AISTRALIA pA TENTSrj A CT, 199 COMPLETE SPECIFICATION FOR A STANDARD PATENT ORtINAL Name of Appicant: FONTERA CO-4 PEATVE GROUP~ LF~' E Actua invetors: FLETCHER, Katt na; CATCHPL .twe; GREY, Joh'-, Ber'ri'm; PR 4CH ARD. Mark Address fo Sei ce A A JFAW, e 11. 60 Marcu.s Carke Stree. Canberra A CT in Autai:2601' utai Inv"nio litle: N NEUTRAL IPfDPL§!TED AND/OR POLAR IPtHID ENRICHD DAjIRY PR ODU/CTS, AND PROCESSES FOR THE-IR PROD.UCTION Tn bowing semem a tio desPiuon Mf tis ne &eN v iiiding the best mlet ad of permiog i knovn ti v NEUTRAL LUIDDEPLETEDWAND/OR POL AR LIFID-ENRICHED DAIRY P PRODUCT S, AND PROCESSES FOR rTHEIIR PRODU ACTION FIELD OF fHE INVENTION Thnresent invention povide airyt produCet and processes for producing these products The dairy products of tke present invention inchade prodiucts suitable for use inm fn them jas. H ACKGR OUN ) COMMercallyR naVaiblersft formijas are typically produced u.ag nonihuma milk. Hocver the ntritia conposition of human milk differs in some respecis oC that of o:-hm~tan mil ;k (such as cow. shee,. oat. Non*htman who:e irlk such as cow, goat or sheep iknot, contains a higher poportion of sarusated ary acds than human mrlk and ja ciein Pieic acid and alpha-4inolenic Ad, pyusaturated ay acids that are essental for normai anf growt and d.ev2loment, A, butyrc acid which is found in miik tat may cause ptad vomir in infntis, Theefre standard inat formuflas are typincaly produced uin~lg l(j:o wr\ air &Xduers such as skin milk. Using a reduced dainy product means undesirable components 0n iulk Ca are nlneJuded n the infant ibrmula bt a als mcens that phosphor.pd and (gycOlphingolipid levels are signIficand lower than these in human mil. a rover the last 5-10 years As Shown increasing and (gcsphingoipid evrsin infant fouations to [evels found in human mink 'pairiuarly gaim~eide G&A 5 gangiosride Git ceramudes and sinomein)1 may la 0o prevention of infections 1y moldiyng gut intestnal flora and competitively biding ant;gens; prevention of he development of aiiergies;and opftmaj Oeural deve'opmenf~t. it ;s thieefore desirable to prdc an inatformulia containing su;fficiert lees of decsirhble lipids whij minimilsingL o; eilminadmg u:ndessimbleingrediens One means urenly us oa ve his i- to add ipid-containing extrats and otlherz i*rmvidual ngredents to a base forrniaiion herby' producing a" irkfa formula wih the d~.e:rd nutritionls pfile 'e lKpid extacts may be produced using. onlventiOna extrac.tion so:vs (Lor example WO 94/189 describs a meato or enraedag: sphingomyelin from a pnooliidcontin fat concenrrate usA solvent extraction weChni ns Such lixrsaets RUN expensive to rodu,2C Futhrtro, extcimsive: .oxie' oand safety :tUdis are required bfooze regulatory approval in some jurisdictons owl ;eMOtie for thair :se. Anteens used to acrh:eve this goal is to inciude buttermilk in ifant formnulas. Io numl IS ie aotteocs bys-Dredue U5i Sire7! proUeecd d,~ring ne ofitree pro CCSseS' (1) itiona! biter manuacture ussag eter A he UrtzA buttermaking or batch btemaking process; (2) Tradiional production of butter-oil also Lnown as anhydrous milk fat or AMF) frosn cream as sot in Fg.ure I; (3 Prouctio;n of :;;eryil: frM; cream lsing a so -seva process as hnown in :igure 2 wecre th e buttemilk is produced by blending the secondary ski; and bet. streams to~goher, hnant formulas containing buteimilk contain lesser amounts o: ndeksiabie componientIs of milk tat shan non-uimanj i . bu ii r levels of phosphol'ipids and (glyco)sphingolipidsthan reduced-fat Oairy product However, the levels of these esirabe ipids ase ;ot high.ogh o e o be in formula in order to achieve phospholipid and (glyccj:phingoipid levels similar to those envhman mni k, Supereritica; exxraction; using; carbon dioxide as the sCJVenft is knownv to extract neutral lipids trom butwailk powdersA Astaire J C, Ward R. German L B3 and Jimenez Wlores f. (2003) Conwntrarian af ! axar'MGM Lipids pom ;utxermilk by Micrq||trtH n S;uprrtim!~ ?!id Srsscian .1. Dairy Sc:. 86, 2207-2307 desci ibes the supreriicai extraction of buitennilk using carte *ioxid: as the solvent to produce a product rich inl proi, and enhanced in levels of olar pidsJ. Ho- wever the etermilk powder so produced si hes ow levels of polar lpids. a; a maximum of 2% of the dry powder mass and is thot unstale 10r n toula Cone possibe way tc provide a sirable product wo hl he o seOparate the protein omponenis rom she lipid componerts in a dary product. Dimethyl ether (DME) has previousjy been used in the extracion. of jipid om raw egg yalk (Yano el A US 41j57,404) and dried egg powder (Yano e; al S 4,234.619). The p ocess causes the ;racination ox tAWe and protein omponcms in to Sate fl am in US 4, 157,404, Yano