Effect of Bread Making Process on Aflatoxin Level Changes

Transcription

1 Journl of Chemil Helth Risks (14) 4(4), 1 7 Journl of Chemil Helth Risks ORIGINAL ARTICLE Effet of Bred Mking Proess on Afltoxin Level Chnges Jfr Milni 1 *, Seyed Smn Seyed Nzri, Elmir Bmyr 1, Gisou Mleki 1 Deprtment of Food Siene nd Tehnology, Sri Agriulturl Siene nd Nturl Resoure University, Sri, Irn Deprtment of Chemistry, University of Mzndrn, Bolsr, Irn Deprtment of Food Siene nd Tehnology, Ferdowsi University, Mshhd, Irn (Reeived: 1 August 14 Aepted: Otoer 14) KEYWORDS Afltoxins Bred Dough HPLC Yest ABSTRACT: Whet flour is ommodity with high risk of fltoxins (AFs) ontmintion. During the red mking there re mny proesses tht n ffet the AFs stility. The effet of red mking proess using different yest types on AFs levels ws investigted. For this purpose, stndrds of AFs inluding B nd G were dded to flour nd then red loves were prepred. Three types of ommerilly ville yest inluding tive dry yest, instnt dry yest nd ompressed yest were used for dough preprtion. AFs levels in flour, dough, nd red were nlyzed y high performne liquid hromtogrphy (HPLC) with fluoresene detetor. The results showed tht mximum redution in fltoxin levels oserved during first proof while the lest deline ws seen for the king stge. The order of AFs redution in red mking proess ws AFB 1 >AFB >AFG 1. Furthermore, the results indited tht the most effetive yest for AFs redution ws instnt dry yest. INTRODUCTION AFs re seondry metolites of Aspergillus flvus, A. prsitius nd A. nominus. They re rinogeni nd mutgeni in humns nd nimls [1]. These myotoxins n enter in humn food hin y diret onsumption of ontminted grin or proessed food suh s met or other niml produts fed with ontminted food. Although there re wys to prevent fungl growth nd fltoxin formtion, so fr no wy hs een hieved to ontrol myotoxins in food []. AFs hve 18 isomers inluding B 1, B, G 1, G, M 1, nd M. Among them fltoxin B 1 is onsidered the strongest heptorinogen gent []. Bred is mjor nutritionl omponent nd red mking is one of the oldest food proesses, known nd prtied for thousnds of yers, ll over the world [4]. Corresponding uthor: Jmilny@yhoo.om (J. Milni).

