Enrichment of Chicken Nuggets with Microencapsulated, 2201

Seediscuss ions,stats,an dauthorpr ofilesforthis publication at: https://www. researchgate .net/publicatio n/28300984 1 Enr ichm ent of C hicke nN ugge tsw ith Micr oe nca psula ted Om ega -3F ishO il:Ef fec tof Fro zen Sto rag eTim eo n Oxida tiv.. .. Article  in  Food andBio processT echnology ·January 2016 DOI:10.1007/s11947-015-1621-x CITATIONS 3 READS 188 4authors , including : Someofth eauthorso fthispubli cationarea lsoworkin gonthes erelatedpro jects: FLAVOPRO T:Flavour ofanimal proteinhy drolysates  Viewproje ct Developm entand optimizati onofadire ctextractio ndevice formonitor ingodorta intindry cured ham  View project EstefaníaJ iménez University ofExtrema dura,Sp ain,Cácer es 16  PUBLICATIO NS  63  CITATION S  SEEPROFILE Trinidad Pérez-Pala cios Universid addeEx tremadura 106  PUBLICATIO NS  699  CITATION S  SEEPROFILE JorgeRuiz Carrascal Universid addeEx tremadura 316  PUBLICATIO NS  4,319  CITATION S  SEEPROFILE Allcontentfollow ingthispagewa suploadedby J orgeRuizCarra scal on20Janu ary2016. Theuserhasr equestedenha ncementof thedownload edfile. ORIG INA L P APE R Enr ichme nt of Chic ken Nugg ets wit h Micr oe ncap sula ted Ome ga-3 Fis h Oil: Effe ct of Fr oze n Storag e Tim e on Oxida tive Stab ilit y a nd Sen sor y Qua lity Este faní a Jimé nez- Mart ín 1 & T rinid ad Pér ez-P alac ios 1 & Jorg e Ruiz Car rasc al 2 & T er esa An tequ era Roj as 1 Rece ive d: 24 June 20 15 / Acce pted : 5 Octobe r 2015 / Pu bli shed on line : 17 Octo ber 20 15 # Spr ing er Scien ce+ Busi ness M edia New Y or k 2015 Abst ract Th is work stud ies fo r the fi rst t ime th e elab oratio n of frozen chicken nuggets enric hed with microca psules of omega-3 fatty acids using fish oil. Three types of chicken nugg ets we re prep ared: co ntro l (C), en riched in bulk fish oi l (BFO), and with added microencap sulated fish oil (MFO). Ef fect of leng th of froz en st orag e after pre- fryi ng and befo re domestic frying was studied. The pre-fried nuggets were stor ed duri ng 24 h at refr iger atio n tempe ratu re (0 – 2 °C ) (T0) or du ring 1 mo nth (T 1M) or 3 mont hs (T 3M) in a do mest ic free zer at − 18 °C be fore fr ying . Length of froz en stor age afte r pre- fryi ng and be fore do mest ic fryi ng prom oted li pid an d pro- tein oxidative reactions in omega-3-enriched nug gets. Micr oenc apsula tion sh owed a pr otec tive ef fect ag ains t lipid and protein oxidation, especially during the first month of stor age. In MF O, sens ory trai ts were no t affe cted by enri ch - ment . In BFO-T 0, a higher ju icin ess an d salti ness an d a less inte nse meat fl avor in co mpar ison wi th C-T0 and MF O-T0 was fo und. T ime of froz en st orag e did no t infl ue nce the sen- sory quality of chicken nuggets enriched with omega-3. Micr oenc apsu lati on seem s to be a promi sing me thod fo r en- rich ment of pr e-fr ied froz en meat pr oduc ts with fi sh oil, im - proving the oxid ative shelf life and pres erving the sensory qual ity ch aracte rist ics of th e enrich ed prod ucts . Keyw ord s Microe ncap sula tion . ω -3 . Chic ken nu gget s . Sens or y cha ra ct er is ti cs . Oxid atio n Intr od uct ion Omega-3 polyunsaturated fatty acids (PUF A) have been wide ly inv es tiga ted d ue to th eir effe ct s on hum an he alt h. Some of the ad vant ages th at have be en rela ted to thei r inta ke incl ude th e redu ctio n in th e risk of card iova scular and ch roni c dise ases an d tumors (S ioen et al . 2006 ). T he only fo od sour ces for l on g- ch ai n o me ga -3 P UF A, suc h a s e ic os ap en ta en oi c aci d (EP A, C2 0:5n -3) an d docosa hexa enoi c acid (D HA, C22: 6n-3 ) are fish an d seaf ood, in w hich pres ence in actual di ets is no t enou gh to reac h omeg a-3 PUF A reco mmen ded in take (S ioen et al. 2006 ). Thus, a poten tial strat egy to inc rease the con- sump tion of EP A and DH A coul d be th eir in clus ion in well- acce pted pr oduc ts (Sioe n et al. 2006 ). In that sens e, nume rous studies have been conducted applying different sources of omega-3 fat ty acids to improve the lipid pro file of variou s food s in order to make them he alth ier (Jim enez-C olme nero et al. 20 01 ). Som e altern ati ves to the us ual dire ct addi tion of the so urce of EP A an d DHA have be en assa yed, as ge ll ed oil- in-w ater em ulsi ons or konj ac matr ix gels , used as fa t replac ers (Sal cedo -San dova l et al. 2013 ; Poyat o et al. 20 14 ). Which ever th e source to be used, th e main ch alleng e of enri chment with omeg a-3 PUF A is thei r high su scept ibil ity to oxidation (Jacobsen 1999 ). However , the way in which thes e bioa ctiv e comp ound s are ad ded to the enri ched pr oduc t coul d infl uenc e the de velo pment of oxidat ive reacti ons an d their adverse con sequences. The brea kdown of the ω -3 PUF A chai n during ox idat ion imp lica tes nutr itio nal lo ss and det rimen tal ef fect on se nsor y , with an unac cept able ra ncid ity and fi shy of f-fl avor (T an eja and Zh u 2006 ). A p ossi bl e tool to prot ect om ega-3 PUF A fr om oxid ation is mi croe ncap sula tion , * Est efaní a Jimé nez -Mar tín esji ma04 @alu mno s.un ex. es * T e resa An teq uera Ro jas tant ero@ unex .e s 1 Dep artm ent o f Food T ech nolo gy , Sch ool of V ete rina ry Scie nces , Res earc h Inst itut e of Meat and M eat Pr oduc t, Uni versi ty of Ext remad ura, 1000 3 Cá cere s, Spai n 2 Dep artm ent of Food Sc ienc e, Un iversi ty of Copenh agen, Rol ighe dsve j 30, 19 58, Fred eriks ber g C, Denm ark Food Bi oproc ess T echn ol (2016 ) 9:285 – 297 DOI 10 .1007/ s119 47-01 5-16 21-x whic h limits th e advers e food proc essi ng enviro nmen t such as the co ntac t with wat er and oxyg en by ke epin g bioact ive func - tional food component s within protective matrix structures (Onw ulat a 2013 ). Fish oi l is curren tly microe ncap sula ted by dif ferent me thods, sp ray dryi ng bein g the most po pular on e (Gha rsallaou i et al . 