Correspondence should be addressed to Muhammad Waseem Mumtaz; waseem and Nazamid Saari;

Transcription

1 Sitifi Wrld Jural Vlum 14, Artil ID , 11 pags Rsarh Artil RSM Basd Optimizati f Chmial ad Ezymati Trasstrifiati f Palm Oil: Bidisl Prduti ad Assssmt f Exhaust Emissi Lvls Muhammad Wasm Mumtaz, 1 Hamid Mukhtar, 2 Farq Awar, 3,4 ad Nazamid Saari 5 1 Dpartmt f Chmistry, Uivrsity f Gujrat, Pakista 2 Istitut f Idustrial Bithlgy, GC Uivrsity, Lahr 0, Pakista 3 Dpartmt f Chmistry, Uivrsity f Sargdha, Sargdha 100, Pakista 4 Cllg f Pharmay, Salma bi Abdulaziz Uivrsity, AlKharj 11942, Saudi Arabia 5 Faulty f Fd Si ad Thlgy, Uivrsiti Putra Malaysia, 4 Srdag, Slagr, Malaysia Crrspd shuld b addrssd t Muhammad Wasm Mumtaz; wasm pk56@yah.m ad Nazamid Saari; azamid@putra.upm.du.my Rivd 28 Fbruary 14; Rvisd 3 Ju 14; Aptd 11 Ju 14; Publishd 5 August 14 Aadmi Editr: Stamatis Kalligrs Cpyright 14 Muhammad Wasm Mumtaz t al. This is a p ass artil distributd udr th Crativ Cmms Attributi Lis, whih prmits urstritd us, distributi, ad rprduti i ay mdium, prvidd th rigial wrk is prprly itd. Currt study prsts RSM basd ptimizd prduti f bidisl frm palm il usig hmial ad zymati trasstrifiati. Th missi bhavir f bidisl ad its blds, amly, POB-5, POB-, POB-, POB-, POB-, ad POB-100 was xamid usig disl gi (quippd with tub wll). Optimizd palm il fatty aid mthyl strs (POFAMEs) yilds wr dpitd t b 47.6 ± 1.5, 92.7 ± 2.5, ad95.4 ± 2.0% fr hmial trasstrifiati atalyzd by NaOH, KOH, ad NaOCH 3, rsptivly, whras fr zymati trasstrifiati ratis atalyzd by NOVOZYME-435 ad A.. lipas ptimizd bidisl yilds wr 94.2 ± 3.1 ad 62.8 ± 2.4%, rsptivly. Distit dras i partiulat mattr (PM) ad arb mxid (CO) lvls was xprid i xhaust missis frm gi pratig bidisl blds POB-5, POB-, POB-, POB-, POB-, ad POB-100 mparativ t vtial ptrlum disl. Prtag hag i CO ad PM missis fr diffrt bidisl blds ragd frm 2.1 t 68.7% ad 6.2 t 58.4%, rsptivly, rlativ t vtial disl, whras a irrgular trd was bsrvd fr NOx missis. Oly POB-5 ad POB- shwd tabl rdutis, whras all thr blds (POB- t POB-100) shwd slight iras i NOx missi lvls frm 2.6 t 5.5% mparativ t ptrlum disl. 1. Itrduti At prst, th wrld is lkig fr apprpriat sluti f rgy riss as its dmad is salatig day by day du t rapidly grwig ppulati ad glbal idustrializati. Currtly, fssil basd rwabl rsurs lik ptrlum, al, atural gas, ad s frth ar mstly fulfillig th rgy rquirmts f th wrld but thir limitd availability is ivrsly prprtial t th risig rgy dmads. Bsids, th sumpti f ths fssil fuls is itimidatig th whl wrld with virmtal impats lik glbal warmig ad dplti f th z shild [1]. Typially, th disl gi xhaust (CO, NOx, ad PM) has b atgrizd as arigi ad svral studis rvaldaaugmtdhalthriskliklugaruplg trm xpsur t disl gi xhaust [2]. Th mai arigi ffts assiatd with disl xhaust ar attributd t st partils ihalati as varius mutagi ad arigi agts (.g., plyyli armati hydrarbs) ar kw t b absrbd as rgai phas partiulat mattr (PM) surfa. PM that ar radily ihald havig mdia dyami diamtr, that is, μm, rsult i dpsitiialvlarrgiflugs [3 6]. Rsarhrs ad virmtalists ar xtsivly mphasizig fr sarh f altrativ ad rwabl fuls that a lad t lar virmt. Althugh diffrt utris thrughut th wrld ar fusig psttratmt thlgis fr simultaus PM, CO, ad NOx rdutis, but us f bidisl (big

2 2 Th Sitifi Wrld Jural rwablrgyful)istadfdislfulhasprvdtgiv bst rsults twards lar virmt. Bidisl has sm advatags vr vtial fssil disl, spially i trms f lssr txi missis [7]. Prvius studis rvald that th us f bidisl istad f bidisl r i bldd frm i disl gi shwd rdud lvls f xhaust missis f CO ad PM, ad s frth [8 10]. Optimizati f bidisl prduti thrugh hmial r zymati trasstrifiati prss is athr ara f sidrati. I this txt, diffrt ptimizati mthds hav b dvlpd by th rsarhrs with th mai targt f ffrig high yild ad gd quality f bidisl prdut. Rsps surfa mthdlgy (RSM) is gaiig muh rgiti ths days as a imprativ tl fr th ptimizati f varius bihmial ad thlgial prsss. Usually, RSM is sidrd as a mbiati f mathmatial ad statistial prtls utilizd fr prss dvlpmt, imprvmt, ad ptimizati. RSM is als prvd t b vry hlpful thiqu fr th aalysis f spifi prblms whr a rsps f itrst is iflud by varius prss variabls with th bjtiv t ptimiz th said rsps [11 13]. Althugh palm il is i us as a fdstk fr bidisl prduti, arlir studis ar rprtd th RSM basd simultaus ptimizati f hmial ad zymati trasstrifiati fr prduti f bidisl usig palm il. Thrfr, th urrt study was dsigd with