GRASAS Y ACEITES 67 (2) April June 216, e128 ISSN-L: 17-3495 doi: http://dx.doi.org/1.3989/gy.55152 Effect of the prtil NCl sustitution y other chloride slts on the voltile profile during the ripening of dry-cured lcón R. Domínguez, P.E. Munekt, A. Cittdini c nd J.M. Lorenzo,* Centro Tecnológico de l Crne de Glici, Rú Glici N 4, Prque Tecnológico de Glici, Sn Ciro ds Viñs, 329 Ourense, Spin Deprtment of Food Engineering, Fculty of Animl Science nd Food Engineering, University of São Pulo, 225 Duque de Cxis Norte Ave, Jrdim Elite, postl code 13.635-9, Pirssunung, São Pulo, Brzil c Scuoldi Bioscienze e Medicin Veterinri, Università di Cmerino, Vi Gentile III d Vrno, 6232 Cmerino (MC), Itly * Corresponding uthor: jmlorenzo@cetec.net Sumitted: 6 My 215; Accepted: 3 Octoer 215 SUMMARY: The influence of three slting tretments (tretment II: 5% NCl-5% KCl; III: 45% NCl-25% KCl-2% CCl 2-1% MgCl 2 ; IV: 3% NCl-5% KCl-15% CCl 2-5% MgCl 2 ) on the formtion of voltile compounds throughout the process ws studied nd compred to those of control lcón (tretment I: 1% NCl). There ws n intense formtion of voltile compounds throughout the processing, prticulrly during the dry-ripening stge. The most undnt chemicl fmily in ll the formultions, in the finl product ws hydrocrons followed y ldehydes. The totl voltile compound relese ws more intense in the control lcóns (1164 AU 1 6 g 1 dry mtter) thn in lcóns from formultions II, III nd IV (817 891 AU 1 6 g 1 dry mtter). The lcóns from formultion I showed the highest mounts of ldehydes. The lcóns from formultions I nd II presented the highest mounts of hydrocrons. The min conclusion is tht the replcement of NCl produces chnges in the voltile profile nd could e ffect the rom of lcón. KEYWORDS: Clcium chloride; Dry-cured lcón ; Mgnesium chloride; Potssium chloride; Voltile compounds RESUMEN: Efecto del reemplzo prcil de NCl por otrs sles en el perfil de volátiles durnte l mdurción del lcón crudo-curdo. Se estudió l influenci de tres trtmientos de sldo (trtmiento II: 5 % NCl-5 % KCl; III: 45 % NCl-25 % KCl-2 % CCl 2-1 % MgCl 2 ; IV: 3 % NCl-5 % KCl-15 % CCl 2-5 % MgCl 2 ) en l formción de compuestos volátiles durnte l elorción de lcón, en comprción con un control (trtmiento I: 1 % NCl). Huo un intens formción de compuestos volátiles durnte el procesdo, principlmente durnte l fse de secdo-mdurción. L fmili químic más undnte en el producto finl fueron los hidrocronos, seguidos por los ldehídos. L lierción de volátiles fue más intens en los lcones control (1164 AU 1 6 g 1 mteri sec) que en los otros lcones (817 891 AU 1 6 g 1 mteri sec). Los lcones de l formulción I mostrron ls myores cntiddes de ldehídos, y los lcones de ls formulciones I y II presentron los myores contenidos de hidrocruros. L principl conclusión es que el reemplzo de NCl produce cmios en los compuestos volátiles y por lo tnto podrín fectr l rom del lcón. PALABRAS CLAVE: Cloruro cálcico; Cloruro mgnésico; Cloruro potásico; Compuestos volátiles; Lcón crudo-curdo Cittion/Cómo citr este rtículo: Domínguez R, Munekt PE, Cittdini A, Lorenzo JM. 216. Effect of the prtil NCl sustitution y other chloride slts on the voltiles profile during the ripening of dry-cured lcón. Grss Aceites 67 (2): e128. doi: http://dx.doi.org/1.3989/gy.55152. Copyright: 216 CSIC. This is n open-ccess rticle distriuted under the terms of the Cretive Commons Attriution-Non Commercil (y-nc) Spin 3. Licence.
