Improvement of Nutritional and Quality of Lasagna Supplemented with Scenedesmus obliquus

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1 Int. J. Phrm. Si. Rev. Res., 46(2), Septemer - Otoer 217; Artile No. 8, Pges: Reserh Artile Improvement of Nutritionl n Qulity of Lsgn Supplemente with Seneesmus oliquus Frouk K. El-Bz*1, Azz A. Omrn2, Ahme M.S. Hussein3, Emn A. Irhim1 Deprtment of Plnt Biohemistry, Ntionl Reserh Centre (NRC), 33 El Bohouth st. (former El Thrir st.), Dokki, Giz, Egypt. 2 Deprtment of Crops Tehnology Reserh, Foo Tehnology Reserh Institute, Agriulturl Reserh Centre (ARC), Giz, Egypt. 3 Deprtment offoo Tehnology, Ntionl Reserh Centre (NRC), 33 El Bohouth st. (former El Thrir st.), Dokki, Giz, Egypt. *Corresponing uthor s E-mil: f_elz@hotmil.om 1 Reeive: ; Revise: ; Aepte: ABSTRACT The present work ime to investigte the effet of mixing mirolge Seneesmus oliquus with whet flour (72% extrtion rte) on enhning the nutritionl hrteristis, phytohemil ompouns n ntioxint tivity of lsgn pst. Chemil nlysis showe tht, S. oliquus power h high ontents of protein, rue fier, fts n sh (52.43, 4.53, 2.2 n 6.13%, respetively). Also, it ontine high mounts of phytohemil ompouns, onsequently hrterize with high ntioxint tivity. Furthermore, it ws sfe s n ele soure, where its lethl toxiity ws 1 g/kg mie oy weight orresponing to 78 g/7 kg humn oy weight. Results lso showe the inrese of protein ontent with perentges 11.2 n 14.5% in lsgn pst of ontrol n 7.5% lge, respetively. Phytohemils n ntioxint tivity inrese with inresing lge perentge ut ooking erese ll phytohemils ompre to the unooke lsgn pst. Cooking losse ws not signifintly ffete with inresing lge in lsgn pst while weight gin erese ompre to ontrol smple. Texture nlyses showe tht, firmness of lsgn pst inrese with inresing the ition perentge of lge s ompre to ontrol smple. Cohesiveness, hewiness n springiness vlues erese in ll lsgn pst smples ompre to whet lsgn pst. Hunter olor prmeter inite tht, lightness of ooke lsgn pst ws higher thn rie lsgn pst. Also, reness (* vlues) of lge lsgn pst h negtive * n positive yellowness (*) ue to the hlorophyll ontent. Lsgn pst of 2.5% lge h higher olor, tste, texture n overll eptility sores ompre to other higher lge ontent. The results reommene using whet flour with 2.5% S. oliquus to improve the nutritionl qulity of lsgn pst. Keywors: Seneesmus oliquus, Phytohemils, Lsgn pst, Qulity hrteristis, Texture evlution. INTRODUCTION F resh wter mirolge, exeeingly istriute in rivers, lkes n polr wters exhiit vrie rnge of morphologil, ellulr, struturl n iohemil omposition.1 Among these mirolgl speies is Seneesmus, tht elongs to Spheropleles orer of Seneesmee fmily whih is frequently preominnt in freshwter lkes n rivers.2 Vrious speies of this genus re use worlwie for ifferent purposes ue to their potentil to pt to hrsh environmentl onitions, grow rpily n ese of ultivtion n hnling.3,4 Seneesmus sp. hs een use in lot of iotehnologil purposes tht epen on its high nutritionl ontent n iotivities. 5, 6 Enormous power for greter iversity of mirolgl speies is use in humn nutrition. Aunnt speies of mirolge ontin proteins of high qulity for humns.7most of ommon mirolgl pigments suh s; hlorophyll, lutein, β-rotene n phyoiliproteins re vlule to humn helth hve ntiner, ntioxint 8-1 n nti-inflmmtoryeffets. Also in mny lgl speies, nutritious polyunsturte ftty is possess essentil ftty is essentil for riovsulr, oulr 11 n neurologil helth. Regrless of this potentil, few mirolgl speies re urrently use in mrket for the humn nutrition. Despite, Seneesmus genus is wiely known s soure of foo for herivorous zooplnkton n in io fuel proution ue to its high oil ontent esies, it hs exhiite the potentil of eing soure of high-vlue omponents with ntiteril hrters. These ntiteril effets hve wie rnge of pplitions in ifferent inustries tht hve not een roly investigte n fully exploite. The nutritious iohemil omponents of Seneesmus sp., initil foo eptility, preprtion, niml n humn experiments inite tht, it my hve goo future in helth foo pplitions. Seneesmus sp. hs previously een suggeste for nutreutil usge euse its onentrtions of essentil mino is, goo ontent of protein, lipi, ftty is, eiospentenoi i, essentil minerls n vitmins. Mirolge hve the ility to enhne the nutritionl ontent of foo onsequently, to fvorly ffet humns helth euse of their originl hemil omposition.14seneesmusontins very high nutritionl qulity n severl toxiologil vlues tht hve not revele ny toxi impts or normlities in experimentl nimls.15 Gross et l.16 rrie out nutritionl stuy y inorporting Seneesmussp. into the iet of hilren (5 g/ily) n ults (1 g/ily). Pst n its prouts re onsiere the min sugroup 17 of mny iets. It is prepre y using ough me from Interntionl Journl of Phrmeutil Sienes Review n Reserh Aville online t Copyright protete. Unuthorise repulition, reproution, istriution, issemintion n opying of this oument in whole or in prt is stritly prohiite. 28