th mai bjtiv t ptimiz bth f th hmial ad zymati trasstrifiati prsss fr palm il bidisl prduti usig RSM as wll as valuati f xhaust missi lvls frm disl gi pratd th sythsizd bidisl ad its blds t hk its mpatibility as a gr ful. 2. Exprimtal Sti 2.1. Cllti f Raw Matrial. All th xprimtal wrk was xutd usig aalytial/rsarh grad hmials ad ragts. Palm il was prurd frm DESCON hmials (Pvt.) Ltd. Davis Rad Lahr, Pakista. KOH, NaOH, NaOCH 3, mthal, tlu, isprpal, phlphthali, starh, ati aid, HCl, sdium thisulfat, -hxa, at, ptassium idid, idi trihlrid, ptassium dihrmat, ad hlrfrm wr btaid frm Mrk Chmials, (Grmay), whil NOVOZYME-435 (lipas aryli rsi frm Cadida Atartia) ad lipas frm Asprgillus igr wr purhasd frm Sigma Chmial C. (St. Luis, MO). A.. lipas (lipas frm Asprgillus igr immbilizd lwatit) was btaid frm Prss Bithlgy Labratry, Dpartmt f Chmistry, GC Uivrsity, Lahr, Pakista Physihmial Charatristis f Palm Oil. Palm il was physihmially aalyzd fr th quality paramtrs iludig rfrativ idx, dsity, aid valu, idi valu, prxid valu, sapifiati valu, ad usapifiabl mattr (%) usig stadard mthds f AOCS Exprimtal Prdur. Bidisl was prdud usig bth hmial ad zymati trasstrifiati f palm il. Tabl 1: CCRD dsig summary f diffrt lvls f rati variabls usd fr trasstrifiati f palm il fr hmial ad zymati trasstrifiati. Fatr Nam Uits Lw lvl High lvl A a Catalyst trati % A b Ezym trati % B a Rati tim Miuts B b Rati tim Hurs C a Rati tmpratur C C b Rati tmpratur C D a&b Alhl : il mlar rati a CCRD dsig fr hmial trasstrifiati f palm il. b CCRD dsig fr zymati trasstrifiati f palm il. Chmial trasstrifiati ratis f il wr atalyzd by NaOH, KOH, ad NaOCH 3 usig spifid rati ditis ardig t tral mpsit rsps surfa dsig (CCRD) [11, 14 16], whras zymati trasstrifiati ratis wr arrid ut with NOVOZYME-435 ad A.. lipas as pr dsripti f Shimada t al. [17] ad Shimada t al. [18]. Fr alkali atalyzd hmial trasstrifiati, th ratiswrarridutialabratrysalratr,that is, thr k glass fabriatd flask fittd with a stirrr, samplig ilt, hillr, dsr, thrmmtr, ad hatig plat [11]. Chmial trasstrifiati ratis i ah bath f il ( grams) wr xutd usig spifid lvls f atalyst trati, mthal t il mlar rati, rati tmpratur, rati tim ardig t tral mpsit rsps surfa dsig (CCRD) at fixd stirrig rat, that is, 0 rpm. Aftr trasstrifiati, purifid bidisl was rvrd frm th glyrl (byprdut) by limiatig thr impuritis suh as rsidual mthal ad atalysts. O th thr had, zym atalyzd trasstrifiati at fixd shakig spd, that is, 0 rpm, with thr-stp mthal additi i rati flask, was arrid ut fllwig th mthd as dsribd arlir by Shimida t al. [18]. Aftr th mplti f th rati, bidisl was sparatd frm th glyrl ad zym was rvrd. (%) was sltd as rsps fr th ptimizati studis ad was dsribd udr-study il mass basis fr trasstrifiati ratis usig th fllwig xprssi: Fatty Aid Mthyl Estrs (FAME s) (%Y) = Mthyl str prdud i grams Grams f il usd durig trasstrifiati rati 100. Furthrmr, th fft f th variabls, that is, atalyst trati (A), rati tim (B), rati tmpratur (C), ad mthal : il mlar rati (D) prtagyildf bidisl, was valuatd thrugh CCRD. Th ivstigatd rags fr variabls A, B, C,adD ar dsribd i Tabl 1. A2 4 full-fatrial CCRD dsig was mplyd durig ptimizati usig fur variabls with fiv diffrt lvls (1)

3 Th Sitifi Wrld Jural 3 ladig t xprimts. Eah xprimt was rpatd thr tims. Th data gratd by xprimts was furthr statistially aalyzd ad usd fr ptimizati f bidisl yild. Th xprimtal rsults wr aalyzd by Dsig Exprt 7 ad SPSS fr ANOVA, rsps surfa plts, ad diagstis hks. Th apprpriat rsps surfa mdls fr ptimizati purpss basd xprimtal bidisl yilds wr hs fr bth hmial ad zymati trasstrifiati th basis f ANOVA with high statistial sigifia, lak f fit tsts, ad high valus f R 2. Furthrmr, sigifia f th sltd mdls ad th idividual ffiits wr tstd by f ad t-tsts [11, 15, 16] Mitrig f Bidisl Prduti. FTIR sptrspi mitrig f trasstrifiati f palm il was xutd usig Itrsp 0-X FTIR sptrphtmtr (Sptri Camsp Ltd., Tudr Hus, Barly Hill Rad, Garfrth, Lds LS25 1DX, UK) quippd with mutabl FTIR liquid lls. Th sptra wr rrdd vr saig rag f 0 t 00 m 1 [19, ] Cmpsitial Aalysis by GC. Fatty aids prfil f th prpard palm il bidisl was aalyzd qualitativly ad quatitativly usig GC/MS (Agilt Thlgis 68N) twrk gas hrmatgraphi systm havig a irt XL massadautijtr.fattyaidmthylstrsfthbidisl prdud wr sparatd usig plar apillary lum (100 m 0.25 mm ad film thikss 0. μm). A 1.0 μl bidisl sampl was ijtd it th lum thrugh split ijti md with a split