Received Accepted

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1 Reserch Journl of Chemicl nd Environmentl Sciences Res J. Chem. Environ. Sci. Vol 5 [3] June 217: Online ISSN CODEN: RJCEA2 [USA] Acdemy for Environment nd Life Sciences, INDIA Wesite: ORIGINAL ARTICLE RJCES Evlution of Edile Cotings Effect on Aneroic Metolism nd Voltile Compounds Production of Fresh Tomto (Lycopersicon esculentum Mill.) cv. Grndell Dávil-Aviñ JE 1, Gil-Chvez GJ 2, Villegs M 2, Olivs GI 3, Ayl-Zvl JF 2, River-Dominguez M 2, Sunil Preek 4, González-Aguilr GA 2 1 Deprtment of Microiology nd Immunology, Fculty of Biologicl Sciences, Autonomous University of Nuevo León, Post Office 124 F, Ciudd Universitri, Sn Nicolás de los Grz, Nuevo León 66451, Mexico 2 Coordintion of Food Technology of Plnt Origin, Center for Reserch in Food nd Development, A.C., Crreter l Victori km..6, Aprtdo Postl 1735, Hermosillo 83, Sonor, Mexico. 3Lortory Technology Lortory of Plnt Origin, Center for Reserch in Food nd Development, Cuuhtémoc, Chih., Mexico. 4Deprtment of Agriculture & Environmentl Sciences, Ntionl Institute of Food Technology Entrepreneurship & Mngement, Kundli, Sonept, Hryn 13128, Indi ABSTRACT Edile cotings hve een developed to reduce posthrvest deteriortion of different crops. Nevertheless,in some cses, when edile cotings re not dequtely selected, they cn cuse neroic respirtion. In this context, this work evluted the effect of crnu nd minerl oil cotings on the neroic metolism nd production of voltile compounds of tomto fruits (cv. Grndel ) t two mturity stges (reker nd pink) over 28 d of storge t 1 C. The voltile compounds nd the neroic metolism ssys included: romtic compounds derived from lipids, mino cids nd crotenoid, s well s, ctivity of lcohol dehydrogense (ADH), lctte dehydrogense (LDH), pyruvte decroxylse (PDC) enzyme nd concentrtions of some metolites such s ethnol, cetldehyde, lctte nd pyruvte during storge. The results of romtic compounds showed significnt increse in hexnl, 6-methyl-5- hepten-2-on nd decrese in 2-methylutnol, hexnol, cis-3-hexenol while mintining 2-hexenl nd pentnl, for ll tretments. Edile cotings did not ffect (p.5) cetldehyde nd ethnol content of tomto fruit t oth mturity stges. PDC ctivity incresed during storge for oth mturity stges. ADH enzyme ctivity ws similr in ll tretments mesured in oth mturity stges. LDH ctivity tends to decrese during storge for oth mturity stges nd the lctte concentrte is mintined during storge for oth mturity stges. Edile cotings hd significnt effect (p.5) on the pyruvte content of tomto fruit t oth mturity stges. However, the production of pyruvte is similr to those of control fruit. These dt revel tht different coting methods not ctive the neroic metolism of tomto fruits, without generting off-flvors, in fruits covered with some type of coting. Keywords: Lycopersicon esculentum, ADH, LDH, Voltile compounds Received Accepted AELS, INDIA INTRODUCTION Tomto (Lycopersicon esculentum) is one of the most widely consumed fresh vegetles worldwide. However, its highly perishle nture limits its posthrvest life [1]. The perishility of tomto requires the development of technologies tht reduce their posthrvest deteriortion nd extend their shelf life [2]. The use of edile cotings in conjunction with low temperture storge ppers to e promising pproch to minimize these prolems nd preserve the freshness of tomtoes [3]. Edile cotings, is semi permele rrier of edile mteril (protein, polyscchride nd/or lipid) nd is pplied to the surfce of fruit produce providing semi permele rrier ginst gses nd vpor, decresing the rte of physiologicl posthrvest deteriortion [4,5]. However, n indequte selection of the mterils nd systems of ppliction, edile cotings could cuse neroic respirtion or desicction of the product ffecting its overll qulity [6]. The rrier formed y the edile coting cn reduce respirtion nd form modified tmosphere inside the commodity, reducing the internl O 2 levels nd cusing decrese in ethylene production, s ws oserved in tomto [7]. In ddition, we hve oserved tht the use of edile cotings does not ffect the nutritionl qulity of tomtoes since it preserves the content of vit C nd other ntioxidnts [8]. RJCES Vol 5 [3] June P g e 217 AELS, INDIA

2 Dávil-Aviñ et l Nonetheless, in order to hve n overll picture of the efficcy of edile cotings to prolong the storge life of tomtoes, their effect on the neroic metolism must to e tested. Edile cotings cn cuse neroic respirtion [9]_ENREF_9, triggering the ctivity of some enzymes like lcohol dehydrogense (ADH), pyruvte decroxylse (PDC) nd lctte dehydrogense (LDH), since under this condition glycolysis is fundmentl for the dpttion to noxic environments [1,11]. This pthwy is predominntly linked to ethnolic fermenttion, involving the production of some metolites from the neroic respirtion, resulting in n undesirle flvor on the fruit [12]. Given the perishility of tomto nd the lck of studies out the effect of edile cotings on their neroic metolism, the purpose of the present study ws to determine the effect of two types of commercil cotings which hve showed good results on posthrvest physiology nd nutritionl qulity [7,8] on voltile production nd the neroic metolism (ctivity of ADH, LDH, PDC enzymes) nd the chnge in concentrtions of some metolites such s