sates tat whil lipids can be exracted Pom raw eg yolks (75% 9OiteUv content e t eroeis re denatured. In US 4,234,619, Yano staes that proteins are not denaired f Vhe eggy i dry, but Yhephosph d can Only 'e extracted WO 2004/066744 describes the extraction of lipids from an aqeous dairy stream usng n er critica extractio where d imethy ether s the sjvent. WO2u 4/Ad66144 also discloses mhat oeitheliperewitieal COD* or snberitieaI dimnethyi ether can~j extract :pi;.b: ieias roem whey protein concentrate ( dairy powders Hiwever, this document doer noi disclose The extractio o lipids frompjwdes rh in milk fglde mfemnbrane material nRmpt' so exract ipids frm dary pwr sears with hgh atose contents (where igh is at least 3r% by nass of the totW A powder) by extraction using iQueMd iMethyl <they have been anmccessfl. t is terefore an oe of mhp venition to prowe improved or aternatsve daiy produas that can be used inos, and/o: to at least provide the publicwith a usefl cHOe. DISCLOSURE OF 4'E INVENTON The present inveniion relates procsses for produchnm daiproducts that have !'xer levet or neuri :ipi o gher eveiS of potar lipids., or bodt These oducts fs n e used as c;tinreiens in infant iorymnias 0ne term UeSweme sous terein means an aq qeouS d '.iry *'gredient separarceoro airy streams contahiig greater tan 60% fat which have heen through phase inversion fioom an oil-in-water to a wae-nil emulsion. Crearm is the refenred starting material fOr tih prodnetion C bets m For example et--serum is prodced during the produeidon of butter- Il (als kown as anhydtrous milk& far or A MF) from cream as shown in tigus 2. Pefierz-albly the d; Ie tt -I is a oVde; thus the invendon provides a procesS fos producing a neual Um-depeted dairy psoduc; Comprining me fAoillig sleps: (1) rovdin adie .ih-a .ow-:actoSe das'y :startin.m. eral (2 subjectins the stalling oaeriat a rear critGca: carbon dioxide extraction process to pr(uce a ppi-depltdi produt, andJ 6 (3) subjecting; the lipid-depjted dairy product o a ligeiddimethyi other extrcton process to produce a lipid enriched faction and a lipid depleted fraction. The invention also provides a lipid depicted dairy prodmt that is enriched in midk "at gobuie mbrane roteins and contains, on a powder Lasis: )/62-79% protein (TN x 6.381. (K) 8-12% lactose, and (c 5-I1% fat ialudig phosphoipcids (39% of fraction). The invention also provides a ipid depleted dairy product that is enrehed nilk fat globuve rnam rane poteinis and contains, n a powder bas: (a) 60-J% protein (TN N &6381 (-) 6-12% aetse, and (c) 5-1 1% rat intuing phosphoiipids (59% of fradion) The nenion also provides a pienrched di- product eomprs:g at least 50% phosp'holiisr an glcoipids handing stantialy redulced ineural lpd. The coeton also provides a VpId enriched d-Iry producQt compr at least 80% tpiospholipids arnd glc.:o)lpids mmnd bein who tialree otfneu:traI lipid. in te decetipton i is speciicaio referene may be made to sbjet mate; which is vt within tho scope of the claims of the current application. That subjct matter should bC ready intiae by a person skilled n the art and may assist W pu-ingr imactice fhet ifnven~ as deined in the claims of dais appIication. Prefrably the rear critical caro dioxide pressure at east 73,2 bar an ihe emperaure is in t range 30412 o 37 K (s.upcriia eion): or to aron diOie pressure isgreater ti-hn oreqal to the v-tapur presue and the temperature is !n the ang 2 7 ro 304.1 K (sueic'al Qe ;n) More prieeWy, He season dioxide pressure i at least 250 ba, and. thi temperature in the sauge 313 to ,35 K. Preferably, the dirnthyi nther is liquefed and pressurised to a pressure at eCast equam to the vapour pressure at the temperatures of te ea orepreferablys at least iW oar greater than the vapour po ssuie Pr.efr-Vay the temperature is in the range 273 - 37.3 K, and more preferably in the rnge 293 --- 3 , The term "high fat" means containing grater than 10% fat ww of solids. prerab;y create wan 15% fat W/w of solidS and mos' potrray greater than 2 0%o fr ww of *olid6s. The erm "owlaiosC means ha the actose content is less than or equal to 30% (n; a >ry weight has') ' ' ''Morf r the tactose coment slss than or o25% (ona KVy wevih nasi Morel4 preferaby. the lactose content K -ls Chan or equal to 20%K (n a dry weiht ais ot p e , th lactose content is less than or equal to '3% (on a dr y weight basis) Pretbrably theace content beta-semm ;s reduced beibre the extraction process. Preferablyv the fctose content is reduced by atrt 2 ''ion .S a uarticulary preferred embodiment of the itrventioni. the lactose consent is further reduced~ by diatiltration duin Q.trafiltration. PreferabIy, t heecise com~en. is teduced to ins than or equa. to 