2 Int. J. Phrm. Si. Rev. Res., 46(2), Septemer - Otoer 217; Artile No. 8, Pges: ny suitle mteril inluing semolin, flour of urum, frin flour, orn, rie, whet or ny mixing of these with wter. In ition, pst n e enrihe, supplemente, fortifie or remine onventionl. Reently, severl ingreients n itives hve een vne n re eing utilize to improve the qulity of pst. 18 It is motivting tht, urum whet (Tritium urum), the fun mentl ingreient in Itlin style pst, shres only to 5% of the worl s whet proution n it in generl tres for higher prie thn the ommon whet (Tritium estivum). To meet the growing pst onsumption, it is neessry to proue pst from unonventionl ommoities ut equte proessing n lne formultions re require to ountert their poor tehnologil properties. 18 Therefore, the ojetive of this stuy is to investigte the possiility of using whet flour with ifferent perents of S. oliquus power to improve the nutritionl qulities of lsgn pst. MATERIALS AND METHODS Smples, ingreients n hemils Bre whet flour (72% extrtion rte, from five strs Compny, Swiss ity, Egypt) n slt were purhse from lol mrkets in Giz, Egypt. All hemils were of nlytil gre. Senile mle n femle mie (23-25 g) were use in the ute toxiity test. Eh group of mie (8 mie eh) ws kept in stinless steel ge, wter n foo were given litium. Animl experiment ws rrie out oring to the Meil Reserh Ethis Committee for institutionl n ntionl guie for the re n use of lortory nimls, Ntionl Reserh Centre (NRC); Ciro, Egypt. Isoltion, purifition n ientifition of S. oliquus The green lg S. oliquus ws otine from Algl Tehnology group, NRC, Dokki, Giz, Egypt. S. oliquus (Figure 1) ws isolte from the River Nile wter y using BG11 mei for lgl isoltion n purifition. 19 S. oliquus ientifition hs een one oring to the keys of ientifition. 19 The strin ws isolte y spreing.1ml of wter smples into petriishes ontining BG11 with 1.5% gr for soliifition. Single olonies of lge were then re-ultivte s nonxeni th ultures (5ml) t 25±2 C for 24h with ontinuous white fluoresent lmp intensity 25Lux. Figure 1: S. Oliquus mirolg BG11 nutrient omposition Nutrient omposition of BG11 ws use 2 for growing S. oliquus s shown in Tle (1). Tle 1: BG11 nutrient omposition Nutrients Mronutrients Conentrtion Interntionl Journl of Phrmeutil Sienes Review n Reserh Aville online t Copyright protete. Unuthorise repulition, reproution, istriution, issemintion n opying of this oument in whole or in prt is stritly prohiite. mg/l NNO K 2 HPO 4 4. MgSO 4.7H 2 O 75. CCl 2.2H 2 O 36. Citri i 6. N 2 CO 3 2. N 2 EDTA 1. Ferri mmonium itrte 6. Mironutrients g/l H 3 BO MnCl 2.4H 2 O 1.81 ZnSO 4.7H 2 O.222 N 2 MoO 4.2H 2 O.39 CuSO 4.5H 2 O.79 Co(NO 3 ) 2.6H 2 O ml/l of mironutrients solution ws e to the ulture Cultivtion of S. oliquus The orgnism ws grown in onil flsk 5 L ontining 2 L BG11 mei for two weeks. After the growth phse, the lgl iomss ws hrveste n inoulte in n open pon with pity of 25 l n ulture with BG11 mei for nother 3 weeks. 2 The lgl iomss ws hrveste using ooling entrifuge t 4 rpm, n then rie t 6 C. The oil ws extrte 21, n then the lge power ws rie in oven t 7 C until omplete ryness. Anlysis of ingreients n lsgn pst Proximte nlysis Protein, ft, rue fier n sh ontents of S. oliquus, whet flour n rie lsgn pst smples were etermine s esrie y AOAC. 22 Totl rohyrte ws lulte y ifferene. Phytohemils n ntioxint tivity Hlf grm of lge n lsgn pst smples were extrte with 1 ml 8% ethnol n shken for 2 h, filtrte n the volume ws me up to known volume. Totl phenoli ompouns Totl phenoli ontent ws etermine oring to Singleton n Rossi 23 using Folin-Ciolteu metho..25 ml of previous extrt ws mixe with.25 ml Folin- Ciolteu regent n.5 ml of 1% soium ronte (N 2 CO 3 ), then the volume ws omplete to 5 ml with 29