ethnol, cetldehyde, lctte nd pyruvte during storge of tomto fruit. MATERIALS AND METHODS Plnt Mteril Fresh tomto fruits (cv. Grndel ) were evluted in this study. The fruits were greenhouse produced in Oregón, Sonor, in the northwestern region of Mexico. Helthy fruit smples were snitized with chlorine (2 ppm) nd lter clssified for size, uniformity nd color ccording to the USDA stndrd tomto color clssifiction chrt 7. Fruits t mturity stges 2 (those presenting color other thn green on less thn 1% of the whole fruit) nd 4 (those presenting color other thn green on 3 to 6% of the whole fruit) were selected. Fruits were divided into two tches ccording to mturity stge nd sudivided into three groups (control, minerl oil, nd crnu). For ech mturity stge, the edile cotings (crnu or minerl oil, FMC Food Tech, Riverside, CA) were pplied mnully using ArtexMR rushes S 111ª No 9 México t rte of 1 liter per ton. One hundred eighty fruits per mturity stge (6 fruits per tretment) were evluted nd stored t 1 C. Finlly, physiologicl nd physicochemicl prmeters, enzymtic ctivity nd metolites were recorded on dys, 5, 1, 15, 21 nd 28 of storge. Physiologicl nd Physicochemicl Prmeters Respirtion rte ws mesured plcing individul fruits in plstic continers (1.8 L cpcity) hermeticlly seled. After 1 h hed spce smple (1 ml) ws tken using hypodermic needle nd injected into Vrin Str 34 CX gs chromtogrph (Vrin, Mexico) equipped with Hye Sep N column (2 mm, 3 mm ID; 8/1 µm prticle size) nd two detectors (the therml conductivity detector (TCD) for CO 2 quntifiction, nd the flme ioniztion detector (FID) for ethylene. Temperture conditions were s follow: 5 C for the column, 7 C for the injector, 17 C for the TCD detector nd 25 C for the FID detector. The concentrtion of ech gs ws quntified y determining the re under the curve nd compred with tht of known stndrds. The results re recorded s ml CO 2/Kg h nd L C 2H 4/Kg h. Skin color ws determined in three points of equtoril re of the fruit using Minolt colorimeter (model CR 3; Minolt corp., Rmsey, NJ, USA). Vlues were otined in CIELAB scle (L*, *, *) nd Hue ngle nd chrom vlues were clculted. Weight loss ws determined ccording to the initil weight using digitl lnce (Mettler Toledo, New York, NY, USA) nd the results were reported s percentge of weight loss. Tomto tissue firmness ws mesured y puncture method, using Chtillon Penetrometer, Model DFM5 with n 8mm dimeter flt hed stinless steel cylindricl proe. Tissue s opposition force ginst the penetrtion ws registered on 3 points in the equtoril region of the fruit nd results were reported in Newtons (N). Enzyme Activity Assys The enzymtic ssys were mesured following the oxidtion of the NADH (nicotinmide denine dinucleotide, reduced form) for unit of time t 34 nm in Spectrophotometer, Cry 5 Bio (Vrin Itly). The lectures in UV visile were registered, nd monitored the chnge of sornce of the NADH. The enzymtic ctivity ws reported in specific ctivity (UI/mg protein). Isoltion of lcohol dehydrogense (ADH) nd piruvte decroxilse (PDC) ws mde using the methodology reported y Chng et l. [13] with slight modifictions. Rw extrction were otined from 2 g of fresh tissue in finl volume of 5 ml of uffer contining Tris se 1 mm (ph 8.), dithiothreitol (DTT) 5 mm nd.5% polyvinylpyrrolidone (PVP). Fresh tissue ws homogenized nd centrifuged (Beckmn Coulter Allegr 64 R rotor C115, USA) t 15 xg for 15 min t 4 ºC. The superntnt ws seprted nd the pellet ws discrded. The superntnt ws used to determine enzymtic ctivity nd protein concentrtion. ADH ws mesured y mixing.8 ml of uffer contining Tris se 25 µm (ph 7.2),.5 ml of NADH 5 mm,.1 ml of rw extrct nd.5 ml of cetldehyde 8 mm. The decline of sornce t 34 nm during t lest 1 min of the rection ws otined. PDC ctivity ws ssyed y mixing 2 ml of phosphte uffer.5 M (ph 6.2),.1 ml of MgCl 2 5 mm,.1 ml of nicotinmide denine dinucleotide (NAD) 1.25 RJCES Vol 5 [3] June P g e 217 AELS, INDIA

3 Dávil-Aviñ et l mm,.1 ml of ADH 13.5 units,.3 ml of rw extrct nd.1 ml of N pyruvte 5 mm. The decline of sornce t 34 nm during t lest 1 min of the rection ws otined [13]. LDH ws crried out ccording to Arulsekr nd Prfitt [14] methodology. Rw extrct ws otined from 2 g of fresh tissue in 12 ml of uffer contining Tris se.5 M (ph 8.), citric cid.7 M, cysteine HCl 1%, scoric cid 1% nd polyethylene glycol (PEG) 1%. The mix ws homogenized nd 8 µl of β mercptoethnol (1 mm) nd.5 g of polyvinylpolypyrrolidone (PVPP) ws dded nd centrifuged t 1 xg for 2 min t 4 C. The superntnt ws otined nd the pellet ws discrded. The superntnt ws used to determine enzymtic ctivity nd protein concentrtion. LDH ctivity ws mesured in the reverse direction y following pyruvte dependent NADH oxidtion t 34 nm in the presence of 4 methylpyrzole to inhiit ADH, nd NCN to lower the ckground rte of NADH oxidtion; the stndrd ssy mixture contined 1 ml of Tris HCl.13 M (ph 8.), 15 µg NADH, 3 µm 4 methylpyrzole, 3 µm NCN, 12 µm N pyruvte, nd.2 ml enzyme extrct. LDH ctivities were reported in specific ctivity (UI/mg protein). The decline of sornce t 34 nm during t lest 3 min of the rection ws otined [15]. Metolites Extrction nd Anlysis The metolites extrction were performed using the methodology previously reported y Good nd Muench [16]. The rw extrcts were otined from 2 g of smple which ws homogenized with 1 