rati 1 : 100. Extra pur hlium wasusdasarrirgas(mbilphas)withflwratf 1.2 ml/mi. Clum v tmpratur was prgrammd frm 1 t 2 C@4 C/mi whil iitial ad fial hld up tims wr st t b 1 ad 5 mi, rsptivly. Tmpratur f th ijtr ad MS trasfr li wr maitaid at 2 ad 2 C, rsptivly. A ltr iizati systm was mplyd fr GC/MS dtti whil saig mass rag varid frm t 5 m/z. Idtifiati f idividual fatty aidsfpalmilbidislwasxutdbymprhsiv mparis f thir rlativ rtti tims with ths f authti stadards f fatty aid mthyl strs (Sigma Chmial C., St Luis, MO, USA). Fr firmati f FAMEs prfil f palm il bidisl, MS sptra f th ukw sampl wr als mpard with MS sptra f th sam frm th NIST mass sptral library prvidd with GC/MS systm. Quatifiati f idividual fatty aids was arrid ut by Agilt Thlgis data hadlig sftwar (Chm Stati 68) ad mpsiti was rprtd as rlativ prtag f th ttal pak ara [11, 21] Exhaust Emissi Lvls Aalysis. Emissi lvls stimati was arrid ut usig disl gi (SD-1110) f pwr HP ad wight 210 kg quippd with a tub wll situatd i villag arby Uivrsity f Gujrat, Gujrat, Pakista. Palm il bidisl as sythsizd i th urrt study alg with its blds was mplyd fr stimati f thir missi lvls. Bidisl sampls wr bldd with vtial fssil basd ptrlum disl ad diffrt blds, that is, B5, B, B, B, B, ad B100 with bidisl prtag 5, 10,,,,,ad100%,rsptivly,wrprpard Mitrig f Exhaust Emissi Lvls. Exhaust missis mitrig frm disl gi, pratd diffrt bidisl blds, was arrid ut frm th mitrig hls alrady prvidd th disl gi staks. All th masurmtswrxutdthriwhthgiwaspratig at thir ptimum lad at diffrt tims t asrtai th missi bhavir. Fr th mitrig f CO ad NOx (NO + NO 2 ) flu gas aalyzr, that is, LANCOM-III (vrsi V1.II, srial umbr basd CTM mthd 034 f US EPA) fabriatd with ifrard ad ltrhmial ssr f disprsiv atur was usd, whras th stimati f partiulat mattr was xutd usig Th Caslla (partiulat samplig systm istrumt) i mplia with ISO- 96 ad BS-35. Cllulsi filtr mdia with pr siz <10 μ wr usd i th istrumt prvidd that it rsultd i quatitativ rtti f PM 10 fr dfiit tim itrvals. Th filtr mdia wr wll assmbld i a lak prf dust lltig prt. Th whl assmbly was td with a stal prb with th hlp f spial tuig t withstad high tmpraturs assiatd with flu gass. Aftr dfiit tim itrval PM rtaid th surfa f th filtr mdia was masurd [11] Ful Charatristis f Palm Oil Bidisl. Ful prprtis f th prdud palm il bidisl, amly,dsity (ASTM D 02), ta umbr (ASTM D 613), pur pit (ASTM D 97), lud pit (ASTM D 20), kimati vissity (ASTM D445),adashtt(ASTMD874),adsfrth,wr dtrmid usig stadards mthds. 3. Rsults ad Disussi 3.1. Physihmial Charatristis f Udr-Study Palm Oil. Physihmial haratristis f palm il usd as fdstk fr th prduti f bidisl wr valuatd whih wr fud t b ± 0.003, 0.88 ± 0.18, ad ± 0.04 mg KOH/g f il, 54. ± 2.51 gi 2 /100 g f il, 1 ± 1.17 mq/kg f il, ± 2.5 mg KOH/g f il ad 3. ± 0.02% fr rfrativ idx, dsity, aid valu, idi valu, prxid valu, sapifiati valu, ad usapifiabl mattr (%), rsptivly Palm Oil Fatty Aid Mthyl Estrs Yilds (%). Basd up xprimtal utputs dvlpd frm zymati ad hmial trasstrifiati f palm il as pr dfid CCRD, mparativ dsriptis f bidisl yilds (%) wr arrid ut ad prstd i Figur 1. Irrgular trds i bidisl yilds wr bsrvd fr bth hmial ad zymati trasstrifiati ratis. Th yild f bidisl prdud thrugh hmial trasstrifiati usig NaOH, KOH, ad NaOCH 3 ragd frm 22.5 t 47.4, 84.0 t 92.7, ad 84.9 t 95.4%, rsptivly, whras it ragd frm 62.5 t 94.2 ad 27.5 t 62.8% fr zymati trasstrifiati atalyzd by NOVOZYME-435 ad A.. lipas, rsptivly.

4 4 Th Sitifi Wrld Jural Palm il bidisl yild (%) Exprimtal ru Palm il bidisl yild (%) usig NaOH Palm il bidisl yild (%) usig KOH Palm il bidisl yild (%) usig NaOCH 3 Palm il bidisl yild (%) usig NVOZYME 435 Palm il bidisl yild (%) usig A.. lipas 3.3. Rsps Surfa Quadrati Mdls fr Optimizati f Bidisl Prduti. Quadrati mdls shw that th mdl is bst fittd fr th xprimtal data ad validatd t b sigifiat with P valus <0.05. Nsigifiat lak f fit tsts als suggstd that quadrati mdls wr bst fittd fr hmial trasstrifiati f palm il atalyzd by NaOH, KOH, ad NaOCH 3 with P valus, that is, , , ad > 0.05, rsptivly, as wll as fr zymati trasstrifiati atalyzd by NOVOZYME-435 ad A.. lipas with P valus, that is, ad > 0.05, rsptivly. Fittss f quadrati mdls was als asrtaid by mputig R 2 ad adjustd R 2 valus. Fr NaOH, KOH, ad NaOCH 3 atalyzd trasstrifiati f palm il, th R 2 valus wr , , ad 47 ad adjustd R 2 valus wr , , ad , rsptivly, whras fr NOVOZYME-435 ad A.. lipas atalyzd