 (2021) 190941 {nthe FTIR specttum. The composite contains lage and well eysalised particles. Beside strong line at 25.4" (2 dheta) there is also present & sual line at 27.4, which corresponds to rate, Ag lines ean haedly be seen in somte eases, because there is very stall anit of Aga lines fare overlapping with the anatase lines Morphology: From the miero-sructural point of view, the Agent phene-TiOy composite is homogeneous and contains one dimension (20 nm particles of TO» anatase and of Ag (2-5 nm), thet are uni formly distributed in the graphene network (Rgure 5. fram Supple smenary file). 8.2. Characterization of new prepared and reused Ag graphene Ty PLA ‘ms ‘The macroscopic view ofthe new prepared composite PLA Civble 1) ilustratesflns where the Ag graphene TiO, composite nanoparticles are relatively uniform embeded in the PLA matrix, The reference na {eral is polypropylene-based film blue colored. The SEM images of PLA sample Table 1) shows that the surface seems tobe quite uniform. In some areas there are micrometer sized defects nt the surface. Some defects are also detected in PLAO.5% SEM micrographs, Sone of those defects are atthe surface, while most of them are buried inthe PLA. SEM rcrographs of PLA‘ show that some defects appentpartaly at the surface and partially ae embedded in the PLA muattix. No changes ofthe uaeroseopically vew and SEM images ofthe reused PLA-based Mins ceurred ‘The tensile strength ofthe PLA based films is included in Table 2 Among the Aggraphene-TiyPLA films, che highest resistance has PLAO.596 film (MD modulus is 3671 MPa and GD modulus 3751 MPa). Thus, if the two types of modulus are averaged, the resistance of the PLAOS596 fil is 4586 and 21096 respectively, higher than that of PLA and DLASSG respectively. The differences between the values of the tensile strength for new and recovered PLA asd flms are not statist cally significant. The presence of the composite ina certain percentage (C84) reduces the mechanical resistance of the PLA.based ln. The same trend was showed also by Li ctl. (2017) and ean be explained by the fact chat the eomposie's nanoparticles added into the PLA form cracks or eavices, which decrease the tensile strength of de film (Li eval, 2017). Thesame trend was observed also by Sivaanornlert el (2020) ater adeition of polybutylene succinate co.adipate)(PBSA) and ‘hymil ino the PLA matrix. The presence of PBSA and thymol cased Aeercase of the mechanical resistance of PLA fins ‘WUP decreased frm 151.1 +7.47 mi? 24 for unmodified PLA to 1135 48.52 m « 24f for PLAS, lis observed a diference between PLA and PLA modified with composite, so the presence of composite confers superior water vapor barrier properties to the PLA matrix by restricting water molecules to permeate cough the lm (Isble 1) Ifthe permenbilty of PLA based materials is analyzed, i can be sen that the Towest level of barrier against oxygen is displayed by the unmodified PLA and inereases with dhe composite’s content. The lowest permeable {0 oxygen are PLAO.S% and PLA3¥ respectively (Table 1). The WWP values ofthe reused PLA-based fins are aso inched in Table 1, Those corresponding to reused PLA and PLAO,S9 respectively are statistically non-different from those of the new prepared fils, revealing tha the structure and morphology ofthe flm was not changed during 17 days of| Storage at 4 °C. The permeation to gases is limited when the composite particles fromthe matrix are wel dispersed, thus providing a substantial tprovement of de gas baie. The theory that explains this Behavior i focxsed on tortous path around the composite particles reducing the Aifvsion coefficient and forcing the permeant gas to travel longer path (o diffuse through the flm (Silvestre et al, 2011, Li et al, 2017). However, Chi etl (2019) showed thatthe permeability 10 gases of PLA decreases by addition of TiO, and Ag respectively. Suwanamnomert tal. (2020) showed that by addition of poly(butylene succinate co ‘adipate)(PBSA) and thymol into the PLA mati, the WVP is reduced by ~ 284, Moreover, by doubling the thy content into fn from co ‘Tablet rod hemi 969 (2021) 190941 Characteristics ofthe Ag graphene-TIO, PLA fmsnew prepared and reused (MD-machine ection, CD ros dreton, WYP- water vapor permeability, GP- grease ‘pemienblty, OTR- oxygen ranaission rte, XOH -ethanoD, Packaging material | macroscopic | SEMUimage | Tensestengh (Pa) | WWP” | GP°G@) | OTR™ ‘Giba mean Cow?) sew (x2 | see | ems! | aoa) HoH | tens Be | RE | WR) [aE FAIS FE |S] aA] a] TE TE aT us | 26s | some | cise | 20s | soe soe | ar (raves 0.3%) ganz | ass | ose | qos FARE fl Wee] Sas] sar | sss | soon | sane | sas [soe | +06 | +00 | soe "he stata igalcance was established for each parte ai a fncion ofthe package Fp; oes te same column With He same Tete indeate aon slgnlicantdiference and means inthe sae column with eliferent lett ndeate sgaieant (p< 0.05) iference. 