3 Int. J. Phrm. Si. Rev. Res., 46(2), Septemer - Otoer 217; Artile No. 8, Pges: istille wter. After inution in rk t room temperture for 3 min, the sorne of the mixture ws mesure t 725 nm ginst the lnk. Glli i ws use s stnr. Chlorophyll ontent Chlorophyll, hlorophyll n rotenoi ontents were mesure using spetrophotometer oring to 24 Yng et l. Alge n lsgn pst (.25 g) were homogenize with 5 ml of 85 % queous etone for 5 min, then entrifuge n the superntnt ws me up to known volume with 85 % etone. The sorne ws mesure t 452.5, 644 n 663 nm. Antioxint tivity Amino is etermintion Amino is omposition of S. oliquus power n rie lsgn pst (ontrol n 2.5% lge) ws etermine using mino i nlyzer (Biohrom 3) oring to the metho outline in AOAC.22 An liquot smple were weighe n igeste with 25 ml of HCl (6 N) t 11 C for 24 h. Then, HCl ws remove y evportion; the remining soli frtion ws issolve y.2 N soium itrte uffers (ph 2.2). One ml of the solution ws filtere through.45 µm. The stnr mino is (onsist of 17 mino is) were trete s the sme s of the smples. Amino is were expresse s g/1 g protein on ry weight sis. Ftty is etermintion ) DPPH free ril svenging tivity The ntioxint tivity of smples ws etermine 25 oring to Brn-Willims et l. using previous extrt se on the ril svenging ility in reting with stle DPPH free ril. Briefly, 2.4mg of DPPH in 1 ml ethnol ws prepre n 3.9 ml of this solution ws e to.1 ml of extrt. The mixture ws shken vigorously using shker for few seon n llowe to stn t room temperture for 3 min in the rk. Then the sorne ws mesure t 515nm. The DPPH ril svenging perentge ws lulte using the following eqution: DPPH ril Svenging tivity (%) = A - A1 x 1 A Where, A= Asorne of ontrol (ontining ll regents exept the test ompouns). A1= Asorne in the presene of the teste extrts fter 3 min. ) Totl ntioxint pity One ml of S. oliquus n lsgn pst with lge ethnoli extrts ws mixe with 3 ml of regent solution (.6 M sulfuri i, 28mM soium phosphte n 4 mm mmonium molyte). The tues were ppe n inute t 95oC for 9 min. After ooling, the sorne of eh smple ws mesure t 695 nm. Stnr series onentrtions of sori i (µm/ml) were trete s the smple.26 Aute orl lethl toxiity test for S. oliquus Aute orl lethl toxiity test of S. oliquus power ws 27 rrie out oring to Goomn et l. The 24 h mortlity ounts (if ny) mong equl size groups of mie reeiving progressively inresing orl ose levels were reore. Oil ws extrte from S. oliquus with n-hexne. The methyl esters of ftty is were prepre oring to the metho of AOAC.22 Ftty is were nlyze y gs hromtogrphy (Agilent HP 689) n reporte in reltive re perentges. The ftty i methyl esters were ientifie y ompring their retention times with known ftty i stnr mixture. Pek res were utomtilly ompute y n integrtor. The ftty is omposition ws expresse s perentge of totl ftty is. Lsgn pst proessing Control smple of lsgn pst ws prepre using 1% whet flour (72% extrtion rte). Lsgn pst of lge power ws prepre t three onentrtion levels using 2.5, 5. n 7.5% lge power with whet flour (72% extrtion). 1 g whet flour n 1.25 g slt were mixe with efinite mount of wter (46-48 ml). The ough ws shpe y using mnul pst mker of Titni rn (single unite stel pst mhine, Itly) n then ut into lsgn sheets with 1.5 mm thikness, 7 mm with n 13 mm length verges. Lsgn pst smples were rie t 45±5 C overnight, then pke in polyethylene gs n kept for further nlysis t 4 C, then the most epte lsgn pst of lge power in pnel test (2.5%) n ontrol smple were exmine for mino n ftty i ontents. Cooking qulity of lsgn pst Cooking tests of lsgn pst (weight gin n ooking loss) were performe in triplites s following: 1 g of smples were ooke for 8-9 min in 2 ml of oiling wter; the pst ws then oole in ol wter for 1 s, rine n weighe immeitely. Cooking loss % (the mount of soli sustne lost in the ooking wter) n weight gin of ifferent pst smples were etermine s esrie y Li28 then lulte oring to the following equtions: Interntionl Journl of Phrmeutil Sienes Review n Reserh Aville online t Copyright protete. Unuthorise repulition, reproution, istriution, issemintion n opying of this oument in whole or in prt is stritly prohiite. 3