trasstrifiati f palm il, th R 2 valus wr ad ad adjustd R 2 valus wr ad , rsptivly. Figur 1: Cmparativ dsripti f bidisl yilds (%) rsultd frm xprimts xutd udr rati ditis dfid by CCRD fr hmial ad zymati trasstrifiati f palm il. Optimizd yild (%) f palm il bidisl NaOH KOH NaOCH 3 NVOZYME 435 A.. lipas Catalysts ad zyms usd fr trasstrifiati Errr bars: +/ 1 SD Figur 2: Optimizd palm il bidisl yild (%) ad ± stadard dviati fr hmial ad zymati trasstrifiati. Basd up sltd quadrati rsps surfa dsig th ptimizd prditd bidisl yilds wr usd fr th validati f atual xprimtal bidisl yilds, amly, 47.6 ± 1.5, 92.7 ± 2.5,ad95.4 ± 2.0%(Figur 2)frhmial trasstrifiati usig NaOH, KOH, ad NaOCH 3,rsptivly, whras 94.2 ± 3.1 ad 62.8 ± 2.4%, rsptivly, fr NOVOZYME-435 ad A.. lipas atalyzd trasstrifiati f palm il (Figur 2). Exprimtal bidisl yilds wr dpitdtbwlliagrmtwithprditdvalus Optimizd Rati Paramtrs. Fr NaOH atalyzd trasstrifiati f palm il, th highst POFAME s yild, was btaid usig NaOH (0.5%) ad mthal t il mlar rati (7.5 : 1.0) at rati tmpratur f 52.5 Cfr75miuts (Tabl 2). Wh trasstrifiati rati usig KOH as a atalystwasd,maximumpofame syildwasahivd by dutig th ratis fr miuts usig 0.75% KOH ad 6 : 1 mthal : il mlar rati at 45 C. Optimizd rati paramtrs fr NaOCH 3 atalyzd trasstrifiati f palm il, ffrig ptimum POFAMEs yild, wr fud t b 0.75%, 6 : 1 mthal t il mlar rati, 45 C rati tmpratur, ad miuts rati tim. Th highst POFAMEs yild was btaid fr NOVOZYME- 435 atalyzd trasstrifiati f palm il usig 1.0% NOVOZYME-435 trati, 6 : 1 mthal : il mlar rati, 32.5 C rati tmpratur, ad hurs rati tim, whras fr A.. Lipas atalyzd trasstrifiati th ptimal POFAMEs yild was rrdd by usig % A.. lipas trati, 9 : 1 mthal : il mlar rati, C rati tmpratur, ad 96 hurs rati tim (Tabl 2) FTIR Sptrspi Mitrig f Trasstrifiati. FTIR sptrspi aalysis was prfrmd fr mitrig trasstrifiati ratis f palm il. FTIR sptra f palm il ad palm il drivd bidisl wr rrdd. Th prs f IR bads i th rgi m 1 fr CH 3 asymmtri bdig ad fr O CH 3 strthig i all bidisls. IR sptra larly dpitd th vrsi f palm il (i.., triglyrids) t fatty aid mthyl strs, as ths IR bads wr t prst i th part il (i.., palm il) IR sptra as shw i Figur 3. Th rgi m 1 fr O CH 2 grups i glyrl (mity f TG, DG, ad MG) was fud t b i IR sptrum f palm il, whil i bidisl sptrum this bad was abst. Furthrmr, th rgis m 1 fr C=O strth ad m 1 fr symmtri C H strthig wr prst i bth th palm il ad th bidisl IR sptra. Ths fidigs ar i agrmt with ths f ur prvius

5 Th Sitifi Wrld Jural 5 Tabl 2: Optimizd rati paramtrs fr bidisl prduti usig hmial ad zymati trasstrifiati f udr-study fdstk. Catalyst/zym C/E trati Rati tim Rati tmpratur Mthal : il mlar rati NaOH 0.5% 75 Mi 52.5 C 7.5 : % KOH 0.75% Mi 45.0 C 6:1 92.7% NaOCH % Mi 45.0 C 6:1 95.4% NOVOZYME % Hrs 32.5 C 6:1 94.2% A..Lipas % 96 Hrs.0 C 9 : % Tabl 3: Rsps surfa quadrati mdl aalysis f varia (ANOVA) tabl fr hmial trasstrifiati f palm il. Sur df SS (MS) a SS (MS) b F valu SS (MS) (P valu) a F valu (P valu) b F valu (P valu) Mdl (5.45) (57.09) (8.) 14. (<0.0001) (<0.0001) 6. (0.0005) A-atalyst trati (12.54) (436.91) (27.05) (<0.0001) 363. (<0.0001).06 (0.0004) B-rati tim (7.72) 0.74 (0.74) (15.33).98 (0.0004) 0.61 (0.4463) (0.0042) C-rati tmpratur (0.77) 3.23 (3.23) 7.17 (7.17) 2.08 (0.1694) 2.69 (0.1221) 5.32 (0.0358) D-alhl : il mlar rati (19.21) (32.67) 7.66 (7.66) (<0.0001) (0.0001) 5.68 (0.08) AB (0.092) 7.02 (7.02) 2.86 (2.86) 0.25 (0.6252) 5.84 (0.0288) 2.12 (0.1662) AC (0.42) (22.09) (11.53) 1.14 (0.26) (0.0006) 8.55 (0.0105) AD (2.17) (17.64) (0.0025) 5.89 (0.0282) (0.0016) (0.9662) BC (0.66) 2.10 (2.10) (11.39) 1.79 (0.03) 1.75 (0.57) 8.45 (0.0109) BD (0.47) (23.52) 0.41 (0.41) 1.28 (0.2748) (0.0005) 0. (0.5896) CD (0.21) (18.49) 5.00 (5.00) 0.58 (76) (0.0014) 3.70 (0.0734) A (0.) (222.79) 0.11 (0.11) 2.16 (0.1621) (<0.0001) (0.75) B (2.81) (23.47) 0.69 (0.69) 7.65 (0.0144) (0.0005) 0.52 (0.4839) C (0.58) 3.86 (3.86) (0.015) 1.57 (0.2288) 3.21 (0.0934) (0.9176) D (23.45) (19.82) (27.13) (<0.0001) (0.0010).12 (0.0004) Rsidual (0.37) (1.).23 (1.35) Lak f fit (0.31) (1.63) (1.78) 0.64 (0.7415) 4.64 (0.0521) 3.69 (0.0813) Pur rrr (0.48) 1.75 (0.35) 2.42 (0.48) Cr ttal SS (MS) = sum f squars (ma squar). Mdl a rprsts quadrati mdl basd xprimtal rsults f KOH atalyzd trasstrifiati f udr-study fdstk. Mdl b rprsts quadrati mdl basd xprimtal rsults f NaOH atalyzd trasstrifiati f udr-study fdstk. Mdl