2 J. Milni et l / Journl of Chemil Helth Risks 4(4) (14) 1 7 The min miroil hllenges ssoited with red re fungl infetions nd susequent ontmintion with their derivtives suh s fltoxins inluding B 1, B, G 1 nd G (AFB 1, AFB, AFG 1 nd AFG ). The eonomi prolems rising from fungi nd miroil toxin hve reently triggered extensive reserhes [, 6]. Although the temperture used in prepring red proly did not influene reduing fltoxins, ut tivting enzymes used during the fermenttion lone or in omintion with het n result in reduing fltoxins. In other words, despite of the het resistne of AFs, these fermenttion sensitive toxins re eliminted during the king [7]. The effets of vrious ooking tretments on reduing of fltoxin toxiity were studied vi simultneous nlysis of AFB 1, AFB, AFG 1 nd AFG using high performne liquid hromtogrphy (HPLC) with fluoresene detetor []. It ws found tht omintion of the moisture ontent nd itri id onentrtion, signifintly ffeted the fltoxin in the extruded sorghum [8]. Moreover, the reovered fltoxin deresed with rise in moisture ontent nd id onentrtion. AFB 1 ontmintions were investigted in 7 mize smples olleted during three yers nd the fltoxin in mize smples vried ross the yers [9]. Contmintions of griulturl nd food produts y myotoxin hve een extensively studied in Irn [1-14], however, there is still lk of dt showing the effet of proess on fltoxin level in the red produed from ontminted flour. This study ws designed to evlute the impt of different stges of red preprtion on AFB 1, AFB nd AFG 1 in dough nd red the effet of used yest type inluding tive dry, instnt dry nd ompressed yest on destrution of these fltoxins. MATERIALS AND METHODS HPLC onditions Chromtogrphi seprtion nd detetion ws rried out using (Knuver Tehnology, Germny) nd fluoresene detetor djusted t exittion wvelength of 6 nm nd emission wvelength of 4 nm. The seprtion ws hieved on ODS C18 ( 4.6 mm, 1µm) olumn from Merk (Drmstdt, Germny). Contminted flour A vil ontining 1 mg of fltoxin B nd G ws dissolved in 1 ml hloroform; this solution ws dded to g whet flour. This mixture ws stirred so tht the solvent evported [1]. Bred Bking To prepre dough, 1 g flour, g ker's yest, 1 g sodium hloride nd 6 ml wter were mixed for five min, then, the dough fermented t C for 1. h to e developed. After dividing the dough, the fermenttion ws llowed to ontinue for 1 min t C. Then it ws ked for min t ± C. Afltoxins extrtion from flour To extrt fltoxins,. g of ontminted flour ws mixed with 1 ml methnol (HPLC grde). The mixture ws gitted on shker t 11 rpm for min. Then, the slurry mixture ws entrifuged t 6 rpm for min. (Hermle Z A, Germny). The superntnt, onsisting of extrted fltoxins ws olleted nd loted in rotry evportor (RV 1B S99, IKA) under vuum t 4 C for - min. The solid residue ws dissolved in. ml methnol (HPLC grde), filtered y. miro-liter filter nd stored in - C freezer [16]. Afltoxins extrtion from dough To extrt fltoxins from dough, 1. g dough ws diluted with ml distilled wter nd stirred y mixer. The mixture entrifuged t 6 rpm for 1 min. Other extrtion proedures were similr to those of fltoxin extrtion from flour.

3 AFB (µg Kg -1 ) AFB 1 (µg Kg -1 ) J. Milni et l / Journl of Chemil Helth Risks 4(4) (14) 1 7 Afltoxins extrtion from red The ked red smples were ompletely dried fter one dy nd then rushed y mill (Sunny, No. SBG - 4) for five min to rete fine or medium-size powder. g of the powder ws mixed with 1 ml methnol. It ws pled on shker t 11 rpm for min. Other extrtion proedures were similr to wht previously explined [16, 17]. Afltoxins nlysis y HPLC Determintion of AFB 1, AFB nd AFG 1 levels in ontrol nd smples were rried out y HPLC using fluoresene detetor (exittion t 6 nm, emission t 4 nm) nd n ODS C18 olumn ( 4.6 mm, 1 µm). The moile phse ws mixture of deionized wter, etonitril, methnol, nd id eti (9:9:9:1 v/v respetively). The flow rte ws 1 ml/min [16]. STATISTICAL ANALYSIS Dunn s method, together with one-wy nlysis of vrine (ANOVA), ws performed to evlute hnges in the levels of fltoxin. A P-vlue of less thn. (P<.) ws onsidered to show sttistil signifine. All nlyses were performed in SPSS softwre progrm version 16 (SPSS In., Chigo, IL, USA) nd the grphis were done in Exel. RESULTS Figure 1. The effet of red mking proess on AFB 1 using three types of yest Effet of red mking proess on AFB Figure shows the effet of red mking proess using different yests types on AFB. 1 1 Ative dry yest Instnt dry yest Compressed yest Ative dry yest Instnt dry yest Compressed yest Effet of red mking proess on AFB 1 The effet of red mking proess on AFB 1 is shown in Figure 1. There ws signifint differene mong the flour, primry dough, finl dough, nd red prepred y tive dry, instnt dry, nd ompressed yest (P<.).The most differene ws oserved fter the first proofing for ll three types yests. In other words, king stge hd the lest effet on redution of AFB 1. Figure. The effet of red mking proess on AFB using three types of yest There ws signifint differene etween the flour, primry dough, finl dough, nd red ontining tive dry, instnt dry, nd ompressed yest (P<.). The most redution ws oserved fter the first proofing for ll three types yests. Furthermore, it should e mentioned tht, AFB ws not deteted in the red. Effet of red mking proess on AFG 1 Figure shows the effet of red mking proess using different yest types on AFG 1. As n e seen, there is

