V61．5，No．1（1953） ∼i
STUD王ES■ON SO■yBEAN CARBO員yD．RATES．
丑．The Carbohydrate Content of Defatted Soybeans．
BySin，itir6KAWAMURA
（Laboratory？fBiologicalChemistry） （Received May18，1953）
INTRODUCTION
Con転afytOOilandprotein，Carbohydrateof soybeans received comparatively
Slight attenion by chemistsu
Earlierliteratureis here reviewed chiefl皆 after WE‡iMER（1）and MATAGRIN（2）． Theearliest
by L飢7ALLOIS（3・4），Who reported the existencein soybeans of9−11％noncrystata11iz・
able sugar m声t？rial（une matiらre sucr6eincristallisable），Which produced atetic acidand mucic acidinstead of saccharic，0Ⅹalic，and cassoic acids on oxidation with
nitric acid（2，P・126〉．Soybeans were thus considered to contain not only“cas占in”but
also sugar very similar tolactose（ingiving rise to mucic acid on oxidation），and
aqueous extract of soybeans wa声really、taken as〃vegetablemilk〃lThis noncrys・
tallizable sugar materialmight be called“galactan”／see STREET and BA王LEY（5））．
MEISSLand BlbcKER（6）reportedin1883the foliowlng analysis on soybeans：m6istufe
lO％，StarCh 5％，Sugar12％，dextrinlO％，a平d cellulose（％not given）′（th拍u盆h
WEHMER（1））AIso HANAUSEKand HARZ（7・S）d占tectedstafchin1883−85，butlユnder the
microsc？peatagreatenlargement，andsampleslice苧Shouldbeimbibedinoilt6see
clearlythe90lor reactionwithiodinelIn1885Harz〈S）“foundthatwhenthebeansdo ト
not rlpen thorough1yor wh占ntheyare allowed to rlpen after the vines ar占cut，
whereasifthebeansar’e thor’Oug’hlyr享penedtheyarepracticallyfreefromstar℃h”， according・tOSTREETandBAILEY（5）．In1886 thel）rdportionJOflO％ofdextfIinrepbr・tedbyMEISSLand′′BbcKER（C）：Jirl：咄eir・・革nalysis −Of soyb占an岳●con畠isted・r・ather，Of′inctystallizablesugar∴（PrIObablyrg幻ac七an）｝′，、As、tO／the existence・Ofstar℃hfmenti、onedaboveBLONDEL（1888）（11）wasalsosuccessfulinf如dipg positiveiodine′reaCtion，WhileINOUE（1Z）；P衰INS王加く18ノ）and BELTZER（13）ノfound・tbat star・ChWaS・absentinsoybeans巾
Much suCrOSe WaS foundin soybeans＜by．MoRAWSKIand STINGL（15），and．by SdHULZEandFRANK如RT（1や）．．▲Accordingto PRINSEN−GEERLINGS（1子）：SugarIisabsent．in fr・eSh beans．，We owe to TANRET（1S）inknowlng・that soybeans contain stachyose≒aS wellas sucr・OS占．，kaffino占ei占去1so containedinsoybeans（汚）．
LiterIatur：esonlhemicelh1losesin℃1udingthose −1909）willbe discussedlater。
Tech．．Bull．Kagawa Agr．，Co11．
2
′ヽ OSIMA（1S）found thatJapanese soybeans contained a traceor・nOne Ofr・educing s喝ar，mOr＄：1han“4％df■sヮc竺■ds？，abbut5・％ofgalactanconsistihg′0′fethanol・SOluble
andinsoluble matter，a traCe Of starch alsoin mature soybeans，SOme pentOSan，
and muchless．叩寧t！lylpentosan．
Quantitative data on the carbohydrate content of soybeansin theliterature
maybesummar・izedasfollowsナ（Cf，DAUBERT（ZO））‥ SrREEで （5） BuRREL （22） FiT・St autbo工● Reference 0．0053−0．229 4．0 ・−6．39 0．．86 −−1．12 2．39・・−553 0 −−0．1（； 2．51 −3．24 1“15 −2一．06 1 3 1 03ノ1 3044 Reducing sugars Sucl・OSe Raffinose Stachyose Dex也in Starcb Araban Galactan Ce十5 a r t
Thesearebased叩Ⅴ卑riousassumptionsandwillbe crIitically reviewedlater
in pertainingSeCtions．
Her・eSOmeStudiesare亨・eViewedr・elatingtothechangeofcar・bohydratebyh占at−
ing・．TAKEMOTO（23〉found that heating of soybeans at1700for251−30min・decreas・ed
moisturIe，nitrogen−freeextract，andcr・ude fat，and thatheating・inducedthedecrease in allof totalsugar，reduci咤＄ugar，SuCrOSe，StaChyose，gumS，and galactoaraban owingtocharT・ing・AccordingtoKAWANO（24）totalcarbohydratedecreasedbutreducq
1ng Sug・ar・increased slowlybyr・alSlng・rpr・eSSurefrom atmospheric to overpresssure
ofo．5・−2．Okg一・KAMIKUBO（25）showedthathydrolysisofsoybeanmealbyboilingWith
20％hydrochlorI蔓cacidchangedcarbohydrat？（30％ofmeal）t6iluminsubstance20，
for・micacid5，1evu1inicacid13，andmelaninpigmentl％」
InthisreporIt the author，s experiments aredescr・ibed whichhavebeen carried
outtoknowthecarbohydrate．contentofdefattedsoybeansl・The experiments onthe
sugar・SOfsoybeanshavebeenrepor・tedformerly・（26）
EXPERIMENTAL
Many analyses hitherto perfor・med by the author show discr・epanCy between the■values fornitrogen・freeextract andforreducingsugarISafterhydrolysisofsoy−・ beans．AllcarbohydratesglV占reducingsugarsafter・hydrolysis小Theconditionsofhy・