2007 ). Spec if ic all y , micro en ca ps ul at io n of fish oil by spray drying usin g multilayered emulsions of leci thin-c hitosa n combin ed with carb ohydra tes as co atin g ma- teri al has be en repo rted to ha ve succ essf ul ef fect s on protec - tion fr om oxidat ion (Jim énez -Martí n et al. 2015 ;K l i n k e s o r n et al. 20 05 ;S h a we ta l . 2007 ). Diet ha bits are ch anging in ac cordan ce with a life styl e that is focu sed on time -savin g, and cons equent ly , there is a grow- ing dema nd for “ read y-to-h eat ” prod ucts, su ch us breade d pre- frie d froz en ones (M iran da et al. 20 10 ; V arel a et al. 2008 ;X i a o et al. 201 1 ). Th e popu larity of these co nven ience pr oduc ts, amon g whic h chic ken nu gget s are th e most po pul ar (Bar but 2012 ), makes th em a promis ing stra tegy t o incr ease the inta ke of om ega- 3 fatty acid s usin g enri chme nt meth ods. This wo uld be of grea t inte rest for heal th-con cern ed cons umers, si nce it woul d allo w to reac h the reco mmenda tions fo r omeg a-3 in - take su gges ted by food au thorit ies; in turn , it woul d cons titute a pote ntial op port unit y for the food in dustry . In relation to omega- 3 microenc apsulati on, most studies have be en focuse d on studyi ng the microc apsule ch arac teri s- tics , wherea s the us e of these mi crocap sule s for en rich ment of food in ω -3 P UF A has rare ly been cons ider ed, and on ly some bake ry prod ucts (San thanam et al . 2014 ;G ö k m e ne ta l . 201 1 ; Umesha et al. 2015 ), beverages (Rubila r et al. 2012 ;K o l a n o w s k i et al. 2007 ; Ilyasoglu and El 2014 ), dairy products (Bermúdez-Aguirre and Barbosa-Cánovas 2012 ;T a m j i d i et al. 2012 ; W an et al. 201 1 ), and meat products (Jos quin et al. 2012 ) hav e been s tu died , and n o exa mple s have b ee n found in the scientific literature of pan-fried convenience meat products enriched with ω -3 microcapsules Ther efor e, the ob jectiv e of this stud y was to st udy fo r the firs t tim e a conv enie nce fr ozen pan-fr ied meat produc t (chick - en nuggets) enri ched with fish oil omega-3 microcapsules, inve stig atin g the ef fect of time of frozen storag e on the ox ida- tive st abilit y and sens ory proper ties in comp arison to bu lk fish oil ad dition . Mat eria l an d Metho ds Exp erimen tal Desi gn Three dif ferent types of chicke n nuggets were produced: a cont rol ba tch (C), a batch en riched w ith bu lk fish oi l (BFO) , and a batc h enrich ed with mi croe ncap sulat ed fish oi l (MFO). Fis h oil (k indly pr ovided by Biomeg a Natura l Nutrie nts S.L. , Gali cia, S pain ) was used as so urce of ω -3 F A (6.9 % EP A, 24.6 % DHA) . Form ulat ion an d manu fact ure of th e chicke n nugget s of the cont rol ba tch was ma de foll owing th e meth odol ogy op timi zed by Medi na et al. (Med ina et al. 20 14 ) usin g chick en brea st as the me at raw mate rial . As for th e enrich ed batc hes, form ula- tion of th e batt er was mod ifie d by the addi tion of 0. 5 % ( w / w ) of fi sh oil in th e BFO ba tch an d by the ad dition of 5 % ( w / w ) of mult ilayer ed micr ocap sules of fish oi l optimi zed in a pre- vious work ( multilayered micro capsules pr epared with lecithin-chitosa n and maltodextrin conta ining 10 % of fish oil) (J iménez -Mart ín et al. 2015 ). T hus, th e enrich ment level in EP A + DHA in B FO and MFO nu ggets wa s 150 mg per 100 g (e noug h to exceed the mini mum le vel re quir ed by th e Euro pean Union le gisl ation to label a food as “ high in ω -3 fatt y acids ” : at least 80 mg of the su m of EP A and DHA per 100 g an d per 100 kc al) (EU 2010 ). Chicken breast was purchased in a lo cal market (Mer cado na, Cá cere s, Spa in) an d minc ed usin g a meat mi ncer (SE B IBERIC A, Barc elon a, Spain) . Then , it was mixe d and furt her minc ed with the ing redi ents in a dome stic ki tchen pr o- cess or Ther momi x (V orwe rk W uppe rtal , German y) acco rdin g to each formulation. A commercial preparation (Procavi; ANVIS A, Madrid , Spain) co ntai ning soya fl our , salt , flavor - ings , soya pr otein, mi lk protei n, dext rose , spices , pent asod ium trip hospha te, tetr asodiu m diphos phate, lo cust be an gum, guar gum, and so dium asco rbate (6 0 g/kg meat ) was adde d. The obt aine d batter wa s stor ed at 0 – 2 °C duri ng 24 h, and it was subs eque ntly po rtione d to obtain ea ch chicke n nugget (4 × 4 × 1 cm, 25 g). Chi cken nugg ets wer e coated fi rst with a com- mercia l stick solution con taining wheat starch , wheat corn, guar gum, and powdered turmeric (A vigum, ANVISA, Madr id, Sp ain) di ssolve d in cold wa ter (2 5 g/L) an d subs e- quen tly with br eadcrumb s (35 g/kg nu gget). Afte rwards , all nugg ets were imme diat ely subjec ted to a pre-fr ied proc ess in sunf lowe r oil at 180 °C duri ng 10 s and imme diat ely remo ved from th e fryer an d placed on pa per towel to remo ve excess of ext erna l oil. Then , all the pr e-fr ied nugg ets were stor ed in a tray co vere d with a plas tic wrap du ring 24 h at refr igerat ion tem pera ture (0 – 2 °C). Afte r this re frig eratio n period , part of th e pre- frie d nugg ets of each batc h were imme diatel y deep-f ried and ana- lyze d (T0) , while th e rest were st ored du ring 1 mont h (T1M) or 3 