rprsts quadrati mdl basd xprimtal rsults f NaOCH 3 atalyzd trasstrifiati f udr-study fdstk. T (%) 100 (05.7) (2931.8) (2847.5) Wavumbrs (m 1 ) (1764.6) (1447.5) (12.0) (1196.9) (1110.2) (838.0) (798.5) Figur 3: A typial FTIR sptrum f palm il basd bidisl. studis [19, ] as wll as with sm thr publishd wrks [14, 22, 23] ANOVA fr Sltd Rsps Surfa Quadrati Mdls. ANOVA fr rsps surfa quadrati mdl (Tabl 3) larly dpitd that th P valus fr all thr mdls (a, b, ad ) wr lss tha th lvl f sigifia 0.05, maig that th quadrati mdls wr sigifiatly fit fr th xprimtal rsults f trasstrifiati f palm il usig thr diffrt alkali atalysts. Fr mdl a, th mai ffts, that is, atalyst trati (A), rati tim (B), ad mthal t il mlar rati (D), wr fud t b sigifiat with P valus lssr tha 0.05, whil fr mdl b mthal : il mlar rati (D) ad atalyst trati (A) wr amg th mai sigifiat ffts, whras fr mdl all thmaifftswrfudtbsigifiat. Amg all first rdr itrati trms, atalyst trati mthal t il mlar rati (AD) frmdla,ad fr mdl b atalyst trati rati tim (AB), atalyst trati rati tmpratur (AC), atalyst

6 6 Th Sitifi Wrld Jural trati mthal t il mlar rati (AD), rati tim mthal t il mlar rati (BD), ad rati tmpratur mthal t il mlar rati (CD), whras, fr mdl, atalyst trati rati tmpratur (AC) ad rati tim rati tmpratur (BC) wr dpitd t b sigifiat. Amg quadrati trms rati tim (B 2 ) ad mthal t il mlar rati (D 2 ) wr asrtaid t b sigifiat fr mdl a, whil fr mdl b atalyst trati (A 2 ), rati tim (B 2 ), ad mthal t il mlar rati (D 2 ) wr sigifiat quadrati trms with P valus lssr tha 0.05, whil fr mdl ly mthal t il rati (D 2 ) was sigifiat quadrati trm. O th thr had, ANOVA fr quadrati mdls d ad ad fr zymati trasstrifiati f palm il atalyzd by NOVOZYME-435 ad A.. lipas, rsptivly, ar dsribd i Tabl 4.Th P valus fr tstig th mdl sigifia fr mdls d ad wr lss tha th lvl f sigifia 0.05, maig that th quadrati mdls wr sigifiatly fit fr th xprimtal rsults f zym atalyzd trasstrifiati f palm il. Fr quadrati mdl d, th mai ffts, that is, mthal t il mlar rati (D) ad zym trati (A), wr fud t b twrthy, whil fr mdl th mai ffts, that is, zym trati (A), rati tim (B), rati tmpratur (C), ad mthal t il mlar rati (D) wr fud t b sigifiat. Amg all first rdr itrati trms, zym trati rati tim (AB), zym trati rati tmpratur (AC),ad zym trati mthal t il mlar rati (AD) wr sigifiat fr mdl d, whras frmdlzymtrati rati tim (AB), zym trati rati tmpratur (AC), ad rati tim rati tmpratur (BC) wrfudtb sigifiat first rdr itrati trms. Similarly, fr mdl d, P valus fr quadri trms, that is, zym trati (A 2 ) ad rati tim (B 2 ), wr lssr tha 0.05 whih asrtaid thir sigifiat tributi; th thr had, fr mdl amg quadrati trms, rati tim (B 2 ), rati tmpratur (C 2 ), ad mthal t il mlar rati (D 2 ) wr asrtaid as sigifiat quadrati trms. Rsps surfa plts (Figurs 4(a) 4()) summarizd sigifiat rsps surfa tributis frm diffrt rati paramtrs fr th ptimizd prduti f palm il bidisl. Rsps surfa plts (Figurs 4(a) 4()), shwig th fft f diffrt rati paramtrs th % yild POFAMEs, dpitd that, fr a iras i lvls f atalyst (NaOH) trati up t 0.5%, mthal t il mlar rati up t 7.5 : 1, rati tim up t 75 miuts, ad rati tmpratur up t 52.5 C, th vrsi f palm il t palm il fatty aid mthyl strs irasd ad rahd t maximum valu, whras byd ths lvls f rati paramtrs a drasig trd i % yild f POFAMEs was bsrvd. Impat f sigifiat first rdr itrati btw atalyst (KOH) trati ad mthal t il mlar rati (Figur 4(f)) dpitd highst POFAMEs whil prfrmig bas atalyzd trasstrifiati ratis f palm il t palm il bidisl usig 0.75% KOH trati ad 6 : 1 mthal t il mlar rati. Highr % yild f POFAMEs was prurd via KOH atalyzd trasstrifiati f palm il usig highr KOH trati mparativ t NaOH atalyzd ratis. Similarly, impat f sigifiat first rdr itratis was als asrtaid by th rsps surfa plts (Figurs 4(g)-4(h)), shwig th fft f rati tim ad rati tmpratur, atalyst trati ad rati tim NaOCH 3 atalyzd trasstrifiati f palm il fr ptimizd prduti f POFAME s; th rsps surfa plts dpitd that maximum POFAME s yild was ahivd wh trasstrifiati f palm il was prfrmd usig NaOCH 3, that is, 0.75% fr a tim prid f miuts at 45 C, whras byd ths lvls lssr POFAME s yild was bsrvd. Rsps surfa plts (Figurs 4(i) 4(k)) dpitig sigifiat tributis f first rdr itratis fr zym (A.. lipas) atalyzd trasstrifiati f palm il rvald that maximum POFAMEs yild was rivd by prfrmig zym atalyzd trasstrifiati f palm il usig % A.. lipas trati ad mthal t il mlar rati 9 : 1 fr a tim prid f 96 hurs