drdlysismayaffectoptimalyieldsofmonosaccharides，SOmeOfwhichmaybeliable
to further・decompositiontononreducingsubstanCeS・Allmonosaccharidesmaydiffer in the extent of reducing the Fehling，s solution・Moreover・，nitrogen−free extract may contain noncarbohydrate compounds，SuCh as organic acids，tannins，COloring
matters，and others．
However，allthese＿COnSiderations may not sufficeinexplainingrathergreat differencesalwaysencounter・edinthese twovalues，nitrogen・free extract and total
Vo 3
The conditions of hydr・01ysisindetermlnlng tOtalsugars afterhydrolysis were Studied（Experiments No．1and No．2）．
Thenroughfractionationof car・bohydrates was tried bysoIvent extractions一丁 ether・，COld water，hot water，ethylalcohol，2％hydrochloric acid，and80。％sulfuric acid・−aCCOrdingto themethodofWAKSMANandSTEVENSく27），aSSuggeStedbyKLEIN（28） （Experiments No．3）．Supplementary experiments on carbohydrates soluble阜n cold water（ExperimentsNo．4）andsolubleihhot90％ethanol（ExperimentsNo．．5）follow、 These experiments（Nos．2to5）were applied on the same sample defatted meal．
Separate samplesserved for experimentsin determinations of galactose and galactanbytheoxidationtomucic
furalyield（Experiments No．7）．
EXPERI∬EⅣTSⅣ0〝トTheCoれdibmsof恥drolysis五n也eDeterminationofTotai RedllCing Sugars afterⅡydrolysis．（。Ⅰ）
Material．The defatted mealwas obtained by extracting oilfrom partially
defatted soybean flakes（T6setu）with naphtha（boiling range60・900）at＝50−550．This
COntained moisture13．28，aSh4．68，Crude fat2．00，Crude fiber5。12，Crude protein 46．57．％on analysis．．Ther・efore，it contained28．41％nitrogen−free extract，Whileit COntained only17．20％’（ノaS glucose）r・educing sugar・S after hydrolysis with2“27％hy−
drochloric acid．
Method．The standard method ofhydrolysisis to add20ml．25％hydrochlor・ic acid and200ml．water to5g．solid sample（this willmake the concentration of hydr・OChlor・ic acid about2．27％）．Here（A）concentration of hydrochloric acid was ）ariedasl．5，3，and6％．To5g．defattedmea1200ml．hydr・OChlor・ic acid atone of these concentrations was added，and themixture WaSkeptinboiling”Waterbath for lhr．（B） Themixture of5g．defatted meal，200ml．water・，and20ml．25．％hydro・ chloric acid（Standard concentration）was heated forl，2．．5，Or5hrs．The result−
ingreducingsugars were determinedby the BERTRANDmethod．
Result．The reducing sugars formed are expressed as glucose。See TableI．（A） The r・educingsugars afterhydroly＄1Sincreasein par・allelwith the concentr・ation of l hydrochlor・ic acid used forhydrIOlysis．But toohigh aconcentr・ationofhydrochloric
acid（SuCh as6％）willgive rise to hydrolysis of car・bohydr・ateS
which are to beincludedin crude
fiber．．（B）The effect of time WaSSmallinthe r・age（1to5hrs岬） experimehted．When（A）and（B） ar・e COmpared，the valuein（B） TableI．Reducing Sugars after Hydolysis．
HCl，％ ‡hrs”I a弓glucose，％for meal
forlhr・．is higher thanthe value in（A）forlり50r・3．％hydrochloric acid．．Two sets ofexper・iments were made on dif■
ferentdays．The chief reasonof this difference may be the temperature ofwater
4 TechlBulil些agawaAgr・・Coll・
experimentundef the sameICOndノitions，perhaps′because the pleVious value was ob・ tained after hydrolysis over a boiling−Water bath andnot byimmersionin boも1ing
＝Wate‡．
EXPER‡MEⅣ甘S NO．2．甘he Condi七五0耶S OぞⅢy血0旦ysis主耶Lhe かe七erminaも互0隕Of Toね呈耽edu￠illg′Sl堵a町S a戯e好調ydyolysi臥（丑）
肋terial．−Defattedsoybeanmeal，thesamewiththatusedinExperimentsNos・3
to5．ThiscontainedlO．95％moistnr・eand30．．85％nitrogen−freeextract（TableIII）・・ Method．Five g samp】eTWaS hydrolyzed with200mi12％hYdrochloric acidor
220ml．2．27％hydrochloric acid for2け5hrs Successive hydrolysis was triedルA5g・
portionof thesam卓1ewasdigested with200耶1・2％hydrpchloric acid for’2．．5hrs・ The hydrolyzate solutionwas analyzed（No小4）The residue董rom thissolutionwas heat芦dwithlOOml．．2％hydぎOChloric acid for2hrs・・、The hydrolyzate was analyzed
（No．5），and the residue from thissolution was、againheated with150ml．2％hydro−
chloric acid for2hrs．The hydrolyz卑te filtered together with washings was ana・
1yzed（No．6）．
Result．As showninTableIIthe result was veryirregular．The volume（200
0r220ml．，）andconcent‡ation（20r2ル芦7％）didnotinfluence，壷henvaヱiedwithin14％・
The