ml of uffer of HClO 4 (6%). These extrcts were centrifuged t 5 rpm during 3 min t C in centrifuge (Beckmn Coulter USA). The ph of the superntnt ws djusted t 4.5 using ph Meter TM 5 (Beckmn instruments, Inc., Fullerton CA, USA) nd mintined during 5 min in oiling wter nd centrifuged under the conditions previously descried nd then filtered. The metolite`s extrction were performed y triplicte nd the quntifiction twice, monitoring the oxidtion of the NADH t 34 nm during 5 min in Spectrophotometer Cry 5 Bio. The pyruvte concentrtion ws mesured using the rection mix of 1 ml of uffer of triethnolmine.4 M (ph 7.6), 2 ml of deproteinized smple, 5x1 3 M of NADH, nd 1 mg protein/ml of LDH in finl volume of 3 ml [17]. The concentrtion of lctte ws mesured using the rection mix of glycine hydrzine uffer, contining.4 M hydrzine, nd 1M glycine (ph 9.5),.5 µl of deproteinized smple, 5x1 2 M of NAD nd 5 mg of protein/ml of LDH, in finl volume of 1.1 ml [17]. Off-flvor Anlysis Ethnol nd cetldehyde determintion were performed using the method previously descried y González Aguilr et l. [2]. 1 g of smple were plced in 2 ml continer nd incuted in wter th (Precision Scientific, Chicgo, IL, USA) t 7 C for 15 min. Afterwrds, 1 ml hedspce smple ws injected into Vrin Str 34 CX gs chromtogrph (Vrin, Wlnut Creek, CA equipped with 2 m x 1/8 inch Chromosor stinless steel column, pcked with 8/1 Am mesh Porpck 11). Ethnol nd cetldehyde in wter solution were used s stndrds for pek identifiction nd quntifiction. Arom Voltiles Concentrtion of tomto voltile compounds were determined y gs chromtogrphy (GC) using solid phse microextrction (SPME) technique. Tomto juice from six tomtoes per tretment were otined with food processor (Turmix). Then 2 ml of juice ws plced in 2 ml PTFE vil, frozen in liquid nitrogen nd kept t 7 C until nlysis. An liquot of 2 ml of thwed tomto juice ws plced in 4 ml vil contining.7 g of sodium chloride nd stirred while SPME fier (65 m, PDMS DVB, Supelco USA) ws exposed to the hedspce of the smple for 1 h t room temperture (25 C). The fier ws desored y splitless injection for 1 min into the injection port of n Agilent 782 gs chromtogrph [18]. Chromtogrphic conditions were s follows: 2 C injector temperture, splitless mode, 3 C flme ioniztion detector (FID) temperture, nd DB Wx cpillry column (J&W Scientific, 6 m,.25 mm i.d..25 rm film thickness). Oven progrm prmeters were: 35 C initil oven temperture held for 5 min, incresed to 5 C t 2 C min 1, incresed to 2 C t 5 C min 1 held for 5 min. Helium ws used s the crrier gs with liner velocity of 3 cm s 1. Identifiction of voltile compounds of interest ws mde y mtching their spectr with those of the NIST 98 MS lirry nd y compring the retention times ginst_enref_17 those of high purity stndrds (Sigm Aldrich nd ChemService) [18]. All vlues represent the verge of triplicte smples consisting of six tomtoes. Sttisticl Anlysis The dt were nlyzed s rndomized complete lock design using the GLM procedure of the Numer Cruncher Sttisticl System version 6. softwre (NCSS, LLC). Storge time ws chosen s the locked fctor to oserve the effect of the edile cotings on tomto physiologicl nd physicochemicl prmeters. Differences etween tretments were determined using Tukey s comprisons test. p.5 ws considered significnt. RJCES Vol 5 [3] June P g e 217 AELS, INDIA

4 RESULTS AND DISCUSSION Physiologicl nd Physicochemicl Prmeters Tle 1 shows the initil dt qulity prmeters of tomtoes in two stges. At the eginning of the study, CO 2 production ws reduced y 38 nd 46% when pplying the minerl oil coting on reker nd pink tomtoes, respectively. In ddition, erly during the study, the minerl oil coting showed suppression of ethylene iosynthesis t oth mturity stges. At the end of storge, minerl oil cotings delyed color chnges nd reduced weight losses for 7 nd 46% t the reker nd pink stges, respectively. In generl, respirtion rte, color, weight loss nd enzyme ctivity were positively ffected y minerl oil coting in oth mturity stges. No effects on firmness, titrtle cidity, nd ph were found y coting ppliction. These dt were pulished in Dávil Aviñ et l. [7], concluding tht minerl oil coting could e good lterntive to preserve qulity nd extend posthrvest life of tomto fruit. Tle 1. Initil dt of qulity prmeters of tomto fruits (cv. Grndel ) t pink nd reker mturity stges treted with minerl nd crnu cotings. Tretments Prmeters Breker Pink CO2 (ml/kg.h) Ethylene (µl/kg.h) L* Control * * Firmness (Newtons) ph Titrtle cidity (%) CO2 (ml/kg.h) Ethylene (µl/kg.h) 1.3 L* Minerl Oil Wx * * Firmness (Newtons) ph Titrtle cidity (%) Dávil-Aviñ et l CO2 (ml/kg.h) Ethylene (µl/kg.h) L* Crnu Wx * * Firmness (Newtons) ph Titrtle cidity (%) Enzyme Activity Figure 1 shows the chnge in the ADH ctivity of tomto fruits treted with minerl oil coting nd crnu coting in two mturity stges nd over 28 dys of storge t 1 C. For fruits in reker (p=.24) in ADH ctivity; control, minerl nd crnu coting were 5.94, 5.22 nd 6.8 UI/mg protein, respectively. On the other hnd, the ppliction of cotings hd significnt effect in ADH ctivity, hving 5.44, 5.5, nd 8.14 UI/mg protein in control, minerl nd crnu cotings. However, no significnt differences were oserved etween control nd treted fruits. The chnges in the ctivity of PDC of tomto fruits treted with edile cotings re shown in Figure 1. In generl, it ws oserved n incresing ehvior for PDC during the storge in oth mturity stges, showing significnt differences within the tretments (p.5). For reker tomtoes ctivity ws 32.19, 4.11 nd 37.9 UI/mg protein for control, minerl oil nd crnu coting, respectively, nevertheless: there ws no difference etween crnu nd minerl cotings. The ctivity of PDC in pink stge showed vlues of 37.33, nd 3.69 UI/mg protein for control, minerl nd crnu coting tretments, ll three groups were sttisticlly different. Figure 1 shows LDH ctivity in tomto fruits treted with minerl nd crnu coting in two mturity stges, during 28 dys of storge t 1 C. In generl, LDH ctivity tended to decrese during the time of RJCES Vol 5 [3] June P g e 217 AELS, INDIA