2 R. Domínguez, P.E. Munekt, A. Cittdini nd J.M. Lorenzo 1. INTRODUCTION Dry-cured lcón is trditionl cured met product mde in the north-west of Spin from the fore leg of the pig which is cut t the shoulder ldehumerus joint, following very similr mnufcturing processes to those used in the production of drycured hm s descried y Purriños et l., (211). In Glici (NW Spin), this product hs een wrded Geogrphiclly Protected Identity (G.P.I.) (Officil Journl of the Europen Communities, 21). Slt is n essentil ingredient in dry-cured lcón due to its contriution to the wterholding cpcity, prevention of microil growth, nd reduction in wter ctivity which fcilitte the soluiliztion of certin proteins nd confer typicl slty tste (Lorenzo et l., 28; Purriños et l., 211; Purriños et l., 213). Moreover, the slt ffects some chemicl nd iochemicl rections such s proteolysis, lipolysis nd lipid oxidtion, which contriute to the development of texture nd typicl flvor of dry-cured lcón (Grrido et l., 212; Purriños et l., 212). In recent yers different studies hve egun to show tht met consumption is eing more nd more influenced y helth nd nutritionl considertions. The men dily sodium intke of the Europen popultion rnges from out 3 to 5 g (8 to 11 g NCl) (EFSA, 25). On generl sis, it hs een estlished tht the consumption of more thn 6 g NCl/dy/person is ssocited with ging nd increse in lood pressure. Therefore, limiting dietry sodium intke should e chieved y restricting dily slt (sodium chloride) consumption to less thn 5 g per dy (WHO/FAO, 23). In the cse of Spin, in 28 the Spnish Food Sfety nd Nutrition Agency strted slt reduction pln with certin specific gols which would enle its intke to go down from the current vlue of 9.7 g/dy to n intke of less thn 8. g/dy y 214. Due to the incresed knowledge out the links etween sodium intke nd coronry hert diseses, consumers demnd for low-slt met products with the sme qulity s conventionl ones hs incresed. To this regrds, the prtil sustitution of NCl y mixture of slts (potssium, clcium, nd mgnesium slts) ppers to e the est lterntive to reduce the sodium content in dry-cured lcón (Lorenzo et l., 215). However, the effect of the prtil NCl replcement y other chloride slts on the voltile profile of dry-cured lcón is unknown. The im of this work ws to determine the influence of prtil NCl replcement y mixture of KCl, CCl 2 nd MgCl 2 on the genertion nd relese of voltile compounds to the hedspce of dry-cured lcón for their possile contriution to the flvor of this product. 2. MATERIAL AND METHODS 2.1. Smples Fifty-two fresh lcón pieces with n verge weight of 4.61±.47 kg ech were otined from locl slughterhouse in the re of Ourense (Spin). Four of the rw pieces were smpled nd nlyzed in order to chrcterize the rw mteril. The remining forty-eight lcón rw pieces were sumitted to the trditionl lcón processing (Purriños et l., 211). The lcón smples were rndomly divided into four tches with twelve lcóns in ech tch. Lcóns from the first tch were slted with the trditionl NCl (1% NCl, tretment I) nd were used s the control of the voltile compounds, wheres the other tches were slted in the sme wy ut with prtil replcements of NCl y other slts. So, the second tch ws slted with 5% NCl nd 5% KCl (tretment II); the third tch with 45% NCl, 25% KCl, 2% CCl 2 nd 1% MgCl 2 (tretment III); nd the fourth tch with 3% NCl, 5% KCl, 15% CCl 2 nd 5% MgCl 2 (tretment IV). The slting stge ws crried out etween 2 nd 5 C nd the reltive humidity (HR) etween 8 nd 9% for totl of 5 dys. After the slting stge the pieces were tken from the hep, rushed, wshed, nd trnsferred to post-slting room where they styed for 14 dys t 2 5 C nd round 85 9% HR. After the post-slting stge the pieces were trnsferred to room t 12 C nd 74 78% HR where drying-ripening process took plce for 84 dys. Smples were tken fter the slting, postslting, nd drying-ripening stges. In ech smpling point, totl of 4 lcón smples from ech tch were nlyzed. The Lcón pieces were skinned, deoned, nd the triceps rchii muscle ws extrcted. The smples were vcuum pcked nd stored t 3 C for no longer thn four weeks until nlysis. 2.2. Chemicl composition nd ph vlues Moisture, ft nd protein contents were determined ccording to the Assocition of Officil Anlyticl Chemists (25) in triplicte. The moisture content ws determined y drying in n oven t 15 C±2 C; the nitrogen content ws determined y the Kjeldhl method nd the protein content ws estimted y multiplying the nitrogen content y 6.25; the ft content ws determined y the Soxhlet method using petroleum ether s solvent. The ph of the smples ws mesured using digitl ph meter (model 71 A+, Thermo Orion, Cmridgeshire, UK) equipped with penetrtion proe. 2.3. Minerl composition The quntifiction of minerl elements (N, K, C nd Mg) ws performed y inductively coupled plsm-opticl emission spectroscopy (ICP-OES), Grss Aceites 67 (2), April June 216, e128. ISSN-L: 17-3495 doi: http://dx.doi.org/1.3989/gy.55152