ficientlycontrolled，and the paralleltitrationvalues（mi．．KMnO4）agreed withina
few％・・Tberefore，thisdiscrepancy幣1§tbeattributedtothedifference、Ofthem左n−
ner of heating，eSp声Ciallythete町peratureOfbathandtheふayofk？ePingth違flask inthe bat王トimme羊Sion of the flaskin boiling water willnecessarily acc占Ierat壱
hydrolysisipcomparisonwithholqingt＊eflaskoverthebath AIsofromthe？0責sid−
erationofs誓Cqrlnguniformheating，thetimeofheatingmaywellbも1占ngthened巌m
standaTd2．5如s．to5brs
Table甘．Reducing Sugars afreF Hydrolysis． as gluqose Sample HCl，ml． ％fo土meal】％forSumNos 4−6 Meal Meal Meal Meal
Residue f工Om No．4 R占sidue’f工Om No5 16．56 15．．42 15‖89 17．47 2．51 0．81 84い0 12“1 3…9 Sum of Nds．．4−6 2080 】 100小0
The thr・ee SuCCeSSive hydr・01yses gave the value totalingtO20＼，80％．．Thisis a COnSiderably high value・But、n主trog■en−free−eXtraCt COntent（30…85％）is not yet reached∴The three values for・Nosレ4−6show that the thirId hydr・01ysisis not neces− Saf’y・・The second hydr’01ysis maybe advisable，but the comparativ声Values canwe11
be′Obtained bya声ing■1e：hydrolysiswith2．％hydrochlor・ic acidfor，5hrs．Ofc9urSei．t is necessarIy tO WaSh wellノthe hydrこ01yzed mixtur・elaftef・＿filtrIationn
l
Vol・5，Noml・′（1、953） 夢
認Ⅹ野雷革Ⅰ施EⅣTS・N0．3．．′Fracti￠n埠二D申叩甲五nati叩ま呼．C軍ゆやhydra無S？f Soy桓9年恥
The method of WAK？MANqnd STEVrENSく2：）wa琶aPplied to＼d寧fat七red′SOybe叩S巾
This methodw卑SCOnSiderIedtoshow、SeParateamPutS′0freduQ享ngJSugar，nOnreduGi喝 Pl短0？aCChar・ides，hやmicellulose（tog・ethe亭 With g−umS），and、Cellulose c．ontained∴in soybeans
Material．・Naphtha−defatted soybean flakes wer・e pulverized to pass a2 mm． Si声Ve．．The analysis gave the values showr）inTableIII．
TableIII．Analysis of Defatted Meal
Usedin Experiments Nos．2to5“
、l、Ill く・‥い．：．＝…！
ト、こ：、．・∴∴
丁い．・トト、 ％ 53小34 5．．56 0．．68 5∼77 34．．65 クら 10．95 47．．50 4り95 01ミ1 51−1 30．．85 Moistu工e Crude protein Ash C工√ude ねt CTude fiber Nitrogen−freeextrpac七Method。The sample（5−6g．．）was extr・aCtedsuccessively with（i）ether・，（ii）cold Water・，（iii）hot water：，′and（iv）e坤ylalcohol，and′fuざ∵叫声r（Ⅴ、）hemicellulose，（Vi） Celluiose，and（Vii）1ignin were to be determin竿d．
（i）EtherIeXtrIaCtion．l−The sample was extracted with etherin theSoxhlet ap−
Pai・atuS for・16hrsLThe r；esidualsolvent was evapor・ated frommiscell．a andtheether extr．act（tと−ude fat）wa
（ii）Cold Yater eXtr坤ioh‖−Theァ・eSidue fr・OmLether extract was freed ftom r・eSidualether・，trranSfer岩：edtoぇ300ml．．。Erlenmeyqrfiask，addedwith．150′ml．．distilled watek；；岬d theihixtur・e WaS allowed to stand at room temper・atur・e（av．about190）
for・20‘hrs．．The extract，filter・edthr・Oug，h weighedpaperI（thisfiltrationwasvefIySlow）， wasputintoa250ml．．volum6tticflask；arliquotsofwhithwereusedfofdetefhl、inatidns Of totalsolid matter’，aSh，reducingSug早r’befor−e and after hydrolysIS，and nitro宮en・・
（iii）Hot water・eXtraCtion．−To the r・eSidue from cold・Water・eXtraCt，150ml． Water WaS added，争nd themixture Ya亨kept fOr’3hrs小atlOOOin a bdiling’−Water bath．．The extr・aCt
in（ii）．．This fiitr・ation was much easier・than the・filtr・ation of cold−Water・eXtraCt．
Thefiltr・atetaken to a250ml．宣olumetr・icflask壷a亭analyzed for・tOtalsolidmatter， ash，nitrog・en，r・educing・Sug・arbefore and after hydrolysIS
（iv）AIcoholextr・aCtion。−The r・eSidueinsolublein hot waterIWaS heated with 150ml・・95％ethylalcoholon a water・bath／for・3hr・S‖The extr・aCt filtered through 6rlg・1nalfilter・papdr・waS putin a flaskto wei畠・htotalalcoholextract
（Ⅴ）DeterIminationof＝hemicelluloseけortreatmentwith2％hydrochlorIiとacidl・・ Dr・iedresidueinsolubleinethylalcohoIwastr・anSferred・Withl蜘aterItO、theor・lglna1300 ml．．Erlenmeyer・flaskいSufficient hydrIOChloric acidsolution was addedto make the Cbncentr・a．tion
6 Tech・Bull・KagawaAgr・Coll・
takeninto a500ml．flask，and thererefor・eits volumewasmeasured and25ml．．was used for analysis of reducing・Sugar・by the Bertrand method・This reducing sugar expressed as glucosewasmultiplied byo．9togivethevaLueof“hemicellulose．”This point was cr・iticized by HAW工．EY and NoRMAN（29）．Theresidue from the treatment
with2％hydrochlor・ic acid was dr・ied and weighed．