6, WP is reduced by 6%. The explanation i given bythe hydrophoble ‘character of thymiol that increases the barrier against water vapors (Soshirvanl et aly 2017. They showed similarly behavior also inthe ‘case of oxygen penetration. The barrier for oxygen increased after Including PRSA and thymol into PLA matrix. ook eal. (2018) deter ined the Oxygen Transmission Rave (OTR) for PLA based fms mod fed with 03 and ole acid and they have established that the OTR is ‘reduced as the T content inereases up (0 0.5%, A higher content of Tin the fm matrix (196 and 396 respectively) do nor significantly langes the OTR. They have obtained better results when the oleic acid was Inluded, asa result of the good dispersion of TiO ito the fatty ac. ‘The permeability to grease increased from 0.3725 + 0.0029 for PLA 10 0.7332 + 0.009% for PLAS. The permosbilty of PLAS is signi ‘cantly higher than tht of PLA and PLAO.Si The permenbility to grense ‘of the reised PLA based flns isnot slgnlfealy different that of the new prepared ones (lable D. “The results of the overall migraton are also presented in Table 1. The highest degrees of migration inthe 1026 ethanol solution that mimics aqueous foods were observed for the unmodified PLA and reference packaging. In contrast, the lowest migration occurred fom PLAO.SY ‘and PLASM packaging. Identical values forall tested materials were ‘obtained forthe overall migration index both in 9596 ethanol at 20 °C ‘and 40 °C respectively, and for isooctane! at 20 °C. Differences appear ‘when migenting into fsooetanol at 40 “C, the highest migration values being obtained fr PLA and PLAO.5%. Thus, PLAS proved to be the less soluble inthis simula, Let). (2077) obtained similar resus fo the ‘migration of Ti and Ag respectively, ftom PLA based films into acetic acid and edhanol respectively. AS the migravion tine elapsed, the ‘amount of the migrated particles increased gradually untl the eq rium is reached. An acidic solution shall more easily transfer Ti or Ti0> Into the fod. Permeability of water vapor (16.2 0.28 m? x 24h), grease (0.049 0.000156), oxygen (124 + 24.5 ml x day) a5 well as the tensile strength (MD = 5242.25 + 34.2 MPa, CD = 5862.52 + 17.8 MPa) ofthe ‘reference polypropylene (PP) commercial fl designed forthe tempo- raily food wrapping have been also determined. The reference bag, ‘exhibited significantly lower WVP, grease and oxygen permeability ‘compared to the PLA-based films. The diferences results from the -various polarities of PLA based fms in comparison with polypropylene reference bag. The PLA films have more polar character than (Suvanamomalet ets, 2020) The results of the films antibacterial activity are presented in ‘Table 8.2 from Suplenientaay’ ater. All packages showed ihibicory activity agains the tested bacteria, PLAO.SMs and PLA: have signif cantly higher inhibitory stivity than PLA against Eco. The same result was obtained By Let al. (2017) namely, the PLA Ans containing & ‘certain peroent of Ti and/or Ag presented a higher antibacterial activity {hat neat PLA. The ability ofthe fils to inactivate the bacteria increases with the increase of he composite content. Even if the Hteratare men: ‘dons that the mmininmm amount of nanoparticles required to ill the bacteria is 3H) (Pascual and Diex Vicente 2014, our results proved that a valuable inhibitory activity was also achieved by incorporating only 0.594(w) composite into the PLA matrix. This is because the nano: composite particles display very small size and are able to generate more lectron-hole pairs and the apoptosis process occurs more rapldly. The growth of Salmonella enercais extremely ess inhibited by PLAO.5%, PLAS: and by reference. However, an improved Inhibitory effet is observed inthe ease of unmodified PLA. PLADS9% and PLAS exert increased inhibitory action against Lis teria monocyogenes in comparison with unmodified PLA and reference. All the tested packaging materials showed reduced inkibitory activity against Pseudomonas aeruginosa In contrast, PLAG.S% and PLAS% were showin to have inactivation potential ageinst Pseudomonas fluorescens to significantly greater extent then unmodified PLA and reference. 13, Storage of cunt cheese in new and reuse Ag grphene-TiO> PLA fil packages 3.3.1. Organoleptic changes ofthe curd cheese stored in new and used packages [At the beginning ofthe experiment (day 0), the cheese had plensant appearance without leaks, white color, homogeneous consistency and Aavor specific to the fresh curd cheese (Fable 2). ‘The characterises ofthe cheese kept inthe newly prepared package. After three days of storage, no difference in organoleptic parameters as & fanetion of the packaging type was observed. After 17 days of storage, the PP cheese bind a slightly whey leakage atthe surface. I fas been » lite mottled, had a white color without changes and the flavor was strougly stinging, sour. In PLA, the external appearance of the cheese showed a weak whey leakage at the surface, iC was mote a