Effect of the prtil NCl sustitution y other chloride slts on the voltile profile during the ripening of dry-cured lcón 3 ccording to the procedure descried y Lorenzo et l. (215). The finl vlue for ech element ws otined y clculting the verge of three determintions. 2.4. Voltile compound profile The extrction of the voltile compounds ws performed using solid-phse micro-extrction (SPME). A SPME device (Supelco, Bellefonte, PA, USA) contining fused-silic fier (1 mm length) coted with 5/3 μm thickness of DVB/CAR/PDMS (divinylenzene/croxen/polydimethylsiloxne) ws used for HS-SPME extrction. The muscle smples were ground with commercil grinder, 1 g portion ws weighed into 24 ml vil nd the vil ws screw-cpped with lminted Teflon ruer disk. The fier ws inserted into the smple vil through the septum nd then exposed to the hedspce. The extrctions were crried out in n oven to ensure homogeneous temperture for smple nd hedspce. The fier ws conditioned prior to nlysis y heting it in gs chromtogrph injection port t 27 C for 6 min, following the mnufcturer specifictions. Extrction ws performed t 35 C for 3 min. Before extrction, the smples were equilirted for 15 min t the temperture used for extrction. Once smpling ws finished, the fier ws drwn into needle nd trnsferred to the injection port of the gs chromtogrph mss spectrometer (GC MS) system. A gs chromtogrph 689N (Agilent Technologies Spin, S.L., Mdrid, Spin) equipped with mss detector 5973N (Agilent Technologies Spin, S.L., Mdrid, Spin) ws used with DB-624 cpillry column (J&W scientific: 3 m,.25 mm id, 1.4 μm film thickness). The SPME fier ws desored nd mintined in the injection port t 26 C for 8 min. The smple ws injected in the splitless mode. Helium ws used s crrier gs with liner velocity of 4 cm s 1. The temperture progrmme ws isotherml for 1 min t 4 C, rised to 2 C t rte of 5 C min 1, nd then rised to 25 C t rte of 2 C min 1, nd held for 5 min: totl run time 49.5 min. Injector nd detector tempertures were oth set t 26 C. The mss spectr ws otined using mss selective detector working in electronic impct t 7 ev, with multiplier voltge of 1953 V nd collecting dt t rte of 6.34 scns s 1 over the rnge m/z 4-3. The compounds were identified compring their mss spectr with those contined in the NIST5 (Ntionl Institute of Stndrds nd Technology, Githersurg) lirry, nd/or y compring their mss spectr nd retention time with stndrds (Supelco, Bellefonte, PA, USA), nd/or y clcultion of retention index reltive to series of stndrd lknes (C 5 C 14 ) (for clculting liner retention index, Supelco 44585-U, Bellefonte, PA, USA) nd mtching them with dt reported in the literture. 1 The results re expressed s AU (re units) 1 6 g of dry mtter. 2.5. Sttisticl nlysis All sttisticl nlyses were performed using the IBM SPSS Sttistics 19 softwre (IBM, Corp, 21). The normlity ws ssessed using the Kolmogorov- Smirnov test, nd the Levene s homogeneity of vrince test ws pplied to exmine the equlity of vrinces. After verifiction of norml distriution nd constnt vrince of dt, significnt differences were determined using one-wy nlysis of vrince (ANOVA). A Duncn s test ws performed to compre the men vlues for processing time (, 5, 19 nd 13 dys) nd prtil sodium replcement t significnce level of P<.5. 3. RESULTS AND DISCUSSION 3.1. Effect of the slting tretments on physicochemicl properties nd minerl content The results of chemicl composition nd ph vlues t the end of dry-cured lcón sumitted to four different slting tretments re shown in Tle 1. Moisture content ws significntly (P<.5) ffected y NCl replcement y other slts, since lcóns sumitted to formultions IV presented the highest vlues. These differences could e due to the quicker penetrtion of the slt mixtures contining KCl tht would hinder the exit of wter from the inside of the met (Aliño et l., 29). This finding is in greement with those reported y Wu et l. (214) who oserved significntly (P<.5) higher moisture contents in con smples slted with 3% NCl nd 7% KCl compred with smples slted with 1% NCl. However, Armenteros et l. (212) did not find significnt differences mong slting formultions on the moisture content of dry-cured hm. On the other hnd, the protein nd intrmusculr ft contents did not show significnt differences mong tretments. Regrding ph vlues, the NCl replcement y other slts induced significnt (P<.1) differences mong tretments. In this study the lcóns slted with lower NCl concentrtions (formultions III nd IV) presented the lowest vlues of ph, while the highest vlues were otined in lcóns from formultion II. Conflicting dt re ville in the literture out KCl, MgCl 2, nd CCl 2 inclusion in dry-cured met products regrding this spect. To this regrds, Gimeno et l. (1999) studied the effect of mixture of NCl (1 g kg 1 ), KCl (5.52 g kg 1 ), MgCl 2 (2.35 g kg 1 ), nd CCl 2 (4.64 g kg 1 ) to prtilly sustitute NCl nd reported greter ph decrese in the reduced NCl formultion compred to the trditionl one. However, Znrdi Grss Aceites 67 (2), April June 216, e128. ISSN-L: 17-3495 doi: http://dx.doi.org/1.3989/gy.55152