（Vi）“Cellulose”．−Tooneweighedportionoftheabovedr・iedr・eSiduelOml．80％ sulfuric acid was added，and the mixtur・e WaS allowed to stand for2hrs．．at roo血 temper・aturIetOChangecelluloseintohydr・0ⅩyCellulose・・Water・（．150ml：）wasaddedto this，and themixture was boiled for5hrIS．．tO hydr・01yze hydroxycelluloseinto glu− cose．，Theglucosemultipliedbyo．．9is＝ce11uloseけ・Thismethodis thesamewiththat usedfordetermlnlng・maXimumIeducingISugar・yieldinsacchar・ificationofwood（3O・31）
（Vii）“Lignin”．−Theresiduefromcellulosesacchar・ificationcontainslignin・The weightof（thisresidue）−−（ashinthisresidue）−（nitrogeninthisresidue x6．25）corres” ponds to〃1ignin：，TAKAHASIandYAMAYA（S2）establishedthepr’eSenCeOfaboutl．．36％ oflig・ninin seedcoat（Prabouto．0087．％forsoybeans）．，SASAKI（33）found onlya−trace oflignlninsoybeans．Therefor・etheauthor・madenodeterminationoflignin・
Result．The result of two parallelanalysesi？g・lVeninTableIV．
TableIV．Fractionation of Carbohydrates Of Defatted SoybeanMeal． Sample i．Ether ext． iiりCold H20ext． Ash Reducing sugar Total＄ugar N
iii．Hot H含O ext． Asb Reducing sugar Totalsugar N iv．AIc…eXt． Residue fT・Omiv． Ⅴ．“Hemicellulose’， Residue from2％ HCI treatment Vi．．＝Celluloseり Residue from vi。
Vii‖“Lignin”
EXPERIMENTSNO．4．DeterminationoftIleCarbohydrateSoltlblein ColdWater． The determination of car・bohydrate（reducing sugars after・hydrolysis）in cold−WatereXtraCtgaVenOreliablevalueinExper・imentsNo・30Thusitwasr・ePeated・
Ether extraction was omitted．
肋thod．No．1：Defattedmealsample（5g．）wastreat6dwith150mll・Waterfor・24 hrs．，andthefiltrate was hydrolyzedwith2％hydr・OChloricacidfor2．．5hrs・Neutral− izedandthenfilteredsolutionwasanalyzed．No．2：To5g・Sample50ml・WaterWaS
Vol．5，No．1（1953） 7
added，afterI16hr・S．thecold・Water・・eXtraCtLWa畠＼Separatedby・SqueeZingthroughcloth
withmuchwashingwater・，andthenbycentrif．uging．Thecentrifugedsolution（144ml一．）
wasagainfiltered，aSthereappearedsorpepr．ecipitatewhen allowed to stand over・ night．．Thiswassaccharified with2％hydrochlor・iG aCid for2。5hrs．No．3：Tothe similar・SOlution（146ml．）as No”2was added drobwise aboutlml．of the E≒bach reagent（■picri（’acidl．5g．・and citricac蔓d3g・madeupto150miリWith wat9r・），Jhe filtrate separ・ated from the precipitated、prOtein by this reagent was sa＄Charified
with2％hydrIOChloric acid for2．5hrs．No；4：To5g．sample was added50，ml．satu・ rated picric acid solution（aboutl．．5％）．After16hrs．th畠SOlutioh filtered（thislrfil− trationwasveryeasy）together with vよshing with satur・ated picric acidwas哲C・ char・ifiedwith2％hydr・OChloric acidfor2．5hr・S．This method was f・eCOmmendedby thelate Dr・．Bunsuke SuzuKIas the means not to dissoIvやPrOtein and to、eXtraCt
Sugar’S．
Result．AsseenfromTableV，COld−Water・・SQluble carbohydrate was only2．3−2．6 ％when determined by the ordinarymethod，Whileit was4．4％and as high as7．5％ whendetermined byexcluding theeffect of protein．Especially the extraction ofsu・ garWithsaturated picricacidwasverysatisfactory．Theresult of Experiments No．
TableV．Carbphydrate SolubleinCold Water．
As glucose，％fo工・meal Method
Fu11descriptionfor the methodsis givenin the text
5（7．′2％）obtained by direct alcoholextr・aCtion agrees wellwiththehighest value （7．5％）obtained herIe
EXPERIMENTS NO．5．，DeterminationoftheExtractObtainedbyDirec七Al℃OhoI Extra七tiom．
The method of WAKSMANandSTEVENS（27〉a占9riginally proposedis not suit・ able to the sample such as defatted soybeanmealr・ichinwaterextract andinwater・ solubleprotein．Thedir・eCteXtraCtionof soybeanmealswithethanoIwasconsideredto be superior・．MonosaccharIides（Only a trace was detectedin soybeans，TableIV）and oligosaccharides dissoIveinhot90％ethylalcohol，Wher・eaS hemicellulose does not．
Mdhod．Sample defatted meal（10g．）with50ml．90．％ethanoIwasrefluxedfor 3hrs．Ther・eSidue was againheatedwith50ml．90％ethanolfor2hrs．andthesecond residue was again heated withethanolfor2hrs．．Thelast residue was wellwashed with90％ethanol．．Totalextracts and washings weremadeup to250ml．．with water． Aliquots were taken for analyses of totalsolid matterI，aSh，nitrogen，and reducing SugarS before and afterhydr・01ysis．
Tech．Bull．Ka女aⅥ閣Agr；．Co11． TableVI．Compositi（肌Of EthanoIExtract
O’f Defatted SoybeanMeal．
8
Summiry〆Exl，eYimenis．，Nos小3−5．Seethesummarizedtable，TableVII．
TableVII・Compositionof DefattedSoybean