months (T 3M) in a domest ic free zer at − 18 °C befo re deep fr ying . Nugget s were de ep-f ried at 18 0 °C in sunf lowe r oil un til th e prod uct reache d 63 °C of in ternal temper ature. Thus , ni ne grou ps of fr ied nugg ets were ob tained : C-T0 , C- T1M, C-T3M, BFO -T0, BFO-T1M, BFO -T3M, MFO-T 0, MFO -T1M, an d MFO- T3M. T empera ture in th e co re of nugg ets during fr ying was re - cord ed usin g a thermo mete r prob e (T esto 73 5-2, L enzk irch , Germ any) . Nine nu gget s were frie d at the sa me time . The oil was repl aced ever y four fryi ng sessio ns. Afte r each deep fr y- ing, nu ggets were dr ained an d placed on pa per towel fo r re- movi ng extern al oil. Ox idat ion assays , vo latile comp ounds , 286 Foo d Biopr oce ss T echno l (20 16) 9:28 5 – 29 7 and sens ory test were an alyzed on the fr ied nugg ets. For vol- atil e compou nds and oxid ation as says , whole nu gget , includ - ing me at and crus t, was minc ed and analyz ed by duplic ate. Met hod s Sen sory Ana lysi s All sess ions were do ne in sensor y panel ro oms with th e con- ditions specified in UNE-EN-ISO 8589:2010 Regulation (UNE -EN- ISO 20 10 ). Pa nel room s were eq uipped wi th whit e fluo resc ent light ing (2 20 – 230 V , 35 W). Nu gget s were se rved hot on whit e plas tic plates to pane lists, ma rked wi th rand om thre e-di git code s. The panel sess ions we re held arou nd 1 – 2h befo re lunc htime. S alt-fr ee crac kers and a glas s of wa ter at room t empe rature we re provid ed to each pane list to rins e be- twee n samp les. Quan tita tive Des cripti ve Chic ken nu ggets we re asse ssed by a traine d panel of 15 mem- bers using a de script ive an alysis me thod. Th e paneli sts we re trai ned i n four se ssio ns for 1 h. In th e first se ssio n, pane lists test ed chicke n nugg ets an d were aske d to expres s terms th at desc ribe th eir pers onal obse rvat ions . In the second se ssion, th e redu ndan t desc ript ive te rms wer e elim inated an d the samp les were test ed to in clud e new at trib utes . In th e thir d sess ion, all t h ea t t r i b u t e ss e l e c t e dw e r eu s e di na nu n s t r u c t u r e ds c a l e o f0 – 10. At the fo urth se ssio n, initia lly , the di ffere nt attr ibutes we re eval uate d indi vidu ally by pane lists in unkn own repres entati ve samp les, an d then, pane lists ’ re sponse s were anal yzed coll ec- tive ly . Data were ga ther ed and anal yzed by anal ysis of vari - ance (A NOV A), an d pane list devi ations we re assess ed to de- term ine wh ere ad diti onal traini ng was ne eded . Nine teen se n- sory tr aits of nugg ets groupe d unde r appe aran ce, text ure, od or , and fl avor were asse ssed . Thei r defi niti ons an d extr emes ar e expl aine d in T able 1 . Ques tions we re presen ted to asse ssors in the no rmal perc eption or der , as follow s: visu al anal ysis , tex- ture , tast e and flav or . Sens ory trai ts of the sa mple s of chicke n nugg ets were asse ssed by ea ch pane list in a 10-cm un stru c- ture d line . Hedo nic T est A hedonic te st was carried out using an uns tructur ed nine- poi nt line sc ale with he donic insc riptio ns at the lo wer extr eme “ di slike d extr emel y ” an d upper ex treme “ li ked ex trem ely ” of the sc ale. Th irty untr ained co nsum ers (stu dents an d staf f of the uni vers ity) volu nteere d to evalua te hedo nic scor e of the chic k- en nugg ets. The test to ok place in th e tastin g room s, under th e same co ndi tions as th e quanti tative -des crip tive anal ysis. Thr ee fried chicken nuggets were evaluated in each session, and samp le orde r was random ized ac ross cons umers. Th irty sam- ples fr om each ba tch were an alyzed . V olat ile Co mpou nds V ol atil e compou nds were an alyz ed by he adsp ace soli d-phas e micr oext ract ion (H S-SP ME) fo llow ing th e meth od desc ribed by Garcia-Esteban et al. (Garcia-Esteban et al. 2004 ) with some modi ficati ons. Each chic ken nugget was groun d, and 5 g were weigh ted into a 20-mL glass flask sealed with an alum inum cap and a se ptum. Sea led vi al was co ndi tioned in a ther most atized wa ter bath at 37 °C fo r 30 min. SPME wa s carr ied ou t by using a cros s-li nked divi nylb enze ne/c arbo xen/ polyd imethyl siloxane fi ber , 50/30 μ m t hi ck and 2 cm long (Sup elco, Bel lefont e, P A, USA), co nditio ned pr ior to us e by heating in the injection port of a gas chromatograph (GC) syst em under th e cond itio ns re commen ded by the ma nufac- ture r (at 27 0 °C for 1 h). Th e fiber was th en insert ed into th e samp le vial th roug h the sept um and expo sed to the head spac e for 30 mi n at 37 °C. The SPME fi ber was deso rbed an d main- tain ed in the inje ction por t for 30 min. Anal yses were per - form ed using a Hewl ett – Pack ar d 6890 ser ie s I I GC c ou pl ed to a mass sele ctive (MS ) detect or (Hew lett – Pack ard HP 597 3) (W ilming ton, DE, US A). V ol atil es were sepa rated us ing a 5 % phen ylmethy l silico ne (HP-5 ) bonded -pha se fuse d silica ca p- illary column (Hewlett – Packard, 50 m × 0.32 mm i.d., film thic kness 1.05 μ m), op erat ing at 6 ps i of colu mn head pres - sure , resu lting in a fl ow of 1.3 mL /min at 40 °C. The in jectio n port was in spli tless mode. Th e temp erat ure pr ogra m was iso- t h e r m a lf o r1 0m i na t4 0° C ,r o s et o2 0 0° Ca tar a t eo f 5 °C/m in and th en rose to 25 0 °C at a rate of 20 °C /min , and hel d for 5 min. The tr ansf er line to th e mass sp ectr omet er was main tained at 280 °C . The ma ss spectr a were ob tained usin g a mass se lect ive dete ctor by elec tron ic impact at 70 eV , a mu l- tipl ier