30% (on a dir weigh basis) before the near Ctical carbon dioxide step. More pferably, the lactose content s educed to ess than or equal to 25% (n a dry wer itical carbon dioxide sep. MAore: prefenab.:, the ;actose content is wedued to less than o~r equal o20% (on a dr weight basis) before the Pear criical aron dioxide step, Most preferably the lactose contet is reduced to less than or equal to 1 0% (n a dI wight hosts neer -n.e n ar criticall ca-rbon diox,,ide st-ep. Q7 1 5: 8 Prefervably the hkig-a low-latose dai::y saa;ting mateial is !acteoe-educed )etasemm. Aternatively the high-at Iow-acto:se iy starting niaerial is lactose-reduced buttermilk Preibrably. the high-tat Eovniaetose dairy staruing material is: spray dried before the near crical carbon dioxide step. Ater:natively high-;at Lowactose dairy startcg materaf i freze-dred or vacu dre before the nCea CiticlJ carbon dioxide tp. The lipid en icbed fc oreferaby contains at least 50% phospho:indr and glycolipids and is substantill reduced in neutral lipids. More preferably, the ±ipd enriched fraction .0omains t lezst 8 O'% pOhph apds and giyeolipids, and :s substania3 y free of neutral Upids. In a prefoerred ebodimnt the lipid depleted fraction is riched in milk Lat globe 2mmbne proteins and contalns, on a powder basis 6x/9% (TN x pol X-12% !a'ose 4 5-1 fr invcding phospholipids (4-9% of actionn, mo prefevably t '2% protei N x 6.3) 9% lactose, 8% fat including phshipid (6% If f 0 5% ash *Rand 4% rmisture Described a process p oig a ipid enriched tractorn and a :ipi ddepleted 1 rction comrprising nh ' ilowig steps )prvig a high-o d crOtarting mateiw; and (2) subiecti te staring material Yo a liqu id dimethyi ether extract process. The lpid depVere. hrcho ctnt, al of the m0" fat globule membrane roteins ad therefoie could be used in nnz formulas to deliwr health benefts that are more strongy associae wi-~ .th milk far globule memranare prosein coemponents. Prefrablhe dimethyl ethel is liquefied and ressur:sed to a pressure a; least tqua: to pe or persiue at the temperature or the extracticit, and more preferably is at teast 10 var reter man the, vhpu pres . Preepeau is m h 273 - 373 K. a r prefery inthe range 293 -- 353 K. The e high t means conainng greater than 10% fat w/w of solids, preferaby Irete tan 15% fat w/w\ 0f solids and rns preerbl greater than 20% fas w/w. of saolid,-Adss.'io Th 1t "low-aCtosC" men that the atoKe caomei less thn or s t r (on C arowwn 'ass t 14cre oreterariv. ne :acto~se coerini s less than or reala to '25 (on a y weg.ht bas'is).ore preferabl the lacore content 5 ls than or equl so 20% (on a y gt *) O porerersay. the actos content is less thanor eqju o 10% (on a dY 'weight bMsis0 Se acse content of sea-sermmi reduced hefere the ertrnction process. Preiraly' the lacrose cont ;s rednce in be rm or toe stati nateial by ultratttration, in a particularly preferred embodiment, the lactose cnitet is farther SeCIeed by d~ jitation during uhrafijrsatio: Preferably the hg->at low~actore dai sartin *material s Iactose-reciuced betr-serum. Preferably the high-fa low;-:'atose airy~ starn mate.rial is spray cxie before the liqiuid dimethy'i ethe xa ion step d Aterntivelyigh-ft lowx.-lactote dr start e is tree-dried or vacom dried bee the iquid dimet.y ether extraction 0ep. In nc- cmbodimset, the lipid enriched action cOnirans at c-ast 40% phosphosipids and glycolipids. This process does not take out the neutral hpds otr wit CO and because DME extracts both polar and neutra! lipids, the DME h etrct will cntain about the a ifn'eural ilpid-to-pEhosp p a in h ed. For examoples o a a ted coninin i eut'tral ligid to 911h9±;h0110i0 ratio of about i the; comcosion of the ivMI 108 ipid extract ona 3% mois ore basis ix 8O 90% &: including phospoipids (35-43% of rtion), 5-9% ash. 25 lactose and 2 o misre. INe hpmo cepcta fuacIon in enritned mn fmlak gflotmmbrane ptemtbOs ann contams, o; o p wde basi;, about 6040% protein (IN a 6.38). 6. 2% iactose 1'j% t including phophoipids (5-9% of fron)L preeaLY 7 p;;otein (TN x 6 38) 9% inciose, 8% fa icuding phosp4oridf 7% of action z% ash and 3% moisture. The leve) of poei in the products described n t pecification is alculted frme low!l nItrogen (N) in a ample muiied by a facr of 6. T i nenon aso provides products produCed by the O cs of epreserf inventIon, and CmpositiOns and infant formulse ccntainin suc producis. The krrm "infant ;nIbiua s usee! herei:; ncludes forrmuas dsiged for :in's 0'2 monthsl, fll rmuas designed for i 6-ts 6-12 months old (flov-on-f:rma) and formuws; desige-: for odd4ers and young chdaen 1'-7 Yews, gnrowimgup miks I ri, poewders). Preforably h ei invention provides an ifan f Aa comprising ()30 - 60 o V'e'os (b' 15 -- 