5 Dávil-Aviñ et l storge in oth reker nd pink stges. For reker tomtoes, the ctivity of LDH ws sttisticlly different (p.5) within the tretment with cotings with vlues of 17.87, nd UI/mg protein for control, minerl wx, nd crnu, respectively. In pink tomtoes the vlues for LDH ctivities were 6.24, 5.77 nd 7.92 for control, minerl wx, nd crnu, respectively. However, there ws no difference etween control nd minerl sed coting in reker or pink stges. PDC ctivity incresed (p.5) with storge reching the mximum (1 12 UI/mg protein) on minerl oil nd crnu cotings, eing significntly higher thn LDH nd ADH ctivity in oth mturity stges. In generl, ADH ctivity (2.5 1 U/mg protein) ws rely ffected y cotings; only pink fruit hd significnt increse on ADH ctivity (from 3 to 2 U/mg protein) fter 1 d. LDH ctivity significntly incresed during the first 1 d in higher extent on fruits coted with minerl coting nd decresed in similr pttern thn controls for oth mturity stges. LDH nd PDC enzymes shre simultneous pthwy competing for pyruvte nd NADH under neroic conditions. In this work, the ctivity of LDH tends to decrese during storge for oth mturity stges while PDC ctivity showed n increse. It is well estlished tht the production of lctte cuses decrese in cytoplsmic ph resulting in the inhiition of LDH ctivity nd the increse of PDC ctivity which is ctivted under low ph [19,2]. On the other hnd, the ctivity of ADH enzyme ws similr in ll tretments mesured in oth mturity stges. However, pink tomto shows the highest vlues of ADH ctivity in crnu coting on dy 1 th of storge. Afterwrds, reched similr vlues thn those otined for reker y the end of storge. In generl ADH ctivity coincides with the production of ethnol which remins constnt thought ll storge time. Breker Control Minerl Oil Wx Crnu wx Pink 3 25 ADH (U/mg protein) c PDC (U/mg protein) c c 4 c LDH (U/mg protein) 3 2 c c 1 Dys t 1 C Figure 1. Activities of PDC, ADH nd LDH (U/ mg protein) enzymes of tomto fruits (cv. Grndel ) t pink nd reker mturity stges treted with minerl nd crnu cotings nd stored for 28 dys t 1 C. Ech vlue is the men of 6 replictes ±SE. Different letters represents significnt difference (p<.5) etween tretments. The use of edile cotings prolong the shelf life of tomto fruit in oth mturity stges, however; some negtive effects such the production of end products cn occur. In generl, the ppliction of crnu RJCES Vol 5 [3] June P g e 217 AELS, INDIA

6 Dávil-Aviñ et l nd minerl coting in reker nd pink tomtoes does not result in ctivtion of neroic metolism which ws mesured y enzymes nd metolites of this process. Metolites The chnges in pyruvte concentrtion of tomto fruit (two mturity stges) treted with minerl nd crnu coting, during 28 dys of storge t 1 C re shown in Figure 2. Pyruvte content ws significntly (p.5) ffected y the edile cotings in oth mturity stges. Minerl coting ws sttisticlly different to crnu coting nd control, hving pyruvte contents of 28.85, 3.5, 33.6 µmol/g fw for control, minerl nd crnu coting, respectively. Control nd crnu coting doesn t show significnt differences.. Stge pink tomtoes showed significnt differences mong edile coting tretments nd control. The increse of pyruvte is consistent with the increse in the ctivity of PDC (Figure 1). This ehvior ws lso oserved y Kder [21] which report tht PDC is ctivted y decrese in ph nd incresed concentrtion of pyruvte. Edile cotings result in incresing the levels of pyruvte during the storge, however; there ws no difference compred with control t the end of storge suggesting the pplictio ion of this cotings did not ctivte the neroic metolism in tomto fruits. These results suggest tht even fter eing coted, tomto fruit mintins eroic conditions nd preferle pyruvte entered the tricroxyilic cid (TCA) cycle. The effect of edile cotings on lctte content ws different depending on the mturity stges of fruits (Figure 2). This effect ws significnt for reker tomtoes (p.5); where lctte content vlues were 25.51, 3.94 nd µmol/ g fw for control, minerl wx nd crnu, respectively. No significnt differences were oserved etween control nd tretment with crnu coting. In ddition, no significnt differences (p=.28) were oserved in pink tomtoes etween controls nd tretments, eing lctte content vlues 29.17, 27.2 nd µmol/g fw for control, minerl wx nd crnu, respectively. Studies on the possile toxicity of the fermenttion end products re minly focused on lctic cid, cetldehyde nd ethnol [22]. Figure 2. Pyruvte, lctte (µmoles/ g fw) contents of tomto fruits (cv. Grndel ) t pink nd reker mturity stges treted with minerl nd crnu cotings nd stored for 28 dys t 1 C. Ech vlue is the men of 6 replictes ±SE. RJCES Vol 5 [3] June P g e 217 AELS, INDIA