voltag e of 1756 V , an d collec ting data at a rate of on e scan ov er the m / z ra nge of 30 – 550 u.m. a. n-Al kane s (Sigma R-87 69) were an alyzed un der the sa me co nditio ns to calc ulat e the li near re tenti on in dice s (LRIs) fo r the vo lati le compo unds. Compounds were identified by comparison with the ma ss spectrum and RI of commercial reference compounds (Si gma-Al drich; St einh ein, Germ any), by comp arison of mass spec trum wi th mass sp ectr al data base (NIS T an d Wil ey librar - ies) an d by comp arison of th eir RI with thos e availa ble in the lite ratu re. Resu lts from vola tile anal yses ar e provid ed in area unit s (AU ). Oxid atio n Conj ugated Dien es Th e meas uremen t of conj ugat ed dien es (CDs ) was dete rmine d according to the procedure described by (Juntachote et al. 2006 ). Frozen chicken nuggets were thawed and minced. Food Bi oproc ess T echn ol (2016 ) 9:285 – 297 28 7 Sub sequ ently , 0.5 g were su spen ded in 5 mL of dist illed wa ter and ho moge nize d to form sm ooth slur ry . A 0.5 mL al iquo t of this su spen sion wa s mixed with 5 mL of extr acting so lution (3:1 ( v / v ) he xane/i sopr opan ol) for 1 mi n. Afte r centri fugati on at 3500 rp m for 5 min, the ab sorb ance of th e supern atan t was read at 23 3 nm. Th e conc entr atio n of CD wa s calcu lated us ing the mo lar exti ncti on coef ficien t of 25,200 M − 1 cm − 1 , and th e resu lts wer e expres sed as microm oles pe r kilogr am of samp le. Thiob arbitu ric Acid Reac tive Subs tanc es Thio barbit uric ac id re activ e subs tance (T BARS ) conten t was measure d by fo llowing the ext raction me thod descr ibed by Salih e t al. ( 1987 ). Eac h nugget was m inced in a kitc hen blen der , and 2. 5 g were ho mogeni zed for 2 mi n with 7. 5 mL of 3.86 % perc hlor ic acid an d 0.5 mL of butylat ed hydrox y- tolu ene. T ubes were ke pt in ice to avoid he at degr adat ion. This homo gena te was fi lter ed and ce ntrifu ged (3 min, 35 00 rpm). The su pernatan t (2 mL) was mi xed with 2 mL of 97 % 1,1, 3,3- tetraethoxypropane (TEP). Immediately , the mixture was heated to 90 °C for 30 min, cooled, and centrifuged again (2 min, 3500 rpm). Absorbance was measured at 532 and 600 nm on a spectrophotometer (Hitachi U-2000, T okyo, Japa n). The me asur emen t at 600 nm is cons idered co ntamin a- tion , and it wa s subt ract ed to th e othe r measur emen t to obta in the fi nal ab sorb ance . The conc entrat ion of TB ARS wa s cal- cula ted as ma londia ldehyd e (MDA) from a st andar d curv e, whic h was de velo ped si mult aneo usly wi th the samp les usin g solu tion s of TEP (Merck , Schc hard t, Germ any). TB ARS we re expr esse d as mg MDA kg − 1 samp le. Pr otein Ca rbonyl s Prote in oxidat ion in chic ken nu gget s was fol lowed by me a- suri ng the format ion of prot ein carb onyls by co nver ting th em to 2, 4 dini trop heny lhyd razo nes (DNP H), and t he deri vati ves were me asur ed spectr opho tome tric ally acco rding to meth od outlined (Olive r et al. 1987 ) with slight modi fications. Ta b l e 1 Sen sory at tribu tes of fri ed chi cken nu ggets Sens ory trai t Def initi on Befo re eatin g V isua l app earan ce Hom oge neit y of the cr ust colo r Hom ogen eity of the cr ust color (les s-mu ch) Hom oge neit y of the st ructur e Hom ogen eity of the st ruc ture ( less -muc h) Odor Int ensi ty Int ensi ty of the ge neral od or (odor less -very inte nse odor ) Fryi ng Int ensi ty of the fry ing od or (odo rless-v ery inten se odor) Fish Int ensi ty of th e fish od or (odor less- very in tens e odor ) Ranc id Int ensi ty of the ran cid odor (od orless -ve ry inten se odor) Dur ing ea ting T ext ure Crus tiness Fee ling of cru sty edge (no t cr usty -ver y crus ty) Oili ness Sens ation of oil cont ent in th e mout h (no t oily- very oi ly) Jui cine ss Per cept ion of j uici ness du ring che wing (no t juic y – ve ry juic y) Hard ness Fir mness perce ption du rin g chewi ng (ve ry tende r-v ery fi rm) T ast e and fl avor Int ensi ty Int ensi ty of the ge neral fl avor (fl avor les s-ve ry inten se flavo r) Meat Int ensi ty of th e mea t flavo r (less -muc h) Oil Int ensi ty of the oi l flavo r (less -much) Salt Int ensi ty of th e salt y tast e (not sal ty-ver y salt y) Spi cy Int ensi ty of the sp icy flav or (not spic y-very sp icy) Ranc id Int ensi ty of th e ranc id fla vor (no t rancid -ver y ran cid) Fish Int ensi ty of the fis h flav or (le ss-m uch) Afte r eating T ext ure Resi dual part icles Sens ation of par ticle s in the mouth aft er swal low ing (less -much) Flav or Afte rtaste Per siste nce of fl avo r afte r swall owing (n on pe rsi stent -very pe rsisten t flavo r) 288 Foo d Biopr oce ss T echno l (20 16) 9:28 5 – 29 7 Chicken nuggets were minced, and then, 1 g of meat was homo geni zed 1:10 ( w / v ) i n 20 mM sodi u m phosp ha te b uff er cont aini ng 0.6 M NaCl (p H 6.5) us ing an ultr aturra x homo g- eni zer for 30 s. T wo equa l aliq uots of 0. 2 mL were ta ken from the homogenates and dispens ed in 2 mL Eppendorf tubes. Prote ins were pr ecip itat ed by cold 10 % tric hloroa ceti c acid (TCA) (1 mL) and subsequently centrifuge d for 5 min at 5000 rp m. One pellet w as treated wi th 1 mL 2 M H Cl (pro tein conc entrat ion me asur ement) an d the othe r with an eq ual vol- ume of 0. 