35% vegtabe .:is .j'd-tim $h..' iOk w (d - whey protein concentrate (c I -- I0I pae prodIce by a process of the present invention. Mr prfaby the invent n provides an infantformula comprsig: (a0 -- g % (b0 -0% veabl oils' I11 (c) 10 - I3% skim I odk (d) 6-- %rotein coneuatre (WC (e) I *- t5% a perc produced by a pr ocess4 rf w pesent invention. The unfan:tformnla may also comprise 2 -- 4% of at least one of the §o oag ( ) vitaminpremix (bmneraj remnix (s) stabilser (f nueonides. In a preferprd embodiment thee infant formula may ne formulated to provide eesween 2700 end 3000 k/f The product .cposc;2ions ande inieam frwuia of the present invejntion may be adminitered to provide booth tenets. For example, the ;ollowing elt hbenets aire eonknmplaied: -- reducing the risk of iection modilying gut nteslin] owrn and compeiAively binng wgiei.ns -preventing inflections present the deveent of all-"gies prevn tii cad: colon ca;cer. 5 mom i1(55:9wi TIhe invenors havc discovered that the ieves of phohoipids and niosies i serum~ Oma it suitable to %e :use~d in the fortitkation of nfan formuas, The snventors have also discowrod that dairy products which arc high in lat but o, in lActosO (icluding lowlactose betaernum) may be processed to reduce the jewels of neutral lipids. or increasee the: level of polar lipids, or both, thus creating products wich are e.v.lore uitbie n the frication of in fanr torrmulas Thee proceseprocessing and extraction techniques which do not S Iave toxiC residues, ther efore f urther processing of the fina ttair produce I nor required Additio~mlly. the use of udtrafdnUadon and near CetidcI extraction wiv t a,:bon dioxide as the Qotvem means it should be easier so obtain regulatory approval tor the use of this productt as there is nimal or no solvent reside in the product compare: wit te use of owentional solvents ruch; a acetone and ethaioI A diiona!iy convention so vents ex~tively denature proteins, making the use of these solvent; unsutable for produc;:g A*~ '6A K 'the tu "day" as used herein means of containio, or Concerning MiR and its products it inldes milk produced by humans, cows. buffalo and goats but is not tjmrted outhese animals. Every substance has its own critical" point at which the iquid and vapour sate of the substance becorne idemi*cal Aove but close to lhe criucal point of a substance,th substoace is in a fuid ste Oa has properties o; both liquids and eases. The had h density simiIfar to a liquid, and vscosity and difusivity similar to a gas. h tm "prr a js usl.ed h it rs to th presure-hemperature region above the critical point o a substance. The "sucritica;" as used herein refers to the pressure Vemperture region eqtia. o o; above the vapour pressre for the iqid, ut; below the criical temperature. Th- term "near critical" as used harei encompasses both 13 "uectc a s'rc a regions, ad r to pressures and temratures noa, Me ctcal POW, BRIEF IDESCRIPTION OF TIE DRAWTING Figure I slows a vcheantic drawing of the tradiiona process for producIg butter-oii from cream and the production of buttermilk ung tis process. Figure 2 Thows a §chematiC drawing of te process for producing butteroil fro:m ceam and the producton of serum dring this pocess. EXAMPLES The following Examples turter illustae ractice of processes described herein and the Examle Exracion of vhey prouliu concettate powvdr This exape shows rhat die extraction of lipidS from po3wer ith 1k:gh concentrations of whey protin eu in very oV yieOs of ipid. Why protein concentrte powdcs extracted w,'o; toe ness critical soLvenr carbco; dioxide, popane, and dimerbyl ethet (DME). The schvent, pressure, teroperature. mass or solds used, masis of solvent used, Tibs 1:Lipid yields for the wxrctk;o; of WPCI soids wsith varsios .sokefts Svent Pr -essure Temocrature Mss t Mss of Mass of Yi V d. br K solids solventr Exr;rc.% % 2 use. k V. solids. ds C 3071 361050 9 0P e 32 s 4 00, 18 9 .5 6 DME 32 14 ~ 26O0.0 10.9 A I .I DV 5 5 323 29.1 0 0 (4 0 26 3. The d ieds are very ow snd a nWcease ir extrac"ion tenperature doe t Inras 3he exmactio; yield to desned evels. Ex "ue2 natc of sit dard 1:eetose' beta serum powiders 'vith supre-rtha This exhale shows hat extraction of neutral uiis is possible from bea scrm powder WiSi standard acse cotenit but ihat be yid is signincantly less tan vih powders whbe:e the rictose cont: em hasbeen reduced The protein and total pospholipid conitent of T funal powder are w Bta strYm powders with the following c sitions were extacsed with superer C a 300 bar and 313 K: batch i tta! poien 294 %, act.:e 42.5 %, tota! fat . moisturee 3. , and ash 6 %; batch 2 aOal prAs 3 %, cOs': 44.6 %, total ft 20,6 %, msie 2 1 % and ash 6.1 %/6, he toml Jt is made