lite, ‘Table2 rod hemi 969 (2021) 190941 Images ofthe urd cheese tre in new an ese packages (PP -plypropvene bag) PLA PLAOSYS PLAS Reference Dave new package ed package DAYS i cea aa Teed package DAY TO _ az = resol package DAV IT - iad ech sed package yellow colored and had a lavor of yeasts and acids. The cheese wrapped Jn PLAO.56 for 17 days had whey leaks and @ mucilaginous film on the surface, the color was yellowish, the appearance in the section was ‘compact and the smell was pungent. The cheese from PLA3Ss had the ‘nian change with regard to aspect and color. It was homogeneous, Inada slightly natte appearance, white color, without changes and the smell was very slightly pungent (abe 2). ‘The characteris of the cheese Ree in the reused packaging. The same ‘observations emerged after 8 days of keeping the cheese in the reused packaging, Le. there are no differences berween the PLA based pack ages, The cheese had a slightly matte appearance, white yellow color ‘and & slightly pungent smell only in the PP packaging. After 17 days of storage, he weak whey leakage at the surface, mowed, brite and ‘rubberized on the edges and deeply yeast-acidie favor can be noted in the cheese stored both in PP and PLA, The results are similar also for LAO 596, butin this ease the favor is beter (very light favor). The best results were obtained inthe case of cheese stored in PLAS, It had no witey leakage at the surface, very reduced brite, white color and pleasant favor (able 2). The overll acceptance of the organcleptic changes of the eurd cheese Is presented in Figure 8.6, from supploasen ‘ary material. The esultsare in agreement withthe Images from Table 2. ‘With regard tothe new prepared package, the preservation activity of, PLAD9% and PLAS was 12.5% and 22.22% respectively, higher than that of PP, whereas the conservative role ofthe reused PLAS was SO and 40% respectively higher than that of reused PP. When, the new and reused packages ate compared, it can be noticed that the active role of LAO 59% and PLAS to preserve the organoleptic characteristes ofthe curd cheese decreased only by 8% and 436 respectively, ater reuse. ‘The correlation coefficient of the organoleptic evaluation between the sample kept in new packaging and that kept in reused packaging (0.89, which demonstrates that the tendeney of organoleptic variation ‘occured in new packaging is similar with that in reuse packaging, ie the cheese isthe preserved the best in PLA 39 ‘The acceptance conditions according to Romanian Standard SR '3604:2008 are those evaluated with the score in the range 4-9 in Table S.2. from Supplementary material. These requirements were accomplished after 10 days of storage in all new and reused packages ‘and moreover, his period cold be extended by 7 days in the case fe ‘and reused PLA 39, because the requested score wns mi ‘lay for shes types of packages. 38.2 wt los ofthe new and reused packages Water lose is at important parameter affecting during storage, transport and marketing. Weight loss of the wrapped ‘cheese refers tthe dehydration ofthe product influenced by the Vapor barrier properties ofthe packaging fl (Chi etal, 2019, Liv, Li etal, loss gradually inereased during che storage time. This would be attributed tothe continuous evaporation of moistre from the cheese to the environment, The mass lose of the cheese packages i presented in Figure $7. from Supplementary material. After 3 days of storage, the differences in weight loss berween the samples were st Hsteally non signfiean. Afr 10 days of storage, the lowest weight lass was determined in PLAS% (0.99%) (p < 0.01) compared to other sam ples where hs been between 1.06% and 1.8%. The differences between the wo valtes are not statistically significant. After 17 days of storage, the weight os increases inthe following order: PLAS < reused PLAS% PLA ~ reused PLA « PLAO.5% ~ reused PLAG. 5%, The weight loss of the cheese from PLAS package is 4.15% (p < 0.01). The values cor responding to reused PLAS, PLA and reused PLA are in the range 7.654% — 8.05%, the differences being not statistically significant. Also, the values of the weight loss corresponding to PLA0.5% and reused PLA0.596 are very close (11.586 and 11.7%, respectively), The mas loss forthe BP packaging was ow (p < 0.01) aseompared to that determined Inthe case of PLA based packages after 17 days of storage, being 1.62% for PP and 0.74% for reused PP respectively ‘The weightloss values ofthe cheese are correlated with dhe WYP values ofthe packaging materials. The lowest mass loss occurred in the package with PLAS, whieh i the less permeable to water vapors. The results are in agreement with those obtained by Siar etal. 2018) tac reported the significantly decrease of the permeation ability of water vapor by adding nanopartles as a result ofthe ehanges in the interaction between molecules and nanoparticles casing a more tortious diffusing ofthe water vapor molecules through