2 % ( w / v ) DN PH in 2 M HC l (car bony l conc entr a- tion meas urement ). Both sa mple s were in cubate d for 1 h at room te mper atur e. Afterw ards, samp les were prec ipit ated by 10 % TCA (1 mL) and wa shed tw ice with 1 mL etha nol/et hyl acet ate (1 :1, v / v ) to remove ex cess of DNPH. Th e pellet s we re then di ssolve d in 1.5 mL of 20 mM so dium ph osph ate buf fer cont aini ng 6 M guan idine HCl (pH 6.5) , stirre d, and ce ntri- fuge d for 2 mi n at 5000 rpm to re move i nsol uble fr agme nts. Protein concentration w as calculated from absorptio n at 280 nm us ing a BSA st anda rd curv e. The amou nt of carbon yls was expressed as nanomoles of carbonyl per milligram of prot ein us ing an abso rption co effici ent of 21.0 nM − 1 cm − 1 at 370 nm fo r prot ein hydr azones . Stat istica l Anal ysis Thre e batche s were prep ared ( n = 3) fo r each chic ken nugget form ulatio n (C, BFO, an d MFO) and subj ected to th e dif ferent cond itions of fr ozen st orag e. For each ph ysicoc hemi cal anal - ysis , five sam ples of chicke n nugg ets from ea ch combin atio n bat ch storag e (C-T 0, C-T1M, C- T3M, BFO -T0, BFO- T1M, BFO- T3M, MF O-T0, MF O-T1M, an d MFO-T 3M) were an - alyz ed in dupl icat e. The effect of enrichment ( pE ), frozen storage ( pS ), and thei r intera ctio n ( pExS ) on oxidat ion parame ters , volati le com- poun ds, and qu antita tive -des cripti ve sens ory da ta were an a- lyze d usin g a two-wa y ANOV A with intera ctio n. When a si g- nifi cant effe ct ( p 0.05 ) was dete cted, pa ired co mparis ons be - tween means were condu cted using the T ukey ’ st e s t . Signi ficant di ffere nces ( p 0.05 ) in the hedo nic test were de - tected using a nonparametric analysis achieved using Friedman ’ s test. The statistics were run using the program IBM SP SS Stati stics v .19. Res ult s and Di scu ssion V olat ile Co mpou nds The GC-M S analys is allowe d the iden tifi catio n of 88 dif fere nt vol atile co mpou nds, 3- meth ylbu tana l, 2-meth ylbuta nal, he xa- nal, 2-methylpe ntane, 3-methylp entane, pentane , 2- propanone, and 2-pentylfur an being the major ones. This profi le basic ally ag rees with prev ious stu dies ab out de ep- frie d batter ed produc ts (Özyur t et al. 201 1 ). Figu re 1 show s the le vel of vo latile co mpound s grou ped in 1 1 main chem ical fa milie s: terpen es, al dehy des, al iphat ic hy- droc arbo ns, ke tones, al cohols , benz enes , fura ns, su lfur co m- pounds, aromatic hydroc arbons, esters, and pyrazines. The most ab unda nt fami lies in all ch icke n nugget s in term s of ar ea uni ts (Fig . 1a ) were alde hydes an d terp enes , foll owed fa r be- hind by al iphat ic hy drocar bons, keto nes, alco hols , benzen es, and fu rans (Fig . 1b ). The mino r vo lati le grou ps (Fig. 1c ), with just li ttle am ount s detect ed in all sample s, were su lfur co m- pounds, aromatic hydrocarbons, esters, and pyrazines. Enri chme nt caus ed signif icant dif fere nces (T able 3 )b e t w e e n bat ches on tota l amou nt of volati le compo unds an d most of the ch emic al grou ps, excl udin g fura ns, su lfur co mpou nds, and este rs. Th e most remar ked change s were tho se af fectin g alco - hol s, keto nes, benz enes, and fu rans , which were de tected in high er amou nts in BFO th an in C and MFO. In addi tion, th e leve ls of alde hyde s and pyrazi nes were also hi gher in BFO and C than in MF O. In meat pr oduc ts as chic ken nugg ets, th e alco hols , ketone s, fura ns, and al dehyde s found ar e most like ly formed through the oxidation of fatty acids (Shahidi et al. 1986 ; Jian g et al. 201 1 ). Thu s, the hi gher le vels of th ese com- poun ds in BFO as co mpar ed to th e othe r batc hes co uld in di- cat e a lower pr otecti on to ox idativ e proc esse s of the fi sh oi l when it is adde d direct ly to th e batt er duri ng manufa cture of the me at prod uct. T ime of froz en storag e signif ican tly af fected th e leve ls of some ch emical grou ps (terpe nes, al dehyde s, alip hatic hy dro- carb ons, ke tone s, fura ns, and este rs) (T able 3 )a sw e l la st h e tota l amount of vo latile co mpou nds. Ho wever , this ef fect di d not fo llow the sa me trend i n C, BFO, an d MFO ba tche s. Thu s, terp ene co nten t incr ease d with fr ozen st orage in C nu gget s (C- TO > C-T1 = C-T3), wh ile no chan ges in the area un its of this chemical fami ly were found in BFO and MFO . Aldehyde cont ent in crea sed with fr ozen st orag e time in BFO an d MFO nugg ets (T0 where as in C ones , C-T1 an d C-T3 showed si milar amoun t and higher tha n C-T0. As for total amou nt of vola tile co mpou nds (Fig . 1a ), froz en stor age had no influ ence on the tota l amou nt of vola tile co mpou nds in the C grou p. On the cont rary , in enrich ed nugg ets, fr ozen st orag e incr ease d the tota l amount on vo lati le compou nds in diffe rent ways . In BF O, the tota l am ount of vo lat ile comp ounds did no t chan ge in T1M with resp ect to T0 bu t increa sed arou nd a 10 % in T3M with re spect to T1M, wh erea s in MFO, it incr ease d arou nd a 50 % in T1M with re spec t to T0 and did not ch ange in T3M with re spect to T1M . Our resu lts agre e with the re sult s of other st udies that rep orted an in crease in aldeh ydes, ke- tone s, hydr ocar bons , esters , an d furans duri ng frozen st orag e of meat pr oduc ts (Ladik os and Lougov ois 1990 ). As fo r the dif ferenc es foun d in the ef fect of froz en st orag e time am ong the ba tche s of chic ken nugg ets, it coul d be relate d to the dif- fere nt type of en rich ment in omeg a-3 PUF A. Some aut hors Food Bi oproc ess T echn ol (2016 ) 9:285 – 297 28 9 have pr evio usly sh own a high er total am ount of vo lati les as a cons eque nce of a high er fat cont ent in meat samp les (Ols en et al. 20 05 ). From th e 88 vo lati le comp ound s initia lly detect ed, 13 were selected (T able 2 ) as indicator s of lipid oxidati on (butanal , hexane, pentanal, hexana l, heptanal, 2,4-hexadienal, 2- pentylfuran, octanal, nonanal, and 2 ,4-decadienal) or Str ecker de gradat ion (2 - and 3- methy lpro pana l and dime thyl disu lfid e) (Ols en et al. 2005 ; Estevez et al . 