up of *n'utral tmist phospholipid5. g a:nglioides, ceramide:: and cerebrotides, such a laC1.yscerainmde. The fat eXiraCtion reenlrs, and masS of pnurSa1;ulipids in Ule extra tar sow" i Le 2. On neuu-al lipids tr.ctd by Supertr1ucal CO as he ter types c fat. and Cspeciaiy phospholipiL are not soluble in this souvent Table 2: Lipid yMds for the extracting of standard lactise beta erm w with CO" .hBatch Mats of C. as' Mass of % e d % Yiusie, so used ofsKphol p d total ipid neutr xtectip uds i a 305 AM e4314 72.3 -- - - - - --- - - - - - - - - - - - - -- - --- - --- -- -- -- - -- - --- - - --- - i s.'4 7 1- 3,0 (15n ~lf6ne powde com;positons alter extCj0on were; uarcn roral proTein 320 %e:et O t. total fst 13. metiur: 3.8 and ash 3 ' batch 2 tob] protein .; 02 os44.2 total fat .3 , mosss e 1 %3 and ash 63 % Ihe powder from batch 2 was tested for whey protein denatuation. It was assumed that e" casein prot~es were not dejaftured. A representati samp o powder 's taken, and Smith water to give approximatey 3 % whey poeis in soluiorM The caseins wvere pcipitd Iat pH 4 6 wit hydrocnjorc cid and removed from senttie, Cnr' IT. composition of the rem:aig soluble h'ey proteins was determi o reverse chase chromatonaeuhy The solubl wvroteis decreased m 3.43 0 of oten n Oe to 839 =. g or p'tiI ik the ehSa powoTt. itir was a very large decrease in natne (undenare.) et Latoglobulin. Denaturation of the protei r:ake% he powder :esststable 'or infni frn nua bhan those products described in Example 3. EXImpe 3:' Etractna of low etose beta seruni owners with superetitiecl CO2 This example shows that extraction of urrai lipids wik greater thaD: 90 % yied i possibic trom low lacaos eta 2mai powders. The reduction in lactose content of the bea serum was carried ou by uHrafiration to a volume cnettion factor of 8. The proteln and total phosphoipid contents of the tnal powder are high .Low lactose beta serIm power wa; *he following com oios wre~ 3 extraotedi wt sup~ererideal CO at 00 bar and 313 K: batch 3 low lactose total protein 4X3 %, L ose 14. % total at 3.%,ms ue 1% and ash 4.8 %; bach 4 ;omal p .2 0 '.8 %. tote: It 3A %, moistr 27 3 %O and ash 4.8 %t The far etracs rsuls, and mass of pospholip~ is oh extract are thown: table 3. ly ne2r' lipids a.rexacted by supemrcrical Co as the other t lype of fat and especially phospholipids, are not solum in ths solvent AS1'2§59 16 Table 3 Lipid ykAN for the extraction of ow lactose beta serum powders with C02 3 698 I :99 0857 6 902 lhe powA- com Op e xt o r ofae 3. %, Itta 'at 18.% &oa ph ooipid 4 itr 4 % and ash 5.7 %; bach 4T rotprtemint~ 'L6 %,. acos [ fl %, toa ';t2 , total pohioiipids :6.8 %, m 4 '% and s 16 % B ad x exrcd with siuperitia C also had enane level of ganliosides at. - 1 % y mass Th .remai ningr differene buete; the total fat in thie residui powder, and the p.kospholipid and gangloside content. is made0 upio mostly ceramides and cerebrosides, especially iesosy icerarnall. Theu powde: from atcheOs 3 (low lactorse) and 4 (veny lowy lactose) wec;e tested tor protein denatnration to ensure at it was uitable for' use in infant sorraua as per" exap 2. Th sonkle whey proteins insreased from i 2.20 g/100 g of pro:ein in the fed t 13 /100 g of proen in the exrracied powder for batch 3; a;nd fromn 12.44 g 10 g of poei ih feed to 12.94 s/ 1 00 g of protein in the exteracted. powder for bh 4.' Thelc of denaturation of the protei and the high proeinan pho'sholpd c:ontens of the1 extractedi ow lactose powder;snmake themii very suitable tMr infant formula. ---- -- -- 7- - -- -- -- --- -- -- - --- -- -- -- --- -- -- --- 5- - -- -- - -- i- -- -- -- - Example 4: Extraction of standard aid !ow sMtose beta sera ; powders with supertritca CO2 faowed by ;seAr critical dimethyl !ther This exampleI shows that the extraction of phuspholipids in ig yiejd ttm bet serum powder t has been Jptracted with suemcritica CO:. is on)'y possible when the those contend of he power was keen reduced; and that dimt;y; ether exacton temer 'Cr induences ame etraction yed. The& zxmple alio shows that it is possi:ie to contol he ina phsphlipds ontnt n teIpwder after etcinbycntrolingth xtractin 4Tperathre. rtilly deft powder barches 2 standardd iateose content. feed m s 318 7 g), 3 4,w lactose, ee mass 2952,6 g) and 'I very ow iact , 10d mass 2668 aa.2 produce n exanmles 2 and . were re-extrcted wri dinethv ec 40 bar and 293 K Usn 12w36 3 3 >828 and Yl 17 kg respecVtcj and then rextraled %w methyl eter at 4: Car and 323 K asing 13.037. 