the polymer ‘matric (Li etal. 2017). The very approprited values ofthe weight loss corresponding t0 the pairs PLA - reused PLA and PLA 0.5% - reused PL.A0.5%, respectively: demonstrates the mininnm changes of theit ‘water vapors permeability afer the Rist use. ‘The correlation coefficient of the weight Toss between the sample kept in new packaging and that Kept in reused packaging is 0.98, ‘showing that te variation tendency was de same in reused packages as in the new ones 9.3.9. pH value and atable acidity of the curd cheese stored in new ond ‘used packages ‘The pH value and testable eldity of the cheese samples are shown In gure 88. from sipplenentory’ material and Fig. 1. The pH of the ‘cheese at te beginning ofthe experiment was 4.069 0.02, det the predominantly aed coagulation (Piero Lago el, 2019), The Tow pl value at dhe Beginning of the experiment bi the absence of proteolysis (Pineiro Lago etal, 2019). The pH of the cheese samples kept in the new packaging reaches ~ 4.55 + 0.03 (p < 0.01) after 17 ‘days of storage, while the cheese's pH in te reused packaging shows & decrease (p< 0.01) in the ist three days and a constancy fom day 30 day 17 (Figure SB. from Supplementary material. The highest pH variation was observed for the cheese stored in PLA, reference and PLASM after 17 day. In contrast, inthe ease of reused packages, the pH rod hemi 969 (2021) 190941 ee eee time (days) Pig. 1, Tuatable acy ofthe cud cheese stored in new ane wed packages. ‘eerease is sharply accentnated in the Mts days in the reference, while the pit remains constant in PLA based packaging. The sante varation was established in the ease of titratable acidity. This parameter de creases overtime for dhe cheese Kept in new packaging and registers & slight inerease in dhe ease of cheese wrapped in the rensed packaging (ig 1, The decrease ofthe titratable acidity was noticed also by ce ‘Moraes et al. (2020) chat stored fresh cheese in the films based on fntimlerobial starch modified with cittic acid, In the case of new peckaging, the most accentuate acidity deerease was observed in the ‘hese kept in the reference, PLA and PLA3%6, while the acidity of the hese in PLAO.5% decrease significantly (p < 0.01) in the Mist 3 days and then remains constant from day 3 to day 17. Normally, the acidity of| cheese decreases as a consequence of lactic acd consumption and ofthe alkalizng effect of compounds generated during the proeins degrada tion (Secowska ot al, 2009). The acidity ofthe cheese in the reused packages increases, dhe most accentuated increase (p < 0.01) being recorded in the cas of the cheese kept in reused PLA, during 17 days of storage. The acidity ofthe cheese samples from the other reused pack ‘ages is approximately the same (ranged between 158 and 165 °T). The Romanian Standard SR 6642008 establishes value 190 °P as the ‘maximum aeoepted value forthe tatable acidity for cheese. This value was not reached by any sample during the 17 days of storage. Youssef cecal. (2015) have showed thatthe lowest increase of the fresh cheese's cdity during of one nonth of storage orcurred in she package based on biacomposite modified with 4% and 8% nan0-Ti0p, Youser et al-(2019) studied the onganoleptial and physical-chemical changes of Ras cheese during storage in chitosan (CS)/polyviny alcool (PVAY bend mi ‘with nano-Ti0. and thee results shoved also a slightly decrease ofthe Ras cheese acidity in both reference materials and CS/PVA with TiOp from day 210 day 3 of storage 3.34. Far content ofthe curd cheese stored in new and used packages ‘According tothe Romanian Standard SM 3664:2008, the cheese with f fat content of minimum 27% Is classified as fatty cheese and that containing minimum 2036 fats semi fatty cheese. The fat content ofthe hese at the beginning ofthe experiment was 29.9 0,463, soca be classified as faty-chees. The fat content continuously decreases during storage 10 values of at least 10.25 + 0.56%6 in the case of rensed PLA cheese (Hg. 2). The most redueed drop took place in the cheese kept in new and reused PLADS% and PLAS respectively, and the most accentuated in new and reused PLA. No significant diferences ofthe fat fat content (4) ae are eee ieee es time (days) Fig. 2 Fa content ofthe cud cheese sored in new a se packages ‘content between new and reused packages appeared. The reducing in ft ‘ontent was observed also by Yousef etl. 2019) who have studied the ‘hanes ofthe cheese properties during storage in bioeomposice mate Finlsbased on chitosan polyvinyl alcohol modified with nano-Ti0. They ‘obtained a non uniformly variation of the cheese fat content as a func tion of the nano-TiO; content into biocomporte matrix and of the storage period. The fet content of the cheese stored in biocomposite material wiht TiO, inereased In 8 months from 28.8% to 29.7% dae to the moisture loss, while in the presence of 1% nano-Ti0s a slightly

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