2005 ;P a r ke ta l . 2007 ; Perez- Palacio s et al. 2013 ;E l m o r ee ta l . 1999 ; Soyer et al. 20 10 ; Sta dtma n 1990 ;X i o n g 2000 ; Huan g et al. 2013 ). All 13 vo lati le compou nds select ed were foun d in the thre e bat ches C , BFO, an d MFO, exce pt for 2, 4-he xadi enal , whic h was only found in BFO nuggets, and 2-4-decadienal, not foun d in C sample s. It is also wor th noti ng the high co nten t of he xane in the fr ied nu ggets of th is study , being th e majo r volatile compound. Other authors have also shown large amou nt of this vo lati le com pounds in coat ed frie d produc ts (Per ez-P alac ios et al. 2013 ), in which it is ascr ibed to lipi d oxi dati on during t he frying pr oces s (Özyur t et al. 201 1 ). Enrichment in fish oil and frozen sto rage caused differ - ence s on the leve ls of the sele cted vo latile comp ounds, ex cept for butanal and 2-pentylfuran, which were not affected by nei ther of th ese two fa ctor s, althou gh they were af fected by thei r in tera ctio n. In fact, inte ract ion of froz en storag e an d te m- pera ture si gnif ican tly af fect ed all sele cted vo latile comp ounds exce pt for 2- methyl butana l and pent anal. Over all, in C , BFO, and MFO nu gget s, the length of frozen storage led to a signi ficant inc rease in most v olatile com- poun ds from li pid ox idat ion an d amino ac id de grad ation, ex - cept for hexane that decreased. Nevertheless , the effect of frozen storage differed among nugget types. For example, the i nc re as e o f hex an al , w hi ch h as b ee n w id ely u se d as o xi - dat ion mar ker in cook ed meat and meat prod ucts (P ark et al . 2007 ), was notice able nota ble in en riched nugget s but not in C ones. A significant incr ement of heptanal, oct anal, and 0 25 50 Total Terp Ald AliH Ket Alc Benz Fur Sulf AroH Est Pyr 100 300 500 700 0 25 50 Total Terp Ald AliH Ket Alc Benz Fur Sulf AroH Est Pyr 100 300 500 700 0 25 50 Total Terp Ald AliH Ket Alc Benz Fur S ulf AroH Est Pyr 100 600 1100 a b b b a a a b b b b a a b b c a b b c a b b a c b a a b b a b b a b c b b a a b c b a a b a a b b a c b a b a a a) b) c) x y y y y x y x y x xy x y x y x y y z z y x y y x x x y y y y x x x x y y y y x x x x y x y y x x x y y y y z x x y y x x y y x x x x x y Fig. 1 Fami lies of volat ile s (in area un its ×10 6 )i nt h ec o n t r o l( a ), enr iche d with bu lk fish oi l ( b ), and e nric hed wi th mic roenc apsul ated f ish oi l ( c ) fri ed chic ken nu gget s, at ini tial time ( T0, da rk gray ) and af ter fro zen stor age for a month (T 1M, medi um gray ) an d 3 mon ths (T3M, li ght gr ay ). Ba rs with dif fer ent lett ers ( a , b , c )w i t h i nt h e same f ormula tion an d family of vola tile com poun ds show sig nifi can t dif fere nce s ( p . 0 5 ) due to fro zen stor age ef fect . Bars wit h diffe r ent le tters ( x , y , z ) amon g fo rmul atio ns ( a , b ,a n d c ) indi cate sign ifi cant di ffe renc es ( p 05) due to enrich ment eff ect. Five ch ick en nugg ets of each of fr ied nugg et batc h ( n =3 ) subj ected to ea ch stor age con diti on (T0, T1M , and T3 M) were a naly zed in du plic ate. To t a l tot al vola til e compo unds , Te r p terp enes, Ald aldehy des, Al iH alip hatic hy droc arbon s, Ket ket ones , Alc alc ohols , Benz ben zene s, Fur f ura ns, Su lf sulf ur com p ou nd s, Ar oH arom atic hydr ocarb ons, Est ester s, Pyr pyr azine s 290 Foo d Biopr oce ss T echno l (20 16) 9:28 5 – 29 7 Ta b l e 2 Selected volatile com pounds (expressed as AU ×10 6 ) related to oxidation in the control (C), enriche dw i t hb u l kf i s ho i l( B F O ) ,a n de n r i c h e dw i t hm i c r o e n c a p s u l a t e df i s ho i l( M F O )f r i e dc h i c k e n nuggets, at initial time (T0) and after frozen storage for a month (T1M) and 3 months (T3M) CB F O M F O IK Compound T0 T1M T3M T0 T1M T3M T0 T1M T3M pE pS pExS 591 Butanal nd c 14.56 ± 0.16 b 28.64 ± 3.56 a 0.12 ± 0.01 b 0.19 ± 0.05 a 0.23 ± 0.02 a nd b 0.15 ± 0.02 a 0.20 ± 0.03 a ns ns * 600 Hexane 148.49 ± 46.93 y 171.24 ± 24.14 141.02 ± 16.26 214.41 ± 21.76 a,x 202.99 ± 10.32 b 158.66 ± 6.13 c 206.12 ± 14.31 a,x 186.27 ± 29.46 b 178.91 ± 11.76 b * *** *** 649 3-Methylbutanal 38.03 ± 7.54 c,y 91.24 ± 14.64 b,z 102.65 ± 31.51 a,y 55.64 ± 8.06 c,x 1 19.87 ± 14.81 b,y 148.8 ± 15.46 a,x 59.01 ± 12.95 b,x 168.09 ± 23.68 a,x 155.61 ± 15.14 a,x *** *** * 659 2-Methylbutanal 17.16 ± 6.44 b,z 43.46 ± 8.25 a,y 46.47 ± 13.54 a,y 25.65 ± 3.67 b,y 60.01 ± 7.31 a,x 78.78 ± 7.59 a,x 36.59 ± 4.19 b,x 77.54 ± 27.86 a,x 88.67 ± 10.65 a,x *** *** ns 698 Pentanal 3.89 ± 1.65 y 4.65 ± 2.39 5.12 ± 1.95 6.05 ± 1.04 b,x 5.25 ± 0.55 b 7.62 ± 0.67 a 3.12 ± 1.76 c,y 5.56 ± 0.56 b 7.73 ± 0.59 a ** *** ns 750 Dimethyl disulfide nd b nd b,z 0.20 ± 0.01 a,y nd b 0.26 ± 0.06 a,y 0.31 ± 0.15 a,x nd b 0.46 ± 0.12 a,x 0.42 ± 0.07 a,x *** *** *** 800 Hexanal 40.97 ± 3.91 a,x 32.98 ± 5.46 b 41.25 ± 3.83 a,x 39.25 ± 3.46 b,x 34.25 ± 3.47 c 49.47 ± 3.61 a,y 35.03 ± 3.75 b,y 31.13 ± 3.00 b 44.60 ± 2.69 a,x ** *** * 901 Heptanal 2.25 ± 0.41 x 1.99 ± 0.42 x 1.82 ± 0.35 y 1.87 ± 0.22 b,x 1.51 ± 0.25 c,x 2.36 ± 0.42 a,x 1.07 ± 0.60 b,y 1.25 ± 0.13 b,y 1.99 ± 0.07 a,y ** ** *** 914 2,4-Hexadienal nd y nd y nd y 0.80 ± 0.02 b,x 0.97 ± 0.01 b,x 1.45 ± 0.02 a,x nd y nd y nd y *** *** *** 995 2-Pentylfuran 13.56 ± 3.95 b 9.16 ± 2.31 c 19.28 ± 2.07 a 14.32 ± 7.76 15.66 ± 1.15 16.79 ± 1.59 13.40 ± 1.30 b 14.34 ± 2.24 b 19.62 ± 1.13 a ns ns *** 