30962 and 65 kg respectivy. The enancfion y %ied resul s are shown in table 4 Table 4 Pspholipid and totai lpid yields from standard and ow m etose powders utgdmethyl ether aer Mupercritical CO2 extraton Batch LA tose DM extrascion DME extraion Overall Jiid v id coridenht v eds )t 29 K ed '., 32 K Tora % V it o % spho Ta pid. Ph;mose pid g -hfsphoh pa p vp'id 2 479 6 6 3-4 5 7 42 1 The.. total fipid 2 e ,,r o coie si n fLiL 'tvels of ga0o' t25% yms for batch 4 'Cf Kh an 1 by Asx *o a..en 4 LA, e prtin conem 10 1 al powe .. incese rektive tJU t.h fee afte; '"rnthy eine ex. (erio) Jhe powder -7- ---- ----- §- -------- --- 18 onpositions afer C02 and dimethyl eer extrttions wer: batch 2 total potin 34,6 %, lact§e 17.1 %, total fa 8.9 41 toal phospholipid; 6.3 %, a.isture 2.7 %, and a:h 6.7 W atch 3 Mopen 44, %9 a 7 %, to;ral ?a .4 . otl phospholiidr 57 %, moistu~nre3.6 ., and ash iW %; batch dl total protein 732 %, lactose 8.7 , total fat 7.6 % moisture 4.3 %, and ash 5 %. Bot powders had significant levels0 ofngl1Isde5, at approximately 0.4 % y mass The remnng difference betwen the totl at i ih residual powder, and the o ipid and gagl!side content. i made up ofm Tie powder frm bates standardd hacrose contem3 low mzcose) and 4 (v1y ow *aclose) after suercrcal C,2 and *ioethyl other extr Veae: wore tested f1r protein t nper example 2. The soluble wey proteins decremad fr 3.43R jW g/ 00 of protin for botch 2 to 800 g/ 00 g ol prohiin in; he DME- exrac:d powd. The .o[ue whey prt-esr ncmasd frm 12.20 g/ 00 g of protein in te foed to 1.23 g/ DO goflmofl n the extraced powder for batch 3': nd h a 1A44 gm00 g of ProteNr in h. feed to 16t g/100 g of protein in the extrace powr for batch '1 Th lack of pIn lenat uration and t h e and pOShpct f h xt lo.t I crose *p;wders rmake them; very sut;ablefo infant formula Etrafl;ctiojn with.. dimty eteJa had the nxpuected fie of in aing th appalem wh-> protch: Solubility, w iniia:ly diminished by the removal of lackose f the feed. Examp5Ve 5: Extraction of staad and low lactose powdvrs wth dimetyl her This e:xaple shows that dhe extreenon baf ot na cip and phosphoipids in ig .- eld An-m bea semm powdr is only possible when the lactose content O the powder hs bec- reduced when using dimchyl ether as dhe Solvm without previously e-ttag he powder with superrrK-al C-and Ih at dimethyl ether extract..n..emperakure inluncens the extraction yieli.The example also [hw a tar it is possible to control the fida op;ospholipids conter in the poder after exactiton y cooling the extraction m'perature. Batch 2 (tranduad actose content, fead raas- 42-6b g) w composition aS nv in example 2 and btchs 3 (low laesnsc. Ceed ass430) 7 .5 )j nd 4 (very low 7H10: 19 cosw food m's,s S204 4 g) h mpJi given i x 3 w vxrc wit dimethyl eter at 40 bar and 27329 K using 1-3.426, 12.666 and 1393k ecvy and he :e .-exacted w i -the 40 bar and 323 K :sing 15727 1 ]673 and 1123 k ctive . Tc xtacton ,i rls arc shown a abic- W Table 5 hosphoipid and !ta fl pik yelds from standard and kow lactose powdes Wig dieli I e r 2'k3 K 2;;d 723 KC Batch Lartose DME- extraCtkm DML extractionl tOverri d~g cei, d % coent yields ai 293 YO yieldsat 323 R % Toa % Tota % Piosh Tota p d Phospn pid Phoisphab nod) Op pid 2, 44 6 ,-.4C 2 L8 0. 27(2 2. The preei cor~' ent o. n!! powdrs inrae relat to rhe feed ater dimety te ex trac0io. The~k powder composif'tions~ Staner, . diethy eher exxau wee batc 2 ta protein 34 8 %,* atoe~ 44 2 %, rosa] fat 16. pThonolipids 8.3 %, moistur 2. %, ad ash 6,2 %;btc 3otai protein 6li %, lactose 113 %6. toml fat N.3 %, phospho''id 6 7 %, mfoiStur" 2. and sh .3 %; batch 4 tota: potei;; 73. %, lastose 8 , oa fat 8.3 %, t.')otalIpospLd 6.8 %, meisture2 6 %, and ash 5.2 %. F'or barthss a--4 no he dffrence henveen the totanlfax and phospolipids content is made up of gangiiosides. KceramrnOdes ared cerebred~des. The pouwder frm hatches 2 standardc: lactote conteno), 3 (low factor) and 4 (very low <aose) after dimerhyl ether extraction wxere tested for 3potein deraturation as per exnmrle 2.Th solnble w'hev nrotwins increased irom; 1343 p/0 @9 gof1 arowe: to 14.38 gGOg for batch ; from 12.20 g 100 g of protein in the bred to li47 g 196 g of proten in te eireacted powder for hatch 3, and from J2A4 g/i.00 g of protein inx thfed to