4 R. Domínguez, P.E. Munekt, A. Cittdini nd J.M. Lorenzo TABLE 1. Chemicl composition nd ph vlues of dry-cured lcón t the end of processing (men ± stndrd devition of four replictes) Slt formultions SEM Sign. I II III IV Moisture (%) 55.28±3.83 53.21±2.92 58.87±.87 c 61.66±.28 c 1.12 * Protein (% of dry mtter) 33.28±2.77 36.76±4.7 31.3±2.91 32.9±3.66.94 n.s. Ft (% of dry mtter) 9.82±2.52 7.65±2.55 6.19±1.57 7.5±2.85.63 n.s. ph 5.96±.8 6.8±.9 5.83±.5 5.92±.5.3 ** Slt formultions: tretment I: control, 1% NCl; tretment II: 5% NCl nd 5% KCl; tretment III: 45% NCl, 25% KCl, 2% CCl 2 nd 1% MgCl 2 ; tretment IV: 3% NCl, 5% KCl, 15% CCl 2 nd 5% MgCl 2 c Men vlues in the sme row (corresponding to the sme prmeter) not followed y common letter differ significntly (P<.5). Sign.: Significnce; n.s.: not significnt; * (P<.5); *** (P<.1). et l., (21) noticed tht the slt mixture (NCl 13.5 g kg 1, KCl 4.2 g kg 1, CCl 2 2.4 g kg 1, nd MgCl 2 2.4 g kg 1 ) did not ffect the ph evolution throughout the processing. Tle 2 shows the minerl content of dry-cured lcón sumitted to four different slting tretments. With regrds to the nturl content of minerls in the lcón, the slt composition t the end of the ripening period inside the lcón is somehow reflected in the slt penetrtion nd diffusion through the lcón. A significnt reduction (P<.1) in the N content ws chieved through the prtil sustitution of NCl y the mixture of chloride slts employed during their production. In our study, the lcón smples sumitted to formultion I presented the highest mount of sodium (2446.6 mg 1 g 1 ), which ws similr to norml level for lcón with these chrcteristics (Lorenzo et l., (23). The prtil sustitution of sodium chloride y the mixture of chloride slts significntly (P<.1) reduced the sodium content nd incresed potssium, clcium nd mgnesium contents. However, lower concentrtions of C +2 nd Mg +2 were found in the lcón in reltion to the proportions of these chloride slts employed during their production. These findings re in greement with those otined y Aliño et l. (29) nd Armenteros et l. (29) who lso oserved the difficulty of divlent ctions to penetrte inside the muscle. This could e explined y the fct tht C +2 nd Mg +2 ctions hve higher chrge density (.5 nd.82 units of chrge/moleculr weight, respectively) tht would increse their difficulty to penetrte inside the lcón (Bles et l., 28). 3.2. Effect of the slting tretments on voltile compounds The SPME technique is not normlly used for solute quntifictions, ut when exctly the sme extrction methodology is pplied, this technique llows for compring reltive mounts mong smples. A totl of 31 voltile compounds were detected in the hedspce of lcón t the end of the dry-ripening process. The compounds were grouped y chemicls nd their liner retention index (Tle 3), comprising 3 cids, 4 lcohols, 4 ldehydes, 2 esters, 15 hydrocrons, 2 ketones nd 1 furn. At the end of the process the voltile compound profile mintined the reltionship hydrocrons>ldehydes>lcohols> cids>ketones>furn>esters. Lorenzo et l. (214) lso reported tht hydrocrons were the most undnt compounds in previous study of drycured lcón. However, other studies found tht the most undnt group of voltile compounds TABLE 2. Minerl composition (ppm) of dry-cured lcón t the end of processing (men ± stndrd devition of four replictes) Slt formultions ppm I II III IV SEM Sign. N 2446.6±18.46 c 1483.91±118.55 738.21±2.34 586.35±19.39 189.5 *** K 565.11±22.22 1915.42±78.12 d 876.8±8.46 1668.94±99.21 c 127.61 *** C 9.35±1.27 9.31±.59 42.87±1.48 33.73±11.8 4.19 *** Mg 35.4±5.35 31.89±3.35 42.73±2.14 36.97±3.7 1.64 * Slt formultions: tretment I: control, 1% NCl; tretment II: 5% NCl nd 5% KCl; tretment III: 45% NCl, 25% KCl, 2% CCl 2 nd 1% MgCl 2 ; tretment IV: 3% NCl, 5% KCl, 15% CCl 2 nd 5% MgCl 2 d Men vlues in the sme row (corresponding to the sme prmeter) not followed y common letter differ significntly (P<.5). Sign.: significnce: n.s.: not significnt; * (P<.5);***(P<.1). Grss Aceites 67 (2), April June 216, e128. ISSN-L: 17-3495 doi: http://dx.doi.org/1.3989/gy.55152