at.0 C, whras abv ad blw ths lvls a dras i POFAMEs yild was bsrvd. O th thr had, wh NOVOZYME-435 was usd istad f immbilizd A.. lipas, th sigifiat first rdr rspss,shwirspssurfaplts(figurs4(l) 4()), rvald that maximum POFAMEs yild was prurd usig NOVOZYME-435 trati, mthal t il mlar rati, rati tim, ad rati tmpratur, that is, 1.0%,6:1,hurs,ad32.5 C, rsptivly. Th POFAMEs yildahivdvianovozyme-435atalyzdtrasstrifiati f palm il was asrtaid t b highr mparativ t A.. lipas atalyzd trasstrifiati f palm il Palm Oil Bidisl Cmpsiti. Majr fatty aid mthyl strs prfil f palm il fatty aid mthyl strs (POFAME) was rvald t sist f myristi aid mthyl strs (C14:0), palmiti aid mthyl strs (C16:0), palmitli aid mthyl strs (C16:1), stari aid mthyl strs (C18:0), li aid mthyl strs (C18:1), lili aid mthyl strs (C18:2), ad lili aid mthyl strs (C18:3) with % mpsitis 1., 41.5, 0., 3., 38.6, 10.6 ad 1.09%, rsptivly (Tabl 5). Palmiti aid mthyl strs, li aid mthyl strs, ad lili aid mthyl strs stimatd i prst study wr mparabltthsrprtdarlir,thatis,42.6%,.5%, ad 10.1%, rsptivly, by Akh t al. [24] Exhaust Emissi Prfil f Bidisl Sythsizd. Plluti-fr air is f th ky rquirmts fr halthy sity [25, 26]. Muh pidmilgial vid has b prvidd by varius rsarhrs rvalig dirt assiati btw halth ffts ad txi xhaust missi lvls. I this rgard, partiulat mattrs, NOx, CO, ad SO 2 ar idrd as th mai fatrs f r. Rdutis i pulmary futis, asthma, lug futi drmts, ad prmaturdaths,adsfrtharrprtdtbassiatd with ths pllutats. Furthrmr, high affiity f CO fr hmglbi t frm arbxyhamglbi is assumd as mai attributr f CO txiity ausig aut pisig ladig t dath [24, 27 31]. Emissis rsultd frm ful burig hav gaid imprta wig t thir advrs halth

7 Th Sitifi Wrld Jural C: r a i ti trat tm p r ys t atu atal r A: i B: r trat at i tim yst atal A: (b) (a) D: m tha ati 5.00 l tr 4.00 : il ml yst ar ra atal ti A: D: m tha m l 4.00 : il ti i t ml a ar ra B: r ti () (d) Yild D: m tha ur l rat : il mp t ml ar ra a ti ti C: r D: m tha 6.00 l : il ml ar ra 3.00 ti (f) Yild Yild () rati t 0.25 ys t atal A:.00 C: r a ti ti tra tm p r yst atu atal r A: C: r at tim i i t tm a pra B: r tur (g) (h) Figur 4: Ctiud.

8 Th Sitifi Wrld Jural B: r.00 a ti tim ti tra 0.25 m zy A:.00 C: r i a trat ti tm m pr zy atu A: r 70 (j) 100 (i) 9.00 D: m tha 6.00 ti l tra : il m lar m rati zy : A i B: r trat a ti m tim zy A: (l) (k) C: r a ti ti tra tm p r m atu zy r A: (m) D: m i tha 6.00 trat l 5.00 : il 4.00 ml 0.25 ar ra 3.00 m ti zy : A () Figur 4: Rsps surfa plts (a) (h) shwig sigifiat first rdr itratis amg diffrt rati paramtrs ivlvd i palm il bidisl prduti via bas atalyzd trasstrifiati ad rsps surfa plts (i) () shwig sigifiat first rdr itratis amg diffrt rati paramtrs ivlvd i palm il bidisl prduti via zym atalyzd trasstrifiati. impats. Currtly muh attti is fusd thrughut th wrld t miimiz th lvls f ths missis by dvlpig altrativ virmt fridly fuls. Durig urrt rsarh wrk bidisl was sythsizd ad th subjtd t xhaust missi lvl studis. Figur 5 shwd sidrabl dras i gi xhaust missis prfil, that is, CO ad PM mattr frm palm il basd bidisl, ad its blds wh mpard with ptrdisl. O th avrag basis % hag i CO missi lvls frm gi xhaust pratig POB-5, POB-, POB-, POB-, POB-, ad POB-100 was fud t b 2.1 ± 0.3, 10.5 ± 0.7, 21.5 ± 2.7, 35.9 ± 2.7, 44.8 ± 3.3, ad 68.7 ± 1.4%, rsptivly, whras % hag i partiulat mattr (PM) missis was rvald t b 6.2 ± 2.1, 31.8±3.9, 44.9±2.3, 46.5±3.2, 55.9±4.5, ad 58.4± 4.0%, rsptivly (Figur 5), mparativ t vtial ptrdisl. O th thr had, a irrgular trd i NOx missis was dpitd; NOx missis frm gi xhaust pratd POB-, POB-, POB-, ad POB-100 wr fud t b highr tha gi xhaust missis pratd

9 Th Sitifi Wrld Jural 9 Tabl 4: Rsps surfa quadrati mdl aalysis f varia (ANOVA) fr zymati trasstrifiati f palm il. Sur df SS (MS) d SS (MS) F valu (P valu) d F valu (P valu) Mdl (1.44) (8.85) 27. (<0.0001) (<0.0001) A-zym trati ( ) (226.00) (<0.0001) (<0.0001) B-rati tim (28.38) 1.55 (1.55 ) 4.08 (0.0615) (<0.0001) C-rati tmpratur (4.07) 2.69 (2.69) 0.59 (58) 7.59 (0.0148) D-alhl : il mlar rati (63.73) (12.62) 9.17 (0.0085) (<0.0001) AB (89.78) 7.56 (7.56) (0.0027) (0.0003) AC 1 6 (6) 6. (6.) 