4 Int. J. Phrm. Si. Rev. Res., 46(2), Septemer - Otoer 217; Artile No. 8, Pges: Cooke lsgn pst smples were use for texturl, sensoril n further nlysis. Tehnology Reserh Institute oring to the metho 3 of Bshir et l. Five prmeters were exmine, i.e., olor, rom, tste, texture n overll eptility using 9-point heoni sle. Physil properties of lsgn pst Color mesurement Sttistil nlysis The olor of rie n ooke lsgn pst smples ws mesure y hn-hel Tristimulus refletne olorimeter Minolt Chrommeter (moel CR-4, Koni Minolt, Jpn). Results reore in the L* * * * h olor system. The otine vlues re presente s mens of triplite etermintions. The totl olor ifferene etween rie n ooke lsgn pst were etermine oring to the following eqution: 2 2 For the nlytil t, men vlues n stnr evition re reporte. The otine t were sujete to one-wy nlysis of vrine (ANOVA) t P<.5 followe y Dunn's new multiple rnge tests to ssess ifferenes etween smples men using Costt sttistil softwre. RESULTS AND DISCUSSION 2 1/2 ΔE= [(ΔL) + (Δ) + (Δ) ] Proximte nlysis of whet flour n S. oliquus power Texturl profile nlysis (TPA) Tle (2) showe the hemil omposition of S. oliquus n whet flour. The results inite tht, S. oliquus power h high ontents of protein (52.43%), fts (2.2%), rue fier (4.53%) n sh (6.13%). While, whet flour (72% extrtion rte) h high ontent of totl rohyrte (86.74%).The urrent results re in greement with the results of Btist et l.31 n Beker32 who foun tht, S. oliquus h 5 to 56%protein n 33 to 64% totl rohyrte, respetively. For whet flour, the present results re lose to the results reporte y Seleem n Omrn.3 The texture of ooke lsgn pst ws etermine y universl testing mhine (Coneteh, B type, Tiwn) provie with softwre s esrie y Bourne.29An luminum 25 mm imeter ylinril proe ws use in TPA oule ompression test to penetrte perpeniulr to 5% epth reltive to the smple height, t 1 mm/s spee test (se on pre-test). Firmness (N), gumminess (N) hewiness (N), ohesiveness n springiness were lulte from TPA grphi. Sensory evlution of lsgn pst Sensory ttriutes of ooke lsgn pst were pprove out y sensory pnelists from Foo Tle 2: Proximte nlysis of S. oliquus power n whet flour (% on ry weight sis) Ingreients S. oliquus power Whet flour (72% extrtion rte) Protein ± ±.41 Fts 2.2 ± ±.9 Crue fier 4.53 ±.5 Ash 6.13 ±.57 Totl rohyrte* ± ±.3.63 ± ±.55 Eh vlue is expresse s men of three replites ±SD, numer in the sme olumn followe y the sme letter re not signifintly ifferent t.5 level. * Totl rohyrte ws lulte y ifferenes. Phytohemil ontents, ntioxint tivity n lethl toxiity of S. oliquus Tle (3) illustrtes the pigment ontents of S. oliquus. The results inite tht, S. oliquus power hrterize with its high phenoli ompouns ( mg/1g). Also, S. oliquus power h 699.9, n mg/1g for hlorophyll, hlorophyll n rotenois, respetively. While, DPPH free ril svenging tivity ws 45.27% n totl ntioxint pity ws µg/ml. These results re in the sme tren with the results of Kent et l.34 who foun tht, Seneesmus sp. h 19 mg/1g hlorophyll, 423. mg/1g rotenois. Moreover, the ute lethl toxiity test revele tht, S. oliquus ws very sfe up to the highest teste ose (1g/kg mie oy weight). This ose orrespons to 78 g/7 kg mn oy weight for humn while the ose of mie ws extrpolte to orresponing estimtes in humn opting interspeies 35 osge onversion sheme. This reflets the highest sfety of S. oliquus. Interntionl Journl of Phrmeutil Sienes Review n Reserh Aville online t Copyright protete. Unuthorise repulition, reproution, istriution, issemintion n opying of this oument in whole or in prt is stritly prohiite. 31

5 Int. J. Phrm. Si. Rev. Res., 46(2), Septemer - Otoer 217; Artile No. 8, Pges: Amino is ontent n mino i sores of lge n lsgn pst Amino i ontents ws evlute in S. oliquus power, lsgn pst of 2.5% lge n ompre with lsgn of ontrol smple s shown in Tle (4). Tle 3: Phytohemil ontents, ntioxint tivity n lethl toxiity of S. oliquus power Constuites Content (mg/1g) Totl phenoli ±16.14 Chlorophyll ± 5.5 Chlorophyll ± 6.1 Crotenois ± 5.5 DPPH free ril svenging tivity (%) ± 1.8 Totl ntioxint pity (µg/ml)* ± 1.1 Lethl toxiity 1 g/kg mie oy weight *Totl ntioxint pity (µg/ml) s sori i. Eh vlue is expresse s men of three replites ±SD. The otine results inite tht, lge power ontins higher mounts of totl essentil mino is espeilly, leuine whih ws the most unnt essentil mino i in S. oliquus power followe y lysine n threonine. Also, ontrol smple of lsgn pst (1% whet) ontine high mounts of totl nonessentil mino is espeilly glutmi i tht ws the most unnt nonessentil mino i in whet. Lsgn pst of 2.5% S. oliquus power ontine higher mount of totl essentil mino is while totl non essentil mino is erese s ompre to ontrol smple of lsgn pst. The results re lose to those finings y Kent et l. 34 for mirolge n ommeril mirolge prouts. Also, Spolore et 11 l. stte tht, protein ontent (oth in mino is ontent n essentil mino i supply for oy) of most lge is higher thn tht of other protein foos. Tle 4: Amino is ontent of