Mealby So！vent Fractionation。
The＄amPlemealcontaindmol占turelO95％，Crudefiber 514％，andN一露eeext 3085％onair−drybasis．
SoIvent lTotalsolid E Cardohydr・atelCr小prOten l Ash
主． iv．．HotII90 ivHot90％EtOH V．〃Hemicellulose” vi”“Cellulose” ‡nsoluble protein 、−ノ︶＼−ノ 飢59貯12訂0863 00317A二6 5 ／し′．＼l ︵ つ上 e 〓山 t 7，亜α 1‖58？ 767 4．08 29日31 1．56 1663？ Sum 1 83．77 1 20‖79 ト虻オ50 ！ 4、95
a Extracted with s左turatedpicric aicd．
01igosaccharides of soybeans are difficult to determinein cold−Water・eXtraCt， andtheyar・etO
No4，Table V，No．，4）or・by dirIeCtalcoholextr・aCtion（ExpeT・iment′SNo．．51）；＜thevalues bbtainedwer・e7A6Ha王idこナ．：19％■i′respeetiveiy岬Th占totalr・占dudngSdgar・S after、hydrol−
ysisamounted to15…4−17．5．％（Or20。8％■after3successivehydrolyses）ofdefatt姐、畠oy− beanhleal（Experim由1鱒No。2）L℡he
amount6dto13L58勉，df／1whiChll．i77％wouldCorr・eSpOnd to olig・OSaCCharides（ExPet・i− mekts
Ihshor・t，defatted′SOyb6anhlea仁和ithld。95％ moisture contained7A6％olig・0− gacch如ides，7．．67％hemicelltilose，and4．，08．％一c占11ulose，tO tained30‖85％■nitrIOgeh−f士・eeeXtraCtゝnd20‖80％totalr・educingsugarsafterthree▲succes− sivehydrolyses．．Th6sevaluesekceptnitrogen−f王・eeeXtr・aCtCOrr・eSpOndto吏・educings璃・ar・白 色xpre占sed asglucose∴ E女中ERI光EⅣTSⅣ0．．烏．．DetermiIlaも主ふもbfGalae七ふseinsoyもeansbyOxida七ioIi：to MふcicAcid”（Incollまbor扇ion☆ithBunpeiMt）RI（：：】））
Whengalactoseorg・alactanis oxidized with hot nitric acid（SpeCific gf・aVity l．．15二），muCicacid forms and cr・yStallizes out小This ToLLENS，rh占thbd壷えsappliedfor
Vdl．5，No．1て1953） 9
defattedsoybeanm6al，S？ybeqnwhey，andprotein・eXtraCtionresidue・
晩terial．．Thisnaphtha−defatted soybean mealcontained moigture13”50，Crud占 proteih47A5，and ash5…07％．．Pr・eparation of soybean protein was carried outl）y extracting withlO partsO．4％soditlm Sulfite andprecipitatingwith，＄ulfuricacid as usual．（S4a）
肋thod− The originalToLLENS甲ethod andVANDERHAARmOdified method
wereused．Inthelattermethod pure
formation of mucic acid crystals小
The whey obtained from50g．defatted flakes arho血七色d to13501ml‖This was conc、entr・a糎d、tlnder，VaCuum、tO 82 ml．．，Put Of which20ml．wasluSed for■determin− ation．WhQnlOml．w早SuSed，it叩aS、insuffi＄ient to‥for甲enOughmucicacid to deter： mlne．
ResultandDiicussio鉄 As givenin Table VIII＞the r．esult w争S higher by the VAN工氾R HAAR method thanby the originalTOLLENS method．Whenthe yield of protein−eXtraCtionresidueisノPut tO be20％ of the defatted meal，the“galactose”
Table VIII．Galactose of Soybeans．
alacte’
≡諾芸霊アmg．．牒浣諾I m㌘夢；。rmeal
Sample Defatted mea1 3い000g． 3000 3．000 ConcdりWbey20．．00ml． （corresponding to50×20／82g． Of defatted meal） Protein・・eXtn．reSidue2．00Og． 鋸町碍 鎗 l l慧遠 以
444 1 733 2 17︵0 0 O10卸 0 （3い43） 258 343content of the residue，1715％，COrrβSPOnds to3．43％’for meal．As ％ for rneal，tOtal g＼alactose4．73．％is thus distributed as whey galactcsel・02％ and residue galactos号 3．43％．The sum of these two valuesis4．45％．
If soybeans contain other substancewhichyields mucic acid on oxidation，these
values arIe Only app争rIent．Soybeanwhey probably containsno such compounds，but prIOtein・eXtraCtionresiduem年yCOntainpolygalactur’Onide，Whichonoxidationwi11give ri畠eto mucic acid not originating■from galactose．
Soybean whey contains reduing sugar・S aS Shownintheauthor’spreviousreport （35）小These should have been formed fr・Om SuCr・OSe，rIaffinose，and stachyose． There− fore，free galactose can existinwhey。Besides，galactosemaybepr・eSentaS Oligosac− Charides（raffinoseand stachyo軍e）inwhey．
The mucic acid・yielding comFOundsin theresidue from soybean・prOtein extrac−
tion cannot contain simple carbohydrates solublein alkaline aqueous solution．They
are pr・eSumablyin the hemicellulose fr・aCtion仙So this should better be expressed as galactan rather・than as galactose．In this experiment theyieldlOf the residue from defattedmealwas not determined．Therefor・e，itis morereasonabletousetheamounts
TechBull・KagavaAgr．．Co恥 10
Oftotalgalactose、andwheyざalactose．astheba軍isfor−Calculation・Ther：eCalculation gives theresult声hownin TableIX・Simple calculation shows′that the yield of f：eSidue should，haverbeen21・6％ofdefatted meal（＝3・71／17・15）・、Thisisveryprobable・
Table＜IX．Galactose and−Galactanin Defatted Soybeans．