1004 Octanal 0.99 ± 0.18 a,x 0.50 ± 0.06 b 0.61 ± 0.19 b,y 0.69 ± 0.17 b,y 0.60 ± 0.19 b 1.26 ± 0.68 a,x nd c,z 0.49 ± 0.14 b 0.71 ± 0.19 a,y ** ** *** 1 101 Nonanal 4.69 ± 1.48 a,x 3.89 ± 1.03 b,x 3.14 ± 0.69 b,y 3.71 ± 0.32 b,x 2.33 ± 0.42 c,y 4.33 ± 0.90 a,x 1.72 ± 0.59 b,y 1.68 ± 0.56 b,z 2.86 ± 0.83 a,z *** * ** 1325 2,4-Decadienal nd z nd y nd z 0.21 ± 0.03 c,x 5.71 ± 2.85 b,x 7.80 ± 0.90 a,x 0.1 1 ± 0.01 b,y nd c,y 4.70 ± 1.41 a,y *** *** *** V alues in the columns with different letters (a, b, c) within the same form ulation show significant differences due to frozen storage effect. V alues i n the column s with different letters (x, y , z) among formulations, indicate signifi cant differences due to enrich ment effect. Five chicken nuggets of each of fried nugget batch ( n = 3) subjected to each storage condition (T0, T1M, and T3M) were analyzed in duplicate, and values are expressed as mean ± standard deviation nd not detected, ns no significant differences, IK linear retentio n index (method of identification: ma ss spectrum and IK identical to a r eference compound), pE p value of enrichment effect, pS p value of frozen storage effect, pExS p value of interaction (enrichment effect × frozen storage e ffect) * p ** p *** p Food Bi oproc ess T echn ol (2016 ) 9:285 – 297 29 1 nona nal in enrich ed nugget s was detect ed in BFO and MF O. This in crem ent was more rema rked in nugg ets su bjecte d to 3 months of frozen storage, especially in BFO batch. 2,4- Deca dien al, an unsatu rate d aldehy de arisin g mainl y from the oxi dati on of lino leic and ar achido nic ac ids, w as only de tected in enri ched pr oduc ts, show ing a more ma rked in crease in BF O than in MF O ba tche s. In addi tion , 2,4-he xadien al was only det ecte d in BFO samp les. Th ese fact s indi cate th at omeg a-3- enriched nuggets tend to show increasin g levels of com- poun ds from poly unsa tura ted fat ty acid ox idatio n throug hout the fr ozen st orage an d that su ch an inc rease is qu ite de pend ent on the way of en rich ment . V olatile comp ounds as Strecker degrada tion markers, 2- and 3 -m et hy lb ut an al , als o r ep or te d a n inc r e as e wit h t im e of frozen storage, which agrees with literature (Olsen et al. 2005 ). Thes e compounds have an impo rtant implica tion in flavor of meat produc ts and have bee n demonstrate d to be originated from degr adation of branched amino acids such as leuc ine and isol eucine (E lmore et al . 1999 ). Dime thyl di sulfid e was not de tected in T0 sample s and was onl y detect ed afte r frozen st orag e, whic h indica tes that du ring freezing and subseque nt frozen stora ge, there are ongo ing chemical changes that allows the further formation of this compound during the subsequent frying. In C nuggets, di- meth yl disulf ide on ly ap peared after 3 mo nths of fr ozen, wh ile in enri ched prod ucts , it was dete cted at the ea rlie r stor age time (T1M ), but it di d not in crease d with furthe r storage le ngth . Disulfide s have been detected in fro zen-store d meat, being implicated in the Strecker degradation of sulfur-containing amin o acid s (cyste ine, me thioni ne) vi a some fr ee ra dical re ac- tions and the form ation of hydrog en sulfide (Hua ng et al. 2013 ). Lipid an d Protei n Oxidat ion CD, TBAR s, and carb onyl s were high ly affe cted by enri ch- ment, frozen storage, and their interaction ( p 01) (T able 3 ). As expe cted, ox idatio n proc ess in th e chicke n nug- get s was reduce d during fr ozen st orag e, but the reac tions were not fu lly st oppe d, whic h agrees wi th lite rature de aling with oxi dati on duri ng froz en stor age (Hua ng et al. 2013 ). Result s for li pid and pr otei n oxidat ion are show n in Fig s. 2 and 3 , resp ecti vely . In all ba tche s, the le vels of CD increa sed (Fig. 2a )f r o mT 0 to T1M and th erea fter decr eased in T3M. Sin ce CD are pri- mary lipid ox idat ion pr oduc ts, th is resu lt indi cates th e forma- tion of CD in sample s subj ecte d to 1 month of fr ozen stor age, whil e in sa mples fr ozen stor ed for 3 mont hs, the de crease of CD seem s to indi cate th at these co mpound s may have be en subs eque ntly tr ansf orme d into ot her prim ary oxidat ion prod - uct s that are pr oduc e later than C D, such as hy drop erox ides . Thus, in all batche s of nuggets frozen sto red for 3 months before frying, it seems that rate of degradation of primary oxi dati on prod ucts in to seco ndar y oxidat ion pr oduc ts is fast er than th eir rate of form atio n. Bots oglo u et al. (Botso glou et al. 2014 ) studie d oxid atio n rate during frozen storage in pork patties enriched with omeg a-3 PUF A and dete cted incr eased CDs af ter 2 months of stor age, wh ich incr eased furt her after 4 mont hs but decl ine d ther eaft er . This cont rast s with our stud y , as decr ease of CD in the chicke n nuggets beg an earlier (af ter 1 month of fro zen stor age). Howe ver , the diffe rences in enri chment and form u- lation of the product should be taken in to account. Comp ari son of ou r data with simi lar st udie s is chal lengin g, as lite ratu re deal ing wit h frozen st orag e of produc ts enrich ed with ω -3 PU F A is scarce an d no record s on effe ct of free zing or frozen storage on pre-fried enriched products with ω -3 PUF A a re a va il ab le . As for TB ARs (Fig. 2b ), in genera l, there was a sign ifi cant incr ease du e to frozen st orag e time, al thou gh this ef fect was quite differen t among batches. Thus, in C nuggets, TBAR val ues hi ghly in crease d with lo nger froz en st orag e time (T 0 e in enriched sam ples, TBAR values slig