Effect of the prtil NCl sustitution y other chloride slts on the voltile profile during the ripening of dry-cured lcón 5 TABLE 3. Voltile compounds (AU 1 6 g 1 dry mtter) in the hedspce of dry-cured lcón slted with different slt formultions t the end of processing. Dt re the verge vlues of four replictes Slt formultions LRI R I II III IV Acetic cid 72 ms,lri 2.47 1.81 1.88 1.31.37 ns Butnoic cid 883 ms,lri 15.58 2.92 22.53 5.16 2.94 *** Nonnoic cid 137 ms,lri.57..67..1 *** Totl cids 19.64 22.1 24.87 5.78 3.18 * SEM Sign. 1-Pentnol 834 ms, lri 21.35 5.59 14.83 23.77 2.71 * 1-Hexnol 925 ms, lri 7.27. 2.89 c 16.63 c 2.59 *** 1-Octen-3-ol 195 ms, lri 24.32 16.61 32.84 c 41.94 c 3.36 ** Benzyl lcohol 117 ms,lri 15.84 12.55 13.29 11.66.85 ns Totl lcohols 64.8 35.82 63.94 93.99 c 6.44 *** Pentnl 736 ms,lri, s 15.64. 13.84 12.5 2.34 *** Hexnl 823 ms,lri, s 68.55 c 4.97 297.1 274.9 59.87 *** Heptnl 933 ms,lri, s 8.32 6.46 12.31 16.2 c 1.29 ** Decnl 1347 ms,lri, s 3.92 c 2.6...53 *** Totl ldehydes 626.52 c 49.38 318.64 299.13 61.48 *** Ethnol, 2-(2-utoxyethoxy)-, cette 1419 ms,lri 1.69 1.27...25 ** 1,2,3-Propnetriol, dicette 1514 ms 1.1.87...17 ** Totl esters 2.79 1.85...39 *** Pentne, 2,3,4-trimethyl- 66 ms 3.81 5.1 2.66 2.7.51 ns Heptne 7 ms,lri, s 2.71 2.26 2.4.63.39 ns Pentne, 2,3,3-trimethyl- 76 ms,lri 12.23 9.38 3.24 3.99 1.25 ** Octne 8 ms,lri, s 14.1 27.82 14.79 16.56 2.45 ns Heptne, 3-methylene- 87 ms,lri 2.63 3.42...52 *** Hexne, 3-ethyl- 86 ms 1.3 1.82. 1.93.32 ns Heptne, 2,2,4-trimethyl- 94 ms,lri 7.29 7.37.. 1.2 *** Heptne, 2,5,5-trimethyl- 924 ms,lri 5.42 6.73...89 *** Heptne, 2,2,4,6,6-pentmethyl- 998 ms,lri 569.66 627.46 376.47 428.29 37.99 * 3-Ethyl-3-methylheptne 13 ms 1.73 2.14 1.64..27 ** Decne, 2,6,7-trimethyl- 1134 ms 48.14 3.57 37.37. 7.54 * Dodecne 12 ms,lri, s 3.97 3.51 2.68 2.45.28 ns Undecne, 5-methyl- 127 ms,lri 5.63 c 4.77 c 4.1 3..4 * Decne, 5-methyl-6-methylene- 1265 ms,lri 3.16 3.95 3.8 1.98.46 ns Tridecne 13 ms,lri, s 1.39 1.42 2.4 1.83.13 ns Totl hydrocrons 65.12 698.82 432.59 459.53 4.89 * 2-Butnone, 3-hydroxy- 765 ms,lri 15.87 c 7.76 4.2 11.86 1.52 * 2-Heptnone 94 ms,lri 3.73 2.73 5.61 3.49.33 ** Grss Aceites 67 (2), April June 216, e128. ISSN-L: 17-3495 doi: http://dx.doi.org/1.3989/gy.55152
6 R. Domínguez, P.E. Munekt, A. Cittdini nd J.M. Lorenzo TABLE 3 (continued) Slt formultions LRI R I II III IV SEM Sign. Totl ketones 2.43 c 1.95 6.42 15.35 1.74 ** Furn, 2-pentyl- 19 ms,lri 4.6 3.8 6.5 5.8.53 ns Totl compounds 1164.38 817.24 873.3 891.36 53.31 * Slt formultions: I: (1% NCl), II: (5% NCl nd 5% KCl), III: (45% NCl, 25% KCl, 2% CCl 2 nd 1% MgCl 2 ), IV: (3% NCl, 5% KCl, 15% CCl 2 nd 5% MgCl 2 ). Sign.: significnce; ns: not significnt; *P<.5; **P<.1; ***P<.1. -c Mens in the sme row not followed y common superscript letter differ significntly (P<.5; Duncn test); S.E.M.: Stndrd error of the men; AU: re units resulting of counting the totl ion chromtogrm (TIC) for ech compound; LRI: liner retention index clculted for DB-624 cpillry column (J&W scientific: 3 m.25 mm id, 1.4 μm film thickness) instlled on gs chromtogrph equipped with mss selective detector; R: Reliility of identifiction; LRI: voltiles identified y compring their LRI with those reported in the literture (Lorenzo nd Fonsec, 214; Lorenzo nd Domínguez, 214; Domínguez et l., 214); ms: mss spectrum greed with mss dtse (NIST5); s: mss spectrum nd retention time identicl to n uthentic stndrd. were lcohols (Wu et l., 215), esters (Bermúdez et l., 215; Gómez nd Lorenzo, 213; Lorenzo, 214; Lorenzo nd Fonsec, 214) or ldehydes (Armenteros et l., 212; Purriños et l., 211; Purriños et l., 212). These different results could e explined y the voltile extrction methods used, since the purge nd trp extrction nd the SPME technique offer potentilly different profiles of voltile compounds (Bermúdez et l., 215). In ddition, there re mny fctors tht ffect SPME fier performnce, such s the choice of sttionry phse nd the extrction conditions. The chnges in the most relevnt voltile compounds during lcón processing re shown in Figure 1. The production nd relese of the voltile compounds incresed throughout the dry-cured lcón processing from 82 AU 1 6 g 1 dry mtter in fresh met to 1164, 817, 873 nd 891 AU 1 6 g 1 DM t the end of dry-ripening of lcóns from formultions I, II, III nd IV, respectively. Our results re in greement with other studies with different met products, such s cecin (Lorenzo, 214), lcón (Lorenzo nd Fonsec, 214; Purriños et l., 212), dry-cured hm (Armenteros et l., 212; Bermúdez et l., 215), dry-cured loin (Lorenzo nd Crllo, 215) nd con (Wu et l., 215). The production of voltile compounds ws prticulrly high during the dry-ripening stge. This fct could e due to the increse in temperture nd reduction in moisture in the course of the process (Lorenzo nd Fonsec, 214). As mentioned ove, hydrocrons were the most undnt chemicl group t the end of the ripening process in dry-cured lcón. They represent 55, 49 nd 51% of the totl voltile compounds in lcóns from formultions I, III nd IV, respectively, nd 85% of the totl voltile compounds in lcóns from formultion II. In this study, heptne, 2,2,4,6,6-pentmethyl, ws the most undnt hydrocron in ll the tches (representing out 9% of totl hydrocrons), followed y decne, 2,6,7-trimethyl nd octne. These results gree with Bermúdez et l. (215) who reported tht the heptne, 2,2,4,6,6-pentmethyl ws the most importnt liphtic hydrocrons in dry-cured hms. Aliphtic hydrocrons with less thn ten crons tom rise minly from lipid oxidtion (Ruíz et l., 22), while those with longer chins could e ccumulted in the ft depots of the niml, proly from feeding. An increse in the totl mount of hydrocrons ws oserved during the process, from initil vlues of 72 AU 16 g 1 DM in fresh met to 65, 698, 432 nd 459 AU 1 6 g 1 DM t the end of the dry- ripening of smples from formultions I, II, III nd IV, respectively (Figure. 