8.70 (0.0099) (0.0007) AD (77.00) 0.42 (0.42) (0.0046) 1.19 (0.2924) BC (13.51) 1.69 (1.69) 1.94 (0.1836) 4.76 (0.0454) BD (24.26) 0.16 (0.16) 3.49 (0.0814) (0.5121) CD (3.52) 0.49 (0.49) 0.51 (0.4878) 1.38 (0.2582) A (49.) (0.055) 7.14 (0.0174) 0.16 (0.6993) B (110.22) 2.13 (2.13) (0.0012) 5.99 (0.0271) C (5.34) 1.95 (1.95) 0.77 (0.3945) 5. (0.0331) D (3.73) (52.88) 0.54 (0.47) (<0.0001) Rsidual (6.95) 5.32 (0.35) Lak f fit (8.31) 4.81 (0.48) 1.97 (0.2358) 4.68 (0.0511) Pur rrr (4.23) 0.51 (0.10) Cr ttal SS (MS) = sum f squars (ma squar). Mdl d = rprsts quadrati mdl basd xprimtal rsults f A.. lipas atalyzd trasstrifiati f udr-study fdstk. Mdl = rprsts quadrati mdl basd xprimtal rsults f NOVOZYME-435 atalyzd trasstrifiati f udr-study fdstk. Avrag % hag i xhaust missi lvls CO x POB-5 POB- POB- NO x Exhaust missis POB- POB- POB-100 Figur 5: % hag i xhaust missis (CO, NOx, ad PM) frm gi xhaust pratd diffrt palm il bidisl blds mparativ t gi xhaust pratd vtial ptrdisl. vtial ptrdisl with % hag, that is, 2.6 ± 0.9, 3.7 ± 1.2, 5.4 ± 1.7, ad5.5 ± 1.8%, rsptivly, whras i as f POB-5 ad POB- NOx missis wr fud t b lssr tha vtial ptrdisl with % hags 2.6±1.0 ad 4.6 ± 2.3%, rsptivly. Grabski ad MCrmik [9] PM Tabl 5: Majr fatty aid mthyl strs f palm il bidisl. Sr.. Fatty aid mthyl str Rtti tims POFAME s 1 Myristi aid (C14:0) ± Palmiti aid (C16:0) ± Palmitli aid (C16:1) 0. ± Stari aid (C18:0) ± Oli aid (C18:1) ± Lili aid (C18:2) ± Lili aid (C18:3) ± Arahidi aid (:0) ± Erui aid (C22:1) rprtd 12% iras i NOx missi lvls usig 100% pur sy bidisl mparativ t th ptrlum disl [9], whras % bld f pur sy bidisl i ptrdisl dpitd ly 2 t 4% ris i NOx missi lvls mparativ t vtial disl. Th irasd lvls f NOx missis v at small sal a gativly impat th bidisl us [9] Ful Prprtis f Palm Oil Bidisl. Th ful haratristis, that is, kimati vissity ((mm 2 /s) C), ash tt (%), lud pit ( C), pur pit ( C), highr hatig valu (MJ/Kg), ad ta umbr fr POFAME, wr dpitd t b 4.31 ± 0.23 mm 2 /s, ± 0.025%, 11.8 ± 1.5 C, 7.49 ± 1. C, ± 0.54 MJ/Kg, ad ± 2.29, rsptivly. Bjami t al. [32] rprtd th ful prprtis, amly, 4.71 mm 2 /s, 16.0 C, ad.0 fr kimati vissity, lud pit, ad ta umbr f palm il bidisl, rsptivly.

10 10 Th Sitifi Wrld Jural Th rsults prstd by Bjuma t al. [33] ar mparabl t th fidigs f urrt rsarh. 4. Clusis Rsults asrtaid th ptimizd prduti f palm il basd bidisl usig rsps surfa mthdlgy. Th ptimallvlsfpalmilbidislviabasatalyzdtrasstrifiati was 95.4±2.0%usigNaOCH 3, whil fr zym atalyzd trasstrifiati th maximum bidisl yild prurd was 94.2 ± 3.1%usigNOVOZYME-435.NaOCH 3 was thrfr prvd t b mst fftiv atalyst i th prst study amg thr alkali atalysts (NaOH, KOH, ad NaOCH 3 )mplyd,whrasnovozyme-435was thmstfftivlipasatalyst.furthrmr,sythsizd bidisl was fud t b gr ful i trms f sidrabl rduti i xhaust missis mparativ t vtial ptrdisl ad it shwd ful prprtis thially i mplia with ASTM D6751 ad EN stadards. Abbrviatis RSM: Rsps surfa mthdlgy CCRD: Ctral mpsit rsps surfa dsig NOVOZYME-435: Lipas aryli rsi frm Cadida Atartia A.. Lipas: Lipas frm Asprgillus igr FTIR Sptrspy: Furir trasfrm ifrard sptrspy GC-MS: Gas hrmatgraph quippd with mass sptrmtri dttr A: Catalyst/zym trati B: Rati tim C: Rati tmpratur D: Mthal:ilmlarrati. Cflit f Itrsts Th authrs dlar that thr is flit f itrsts rgardig th publiati f this papr. Rfrs [1] J. Tikll, Frm th Fryr t th Ful Tak: Th Cmplt Guid tusigvgtabloilasaaltrativful,tikllergy Csultats, 3rd diti, 03. [2] J. L. Maudrly, Txilgial ad pidmilgial vid fr halth risks frm ihald gi missis, Evirmtal Halth Prsptivs,vl.102,.4,pp ,1994. [3] P.T.J.ShprsadR.P.Bs, Cmbustifdislfulfrm a txilgial prsptiv II. Txiity, Itratial Arhivs f Oupatial ad Evirmtal Halth,vl.64,.3,pp , [4] P.T.J.ShprsadR.P.Bs, Cmbustifdislfulfrm a txilgial prsptiv I. Origi f implt mbusti prduts, Itratial Arhivs f Oupatial ad Evirmtal Halth,vl.64,.3,pp ,1992. [5] J. Huisigh, R. Bradw, R. Jugrs t al., Appliati f biassay t th haratrizati f disl partil missis, i Appliati f Shrt-trm Biassay i th Fratiatiad Aalysis f Cmplx Evirmtal Mixturs, M.D.Watrs,S. Nsw, J. L. Huisigh, S. S. Sadhu, ad L. D. Claxt, Eds., pp ,PlumPrss,NwYrk,NY,USA,1978. [6] J. Krahl, J. Bügr, O. Shrödr, A. Muak, adg. Kth, Exhaust missis ad halth ffts f partiulat mattr frm agriultural tratrs pratig rapsd il mthyl str, JuralfthAmriaOilChmists Sity,vl.79,.7,pp , 02. [7] D. S. Baik ad Y. C. Ha, Th fft f bidisl ad ultra lw sulfur disl fuls missis i 11,000 havy-duty disl gi, Jural f Mhaial Si ad Thlgy, vl. 19,. 3, pp , 05. [8] Amria Bifuls Assiati Ifrmati Rsurs (ABAIR), Bidisl: A Thlgy Prfrma ad Rgulatry Ovrviw, Natial Sy Disl Dvlpmt Bard, Jffrs City, M, USA, [9] M. S. Grabski ad R. L. MCrmik, Cmbusti f fat ad vgtabl il drivd fuls i disl gis, Prgrss i Ergy ad Cmbusti Si, vl. 24,. 