htly incr eased only af ter 3 mont hs of froz en stor age. In addi tion , it is worth th e hi gher T BAR valu es found in BFO in comp aris on to C and MFO ba tche s at T0, T1M , and T3M. In th e stud y of Bo tsoglo u et al. (B otso glou et al. 20 14 )p r e v i - ously mentioned, concentrati ons of TBARs in frozen pork pat ties enri ched with om ega- 3 PUF A stored at − 18 °C rapi dly incr ease d duri ng 120 da ys of stor age. Ho wever , the di ffere nt comp osit ion of th e meat and th e specif ic form ulat ion ma kes the co mparis on diffi cult . Then , it seem s that en rich ed batc hes (b oth BF O and MFO) had a mo re stab le behavi or rela ted to seco ndar y lipid ox ida- tion du ring stor age with fewer ch ange s in rate of ox idatio n than C. Ho wever , in BFO, the resu lts indic ate an adva nced stag e of se cond ary li pid ox idat ion ev en in sa mple s not su b- ject ed to fr ozen st orag e, which co uld be rela ted to their fo r- mula tion with the di rect ad diti on of fish oi l. Thus, an in crea sed conc entratio n of PUF A in the batter of BF O ba tch could ac t as a proo xidant fa ctor . An i mbalan ce in the initia l pr ooxida nt- ant ioxi dant eq uili brium in th e batter of th is batch coul d ha ve caus ed so lipi d oxidat ion stat us at T0 that the st eady st ate for seco ndar y oxid atio n was re ache d fast er . This stea dy st ate is the ph ase of ox ida tion pr oces s wher e format ion and deco m- po siti on of oxidat ion pr oduc ts as well as in ter acti ons of oxi- dation compounds with other components are equivalent (Mie lnik et al. 2002 ), and so, no chan ges in oxid atio n inde x are me asured. The re sult s sugg est th at the li pid oxid atio n stat us in fi sh oil- enri ched nu ggets is infl uenced by t ime of fr ozen st orag e, al- thou gh the type of ad diti on seem s to be a more ma rked ef fect, being the enrichment with microcapsules more eff ective agai nst lip id oxid ation th an bulk fish oi l addi tion . The valu es of carbon yls (Fig . 3 ) in the nugget s of this stud y ranged from 3.71 to 6.31 nmol/mg pro tein, quite in 292 Foo d Biopr oce ss T echno l (20 16) 9:28 5 – 29 7 Ta b l e 3 Statistic two-way ANOV A with interactio n from results pres ented in Fi gs. 1 , 2 , 3 , 4 ,a n d 5 pE pS pEx S V ola tile co mpoun ds (Fig. 1 ) T ota l * ** ** Te r p e n e s * * * * Alde hydes * *** ns Alip hatic hy droc arbon s ** ** ns Keto nes *** ** ** * Alco hols *** ns ns Benz enes *** ns * Fur ans ns *** ** * Sul fur com pounds ns ns ns Arom atic hy droc arbo ns ** ns *** Este rs ns * ns Pyr azin es ** * ns ** Lipi d ox idati on (Fig. 2 ) Conj ugate d diene s *** *** ** * TBAR s *** ** * *** Prot ein oxid atio n (Fig. 3 ) Carb onyls *** ** * *** Sens ory analy sis Quan ti ta tiv e-d es cr ip ti ve assa y (Fig . 4 ) V isua l app earan ce Hom oge neit y of the cr ust colo r ns ns ns Hom oge neit y of the struct ure ns ns ns Odor Intens ity ns n s ns Fryi ng ns ns ns Fish ns ns ns Ranc id ns ns ns T ext ure Crus tiness ns ns ns Oili ness ns ns ns Jui cine ss * ns ns Hard ness ns ns ns Resi dual par ticles ns ns ns T ast e and fl avor Intens ity ns n s ns Meat * ns ns Oil ns ns ns Salt * ns ns Spi cy ns ns ns Ranc id ns ns ns Fish ns ns ns Afte rtaste ns ns ns Hedo nic tes t (Fig. 5 )n s n s n s pE p value of e nric hme nt eff ect, pS p valu e of fro zen st orag e eff ect , pExS p val ue of inte raction (e nrichm ent ef fect × froz en stor age ef fect), ns no sig nifi can t dif fere nce s * p . 0 5 , * * p *** p . 0 0 1 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 CB F O M F O CD (µmol/m g chicken nugget) a) a a b c a b b b b 0 0.5 1 1.5 2 2.5 3 3.5 CB F O M F O TBARS (mg MDA/kg chicken nugget) b) a a b b b a b b c z x y x y y x z y y y x xy x y y x y Fig. 2 Li pi d o xi da ti on , det er mi ne d b y C D ( a ) and TBAR s ( b )i nt h e control (C), enriched with bulk fish oil (BFO), and enriched with mic roenc apsul ated f ish oi l (MFO) fr ied chic ken nugg ets, at in itial t ime (T0, dark gr ay ) an d afte r frozen st orage for a mo nth (T 1M, me dium gray ) and 3 mo nths ( T3M , light gr ay ). Bars wi th diff er ent lett ers ( a , b , c ) wit hin the s ame fo rmul ati on show sign ifica nt dif ferenc es ( p ) du e to froze n stora ge effec t. Bars with diffe rent let ters ( x , y , z ) among formu lations indi cate sig nifica nt dif feren ces ( p 0.05 ) due to enr ichm ent ef fect. Fi ve chi cken nu ggets of eac h of fr ied nu gget batc h ( n = 3) subj ected to each sto rage co nditio n (T0, T1M, an d T3M ) were an aly zed in du plicat e 0 1 2 3 4 5 6 7 8 9 10 CB F O M F O Carbonyls (nmol/mg protein) a a b c b y x y c x y z x b y b a z Fig. 3 T otal protein carbonyls (DNPH method) in the cont rol (C), enr iche d with bu lk fish oi l (BFO ), and en ric hed with microe ncapsu lated fis h oil (M FO) fr ied chi cken nu gget s, at in itia l tim e (T0, da rk gray )a n d after frozen storage for a month (T1M, medium gray ) and 3 months (T3M, ligh t gray ). Ba rs with di ffer ent le tters ( a , b , c ) wi thin the sa me for mulat ion show sign ifican t dif feren ces ( p 0.05) due t o f ro ze n s to rage effe ct. Bars wi th diff er ent lette rs ( x , y , z ) amon g formu lati ons in dica te sig nifi can t diffe renc es ( p 0.05) due to en richm ent effe ct. Five c hicken nugg ets of eac h of frie d nugge t batch ( n = 3) sub jecte d to ea ch stor age con diti on (T0, T1M , and T3 M) were analy zed in dupli cate Food Bi oproc ess T echn ol (2016 ) 9:285 – 297 29 3

Enrichment of Chicken Nuggets with Microencapsulated, 2201

UEPA

Tecnologia de Alimentos

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