1). In the slting nd post-slting stges, the contents of hydrocrons decresed nd susequently incresed. This tendency ws previously descried in lcón (Purriños et l., 212). On the contrry, Armenteros et l.(212) reported tht hydrocrons incresed during the post-slting stge nd decresed in the finl stge of the process. In this study, the production of hydrocrons ws not ffected during the slting nd post-slting stges y the prtil replcement of NCl content y other chloride slts, ut their mounts incresed t different rtes up to the end of the dry-ripening stge. Furthermore, the totl content of hydrocrons ws significntly higher (P<.5) in cured lcóns from formultions I nd II thn those from III nd IV. This fct ws minly due to the significntly higher vlues (P<.5) of heptne, 2,2,4,6,6-pentmethyl, ut lso due to higher vlues (P<.1) of pentne, 2,3,3-trimethyl, heptne, 3-methylene, heptne, 2,2,4-trimethyl nd heptne, 2,2,5-trimethyl in formultions I nd II thn in the other ones. In this cse, NCl could hve contriuted to n incresed relese of these compounds. On the contrry, Armenteros et l. (212) Grss Aceites 67 (2), April June 216, e128. ISSN-L: 17-3495 doi: http://dx.doi.org/1.3989/gy.55152
Effect of the prtil NCl sustitution y other chloride slts on the voltile profile during the ripening of dry-cured lcón 7 Are Unitsx1 6 Are Unitsx1 6 35 3 25 2 15 1 5 3 25 2 15 1 5 Acids 2 4 6 8 1 12 Processing dys c Ketones 2 4 6 8 1 12 Processing dys c c Are Unitsx1 6 Are Unitsx1 6 14 12 1 8 6 4 2 7 6 5 4 3 2 1 c Alcohols 2 4 6 8 1 12 Processing dys Aldehydes 2 4 6 8 1 12 Processing dys c c Are Unitsx1 6 8 7 6 5 4 3 2 1 Hydrocrons 2 4 6 8 1 12 Processing dys Are Unitsx1 6 14 12 1 8 6 4 2 Totl compounds 2 4 6 8 1 Processing dys Slt formultion I: (1% NCl) Slt formultion II: (5% NCl nd 5% KCl) Slt formultion III: (45% NCl, 25% KCl, 2% CCl2 nd 1% MgCl2) Slt formultion IV: (3% NCl, 5% KCl, 15% CCl2 nd 5% MgCl2) FIGURE 1. Evolution of voltile compounds throughtout dry-cured lcón processing using different slting formultions. Error rs indicte the stndrd error for ech tretment. Different letters indicte significnt differences mong formultions (P<.5). Plotted vlues re the mens of four replictes. found higher quntities of rnched hydrocrons in hms with the replcement of NCl thn in the control tch (slted with 1% NCl). Despite the high content of these compounds, they proly hd low impct on flvor due to their reltively high odor thresholds (Domínguez nd Lorenzo, 214). In the present study, ldehydes represented out 35 5% of the totl voltile compounds in the lcóns from tches I, III nd IV nd 5.9% of the totl voltile compounds in lcóns from tch III. Regrding the ldehydes, hexnl, derived from the n-6 ftty cids such s linoleic nd rchidonic cids ws the most undnt (Węsiersk et l., 213). It represented etween 8% nd 95% of totl ldehydes. These results gree with those reported y Lorenzo et l. (214) nd Lorenzo nd Fonsec (214) who oserved tht hexnl ws the most undnt ldehyde in dry cured lcón nd it ws considered good indictor of the lipid oxidtion in this met product. Moreover, hexnl ws lso the most importnt ldehyde in dry-cured con (Wu et l., 215), cecin (Lorenzo, 214), hm (Armenteros et l., 212; Bermúdez et l., 215) nd dry-ripened chorizo (Gómez nd Lorenzo, 213). The high levels of stright chin ldehydes, such s pentnl, hexnl, heptnl nd decnl, indicted tht lipid oxidtion occurred ctively during the dry-curing process nd plyed very importnt role in the formtion of hedspce voltiles y cured met products (Wu et l., 215). The mount of ldehydes incresed from initil vlues of 6.45 AU 1 6 g 1 DM in the rw pieces to 626, 318 nd 299 AU 1 6 g 1 DM t the end of dryripening of lcóns sumitted to formultions I, III nd IV, respectively. These results were previously reported y Armenteros et l. (212) nd Lorenzo nd Crllo (215) who noticed tht totl ldehydes incresed during the process of dry-cured met products. However, in lcóns from formultion II, the vlues of ldehydes incresed until the end of the Grss Aceites 67 (2), April June 216, e128. ISSN-L: 17-3495 doi: http://dx.doi.org/1.3989/gy.55152