7 Dávil-Aviñ et l Off-flvor Anlysis Figure 3 shows the chnges in the cetldehyde contents of tomto fruit t two mturity stges treted with minerl nd crnu cotings over 28 dys of storge t 1 C. Edile cotings hd not significnt effect (p.5) on the cetldehyde content of tomto fruit t oth mturity stges. For reker tomtoes, the nlysis of vrince for cetldehyde showed not significnt differences (p=.7) mong tretments. A glol effect showed men vlues of , nd µl C 2H 4O/g fw for control, minerl nd crnu cotings, respectively. The cetldehyde content showed not significnt differences (p=.87) for pink tomtoes; the vlues oserved were , nd µl C 2H 4O/g fw for control, minerl, nd crnu cotings, respectively. In generl the cetldehyde content tends to decrese during storge for oth mturity stges. Figure 3 shows the chnges in the ethnol content of tomto fruits t two mturity stges treted with minerl nd crnu cotings over 28 dys of storge t 1 C. For reker tomtoes, no significnt effect of the tretments on ethnol content ws oserved (P =.82); men vlues for control, minerl wx nd crnu were , nd (µl C 2H 5OH/g fw), respectively. For pink tomtoes, the edile cotings did not hd significnt effect on ethnol content; men vlues were 5.4 3, , nd (µl C 2H 5OH/g fw) for control, minerl wx nd crnu, respectively. No significnt differences were oserved etween minerl wx nd crnu tretments. Breker Control Minerl Oil Wx Crnu Wx Pink.4.3 Acetldehyde (µl C 2 H 5 OH/ g fw) Ethnol (µl C 2 H 4 O/ g fw) Dys t 1 C Figure 3. Ethnol (μl C 2H 5OH/g fw) nd cetldehyde (μl C 2H 4O g 1 fw) production of tomto fruits (cv. Grndel ) t pink nd reker mturity stges treted with minerl nd crnu coting nd stored for 28 dys t 1 C. Ech vlue is the men of 6 replictes ±SE. RJCES Vol 5 [3] June P g e 217 AELS, INDIA

8 Dávil-Aviñ et l Ethnol nd cetldehyde re y products of fruit fermenttion, nd their presence is ssocited with off flvours nd odours, which reduce qulity of produce. Previous reports on the use of edile cotings nd modified tmospheres indicted tht their inpproprite use could led to formtion of odours nd flvours unplesnt to the consumer [23]. Bldwin et l. [24] evluted the effect of two different edile cotings, sed on polyscchrides nd crnu wx, on the voltile content of mngo fruit. Both edile cotings decresed fruit deteriortion, lthough the polyscchride coting incresed the ethnol nd cetldehyde contents s compred to the crnu wx coting, nd control fruits. This increse in offflvor compounds ws ttriuted to the low permeility of the edile coting used [24]. According to the sensory evlution results mde y the l group, it ppered tht those levels were not sufficient to cuse off flvors tht could ffect the cceptility of the product. It hs een reported tht edile coting tretments incresed the voltile concentrtion in fruits, due to the formed semi permele rrier [24,25]. However, the mteril nd concentrtion used to produce the edile cotings cn ffect their performnce nd functionlity. Voltile Compounds The flvor voltiles re formed y severl different pthwys, including the demintion nd decroxyltion of mino cids nd lipid oxidtion of unsturted ftty cids [26,27]. Figure 4 7 shows the res of romtic compounds derived from lipids, mino cid nd crotenoid of tomto fruit t two mturity stges treted with minerl nd crnu cotings over 28 dys of storge t 1 C. The results showed significnt increse in hexnl, 6 methyl 5 hepten 2 on nd decrese in 2 methylutnol, hexnol, cis 3 hexenol while the levels of 2 hexenl nd pentnl were mintined for ll tretments. This trend ws oserved eqully in oth mturity stges; however, the concentrtion of the voltile compounds ws higher (p.5) in pink. Voltile Compounds Derived from Lipids Figure 4 shows the chnges in the pek res of the ldehyde hexnl tends to increse during storge for oth mturity stges. For reker stge tomtoes, the nlysis of vrince for hexnl res showed significnt differences (p.5) mong tretments. A glol effect showed men vlues of , nd (pa) for control, minerl wx, nd crnu, respectively. However, there were no significnt differences etween control tomtoes nd those treted with minerl sed wx while the edile cotings effect on hexnl res of pink tomtoes were not significnt (p=.11). Hexnl concentrtion vlues were , nd (pa) for control, minerl wx nd crnu, respectively. In the cse of trns 2 hexenl, this compound tends to kept during storge for oth mturity stges (Figure 4). No significnt differences mong tretments (p>.5) for oth mturity stges were oserved. For reker tomtoes, men vlues for control, minerl nd crnu cotings were , nd (pa), respectively. For pink tomtoes, men vlues were , nd (pa) for control, minerl wx nd crnu, respectively. Figure 5 shows the levels of the lcohols hexnol nd cis 3 hexen 1 ol tends to decrese during storge for oth mturity stges. The edile cotings effects on hexnol res were different depending on the mturity stges of treted tomtoes (Figure 5). Breker tomtoes showed significnt differences mong tretments (p.5); however, there were no significnt differences etween control tomtoes nd those treted with minerl coting. The vlues oserved were , nd (pa) for control, minerl wx, nd crnu, respectively. For pink stge tomtoes, nonetheless, significnt differences