S. oliquus power, lsgn pst of ontrol n S. oliquus Amino is g/ 1g protein S. oliquus power Lsgn pst (ontrol) Lsgn pst with 2.5% S. oliquus Essentil mino is 2.68± ± ± ± ±.15 Lysine 5.38±.1 2.6± ±.7 Methionine 2.17±.4 2.5±.3 2.5±.6 Phenyllnine 5.19± ± ±.14 Isoleuine 3.53±.7 Leuine Threonine 5.36± ± ±.1 Vline 5.36± ± ±.13 Histiine 1.72± ± ±.6 Nonessentil mino is Tyrosine 3.82± ± ±.9 Arginine 6.49± ± ±.11 Alnine 9.58±.18 Asprti i 3.3± ± ± ± ± ±.3 Glutmi i 1.53± ± ±.83 Glyine 5.38± ± ±.1 Proline 4.24± ± ±.26 Serine 4.39± ± ± ±.6 Cysteine 2.62±.7 Eh vlue is expresse s men of three replites ±SD, numer in the sme row followe y the sme letter re not signifintly ifferent t.5 level. Amino i sores of previous smples were lulte n presente in Tle (5). The otine results showe tht, S. oliquus h higher perent of mino i sores ompre to ontrol lsgn pst n lsgn pst of 2.5% S. oliquus power. Result gree with Kent et l.34 who reporte tht, mino i sores were higher in mirolge priniplly ue to higher proportions of the essentil mino is. Ftty is omposition Tle (6) presents perentges of ftty is of lge, ontrol n 2.5% lsgn pst with lge. The results inite tht, lge h high ontent of ftty is espeilly C8:, C1:, C12:, C13:, C14:, C16:1, C17:, C17:1, C18:, C18:2, C18: 3n3, C18:3n6,C2: nc2:1. From the t in Tle (6) it oul e stte tht, ontrol pst is rih in C16:, C18:, C18:1 n C18:2. The Interntionl Journl of Phrmeutil Sienes Review n Reserh Aville online t Copyright protete. Unuthorise repulition, reproution, istriution, issemintion n opying of this oument in whole or in prt is stritly prohiite. 32

6 Int. J. Phrm. Si. Rev. Res., 46(2), Septemer - Otoer 217; Artile No. 8, Pges: ominnt ftty is were C16:, C18:1 for lge; C16:, results re lose to those finings y Custoio et l. 36 n 34 C18:1, C18:2 for ontrol n 2.5% lsgn pst. The Kent et l. Tle 5: Amino i sores of S. oliquus, lsgn pst of ontrol n lsgn pst mixe with S. oliquus Amino is FAO/WHO Referene vlue* Isoleuine Amino i sors (%) S. oliquus power Lsgn pst(ontrol) Lsgn pst of 2.5% S. oliquus Leuine Lysine Methionine+Cysteine Phenyllnine +Tyrosine Threonine Vline Histiine Totl mino is *FAO/WHO Protein qulity evlution; Foo n Agriulture Orgniztion of the Unite Ntions. Dily requirements for humn ults.1991, Rome, Itly.P. 66. Tle 6: Ftty is ontent of S. oliquus power, lsgn pst of ontrol n lge Ftty Ai(% of totl ftty is) S. oliquus power lsgn pst of 1% whet 2.5% S. oliquus Sturte ftty is C8: C1: 1.21 C12: C13: C14: C15: C16: C17: C18: C2: C22: Sutotl Mono unsturte ftty is C14:1.16 C15:1.19 C16: C17: C18: C2: C22: Sutotl Poly unsturte ftty is C18: C18: 3n C18:3n Sutotl Interntionl Journl of Phrmeutil Sienes Review n Reserh Aville online t Copyright protete. Unuthorise repulition, reproution, istriution, issemintion n opying of this oument in whole or in prt is stritly prohiite. 33

7 Int. J. Phrm. Si. Rev. Res., 46(2), Septemer - Otoer 217; Artile No. 8, Pges: Fr et l.37elre tht, the ition of mirolg to ookies signifintly inrese the protein ontent of fortifie smples s ompre with ontrol. Spirulin my e use in kery prouts for protein enrihment without ny signifint hnges in the texture. 38 Also Fr 37 et l. stte tht, the fortifie ookies with Spirulin t the onentrtions of,.5, 1 n 1.5% (w/w) to the ookies formultion le to signifintly higher protein ontent. Also Ao El-Ng et l.39 stte tht, supplementtion of pst with high protein soure like surimi n mushroom resulte in etter qulity n nutritious. Chemil omposition of lsgn pst Chemil omposition of lsgn pst(ontrol) n lsgn pst tht mixe with 2.5, 5 n 7.5% S. oliquus power re presente in Tle (7). The results inite tht, inresing lge perentge in lsgn pst signifintly inrese protein, sh n rue fier ontents ompre to ontrol lsgn pst. This result oul e ue to, the high ontent of protein n sh in S. oliquus. Regring to ft ontents, Tle (7) showe tht lsgn pst not ffete with ifferent ition levels of lge. While, totl rohyrte ws signifintly erese with inresing level of lge. Tle 7: Proximte hemil ontent of lsgn pst mixe with S. oliquus (%ry weight sis) Lsgn pst Protein Fts Ash.86±.4.88±.2 Crue fier 1.5±.3.4± ±.5.5±.2 Totl rohyrte 86.75± ±.34 Control 11.2± % S. oliquus 12.22±.32 5.