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5 55 g/ 00 g of re 04 0h xck of protein d r ad h high;.rotein content of the! DME3 extracted lo' jiose powderS make them sutale'o a ide range~ of food apihcations, espec.ally sp8t mnrti. Extraction with dimhy- Ctr has had the unexpected efMet of incre asinge apparent whev 3irco - .i. which- 1r sttai diminished by te removai f ate hom the feed. The extration yield of toal piS and phosph osis very Low Ior powder with g e;-actose coments (bajch 2) when Using ity; ether alone s the exir(ction olivent. The gc ? neurlis 4 kes.this owdet less sitable fOr f ant mula Fnample 6:. Lees o phosphdpd4 oand gangsiosimes i; selected dIry prod e;Cm T'e popoipid contents of human mek typicAly ranges froM 200-40 mgL (Jensf n R: (1989) Te i Gore oy n o iion, I Leventhal Ld, Raven Pres L, New 'ork, 157-208). Asccorli to -Harzer 0, Haug M. Dietich I & Gentner PR (1983i Changing patterns of 6.(Aa lk jipids in the course of the scitmown and during the day, Amerwcan .morna! r), Ci/n/ca ANatrib. W7, 612-62. the phospholipid composition of human Ok 36 days postparmu is phosphatiylchomnne (PC)- 24,%, phocsphasidyicthrnoiamine (PE) 27,7%, phosp dysn () 9 3 hosphtidynsitol (D 4% and spingom Table 6 :ihows the phosphoipid contents of: - lo lactose bea-um I powde: (prodct B a -rneusrta iddeplee bet seru:oowder (product C)f and - the :1ipi extract D and residual powder (roduct E) ot product C after DME extractin Thes produacts are derived from boevine milk.
21 Product~ A (bet-semmlf piowder) was prroduced us;;;g the method iostrated inl Fig9e 1. Product B (ow lactose beta-serum owder) was produced by ultrafiltration of ;rot A. Produc? C wads piroducedsinge process as descried in Example 3. Products D and [E were produced using the process as described for bach 4 in Eaapic, except that a single srag DME extraction was carried out at 15C. The HoAN lip contem was measured by a modified Ruse-Gottieb method where the lipid etacss were vacuum evatporated and freeze-dried ef ov-en dry'ing (3ow tempneratutrn drying mAinwiies the phospholipid hydrdysis that occurs ocrin Oven crying due to the presence of ammonia in he pd exuacts) T 'oa phosphoipid cone w cuted bymuiplying the phoshoru ~onent o e' rmodied Rdse-Gottieb t extract by 25.5 (er McDowl AKR (1958) Phosp - pid tn New Zeaand dairy prodcts..um f Dairy, eseach 2z 192 -20 P..- X- u idliial phospolds wr measred by f-P-NF'Y Pan XL. & mzar T OPO0) VarIanonf of lit egYgl;sid compitions of hmsan mniis c ow smlk and .am muias any Jhua Deveopmer 57, 25-31 snow mat iagdiside GD and gan gliC,, GMv3 accout fo abiut 60% f Ith oa gnuesides in h man il and -h- th total gatnloside .4 cnn ofim frnml, ias meased by the ijpidmbound sCAlic c ( tent, IS sigifdanry ow her ha tat of hunan milk The content of gaglsde GD3 pju ganglioside (M in human -k ranges nm 10-i6 oig/L pendingg on tie stage a(Nakano e at, 2001 Sialic in uan milk: .omoshioni and fmsetons. Acta !aarica Taiwanima, 4., li-: ble 6 Thvws the ganglioside GD3 and Gioside G3 content: of product A, product 8, product C, ;-oduer D and 7-du E. The candioside GD3 and eaeghoside GM3 cvels were cakculaed as follOws: samples were dAlved in clorolorm/mthao/waer 6:3:04a filtered. The gngyosides werae ten separated into M3 and G3Lfractions y s ong anion excnge sod phase extaction and s'acnc acid quahed by h Resorcino' method of Svennerholn (Svennerhoin L, 1.95'7 Quanitative estimtio of sai -C acis. ;1. A cometri ;esrinol--hydroclorie acid method. Bochwm Bispevs. Q0 5: Acfo 2:6064l';. The s:ialeacid waey2 wre Wen sed talcite the concentrations of .. MN3 and GD3. Table 6 -PMr hpid compositions of producs A-C mpe v; A R C D E f t ph1sp ;"p * 7 5 66 6 o PhKshatidyveahanolamina 2. / 8 22.6 L0 Ph.paidlerT .2 .3 Phosphatidtvlinesirni 9*. 1 2 .1 0 5 Spliin 1[o flelin 4A 15. 1'; angliosid GD4 4 0587} .6 2.o 102 Exampwe . fnfant iormulas cntiing sekctedi daivy produt Tal 7 shos the percem~age ofa ea product (A B~ C, D and EI ha ned toab added ro infant tormuk on a powvder hasuirs n ode to increase the nota gang!eside (gd~giosie'D p!as ganHtuliOee GM?) content of read's-s0--Red (PTU) isfats rnrmula Thne assumptions are that the asenne lavei of thest comoponents irn utandard infarnt ----- -% -- h-t sh 7ns-; -------- -
---
rpowder is reconstituted to 13% totaO so.ids PO3 1hat the densit of the O r F is 1.6 kgE The beta soern product addhiun res shown in Table 7also increase th individual phosho p cotents5 of the nfant forrni to) lee garaer than Those found in human state fromf measuwad vaues for i.i.BSP. tmlatd rm mean values for standard beta scrm por. o maed These are ehe resuis of another BPC60 esmola * 1k th only C e eo being prodc E wer ahdded PE and Pt levels are s1~lil o han tho-' und i- ""mn (se levels ofthesecoonsi standard infant ff4rmula would probdably compensTi for theseY siotfls.1t [a produce! D) could be me jinsateadhi of' so lcihin wich. .s commer; used to Fanmise infant fornmas. 1310 lip pirojh", VG, Tabe 7 A d ek of p&air lipid cmeponen is s RTF F at 13% Sota s&4dt Phspatd anfl 9 F 6 5 Ph'ospamid nos f 26 2740 2 Pho---------- C, 5 3 7 Sph ngomy~e n l8OS 99 100 10. *agotGD M 16 jt 6 6 Addtd neua ip ( NA . 469 76 (9 7 Efanmpie 8: 1faL foI d Rsadyv-teeftd total 5o2ds 50% Wheyl~-ro-c§:sein ratio =A 64 4 (whey pren eat least 60.0% of toMa protein I Prtaein target" = I Mean of range reported by Jensen (1989). indivdnal pThcsphcorids are calculated based on thi ii canr value- and the pesrcentages repottd byHV e t 18) "\Netri pid*, :s estimatedi as the dtueience between the Fotal lpd and phosphelipid T his as, *howe ver, anl :versstimata as in :ncude the g:ycpid (sog csrsmidss, gsngic;idet). To put the adoed amount of nesutra: lii ir eerpective the ANZF:A range fr fat m RT is 21600iC4ll0i mg/k i . e :mpact of added tra i ipid on the baanced fatty acid prols leo F as emsi!, pediculady wdh prouuts C D arid E.