2, pp , [10] M. E. Tat, P. S. Wag, J. H. Va Grp, ad T. E. Clmt, Exhaust missis frm a gi fuld with bidisl frm high-li sybas, JuralfthAmriaOilChmists Sity,vl.84,.9,pp ,07. [11] M. W. Mumtaz, A. Ada, F. Awar t al., Rsps surfa mthdlgy: a mphati tl fr ptimizd bidisl prduti usig ri bra ad suflwr ils, Ergis, vl. 5,. 9, pp , 12. [12] D. C. Mtgmry, Dsig ad Aalysis f Exprimts: Rsps Surfa Mthd ad Dsigs, JhWily&Ss, Hbk, NJ, USA, 05. [13] H. M. Raymd ad M. C. Duglas, Rsps Surfa Mthdlgy Prss ad Prdut Optimizati Usig Dsigd Exprimts, Jh Wily & Ss, Hbk, NJ, USA, 2d diti, 02. [14] M. Saqib, M. W. Mumtaz, A. Mahmd, ad M. I. Abdullah, Optimizd bidisl prduti ad virmtal assssmt f prdud bidisl, Bithlgy ad Biprss Egirig,vl.17,.3,pp ,12. [15] U. Rashid, F. Awar, ad M. Arif, Optimizati f bas atalyti mthalysis f suflwr (Hliathus auus) sd il fr bidisl prduti by usig rsps surfa mthdlgy, Idustrial ad Egirig Chmistry Rsarh, vl.48,.4, pp ,09. [16]U.Rashid,F.Awar,T.M.Asari,M.Arif,adM.Ahmad, Optimizati f alkali trasstrifiati f ri bra il fr bidisl prduti usig rsps surfa mthdlgy, Jural f Chmial Thlgy ad Bithlgy, vl.84,. 9,pp ,09. [17] Y. Shimada, Y. Wataab, T. Samukawa t al., Cvrsi f vgtabl il t bidisl usig immbilizd Cadida atartia lipas, Jural f th Amria Oil Chmists Sity,vl.76,. 7, pp , [18] Y. Shimada, Y. Wataab, A. Sugihara, ad Y. Tmiaga, Ezymati alhlysis fr bidisl ful prduti ad appliati f th rati t il prssig, JuralfMlularCatalysis B,vl.17,.3 5,pp ,02. [19] M. W. Mumtaz, A. Ada, Z. Mahmd t al., Bidisl frm wast kig il: ptimizati f prduti ad mitrig

11 Th Sitifi Wrld Jural 11 f xhaust missi lvls frm its mbusti i a disl gi, Itratial Jural f Gr Ergy, vl.9,.7,pp , 12. []M.W.Mumtaz,A.Ada,Z.Mahmd,H.Mukhtar,M. Daish, ad Z. Ahmad, Bidisl prduti usig Erua sativa il: ptimizati ad haratrizati, Pakista Jural f Btay, vl. 44,. 3, pp , 12. [21] F.Awar,R.Nasr,M.I.Bhagr,S.Ashraf,F.N.Talpur,ad F. A. Aladduy, Physi-hmial haratristis f itrus sds ad sd ils frm Pakista, Jural f th Amria Oil Chmists Sity,vl.85,.4,pp.321 3,08. [22] M. A. Dubé, S. Zhg, D. D. MLa, ad M. A. Kats, A mparis f attuatd ttal rflta-ftir sptrspy ad GPC fr mitrig bidisl prduti, Jural f th Amria Oil Chmists Sity,vl.81,.6,pp.599 3,04. [23] N.G.Siatis,A.C.Kimbaris,C.S.Pappas,P.A.Taratilis,ad M. G. Plissiu, Imprvmt f bidisl prduti basd th appliati f ultrasud: mitrig f th prdur by FTIR sptrspy, JuralfthAmriaOilChmists Sity,vl.83,.1,pp.53 57,06. [24] C.C.Akh,S.Chag,G.L,adJ.Shaw, Ezymatiapprah t bidisl prduti, Jural f Agriultural ad Fd Chmistry,vl.55,.22,pp ,07. [25]S.Furuta,H.Matsuhashi,adK.Arata, Bidislfulprduti with slid amrphus-ziria atalysis i fixd bd ratr, Bimass ad Birgy, vl.,. 10, pp , 06. [26] T. F. Dssi, M. Ryirs, R. J. Brgr, ad G. B. Mari, Simulati f htrgusly MgO-atalyzd trasstrifiati fr fi-hmial ad bidisl idustrial prduti, Applid Catalysis B: Evirmtal,vl.67,.1-2,pp ,06. [27] D. d Olivira, M. Di Lui, C. Fai t al., Optimizati f zymati prduti f bidisl frm astr il i rgai slvt mdium, Applid Bihmistry ad Bithlgy, vl. 115,.1 3,pp.771 7,04. [28] M.Ezzati,A.D.Lpz,A.Rdgrs,S.VadrHr,adC.J. L. Murray, Sltd majr risk fatrs ad glbal ad rgial burd f disas, Th Lat,vl.3,.9343,pp , 02. [29] G. Tulumi, K. Katsuyai, D. Zmiru t al., Shrt-trm ffts f ambit xidat xpsur mrtality: a mbid aalysis withi th APHEA prjt, Amria Jural f Epidmilgy,vl.146,.2,pp ,1997. [] D. Zmiru, J. Shwartz, M. Saz t al., Tim-sris aalysis f air plluti ad aus-spifi mrtality, Epidmilgy,vl.9,. 5, pp , [31] M. Saz, F. Ballstr, M. A. Barló t al., A mbid aalysis f th shrt-trm ffts f phthmial air pllutats mrtality withi th EMECAM prjt, Evirmtal Halth Prsptivs,vl.110,.3,pp ,02. [32] I. U. Bjami, M. K. Harua, ad O. O. Emmaul, Dtrmiati f ptimal atalyst trati fr maximum bidisl yild frm Tigrut Oil, Lard Jural f Sis,vl.6,. 10, pp , 07. [33] P. Bjuma, J. Agudl, ad A. Agudl, Basi prprtis f palm il bidisl-disl blds, Ful, vl. 87, , pp , 08.

12 Itratial Jural f Pptids BiMd Rsarh Itratial Stm Clls Itratial Advas i Virlg y Itratial Jural f Gmis Jural f Nuli Aids Zlgy Itratial Jural f Vlum 14 Vlum 14 Submit yur mausripts at Jural f Sigal Trasduti Th Sitifi Wrld Jural Gtis Rsarh Itratial Aatmy Rsarh Itratial Itratial Jural f Mirbilgy Bihmistry Rsarh Itratial Advas i Biifrmatis Arhaa Ezym Rsarh Itratial Jural f Evlutiary Bilgy Mlular Bilgy Itratial Jural f Mari Bilgy