％galacto＄e ％galactan ヽふ＋．＿＋ Total Ⅵ7hey Residue 4り73 （：）α 3，這ウ α＝4．73−1．02 あ＝3．71×0．9
STREETahdBAILEY〈5）calculated thedistribution of“galactan”bydetermining mucic acid formed on oxidation with nitric acid．Their fraction褒tion wasbasedoIゼthe
analysis for−thesoybeanmeal，SuCCeSSively extracted withboilir）g95％ethanol，Cold 壷at占randmalt′extract．Thetotalgalactanfoundwa岳4．86％．InthealcohoL・eXtracted materia14．．62％waspr・eSentandinthewater−eXtraCtedandmalt⊥digestedresidue3．05 ％・Thus 4・86」N4い62＝0・・24％’corTeSPOnded to alcoholこSOlubleノ、StlgarS（raffinose and
StaChyose，though STREET and、BAmEY COnSidered only r・affinose二），and 4．62−3．05 ＝rl岬57％towater’・SOlublegalactan（VegetableぎumS and mucilages）・・Galactaninthe
insoluble cell−Wallconstituents amounted to3．05％or62．．7．％of totalgalactan．
EXPERIMENTS NO．．7．．Determination of Pensosanin Soybeansい（In collabora− tion with Kazhharu TAKIZAWA（S8））
Materi’al“ThesahlplewasprovidedbySy（うWaSangye）K．K．＊Thesoybeanwasdefat− tedwithpetr・01eumnaphtha about600，and the soIvent was recover・ed by steamlng； thisisthe commonest commer・Cialsolvent−defatted flake nowavailableinJapan
Meihod．The ToLLENS disti11ation was applied to（a）sample d戎atted meal，（b） Water．eXtraCt Of defattedmeal，Obtainedbyfilter・ing・themixture of5・−20g．sample and asufficientamountofwaterallowedto stand ovemight，（Cr）residue from（b），（d） hot−Water・eXtr・aCt，Obtain畠dfromthemixtureof占amplemealandwatetboiledforlO min・，（e二）theredisuefroIn（dr），（f）methえnolextiIaCt，and（g）residuefrom（f）・
ResultandDiscitssion．As shownin TableX，appar・ent“pentosan”content（ex■・ pressed aspentose）of soybeanswas5．．7・6‖1％，Of which5％wassolubleincoldwater， 30％was solublein hot water・，and35％■was solublein hot methanol．As hexur・Onic acid and evenhexose（especia11y ketohexose）willgive furfuralondistillationwith
Table X．PentosaninDefatted Soybeans．
％as pentose Sum l ％distribution ∼、 （a）Total （b）H20ext （c）Residuefrom（b） （d）Hot H20ext （e）Residue from（d） （f）MeOH ext叫 （g）Residue from（f）
覚The authorisindebtedtoMr．．Sy62；iTAEAⅡASIOf the TurumiFactory，Yokohama，Ofthis
Vol．5，′NoJl（1953）＼＼、 11
12％hydrochloricacid，theappreciationofthesedataisimpossible，before morein− formationis obtained byノothermethod of attack・Water−SOlubleoligpsaccharide con− tainingpentoseresidueisnotyet recordedintheliterature・The author fopnd no pentoseinsoybeanwheybyexamlnlngthesug・arSOfwheybypaperchromatography（26）・
According・tO MIYOSIand HuzII（37），the“pentosan”containedin defatted soy・ beanwas most solubleinmethanoland ethanolamong organicsoIvents；thecarbohy− drate content of soybean pr・Otein extracted from defatted mealwith o。2％sodium
hydroxidewas the highest，When the defatted mealwas treated withalcohol；thus， theremayexistdifferent〃pentosans”solubleandihsolubleinalcohol・
According・tOSTREET and BAILEY（6）there was4t・94％total“pentosan”，alcohol extraction residue contained 4叫11％，and water−eXtraCted and malt−dig・eSted residue contained3．09％pentosan．“Thesmallamount（4．94−4・11＝0・83％）removed byalcohol
isla柑elyduetosubstancesinsolubleinwater，preCipitatedor occludedin thelead
precipitate，Whenthealcoholicextractistakenupwith waterandtreatedwithbasic
lead acetate。Thislead precipitatewas found to yieldCOPper−reducing matter equl・・ valenttoo。5％pentose．Thenatureof thesaccharo−COlloidsisratherobscur’ebutthey arequiteprobablythe veg・etable g：umSand mucilaglnOuS materials presentin seeds
andplant tissues，WhicharedissoIvedina slight degTee by alcohol，and which are removedmore completelybywaterin the r）eXt Stage Of our’analysis・・The removal of fructose as such，Orincomplex，may also offer a partialexplanation・〃Pentosan intheinsoluble cellwallconstituentsamounted to3．09％’or62．5％of totalpentosan．
SUMMARY
Thereis alwaysacorlSiderabledifferenceintheamountoftotalreducir3gSugarS
afterhydr・01ysisexpressedasglucoEeandtheamountofnitrogen・free extract calcu−