4 Amount of Afltoxin (µg Kg -1) Amount of Afltoxin (µg Kg -1) Afltoxin G1 (µg Kg -1 ) Amount of Afltoxin (µg Kg -1) J. Milni et l / Journl of Chemil Helth Risks 4(4) (14) 1 7 no onsiderle differenes etween the tive dry, instnt dry, nd ompressed yest, however, signifint differene in AFG 1 levels ws oserved etween the flour, primry dough, finl dough, nd red (P<.). No AFG 1 ws detetle in red. Effet of yest type on fltoxins during the finl proof Aording to Figure, the highest redution in fltoxins level, espeilly for AFB 1, ws otined in dough ontining ompressed yest. It ws signifintly different from other yests (P<.). Similrly, redutions in smples ontining tive dry yest nd instnt dry yest were signifintly different. 1 Ative dry yest Instnt dry yest 6 Afltoxin B1 Afltoxin B 1 4 Afltoxin G1 1 Figure. The effet of red mking proesses on AFG 1 using three types of yest Effet of yest type on fltoxins during the first proof Aording to Figure 4, the lest redution in the level of AFB 1 nd AFB ws oserved using ompressed yest. Additionlly, s mentioned previously, the redution in G 1 ws not ffeted y the yest type. 1 1 Afltoxin B1 Afltoxin B Afltoxin G1 Figure. The fltoxin redution fter the finl proof Effet of king on fltoxins Aording to Figure 6, AFB 1 nd AFB redution in dough ontining ompressed yest ws the highest. It ws signifintly different from other yests (P<.). However, for AFG 1, destrution ws less thn two other yests Afltoxin B1 Afltoxin B Afltoxin G1 Figure 4. The fltoxin redution fter the first proof Figure 6. The fltoxin redution fter king 4 4

5 J. Milni et l / Journl of Chemil Helth Risks 4(4) (14) 1 7 DISCUSSION Results of this study showed tht proessing of whet flour into red ould signifintly redue the fltoxins level. The lrgest deline in ll stges of the proess ould e oserved etween flour nd primry dough (first proof) euse it ws longer thn finl proof nd performed t temperture loser to optimum for S. erevisie i.e. 7 C. Among ll AFs, the most destrution in the first proof ws seen for AFB 1, ut AFG 1 ws stle t this onditions. The lest deline ws seen for the king stge. The fltoxins hve high deomposition tempertures rnging from 7 C to 6 C. Solid AFB 1 is quite stle to dry heting t tempertures elow its therml deomposition temperture of 67 C [18]. The moisture ontent is ritil ftor on fltoxins intivtion espeilly for AFB 1. The presene of wter helps in opening the ltone ring in AFB 1 [19].The king hd the most destrutive effet on AFB on red prepred using ompressed yest. The order of stges effetiveness on AFs destrution ws s follows: first proofing>seond proofing>king. Proly euse of het resistne of fltoxins, king under the ertin temperture hd little effet on AFs destrution. The most derese in AFs during red mking proess ws for AFB 1, sine AFB 1 hd lower moleulr weight nd smller moleulr size nd proly ws more ffeted y destrutive ftors. The lest deresing AF ws seen for AFG 1. This n e eluidted the smller ltones rings in AFG 1 euse AFs destrution minly ourred on ltones ring rupture. There re some reports on the fte of myotoxins during the fermenttion proess. Erlier work showed tht OTA, AFB 1 nd AFB t.19,.9 nd.9 μg/m1 respetively were degrded in 87 91% y three strins of S. erevisie during fermenttion of wort t C for 8 dys []. Redution of ph during fermenttion, due to orgni ids produed y the yests is likely resulted in derese or degrdtion of AFs [1]. Mny food-proessing opertions (suh s rosting, ooking, frying, nd king) inlude het tretment. Therefore, therml intivting of myotoxins hs een thoroughly studied. Myotoxins differ in their stility under het tretments. Afltoxins re resistnt to therml intivtion nd re destroyed only t tempertures of round ºC []. However, fermenttion n sensitize them due to enzymes, ethnol nd Co prodution. The effet of vrious tretments suh s wshing, heting nd steming on redution of fltoxin toxiity ws investigted. It ws onluded tht onentrtion of AFB 1 is more redued y heting thn wshing tretment []. Redution of fltoxin during het proessing depends on the moisture ontent of the produt. In ottonseed mel with % moisture ontent, n 8% redution in AFs ws reorded following het tretment of 1 ºC for h ut with the sme temperture nd time omintion, only % of the AFs were destroyed when the mel moisture ontent ws 6.6% []. In the light of the dt umulted from severl studies of the effet of moisture ontent on fltoxin degrdtion, it n e onluded tht degrdtion is enhned t higher moisture ontent []. Study on effet of fermenttion period hs shown tht there is signifint flling in fltoxin ontent in Idli up to hr of fermenttion [4]. These results gree with those otined in this study. In investigtion of the effet of yest type on AFs, it ws found tht the ompressed yest hd the lest effet on ll AFs during the first proof. The higher moisture ontent of this type of yest thn two others, leds to less live yest ell in the sme weight, onsequently less destrutive tivity on AFs. Differene in ility of yests for fltoxin redution during the first proof is euse of differene in its mnufture nd preprtion methods. The most destrutive effet mong three types