were not oserved (p=.31), nd the men vlues were 2.3 5, nd (pa) for control, minerl, nd crnu cotings, respectively. While for reker tomtoes, the cis 3 hexen 1 ol res were , nd (pa) for control, minerl oil nd crnu wx, respectively. However, there were no significnt differences in cis 3 hexen 1 ol res etween control tomtoes nd those treted with crnu sed wx. Pink tomtoes showed cis 3 hexenol res vlues of 3.1 5, nd (pa) for control, minerl, nd crnu tretments. Significnt differences (p.5) were oserved etween cotings tretment groups; ut not when compred to control. In generl pentnl compound tends to kept during storge for oth mturity stges. For reker stge tomtoes, the nlysis of vrince for pentnl res showed significnt differences (p.5) mong tretments. A glol effect showed men vlues of , nd (pa) for control, minerl wx, nd crnu, respectively. No significnt differences were oserved etween controls nd crnu wx treted reker tomtoes. Significnt differences (p.5) were oserved etween control nd crnu cotings tretment; ut not when they re compred to minerl sed wx. The pentnl res showed significnt differences (p.5) for pink tomtoes; the vlues oserved were , nd (pa) for control, minerl nd crnu cotings, respectively. No significnt differences were oserved etween control tomtoes nd those treted with minerl coting. RJCES Vol 5 [3] June P g e 217 AELS, INDIA

9 Dávil-Aviñ et l Lipids re one of the most importnt min sustrtes for flvor formtion [28]. Zhng et l. [29] reported tht lipid derived compounds, hexnl nd (E) 2 hexenl were the most crucil compounds contriuting to the difference in voltile compositionl chrcteristics of tomtoes t the fresh nd stle storge phses nd re lso the chrcteristic voltile compounds of tomto. The trends in these compounds is tht hexnl incresed grdully during storge, wheres unsturted (E) 2 hexenl decresed [29]. These results re consistent with those found in our study; we find significnt increse in hexnl nd decrese in hexnol, cis 3 hexenol while mintining (E) 2 hexenl for ll tretments. This tendency ws oserved eqully in oth mturity stges; however, the concentrtion of the voltile compounds in pink ws significntly higher when compred to reker. This could e due to the formtion of the voltile compounds s result of the lipid oxidtion pthwy with involvement of series of enzymes such s lipse, lipoxygense, lyse, isomerse nd lcohol dehydrogense, different expected rtes of production of voltile compounds identified y the numer of enzymtic steps required nd the ctivity of specific enzymes [3]_ENREF_9. 35 Breker Control Minerl-oil Wx Crnu Wx Pink 3 Hexnl (pa /1 5 ) X Dt Trns-2-Hexenl (pa /1 5 ) Dys t 1 C Figure 4. Voltile compounds derived from lipids of tomto fruits (cv. Grndel ) t pink nd reker mturity stges treted with minerl nd crnu coting nd stored for 28 dys t 1 C. Vlues re presented in pa 1 5. Mens within dy followed y sme letters re not significntly different t p.5. RJCES Vol 5 [3] June P g e 217 AELS, INDIA

10 Dávil-Aviñ et l 1 Breker Control Minerl-oil Wx Crnu Wx Pink 8 6 Hexnol (pa /1 5 ) Cis-3-Hexen-1-ol (pa /1 5 ) Pentnl (pa /1 5 ) Dys t 1 C Figure 5. Voltile compounds derived from lipids of tomto fruits (cv. Grndel ) t pink nd reker mturity stges treted with minerl nd crnu coting nd stored for 28 dys t 1 C. Vlues re presented in pa 1 5. Mens within dy followed y sme letters re not significntly different t p.5. The first voltile compounds formed y the ction of HPL re the ldehydes hexnl nd (Z) 3 hexenl derivtives of linoleic nd linolenic cids, respectively. Isomerse enzyme cn convert the ldehyde (Z) 3 hexenl to (Ε) 2 hexenl nd oth cn e converted to the corresponding lcohols y the enzyme ADH [31]. Even if they re not identified these lcohols ADH enzyme product, we cn ssume tht the production is low considering tht the ADH enzyme shows no increse in ctivity t the concentrtion of hexnol continues to increse during storge t oth mturity stges. While the decrese in (Z) 3 hexenl suggests low sustrte in tomto nd consequently low production of (Ε) 2 hexenl, this could explin the evolution of the trends oserved in this study. However, for specified conclusion would e necessry to quntify the concentrtions of linoleic nd linolenic cid contined in tomtoes studied, nd specific enzyme ctivity for these sustrtes. RJCES Vol 5 [3] June P g e 217 AELS, INDIA

11 Dávil-Aviñ et l 3 Breker Control Minerl-oil Wx Crnu Wx Pink 25 2-Methylutnol (pa /1 5 ) Dys t 1 C Figure 6. Voltile compounds derived from mino cids of tomto fruits (cv. Grndel ) t pink nd reker mturity stges treted with minerl nd crnu coting nd stored for 28 dys t 1 C. Vlues re presented in pa 5. Mens within dy followed y sme letters re not significntly different t p.5. On the other hnd, rte mturity stge t hrvest is the second most importnt fctor (fter genotype) influencing flvor qulity of fruits nd vegetles [32], however, the temperture nd tmospheric conditions hve direct effect on the fruit metolism, leding to chnges in the formtion of flvour compounds, ut there re few studies on the effect of modified tmosphere storge on the voltile compounds of tomtoes. Mul et l. [33] conducted studies to descrie flvor nd rom in ripe tomtoes stored t 5, 1, 12.5 nd 2 C. Found tht fruit stored for 2 d elow 2 C were rted y trined sensory pnelists s significntly lower (p.5) in ripe rom, tomto