% S. oliquus 13.34±.11.9± ±.4.56± ± % S. oliquus 14.5±.16.93± ±.6.63± ±.23 *Totl rohyrte ws lulte y ifferenes. Eh vlue is expresse s men of three replites ±SD, numer in the sme olumn followe y the sme letter re not signifintly ifferent t.5 level. Phytohemil ontents n ntioxint tivity of lsgn pst efore n fter ooking the ssoite hlorophylls in ition to, simultneous inrese in pheophytine n pheophorie ontent, minly ue of the therml tretment. Turkmen et l.41 foun tht, ooking resulte in loss of hlorophyll n to vrious extent epening on the type of metho n the vegetle type. Besies pigment iffusion into the ooking wter, pst olor losses my lso e ttriute to the mirolge pigment oxition uring therml tretment.31 Tle (8) showe phytohemil ontents of lsgn pst efore n fter ooking. The results inite tht, phytohemils n ntioxint tivity inrese with inresing the mixing level of lge. On the other hn, ooking of lsgn pst erese phytohemils ompre to rie lsgn pst n this oul e ue to 4 the leh in ooking wter. Sumn et l. mentione tht, rie lsgn smples showe rsti erese in Tle 8: Phytohemils ontent n ntioxint tivity of lsgn pst mixe with S. oliquus efore n fter ooking Lsgn pst mixe with lge t level Lsgn pst 2.5 % 5. % 7.5 % Drie lsgn pst Totl phenoli (mg/1g) 89.41± ± ± ±1.73 Chlorophyll (mg/1g).±. 46.9± ± ±.55 Chlorophyll (mg/1g).± ± ± ± ± ± ± ± ± ± ± ±4.32 Crotenois (mg/1g).72±.2 DPPH ril svenging tivity (%) 9.2±.52 Totl ntioxint pity (μg/ml) * 85.6± ±6.4 Cooke lsgn pst Totl phenoli (mg/1g) 21.71± ± ± ±1.37 Chlorophyll (mg/1g).±. 22.± ± ±.6 Chlorophyll (mg/1g).±. 71.4± ± ±6. Crotenois (mg/1g).7± ±.6 38.± ±.59 DPPH ril svenging tivity (%) 5.4±.22 Totl ntioxint pity (μg/ml)* 76.61± ± ± ± ± ± ±16.7 Eh vlue is expresse s men of three replites ±SD, numer in the sme row followe y the sme letter re not signifintly ifferent t.5 level.*s sori i Interntionl Journl of Phrmeutil Sienes Review n Reserh Aville online t Copyright protete. Unuthorise repulition, reproution, istriution, issemintion n opying of this oument in whole or in prt is stritly prohiite. 34

8 Int. J. Phrm. Si. Rev. Res., 46(2), Septemer - Otoer 217; Artile No. 8, Pges: Cooking qulity Tle (9) shows the effet of ifferent mixing level of lge on ooking qulity prmeters of lsgn pst (ooking losses n weight gin). Results inite tht, ooking losses of lsgn erese slightly in mixe lsgn pst with lge t ifferent mixing levels ompre to ontrol smple. Also, there ws no signifie ifferene etween lsgn pst of ifferent mixing levels with lge. The lge inorportion my weken the struture of pst ue to their proteins re not le to evelop gluten network n steri hinrne interferes in network formtion. Due to the protein network limits the wter iffusion into the entrl zone of pst uring ooking42, weker gluten-protein network filittes wter iffusion into it, reuing ooking time. In ition, sine the gluten-protein network is responsile for retining pst physil integrity uring ooking, weker struture lehes more solis from pst smples into ooking wter, inresing ooking resiues.43, 44 On the other hn, ooking loss vlues in ll smples were less thn 8% whih is the limit vlue onsiere esirle for whet pst s mentione y Dik & Youngs45who stte tht, ooking losses re serious initor for the pst ooking performne y oth onsumers n inustry. Higher weight gin vlues of the smples enrihe with mirolge inite tht, they sore higher mounts of wter uring ooking. The ition of S. oliquus (2.5, 5. n 7.5%) in lsgn pst inue signifint erese of weight gin (66.33, 69.1 n 76.53%) ompre with ontrol lsgn pst (17.12%). Pst prepre with S. oliquus presents weight gin lower thn the ontrol lsgn pst. This result my e ttriute to the ility of S. oliquus to sor wter n retin it in the protein-strh net. Aoringly, weight gin ws lso ffete y the utiliztion of S. oliquus iomss, whih erese from 17.12% in the ontrol pst to 76.53% in 7.5% lge power. Furthermore, these results re in orne with those otine y Zretto & Ros46 who reporte tht, the pst prepre with 2% of mirolge possesse higher wter sorption thn other smples reporte. Although, the lk of ontinuous protein network uses high hyrtion of the strh mteril n inreses the weight of pst. When gluten ws ilute y the sustitution of whet flour to spirulin iomss, the protein network eme weker, filitting the lek of mylose into ooking wter.47 Tle 9: Cooking qulity of whet n lsgn pst with S. oliquus (%) Lsgn pst Cooking loss Weight gin Control 5.62± ± % S. oliquus 5.