1atedby differenceinthe case of defattedsoybeans・Inroundnumbersdefatted meal contains30％of nitrogen−freeextract，While onlyup to20％sugars after hydr・01ysis areencounteredby analysis．．TheconditionsofhydrolysiswerestudiedforIanalyzlng this totalsugarafter・hydrolysis（Oftencalledtotalcarbohydrate，butexcludingcrude fiber）（ExperimentsNo．1andNo．2）．Theconcentr’atrationofhydroch】oricacid（2％or 2．27％）or・timeofheating・（1−5hrs．）influenced theresult scarcely仙Uniformheatingin constantlyboilingwater（not over the boiling・・Waterbath）was found essentialin obtaining comparable values‖Anyhowthediffer’enCePerSistedbetween30％nitrogen・
free extrIaCtand20％r・educing・Sugar・after．hydrolysis・
The proximatefractior7aldeterminationof soybean carbohydr’ate（Experiments Nos．．3−5）gave the resultthat defattedsoybeanmealwithlO”95％moistur’eCOntained 7．．46％01igOSaCCharides，7．67％hemicelluloses，and4”08．％cellulose，tOtaling to19・21％・ Calculationshows that this corr・eSpOnds to6．12．％01igosacchar’ides，6．30％hemicell11lo− ses，and3い82．％cellulose，Of soybeans，basedontheassumptionthataveragesoybeans
COntainlO％moisture and16％fat．
12 Tech・Bu11・Ⅹagawa．Agr・、Coll・ residue．（Exper車ehtsNo▲6）・Itisondifferentsample，bptl・02％“g’alactose’’inwhey
（TableIX）shoutd’corTeSPOndtothegalactoseres、iduein galactose・COntainingoligOSaC・
char・ides．・YuxAWA（2ミー）andSASAKI（21）calculatedtheweig’ht of sucrose and s！achyose
based ontheassumptionthatonlythesetwooligOSaCCharidesar・epreSentinsoybeans・
Itis simple to calculate the contents，ifit were valid that soybeanscontain only
SuCrOSeandstachyoseasoligosaccharidesandthattherewer・e nOOther90％・卑1GOhol−
solublecoInpOundsinsoybeanswhichyieldmucicacidonoxidation・
Apparentpentosancontent（ExperimentsNo・7）ofdefatted s、oybeanswas51・04・ 5小39％・（5．72・6．12inTable Xmultiplied byol・88）・This should comprise hemicellulose fraction of TableVII小
uTERATURE CITED
Theliterature consultedisgivenfirst，■beit anoriginalpaper，OrabstractingラOurnal・C・A・＝ chemicalAbstracts；C．Z．＝ChemischesZentralblat七；N。K。S．＝NipponkagakuSe・ran（Complete＜Ab・ stract＄OfJapaneseChemicalLitefatures）．Thislistisarrangedintheorderdfthefirs七1Citation・
（ユ）C．W玉：Ⅱ丸柑R。I）ie Pflanzenstoffe，1；567−70（1929）
（2）A．MATAGRIN．Lesoラaetlesindustriesdusoうa・Produitsalimentaires，huile8esoja，16cithine v6g6tale，CaSiinev6g畠tale．Gauthier−Villars，Paris（1939）・
（3）A．LEVALLO‡臥（qegenwartein？雪inGlykose Gberfuhrbaren Substanzin S（中hi＄pida・）C・Z・ ∫88P，500；Compt・rend‖90，1293（1β801）
（・4）AいLEVALL。IS．，（tlberdiezuckera坤geSubstanzip demsamenvonsoぅahi＄pid乱）C・Z・1甲I， 662；Compt”rend．98，281（1881）
（5）J．P STREETandE…M．BATLE￥・一Thecarbohydratesandthe enzymes ofthe soy beanuJ・Ind・ EngいChem一．7，防3−8（1915’）． （6）E。MEISSLu∴RB6cEER．・tlberdie．．B甲tandteile derBohnenvonSo5ahispidalP C・Z・1888・619； Monatsll‖i．，Chem．4，349−6g（1g83）． （7）TF．HANAUSEKulHARZl（2），pl12台，1142；Jahr小8b・d・Fortschr小dけPharmakognosie，Pharm：u・ Toxikol．ヱβ∂β−∂4，279 （8）T．F”HANAT3SEXu．HARZ．（i2），ibid”；Ibid・・1885，117・ （9）HARZ．，（5），pU853；Z．allgem”OesterrlApoth・Ver”2B，40（1885）・ （10＝．．STINGI，u．T．MoRAWSEI．ZurKenntnisderSo3abohne・C・Zu18P6，724；Mopatsh・f・Chem・7・ ユ76−90（1886）小 （11）R．．BLOND弘．（2），p．．123，142∴Jl・p壬1arm・etCbimie∫ら587（1888）・ （12）M．王NOVE．Thepreparationandchemicalcompositionoftofu小Bull・Coll”Agrll，TokyolmpUniv・ 2，Noり4，209−15（1895）． （13）PRINSEN。（2），pい142；Chem‖・・Ztgu1896，Jan・29lCf・（17）v （14）F．J．．G．BELTZER。（2），p．58；Revuescientifique，1911，JunelO・ （15） Monatsh．，f”Chem。8，82−4（1887）
（16）EりScTULZEu”S．．FRANKFURrいtIber die Verbreitung des Rohrzuckersin den Pflan2；enund iiber16slicheKohlenhydrate，dieihn begleiten・C・Zl・18・95，II，141；Zいphysiol・Chem・20・511・55
（1895）
（17）PRINSENrGEERLI2qGS。（1）；Chem．−Ztg．20，67（1896）”Cf・．（13）・
Vol．5，▲No．．1（1953） 13 ander・enLeguminosen／）C叫Z・1918，1，634，1208；CoTrlpt．rend．rニヱ55，1526・8（1912）；Bull．soc．chim． （4）ヱβ，176・・82（1913）． （19）Kintare・Os工WA．．（Carbohydratesof soybeans．，）N．K．Sけ（I）8，14；SapporoN8rinGakkaih64，′1−8 （1909） （20．）BいF∴DATIBERrいOtherconstitueIitsthanoilandprotein＄Ofthesoybean”SoybeansandS（tybe？n Products（ed…byKS‖MARKLEY）1，371−81。IntersciencePublisher竿i，甲eYork（1950）．
（20al）MataoYuEAWA．（Carbohすdratesof soybeansandtheirrelationtothe brewing of soy sauce．） JいTokyoChem．Soc．．88，318−88（1917）；J巾Soc．Br声wing，Japan12，血．ヤ，23−34，No．8，1・・30（1917）；
KいS．．（J）4，471
（21r）Sy凸iku SASARI．：（Metabolismof carbohydratein germinationof soybeansい）J．Agr．Chem．Soc．
・Tapan19，69軍−6（1933）；N・K・Sり7，415・