6 J. Milni et l / Journl of Chemil Helth Risks 4(4) (14) 1 7 of yest ws relted to instnt dry yest. This is due to mintenne of enzymes tivity during friting this kind of yest. Moreover, this yest unlike tive dry yest needs less mount of wter for tivtion, so ville wter is enough for instnt dry yest tivity. In the finl proof, the most effetive yest ws ompressed yest. Proly higher Co produing ility nd resulted lower ph is the reson of high destrutive pility t the finl proof. The differene etween tive dry yest nd instnt dry yest in this stge euse of their resemlne ws not onsiderle. During the king, the most derese ws seen for AFB 1 nd AFB using ompressed yest. AFG 1 destrution through the king ws little. It seems tht AFG 1 hs een sensitized to het during fermenttion less thn two others. During the king with ompressed yest, higher moisture ontent of yest nd/or its lower ph my enhne the effet of temperture on AFs destrution. The moisture is n importnt ftor to redue AFs y heting euse the moisture is required to hydrolyze the ltone ring of the AFs during the king []. Previous reserhes showed tht rise in moisture ontent signifintly influened the redution of fltoxin, whih onfirms our findings [8]. CONCLUSIONS The effet of red mking with vrious kinds of yests on fltoxins indited signifint depletion during the first proof due to longer time nd the king stge hd the lest effet on fltoxins. Overll, the ompressed yest during the red mking proess ording to the fermenttion nd king ws the most effetive in reduing levels of fltoxins. ACKNOWLEDGMENTS The uthors ppreite the Sri Agriulturl Siene nd Nturl Resoures University for finnil support of this projet. The uthors delre tht there is no onflit of interests. REFERENCES 1.Brki-Goln R., Aspergillus miotoxins. In: Myotoxins in Fruits nd Vegetles. Brki-Goln R., Pster N., Eds., Ademi Press: 8. pp Bley N Dede C., Duffy A.P., Jolly M.C., Wilson L.N., Flowers T.G., Eonomi risks of Afltoxin Contmintion in the Prodution nd Mrketing of Penut in Benin. MS Thesis, Auurn University, 9, pp hwng J.H., Lee K.G., 6. Redution of fltoxin B1 ontmintion in whet y vrious ooking tretments. Food Chem. 98(1), Plesss S., Pherson L., Bektorou A., Nigm P., Koutins A.A.,. Bred mking using kefir grins s ker s yest. Food Chem. 9(4), Dunnewind B., Vn Vliet T., Orsel R.,. Effet of Oxidtive Enzymes on Bulk Rheologil Properties of Whet Flour Doughs. J Cerel Si. 6(), Vn der Mrel M.J.E.C., Vn der Veen B., Uitdehg J.C.M., Leemhuis H., Dijkhuizen L.,. Properties nd pplitions of strh-onverting enzymes of the α- mylse fmily. J Biotehnol. 94(), Giry B., Girgin G., Engin A.B., Aydin S., Shin G., 7. Afltoxin levels in whet smples onsumed in some regions of Turkey. Food Control. 18(1), Méndez-Alores A., Veles-Medin J., Urin- Álvrez E., Mrtínez-Bustos F., Moreno-Mrtínez E., 9. Effet of itri id on fltoxin degrdtion nd on funtionl nd texturl properties of extruded sorghum. Anim Feed Si Teh. 1, Krmi-Osoo R., Mirolfthy M., Kmrn R., Shet-Boushehri M., Srkri S., 1. Afltoxin B1 in 6