% S. oliquus 5.17± ± ± ± % S. oliquus 5.48± ±2.5 Eh vlue is expresse s men of three replites ±SD, numer in the sme olumn followe y the sme letter re not signifintly ifferent t.5 level. Physil properties eome less green ( eomes less negtive) n more 4 yellow ( eomes more positive). Texture Profile Anlysis (TPA) Color Tle (1) isplys olor prmeters (L,,,, h n ΔE) of rie n ooke lsgn pst prepre from lge power. It oul e notie from these results tht, sustitution of whet flour with lge in the formultion of lsgn pst ffete the lsgn pst olor. Dt in Tle (1) lso inite tht, sustituting of whet flour with S. oliquus signifintly erese the lightness (L) vlues of rie n ooke lsgn pst with inresing level of sustitution where, ontrol lsgn pst reore the highest vlue. However, L vlues inrese in ooke lsgn pst ompre with rie lsgn pst n eme lighten (L vlue inreses). The 14 vlues of lsgn pst re gree with Zouriet l. results who foun tht, firmness of pst ontining A. pltensis h negtive n positive ue to the hlorophyll ontent of these lge. During the ooking of lsgn pst, there is teneny for the olor to Regring, (olor sturtion) n h vlues of lsgn pst were foun to e inrese with inresing level of superstition. Besies, S. oliquus lsgn pst h the highest hue ngle vlues s ompre to ontrol pst. These results re in greement with Zouri et l.14 who foun tht, fter ooking, ll smples of pst enrihe with A. pltens is mirolge were slightly righter n more yellow. Color losses resulting fter ooking of pst pries re expresse in terms of totl olor ifferene (ΔE) etween rie n ooke lsgn pst s speifie in Tle (1). The sustitution with lge with 2.5, 5. n 7.5% h higher olor loss n this my e ue to the therml proess whih hs ffete the olor. Also, pst olor losses rise from pigment lehing into the ooking wter n my lso e relte to S. Oliquus pigment oxition uring therml tretment. Mirolge re reognize s n exellent soure of pigments inluing; hlorophylls, rotenois n phyoiliproteins. This ft expline the reution in olor prmeter vlues.14 Interntionl Journl of Phrmeutil Sienes Review n Reserh Aville online t Copyright protete. Unuthorise repulition, reproution, istriution, issemintion n opying of this oument in whole or in prt is stritly prohiite. 35

9 Int. J. Phrm. Si. Rev. Res., 46(2), Septemer - Otoer 217; Artile No. 8, Pges: Tle 1: Color of rie n ooke lsgn pst mixe with S. oliquus Lsgn pst smples Lsgn pst mixe with S. oliquus t levels Lsgn pst 2.5% Prmeters 5.% 7.5% L Drie 73.97± ± ± ±.33 Cooke 77.41± ± ± ±1.69 Drie.1± ± ± ±.4 Cooke -2.23± ± ± ±.8 Drie 14.73± ± ± ±.17 Cooke 19.46± ± ± ±.11 Drie 14.8± ± ± ±.16 Cooke 19.58± ± ± ±.1 Drie 89.62± ± ± ±.65 Cooke 96.54± ± ± ±.26 ΔE ± ± ±.43 Color Yellow Greenish yellow Greenish yellow Greenish yellow h *L (lightness with L = 1 for lightness, n L = zero for rkness), [(hromtiity on green ( ) to re (+)], [(hromtiity on lue ( ) to yellow (+)], (olor sturtion), h [(hue ngle where = re to purple, 9 = yellow, 18 = luish to green n 27 = lue sle. ΔE is the totl olor ifferene etween rie n ooke lsgn pst. Eh vlue is expresse s men of three replites ±SD, numer in the sme row followe y the sme letter re not signifintly ifferent t.5 level. Texture profile nlysis of ooke lsgn pst The urrent results re in greement with Zouriet l.14who foun tht, firmness of pst ontining Arthrospir pltensis ppere to e higher thn tht of the whet ontrol pst. Besies, there ws erese in ohesiveness, hewiness n springiness vlues in ll lsgn pst smples ompre to whet lsgn pst. Moreover De Mro et l.47 foun tht, repling the whet flour with spirulin le to n inrese of firmness, ohesiveness n hewiness. Tle (11) emonstrtes texturl profile (firmness, ohesiveness, gumminess, hewiness n springiness) of ooke lsgn pst prepre from S. oliquus power. Depening on the present results, sustitution of whet flour with S. oliquus power in formultion of lsgn pst ffete the lsgn pst texturl properties. Vlues of lsgn pst firmness n gumminess inrese with inresing the sustitutions perent of S. oliquus power s ompre to whet lsgn pst. Tle 11: Texture profile nlysis of ooke lsgn pst mixe with S. oliquus Lsgn pst smples Firmness(N) Cohesiveness Gumminess (N).61± ±.1.98±.8.85±.3 2.5% S. oliquus 5.% S. oliquus 2.23± ±.3.57±.3.56± ± ±.3.96±.7.94±.8.75±.2.71±.3 7.5% S. oliquus 2.5± ±.3 Springiness 1.88±.2.54±.4 Chewiness(N) Control.82±.3.61±.8 N= Newton. Eh vlue is expresse s men of three replites ±SD, numer in the sme olumn followe y the sme letter re not signifintly ifferent t.5 level. Interntionl Journl of Phrmeutil Sienes Review n Reserh Aville online t Copyright protete. Unuthorise repulition, reproution, istriution, issemintion n opying of this oument in whole or in prt is stritly prohiite. 36