flvor, compred to those stored t 2 C. The GC nlysis showed tht voltile compounds (hexnl, 2 methylutnol) hd lower concentrtion in the smples stored t 5 C compred with those stored t 12.5 nd 2 C. While the concentrtion of the compounds trns 2 hexenl, cis 3 hexenol ws significntly lower (p.5) in tomtoes stored t 1 C compred to those stored t 12.5 C. The highest concentrtions of voltile hexnl, 2 methylutnol, methylutnol, trns 2 hexenl, 6 methyl 5 hepten 2 one, cis 3 hexenol were otined from tomto fruits stored t 2 C. They found tht incresed production of voltiles in tomto originted stored t 2 C nd tempertures elow 12.5 C suppressed (p.5) the synthesis of voltile nd thus significntly reduces the rom. These results re consistent with those found y Boukoz nd Tylor [34] studied the effect of storge of tomto t different tempertures (6, 21, 35 nd 45 C) found lower levels of voltile t tempertures of 6 C during storge. At 6 C, the voltile methylutnol showed n increse, ut overll the production ws low compred with production t 35 C. While, Díz de León Sánchez et l. [35] studied the effect of storge t 1 C in 5 ripeness of fruit of tomto on the production of voltile compounds responsile for rom of this fruit oserving quntittive nd qulittive chnges on the production of voltile compounds responsile rom fter the sixth dy of storge t 1 C. Among the min chnges, there is n incresed rtio of 3 metilutnl/3 metilutnol nd hexnl/hexnol, nd n increse in the levels of trns 3 hexenol. Díz de León Sánchez et l. [35] reported tht these chnges cn e explined y decrese in ADH enzyme ctivity. Considering these results, the storge temperture used in our study could e influencing the effect of using edile cotings nd production of voltile compounds. However, under these conditions we cn conclude tht the use of edile cotings does not ffect the trends of the voltile compounds studied nd their concentrtions during storge, s seen in the glol effect concentrtions of voltile compounds studied. Voltile Compounds Derived from Amino Acids Amino cids lso represent n importnt source in the formtion of voltile compounds tht contriute to the rom of tomto. The mino cid metolism genertes liphtic, romtic, rnched lcohols, ldehydes, ketones, cids nd esters. This rom iosynthesis pthwy comprises three enzymtic ctivities: minotrnsferse, decroxylse, nd ADH [36,37]. The 2 Methylutnol compound tends to decrese during storge for oth mturity stges (Figure 6). Breker tomtoes showed significnt differences (p.5) mong tretments. 2 Methylutnol res vlues were , nd (pa) RJCES Vol 5 [3] June P g e 217 AELS, INDIA

12 Dávil-Aviñ et l for control, minerl nd crnu cotings, respectively. Significnt differences (P.5) were oserved etween control nd crnu wx tretment; ut not when compred to minerl coting. Sttisticl differences were lso oserved for pink tomtoes, which hd men vlues of , nd (pa) for the control, minerl nd crnu coting groups, respectively. However, no significnt differences were oserved etween coting tretment groups; differences were only evident when compred to control. A result from the literture indictes to lipid nd mino cid metolism s some of the synthetic pthwys for rom voltiles tht were ffected y low tempertures. Díz de León Sánchez et l. [35] report tht during storge t 1 C, there ws significnt increse in 3 methylutnl nd significnt decrese in 3 methylutnol; resulting in n increse in the ldehyde/lcohol lnce (i.e. the 3 methylutnl/3 methylutnol quotient). This is consistent with the decrese found with lcohol 2 methylutnol nlyzed in this study, ecuse, similr enzymtic trnsformtions hve een descried for different mino cids leucine. These results suggest tht t 1 C, ctivity of ADH might e ffected. Activity of this enzyme incresed during tomto ripening nd hs een implicted in the interconversion of ldehyde nd lcohol rom voltiles [35]. In this study, the decresing trend of the compound 2 methylutnol is not ffected y the use of edile cotings. However, the glol effect shows significnt differences etween control nd treted fruit with coting edile ut not etween them, this my e ecuse in generl the fruits treted with coting mintined lower metolism thn the control. However it would e importnt to study the precursors of this compound to confirm the theory. Voltile Compounds Derived from Crotenoids The 6 methyl 5 hepten 2 on levels in tomto tends to increse during storge for oth mturity stges (Figure 7). Breker tomtoes showed significnt differences (p.5) mong edile coting tretments nd control fruits; however, no significnt differences were oserved etween wxes. 6 methyl 5 hepten 2 on res were , nd (pa) for control, minerl wx nd crnu, respectively. For pink stge tomtoes, however, significnt differences were not oserved (p=.86), nd the men vlues were , nd (pa) for control, minerl wx, nd crnu, respectively. 18 Breker Control Minerl-oil Wx Crnu Wx Pink 6-methyl-5-hepten-2-on (pa /1 5 ) Dys t 1 C Figure 7. Voltile compounds derived from crotenoids of tomto fruits (cv. Grndel ) t pink nd reker mturity stges treted with minerl nd crnu coting nd stored for 28 dys t 1 C. Vlues re presented in pa 5. Mens within dy followed y sme letters re not significntly different t p.5. In mny cses the metolic pthwys involved in the synthesis of some compounds re unknown, s is the cse of voltile compounds preceded y crotenoids [38]. However, knowing their chemicl structure nd correlting the high production of certin voltile compounds when there is ccumultion of crotenoids in tomto fruit, one cn predict the possile oxidtive products division of crotenoids nd the possile reltionship etween the voltiles nd their possile precursors. For exmple, it is considered tht the tomto rom voltiles: 6 methyl 5 hepten 2 on, gernyl frnesyl cetone nd cetone re the result of oxidtive rekdown of crotenoids cyclic. Similrly, the α ionone, β ionone nd β dmscenone proly oxidtive rekdown product of cyclic crotenoids nd other terpenoids certinly my exist in plnt tissue ound crohydrtes [38]. RJCES Vol 5 [3] June P g e 217 AELS, INDIA