8 R. Domínguez, P.E. Munekt, A. Cittdini nd J.M. Lorenzo 1 post-slting stge (85.15 AU 1 6 g DM) nd 1 then decresed in the finl stge (49.38 AU 1 6 g DM). These findings re in greement with those reported y Lorenzo et l. (214), Lorenzo (214) nd Lorenzo nd Fonsec (214) who noticed decresed during the finl stges. The decrese in ldehydes in lcóns sumitted to formultion II t the end of the process could e the reflection of their prticiption in other chemicl rections yielding other voltile or non-voltile compounds (Armenteros et l., 212). On the other hnd, hexnl showed differences (P<.1) mong the tches. The lcóns from formultion I hd the highest vlues (68.55 AU 1 1 6 g DM) nd the lowest were presented y the lcóns 1 sumitted to formultion II (4.97 AU 1 6 g DM). The other 2 tches presented intermedited vlues (297.1 nd 274.9 AU 1 6 g 1 DM for the lcóns from formultions III nd IV, respectively). Pentnl ws found in the smples sumitted to formultions I, III nd IV (vlues etween 12.5 nd 15.64 AU 1 6 g 1 DM), while the highest vlues of heptnl were oserved in the lcóns sumitted to formultion IV (16.2 AU 1 6 g 1 DM) in comprison with the other tches (etween 6.46 nd 12.31 AU 1 6 g 1 DM). The formultions I nd II lso presented low vlues of decnl. Armenteros et l. (212) nd Purriños et l. (212) lso found tht dry-cured products with high contents of NCl showed the highest mounts of hexnl. According to these uthors, the higher vlues of hexnl in lcóns sumitted to formultion I might e relted with the fct tht NCl could hve exerted pro-oxidnt effect. This result ws lso reported y Du nd Ahn (25) who noticed tht NCl (less thn 2%) ws pro-oxidnt in met nd met products. Another possile explntion could e tht the replcement of NCl y other slts ffects the degree of proteolysis in ripened mets s NCl inhiits muscle proteses (Grrido et l., 212), nd significnt reduction in such ions in the met systems generlly leds to more intense proteolysis (Estévez, 211). A lrge production of free mino cids nd peptides in lcóns from formultions II, III nd IV could hve contriuted to inhiiting lipid oxidtion nd, hence, the formtion of lipid derived voltiles such ldehydes. On the contrry, Andrés et l. (27) did not oserve vritions in the hexnl contents mong dry-cured hms with different slt contents. Aldehydes re known s the mjor contriutors to the unique flvor of drycured products due to their low odor thresholds (Bermúdez et l., 215). Alcohols were the third chemicl fmily fter the ripening period, they represented etween 4 nd 1% of the totl voltile compounds, nd four different lcohols were identified: 1-pentnol, 1-hexnol, 1-octen-3-ol nd enzyl lcohol. These compounds hve lso een detected in other dry-cured met products (Bermúdez et l., 215; Lorenzo et l., 214; Purriños et l., 212). Among the lcohols, in ll the tches, 1-octen-3-ol ws the most undnt nd represented out 4% of the totl lcohols. Our results gree with those reported y Bermúdez et l. (215), Lorenzo et l. 214 nd Lorenzo nd Fonsec (214), who found tht the most undnt lcohol t the end of the dry-ripened stge ws 1-octen-3-ol. The formtion nd relese of lcohols ws ffected from the eginning y the processing time, s well s y the formultion. In fresh met lcohols were not detected, nd they incresed during cold stges. After the post-slting stge the vlues of lcohols were 131.2, 64.4, 37.9 nd 43.4 AU 1 6 g 1 DM in lcóns from formultion I, II, III nd IV, respectively. Then, totl lcohols of the smples from formultions I nd II grdully declined to the end of the process, while in smples sumitted to formultions III nd IV incresed to the end of the process (Figure. 1). The mount of totl lcohols ws significntly (P<.5) higher in lcóns from formultion IV (93.9 AU 1 6 g 1 DM), followed y lcóns sumitted to formultions I nd III (64.8 nd 63.9 AU 1 6 g 1 DM) nd finlly the lcóns from formultion II (35.8 AU 1 6 g 1 DM). The smples sumitted to formultion IV presented the higher vlues of 1-octen-3-ol, 1-pentnol nd 1-hexnol thn the smples from formultions I nd II. The intermedite vlues were oserved in smples from formultion III. Our results re in greement with Armenteros et l. (212), who found the highest mounts in the smples with replcement of NCl. This result proly ws due to more intense chemicl reduction of corresponding ldehydes y microil enzymes in lcóns sumitted to tretment IV. NCl is known to hve ntimicroil ctivity, nd prtil replcement could explin more intense microil growth nd higher production of these compounds. It is in greement with reported y Lorenzo et l. (215) in previous study, who found tht lcóns from formultions III nd IV presented the highest counts of totl vile counts, slt tolernt flor nd yests. In contrst, Purriños et l. (212) found tht the lcóns with high slt content hd higher mount of lcohols thn in lcóns with reduced slt content. Only three cids were detected through the mnufcture process of dry-cured lcón. Butnoic cid ws the most undnt t the end of the process representing out 8 9% of totl cids. These findings re in greement with the study of Bermúdez et l. (215) in dry-cured hms. However, other uthors found tht cetic cid ws the min cid detected in dry-cured products (Lorenzo et l., 214; Wu et l., 215). During cold stges, the genertion of cids ws ffected y prtil replcement of NCl y other slts. After post-slting stge, the tches I nd II presented higher (P<.5) vlues of cids (17.3 nd Grss Aceites 67 (2), April June 216, e128. ISSN-L: 17-3495 doi: http://dx.doi.org/1.3989/gy.55152