10 Int. J. Phrm. Si. Rev. Res., 46(2), Septemer - Otoer 217; Artile No. 8, Pges: Sensory hrteristis The vlues in Tle (12) inite the perentge of eptility in eh ttriute; olor, rom, texture, tste n overll eptility. Lsgn pst ws prepre y mixing whet flour (72% extrtion rte) with 2.5, 5. n 7.5% lge S. oliquus power. Results in Tle (12) showe tht, sustituting of whet flour with 2.5% mirolge h higher olor, tste, texture n overll eptility sores ompre to other sustitution perents (5. n 7.5%). On the other hn, sustitution of whet flour with S. oliquus mirolge t 5. n 7.5% signifintly erese sensoril prmeters. In ition, rome sore signifintly erese in ll smples ompre to ontrol. It n e notie tht, S. oliquus oul e inorporte up to 2.5 g/1 g of whet flour in the formultion of lsgn pst without ny effet on eptility. These results re lose to those of Zouriet l.14 who mentione tht, A. pltensis my inorporte up to 2 g/1 g of semolin in pst formultion with enhning its sensory qulity ue to the mirolgl 38 effets on rightness of the upkes. Also, Dnesi et l. reporte tht, kery prouts my e enrihe with Spirulin without ny reverse effet on sensory eptne of the prout. Tle 12: Effet of mixing lsgn pst with S. Oliquus on sensory evlution Lsgn pst smples Color Arom Texture Tste Overll eptility Control 8.8± ± ± ± ±.5 2.5% S. oliquus 8.± ± ± ± ±.78 5.% S. oliquus 7.± ± ± ± ± % S. oliquus 6.5± ± ± ± ±1.43 Eh vlue is expresse s men of ten replites ±SD, numer in the sme olumn followe y the sme letter re not signifintly ifferent t.5 level. CONCLUSION The results of this stuy lerly emonstrte the usefulness of mixing S. oliquus power with lsgn pst s vlule foo itive to enhne the nutritionl hrteristis of lsgn pst. Where, lge re suitle soure of protein, unsturte ftty is n pigments with goo soure of essentil mino is n ntioxints effets. The most eptle mixing level with the highest eptility ws 2.5% S. oliquus power. Aknowlegement: This work ws supporte n fune y the projet entitle "Bioiesel proution from lge s renewle energy soure". Funing orgniztion: Reserh Development n Innovtion progrm (RDI), Funing Progrm: EU-Egypt Innovtion Fun, REFERENCES 1. Chu CY, Lio WR, Hung R, Lin LP, Hemgglutinting n ntiioti tivities of freshwter mirolge. Worl J Miroiol Biotehnol 2(8), 24, Guiry MD, Seneesmus meyen, Worl-wie eletroni pulition, Ntionl University of Ireln, Glwy (txonomi informtion repulishe from Alge Bse with permission of M.D. Guiry), Lurling M, Phenotypi plstiity in the green lge Desmoesmus n Seneesmus with speil referene to the inution of efensive morphology. Ann Limnol IntJ Lim 39(2), 23, Pultz O, Gross W, Vlule prouts from iotehnology of mirolge. Appl Miroiol Biotehnol 65(6), 24, Chon-Lee TL, Gonzlez-Mrino GE, Mirolge for helthy foos Possiilities n hllenges. Comp Rev Foo Si Foo Sfety 9(6), 21, Guees AC, Ctrin RB, Helen MA, Clui IP, Frniso XM, Mirolgl n ynoteril ell extrts for use s nturl ntiteril itives ginst foo pthogens. Int J Foo Si Tehnol 46(4), 211, Beker EW, Miro-lge s soure of protein. Biotehnol Av 25, 27, Pngestuti R, Kim SK, Biologil tivities n helth enefit effets of nturl pigments erive from mrine lge. J Funt Foos 3, 211, Amro HM, Guees AC, Mlt FX, Antimiroil tivities of mirolge: An invite review. In A. Ménez-Vils (E.), Siene ginst miroil pthogens: Communiting urrent reserh n tehnologil vnes. Formtex Miroiology Book Series, Nov Siene Pulisher, Wn-Loy C, Biotehnologil pplitions of mirolge. IeJSME 6(Suppl 1), 212, S24-S Spolore P, Jonnis-Cssn C, Durn E n Esmert A 26 Commeril pplitions of mirolge. J Biosi n Bioeng 11(2), 26, Mrkovits A, Conejeros R, Lopez L, Lutz M, Evlution of mrine mirolg Nnnohloropsis sp. s potentil ietry supplement. Chemil, nutritionl n short term toxiologil evlution in rts. Nutr Res 12, 1992, Bishop WM, Zuek HM, Evlution of mirolge for use s nutreutils n nutritionl supplements. J Nutr Foo Si 2, 212, Zouri N, Ai M, Fkhfkh N, Ayi MA, Zorgui L, Ayi M Atti H, Blue-green lge (A. pltensis) s n ingreient in pst: free ril svenging tivity, sensory n ooking hrteristis evlution. Int J Foo Si Nutr 62(8), 211, Beker EW, Biotehnology n exploittion of the green lg S. oliquusin Ini. Biomss 4(1), 1984, Gross R, Gross U, Rmirez A, Cur K, Collzos C, Felheim W, Nutritionl tests with green Seneesmus with helth n Interntionl Journl of Phrmeutil Sienes Review n Reserh Aville online t Copyright protete. Unuthorise repulition, reproution, istriution, issemintion n opying of this oument in whole or in prt is stritly prohiite. 37

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