（22）R・・C・BuRR飢LandA・C二・WoLF鳳Compar・ativestudyofthech？micalc？mpOSitionof5var・ieties Ofsoybe？nS・C・A，84，2090栗；FoodR？SearCh5，109−13（1940）・
（23）Tei2＝e・TAEEnIOrO．（Chang今Of components of soybeans by heating．）N．K．S．7，331；Ry∂syoku
Kenkya（FoodResea工CIl）7．9，6ユ仙6（1933）．
（24）YosioKAWANO・（Changeofcompopentsofthe＄Oybe禦by steam cooking”）N・K・Sこ18，156；
Zybz8gakuZassi（Jl：Brewing）i6’，7牢・・67（1938）
（25）TadasiKA丸首IEtjBO。（Change of carbohydrate during manufacture of aminoaCid condiments．） N．K S，25．1038；J一．FermentationTechnbl．，（Japan）29，51−4（1951）．
（26）Sin’itir・e・KAwA血RA…（Studiesonsoybean car・bohydrates．I。Sugar畠bf soybeans．）Tech．Bull． KagawaAgr。Coll．4，65−11（1952）；C小A．46，9743g．・
（27）S・A・WAESMANandK R．S柑VENS‖Asystemof proximate chemicalanalysis of plant mater・ ials、Ing…Eng．Chemい，Anal．．Ed．2，167−73（1930）；CいA24，2965 （28）G…KLEIN・Handbuchder・Pflanzenanalys （29）LF“HAWLEYandAいG。No㌫fAN．The differ・entiationtofilもmicelluloses．Ind‖Eng．．Chem…24， 1190−4：C“A．26，6124 （30）H・LiゴERSりZellulosever警uCkeiungmitverd血ntenSiure；Sch占11去r・−Tornesch−Verfahren”Angew・ Chem．4」5，369−76（1932）；CいAい26，4946
（31）Bunz8Ro肌SYOandA∴KA冨AfuKA。MokuzainoTbkよoyobiRiyb（Saccharification and utiliza．
tion of woods二）…p．39（1938）
（32）Teiz8TAEAEASTandZyunsinYARfAY・（Ligninofsoybeans・）J・Agr・Chem・Soc」・Japan6■・242・7
（1930）
（33）Sy凸iku SASAI（IandYosimasa YAM：ASITA．（Cellwallconstituent台 Of the soybean。・Ⅰ．Cellwall COn＄tituent＄Of the seedof soybeans．．）J．Agr．．Chem．SocいJapan14，1257−63（1938）；N．K．S．18，15小 （34）BunpeiMoRrい（Galactosein＄Oybean carbohydrates）Thesis for graduating from Co11．Agr．，
NihonUniv．，PreSentedtoSin’itir・b KAWAhfURA（1951）
（34a）Sin’itire＞ⅩAWAMURA・Studies70n themanufactureofsoybeanproteinり，町IExtraction and pre−
Cipitationof protein，Smalトscalemanqfacture of soybean proteinりTech Bull．Xagawa AgrいColl．
4，206・・225（1953）
（35）Sin’itirbKAWA耶RA．（Utilizationof the by−prOducts of soybean・prOtein manufacture．Ⅰ．AIco・
holicfermentaionof the wasteliq110rOf pr・Oteinmanufacture。）J．Agr．Chem．Soc．Japan28，129− 34（1949）；C．A‖44，5059β．
（36）Kazuharu TAEIZAWAn（Pentose of soybeans二）Thesis for graduating from Coll．Agr。，Nihon
Univい，
（37）SigeoMI￥OSIand Sin’itiHuzII”（Soybeanproteinfiber．XI．Carbohydratesofdefatted＄Oybean＄
treatedwith organic soIventsandof the preteinprepared from them forIuSein fiber・）N．K．S．
Tech．Bu11．KagAwa Agr．ColI．
Authと）rIndex of the Literature Cited
14
Mori，B．．（34）
Norman，AいGこ See Hawley，L．F．
Osima，K．（19） Prinsen．（13） Prinsen−Gee工1ingsい（17） Rokusyo，B．，andKamatuka，A．（31） Sasaki，S．（21） −−−・andYama＄ita，Y．（33） Schulze，Eい，and Frankfurt，S．（16） Stevens，K．R．See Waksman，S．A． Stingl，．J’．SeeMorawski，T． 〟andMor・awsl【i，T．（10） Street，J小Pい，andBa皐1ey，E．M．（5） Takabasi，T．．，andYamaya，Z．（32） Takemoto，T．、・（23） Takizawa，K．．（36） TanIet，G．（18） Waksman，SいA．，andStevens，K R．（27） Wehmer，C．（1） Wolfe，A．．C．See Burrell，R C． Yamasita，Y．See Sasaki，S． Yamaya，Z．See Takahasi，T． Yukawa，M。（20a） Bailey，E．M。See Street，，．P． Beltzer・，Fひ．丁．G．（14） Blondel，R．（11） B6cker，F．．See Meissl，E． Burrell，R．C．，and Wolfe，A．C．（22） Daubeれ，B．F．（20） Frankfurt，S．．See Schu12；e，E． Hanausek，T，Fりand Har2；．（7）；（8） Harz小（9）；See Hanausek，T．F． Hawley，L FリandNorman，A∴G．（29） Huzii，S．See Miyosi，S． Inou9，叩・（12） Kamatuka，A．See Rokusyo，B． Ⅸamikubo，T．（−25） Kawamura，S．．（26■）；（34よ）；（訪）；Cf．（鎚）；（36） Kawano，Y．（24） Klein，G．（28） Levalloi占，A．（3）；（4） Liie工S，H．（30） Ma工トkley，Ⅸ．S．Cf．（20） Matagrin，A．（2） Meissl，E：，andB6cker，F．（6） Miyosi，S。，andHu2：ii，S．（37） Morawski，T．．See Stingl，J． −−and Stingl，J．（15） 大豆汲水催物の 研究 正．脱脂大豆の炭水化物含有量 川 村 億 一 郎 脱脂大富を分析すると計算の上では可溶無窒素物が約30．％となるが，うすい酸で加水分解したあと の還元性糖罫をグルコーズとして表わす・と約20．％となる．このときの加水分解に凧、る塩酸の感度 （2％又は2．小27％）や加熱時問（1・−−・5時間）に・よ、つては結果に・大きな差を生じない（実験第1一策2）・ 試料脱脂大豆をエーテル，冷水，熱水，95％熱エタノールで順次抽出し，各抽出液の全国形物，窒 素，衣分，糖分を定盈し，さらにこの後淀に・つきヘミセルロ1−ズ，セルローズ，およびリグこ・ンを定 慶するWAKSMAN，STEVENS（27）の方法を適用した（表ⅠⅤ，実験訝3）．．挽脂大■豆のように冷水で抽 出さかる蛋白質が多い試料ではこの方洪は適当でない．水抽出の代りに飽和ピクリン酸渾液で糖類を 抽出する（実験4）か，又ほ眉凝エタノールで抽出する（実験第5）ことにより糖顆を求める方がよい． 結局脱脂大豆（水分10．95，粗繚絆5仙14，可溶無窒素物30．．85，油分0．61％）は冷水に溶ける糖類7・・亜％， 熱水に溶ける炭水化物1仙58％，ヘミセルローズ7．67％，セルロ1−ズ4．08％を含むという結果（表Ⅶ）を 得た．．即ち炭水化物合計20．79％’と．なった． 前報（26）では糖類についての数倦をまとめたが，そのうちガラクトーズ定量とベントーズ定畳の結 果につき多糖渠に基づく分の数倍（実験第6一帯7）を補い，文献の数倍と比較して論じた． なお大豆炭水化物の研究史を文献（1）￣（埼〉を引用して記述した．