Effect of the prtil NCl sustitution y other chloride slts on the voltile profile during the ripening of dry-cured lcón 9 16.7 AU 1 6 g 1 DM, respectively) thn tches III nd IV (9.9 nd 8.7 AU 1 6 g 1 DM, respectively). Then, the evolution of cids ws different depending on the tches; in the smples sumitted to formultions I, II nd III the cid content incresed to the finl of dry-ripened process. However, the cid content of the lcóns from tch II decresed t the end of the process (Figure. 1) nd is in greement with those reported y Lorenzo nd Fonsec (214) who lso found tht the cids content decresed fter post-slting stge. At the end of the finl stge, were not found differences mong smples sumitted to formultions I, II nd III (19.6, 22. nd 24.9 AU 1 6 g 1 DM, respectively), while the smples from tch IV showed significntly (P<.5) lower vlues (5.78 AU 1 6 g 1 DM) thn the other tches. These differences were minly relted to the differences in utnoic cid content (15.6, 2.9, 22.5 nd 5.78 AU 1 6 g 1 DM for smples from formultions I, II, III nd IV, respectively). In this study, two ketones were detected in the hedspce of lcón. The most undnt ketone ws 2-utnone, 3-hydroxy representing out 7 75% of totl ketones. In contrst, Bermúdez et l. (215) nd Lorenzo nd Fonsec(214)reported tht 2-heptnone ws the most undnt ketone in dry-cured hm nd dry-cured lcón, respectively, while Wu et l.(215) noticed tht 2-utnone nd 2-utnedione were the predominnt ketones in con. At the end of the processing, lcóns sumitted to formultion I hd significntly (P<.5) higher levels of ketones thn those from the other formultions. Lcóns sumitted to formultion III presented 1 the highest vlues of 2-heptnone (5.61 AU 1 6 g DM), while the vlues of 2- utnone, 3-hydroxy were significntly (P<.5) higher in the lcóns from tretment I thn from the other formultions. Our results re in greement with those reported y Armenteros et l. (212), who found the highest vlues of 2-utnone, 3-hydroxy in control dry-cured hms (slted with 1% NCl) thn in dry-cured hms with slt replcement. Only furn, 2-pentyl, ws detected fter the dryripening process (vlues etween 3.8 nd 6.1 AU 1 6 g 1 DM) nd it did not show differences mong tretments. Furn, 2-pentyl, hs een found in other dry-cured met products mnufctured from whole pieces (Bermúdez et l., 215; Lorenzo et l., 214; Lorenzo nd Crllo, 215). This furn is noncroxylic compound derived from linoleic cid nd other n-6 ftty cids, with reltively low threshold nd vegetle romtic note (Fy nd Brevrd, 25). Finlly, esters were only detected in smples from formultions I nd II, ut their mounts were very low (2.79 nd 1.85 AU 1 6 g 1 DM in the lcóns sumitted to formultions I nd II, respectively), nd they represented less thn.2% of the totl voltile compounds. Esters hve low olfction threshold vlues. However, tking into ccount tht the smples hve very low vlues of these compounds, it cn e considered tht they do not contriute to the rom of lcón. In ddition, in previous study (Lorenzo et l., 215), the sensory nlysis showed tht the pnellists gve the highest scores of intensity odor to lcóns slted with 1% NCl (formultion I) nd the lowest vlues were oserved in lcóns slted with 5% NCl nd 5% KCl (formultion II). As discussed ove, the odor of lcóns not only depends on the mount of voltile compounds, ut lso for their threshold. However, the lcóns from formultion I, which hd the highest mount of totl voltile compounds lso presented the highest scores of odor intensity ccording to Lorenzo et l. (215), while the lcóns from formultion II hd the lowest vlues of totl voltile compounds nd odor intensity (Lorenzo et l., 215). Therefore, in this cse it seems tht the mount of totl voltile compounds ws directly relted to odor intensity. 4. CONCLUSIONS The formtion of the voltile compounds significntly incresed during the dry-curing process, prticulrly during the dry-ripening stge. The replcement of NCl y other slts influenced the formtion of the mjority of voltile compounds. At the end of processing, the control smples (slted with 1% of NCl) showed the highest vlues of totl voltile compounds. This fct is minly due to higher vlues of hexnl nd totl ldehydes in this tch thn in the other ones. Therefore, NCl cts s pro-oxidizing nd soluilizing gent, incresing the reltive levels of voltile compounds from lipid oxidtion, mong others. The results otined in this study indicte tht prtil NCl replcement y other chloride slts hs n impct on the formtion of voltile compounds in dry-cured lcón. Finlly, the odor intensity ws directly relted to the mount of totl voltile compounds. ACKNOWLEDGEMENTS The uthors re grteful to The Xunt de Glici (The Regionl Government) (Project FEADER 212/45) for finncil support. 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