7 J. Milni et l / Journl of Chemil Helth Risks 4(4) (14) 1 7 mize hrvested over yers in Irn. Food Control., Cherghli A.M., Mohmmdi H.R., Amirhmdi M., Yzdnpnh H., Aouhossin G., Zmnin F.,. Inidene of ptulin ontmintion in pple juie produed in Irn. Food Control. 16, Heshmti A., Milni J.M., 1. Contmintion of UHT milk y fltoxin M1 in Irn. Food Control. 1(1), Krmi-Osoo R., Mirolfthy M., 8. Nturl zerlenone ontmintion of whet from Golestn provine, northern Irn. Irn J Plnt Pthol. 44: Krmi-Osoo R., Mirolfthy M., Alikri F., 1. Nturl Deoxynivlenol Contmintion of Corn Produed in Golestn nd Moqn Ares in Irn. J Agri Si Tehol. 1, Yzdnpnh H., Mirgli M., Clfpietr F.R., Brer C., 1. Nturl ourrene of fltoxins nd ohrtoxin A in orn nd rley from Mzndrn nd Golestn in north provines of Irn. Myotoxin Res. 17, Reiss J., Myotoxins in foodstffs. XI. Fte of fltoxin B1 during preprtion nd king of wholemel whet red. Cerel hem. (4), As H.K., Crtwright R.D., Xie W., Thoms Shier W., 6. Afltoxin nd fumonisin ontmintion of orn (mize, Ze mys) hyrids in Arknss. Crop Prot. (1), Ventur M., Gomez A., Any I., Diz J., Broto F., Agut M., Comells L., 4. Determintion of fltoxins B1, G1, B nd G in mediil hers y liquid hromtogrphy- tndem mss spetrometry. J Chromtogr A. 148, Betin V., Myotoxins: hemil, iologil nd environmentl spets. In: Biotive moleules. Betin V., Eds., Elsevier Applied Siene: pp: Rustom I.Y.S., Afltoxin in food nd feed: Ourrene, legisltion nd intivtion y physil methodes. Food Chem. 9, Sott P.M., Knhere S.R., Lwrne G.A., Dley E.F., Frer J.M Fermenttion of wort ontining dded ohrtoxin A nd fumonisins B1 nd B. Food Addit Contm. 1, Fndohn P., Zoumenou D., Hounhouign D.J., Mrss W.F.O.,Wingfieldd M.J., Hell K.,. Fte of fltoxins nd fumonisins during the proessing of mize into food produts in Benin. Int J Food Miroiol. 98, Shpir R., Pster N., Control of myotoxins in storge nd tehniques for their deontmintion. In: Myotoxins in food. Mgn, N., & Olsen, M. (Eds.). Woodhed Pulishing Ltd nd CRC Press LLC: Cmridge, Englnd: 4. pp Mnn G.E., Codifer L.P. Jr., Doller F.G., Effets of het on fltoxin in oilseed mels. J Agri Food Chem. 1(6), Um Reddy M., Gull S., Nglkshmi A.V.D., 1. Effet of fermenttion on fltoxin redution in seleted food produts. Food Si Tehnol Nutr. 4(1), Smrjew U., Sen A.C., Cohen M.D., Wei C.I., 199. Detoxifition of fltoxins in foods nd feeds y physil nd hemil methods. J Food Protet.,

8 J. Milni et l / Journl of Chemil Helth Risks 4(4) (14) 1 7