Ft tre =35A-37.6 g/L Carboydat targe" = 721- 72.9 g/L Thee target levels lco, &he ANZFA energy requirements of infant forimu (2700.3C0 id/L). 1 Thdie twen h su of thCe po0n 4arbydrate andl fat levels and the L3% jotal soids carget was umed o the vitOa and mineral prMixes, aandoxidants chin ed to 'rotantse t'nlfaA formula),3an posiby nucedes/mlbsleooss. Ths compone'n' ts typically amounteded to about 3% of the powdeOred ;inIntfomua The oi m- used in preparing fowulas iypically comp.s a.blend of egttable k in orer to achieve a fatty acid nil close to that ofnuma wilk. Vegeablet o that re common used in nfant formula are higi nlic & paie le. h0gh Le sunilower oiL high l safiower oil, coconU oil and y oft rhermore, many of te premium *ods also contain fish/microalga and fIngal :ile as sources of doco:sabexaenoic acid and erS:nbidonic acid resectively> ifnt i o multa (Using beta serum pow V - peduct A): A nutittomnd formula a comprisn a, About 46;4% lactose - Aboud. 2692 oi ui.x (comprising 45% high oleicoal olain, 20% soy oiL, 20% c oil and /1% of either high oleic saffkoe o' o high About I3'% sk im milk powder (SMP' 1 A bo 1 .40 ALACEN 392 (8A% pro.min vey proei conemate) v. About 3. 5% beta scrum powder (product A) f. /Abo'ut 3.00% v mi and mineral prernixes, lec, antioxidants/abilisers optioal comp;ne~nnts e~g. n::clides IBased on Similac Advance wih li:on and Enami LIN with iron levels, noig; that th Enmii . P!Lah. i rn eve vfs were reported as g per 10 e (c g0 ) that a R T enery contend of 2300 kJ/L for this pcdudt has been asumed. *Estimated levels for Similac and Enrfami: oroduct s spectiv&ly iatimt formula 2 (UsingI, iow lctose b Ita serur powder -produc B): A nutritUnl MOM comprise: a. Aboet 47.9 lactoI 'vse b. A'e bout 2 %o'il mix t. A bout % SMP s About 6.I4% ALA CEN 392 e. but .9% low O-c{osC beta serum powder (prodSt B) f. About 3.08% v7tama and mineral prernixes, lecithin anioxidants/stabiliser;, optional components *:.g. nucleKtides In"ant formula 3 (sing - cduct C1) A nUrionm Iorirna cotiom Aboui 47.'% Jaetos o About 26.2 Ai mix c About 1615% SMP About 6.54 A L ACEN 392 e A bot i.7 . produce C 2 About 3. 7% vitam;n and minea prmxI es, Iecithin antioxidmrsstabilisers, otoa componens eng. nucieotides lrsfant ibrula 4 (SIne epod UC' U): A\ nurstional formula comprising a.About 46.34% lactoec u A boel 26.92)% 55jJrnix c. About 16-5 SM? SA bout 6.77% Al A\CEN 392 .About .5% product D f. About 3.''% vitamin and mineral premhixes. lecithin. asioxida;ntstbilsers otional c;OoneCma eg. nucleotides inihint o:'mula a (in prod1 uet F): A r3nsitiontal ~ fomdacmpiig a. Aboui5 '.77% lact:s U, A bo.t 2W.92 *il rd ix .A bo-ut 82%SMPF A About 6.u0% Al.Af GEN 92 c. Abo 4,40% product E f. About 3.68% vianin and rnineral prmies. lecithin. anmioxidamr/sbiisrs, optonad componems .g. nueje& nd&:s Thie ave~ exmple Ore illustrthons of poetCe1 6f the pr!ocesses doeried herefinf nd the invention, It wll beappreciatedi y those ;kied in the art that invention may be carreid 01ut with numru mvanations and miicarons. For example temperatures and nres'rrs '.of exira;L;ons in y CVIarIed as Can he prole n d lactose eontei:s 'd stai maverias. An, it will appciated that the dairy produces of t psent invention may also be ei product os or en t~ologcal or general n uoal benefit in the consume. nc;uding sports uition and iod for the elderly The Lnrm "comprisin asud in UhiS Spiao and claims means consisung ata mn part of".v When~Qv inteprda statements in tosspecistion and claims which include th;e "comnprisng", mother katures besides ihe leatures pr-'acedb' ti tr inl eCht; statement also be present. Related terms such as "comprise" and "comprise" ate to an inprted In similar manner. In t!WS speciteaton where ;ciberenc- has been made to patent specifications, othci externa documients, or ozer sources of information, this is generlly for die pupose of providing na contexi for discussing the features of the invention, Uniess specifically tated rwse reference o u ctem doceni is no t. be consrue asc an - iO ha~t such documents, or sach sources of inforunaaion, in any jurisdiction,. are prior att or form1 part of the common~f gene:'a: kiowltc' in the art