13 Dávil-Aviñ et l Simkin et l. [38], mentioned tht Bouvier et l. [39] identified lycopene specific 5, 6 (5, 6 ) clevge dioxygense (BoLCD) from Bix orelln, responsile for the formtion of ixin dildehyde nd C7 clevge product previously identified s 6 methyl 5 hepten 2 on, nd hs een identified s n importnt contriutor to tomto flvor. Schw et l. [4], report the synthesis of β ionone, gernyl cetone nd 6 methyl 5 hepten 2 on in tomto fruits increses 1 2 fold during fruit ripening. The results of our study showed significnt increse of 6 methyl 5 hepten 2 on, during storge in oth mturity stge, specificlly of 6.8, 7.9, 7.8 fold for control, minerl wx, nd crnu, respectively, nd for pink tomtoes the increse is 2.4, 2.2, 2.3 fold for control, minerl wx, nd crnu, respectively. 6 methyl 5 hepten 2 on is compound derived from lycopene, compound nlyzed, ut dt not shown, however for reker tomtoes, no significnt effect of the tretments on lycopene content ws oserved (p=.61) nd for pink no significnt differences of lycopene content were oserved etween minerl wx nd crnu tretments. This my relte to which there is no tretment effect in this compound. CONCLUSIONS Bsed on the results oserved on the effect of edile cotings on the ctivity of the min enzymes nd metolites present in the neroic pthwys, long with orgnoleptic evlution through sensoril nlysis out cceptnce/rejection of treted tomto fruits, (Rndom popultion s testers, dt not shown), we cn suggest tht the use of edile cotings sed on minerl wx nd crnu wx do not hve significnt effect on the flvor of tomto fruits. REFERENCES 1. Nrnjo RDDP, Otiz S, Srgusti AC, Broni V, Crrnz AdV, Perlt IE, Vlle EM, Crrri F, Asis R (216). Hydrophilic ntioxidnts from Anden Tomto Lndrces ssessed y their ioctivities in vitro nd in vivo. Food Chem. 26 : González Aguilr GA, Vlenzuel Soto E, Lizrdi Mendoz J, Goycoole F, Mrtínez Téllez MA, Villegs Ocho MA, Monroy Grcí IN, Ayl Zvl JF (29). Effect of chitosn coting in preventing deteriortion nd preserving the qulity of fresh cut ppy Mrdol. J. Sci. Food Agric. 89(1) : González Aguilr GA, Ayl Zvl J, Olivs G, de l Ros L, Álvrez Prrill E (21). Preserving qulity of freshcut products using sfe technologies. J. für Verruchersc. Leensmittel. 5(1) : Espino Díz M, Molin Corrl F, Sepulved D, González Aguilr G, Olivs G (216). Alginte cotings contining high levels of isoleucine improve romtic nd stndrd qulity in fresh cut pple. Eur. J. Hortic. Sci. 81(3): Espino Díz M, De Jesús Ornels Pz J, Mrtínez Téllez MA, Sntillán C, Bros Cánovs GV, Zmudio Flores PB, Olivs GI (21). Development nd chrcteriztion of edile films sed on mucilge of Opunti ficus Indic (l.). J. Food Sci. 75(6). 6. Pérez Ggo M, González Aguilr G, Olivs G (21). Edile cotings for fruits nd vegetles. Stewrt Posthrv. Rev. 6(3): Dávil Aviñ JE, Vill Rodríguez J, Cruz Vlenzuel R, Rodríguez Arment M, Espino Díz M, Ayl Zvl JF, Olivs Orozco GI, Heredi B, González Aguilr G (211). Effect of edile cotings, storge time nd mturity stge on overll qulity of tomto fruits. Am. J. Agric. Biol. Sci. 6(1): Dávil Aviñ JE, Vill Rodríguez JA, Villegs Ocho MA, Tortoledo Ortiz O, Olivs GI, Ayl Zvl JF, González Aguilr GA (214). Effect of edile cotings on ioctive compounds nd ntioxidnt cpcity of tomtoes t different mturity stges. J. Food Sci. Tech. 51(1): Cisneros Zevllos L, Krocht J (23). Dependence of coting thickness on viscosity of coting solution pplied to fruits nd vegetles y dipping method. J. Food Sci. 68(2): Kto Noguchi H, Morokum M (27). Ethnolic fermenttion nd noxi tolernce in four rice cultivrs. J. Plnt Physiol. 164(2): Zhou R, Li Y, Yn L, Xie J (211). Effect of edile cotings on enzymes, cell memrne integrity, nd cell wll constituents in reltion to rittleness nd firmness of Hunghu pers (Pyrus pyrifoli Nki, cv. Hunghu) during storge. Food Chem. 124(2): Tdege M, Dupuis I, Kuhlemeier C (1999). Ethnolic fermenttion: new functions for n old pthwy. Trends Plnt Sci. 4(8): Chng LA, Hmmett LK, Phrr DM (1982). Ethnol, lcohol dehydrogense, nd pyruvte decroxylse in storge roots of four sweet potto cultivrs during simulted flood dmge nd storge. J. Am. Soc. Hort. Sci. 17(4): Arulsekr S, Prfitt D (1986). Isozyme nlysis procedures for stone fruits, lmond, grpe, wlnut, pistchio, nd fig. HortScience (USA) 15. Hoffmn NE, Hnson AD (1986). Purifiction nd properties of hypoxiclly induced lctte dehydrogense from rley roots. Plnt Physiol. 82(3): Good AG, Muench DG (1993). Long term neroic metolism in root tissue (metolic products of pyruvte metolism). Plnt Physiol. 11(4): RJCES Vol 5 [3] June P g e 217 AELS, INDIA

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