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AH mmnIcu 0? 9694111.”. BICLCCICLL 8131‘343 F033 YdTIIILfiIOXAL

hem hm

.8 ”$3313

Surname! to flu School or emanate Studio: of nichigam
State University of Agricultun and Applied Science
in partial fulfillment of the raquiranantl
for th- dagrao or

HASTIR G‘ SCIJ-ICE

Depar‘hnant of Chmistry

1955

57$ [/37 r!
2:5. 10/“?
0*

mmamtm

the nth» via)»: to oxpr-eu 111: linear.
tppmittion to Dr. John C, Speck Jr. for w.-
cdvioo mi oncoumgament during the cow of

thin research and the 13th of this
lama-om.

11%

”Hi“.

£3 1.321313? {1‘ 3&1: BIC-LOGICAL 31f7f.£{3 PW. MitTHYLGL‘IOKAL

3.?

Bane Starr!

.33 37?: 3131‘

macaw to the School of (infinite Stain" of rdcmgan
5&8. Univaflity of kgricultm- mi Applied Science
in mun Want of tha new:
for tho dame of

mm or 361.41%;

Expat-twat of Chemistry

I.» 1935

Approvad __

RESI’RQCI

Sm]. biologist]. 13m have been) emined for th- preleaoo of
mthfiglyom. T1» methods employ“! for this mveoufiuw after
anal adv-mugs. our the: pruden- uchniqma and to:- dawning
uflvlghonll may m quite umitiw an! spawn, AM the: permit
animation of hidoglnl am under that WW conditional.

In post 0! than expert—nu, tho uterifl under investigation Ill
13139111111011. and tho dams. callootod. T111. dictflhh m .181»?
“and with emu-epic aid-emu acid m and the noncon-
mum at nethylglyonl deter-ind by Amt-w}, or m: W1
WI patent vars confined to than 011m: and the «sum I1»-
turo outlined for amylase“ by piper Miami.

I in mthylglyonl 6011141 be found in 13:33:“ 10;», yea” {amen-
tauon 3111.1:ch , r“ liver It at blood. Ilsa adding: of very 10!!
«mutations at uthylglml he that» 333m; was (no ltudiad.
”WWW“ mama manna-gimmwemudy
inhibited with Mow-tic acid.

Ismm plan“ fun the been mined. Th- older lawn warn
{www.mmchmuwcmmpmmm to
field 3 {Imam ”pound. Ham, 8 what-um no meat mick:
Apparently for" an on». mu 'm' can be converted to a ”new
niobium complex, but tho R! value for the 'm' is considerably
mats: flan am. for mthylglyam.

1v

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KISI’CRIC :‘J. INTROEIICTICR

HISTORICAL ’TROZXICTIOH

The now obnoloto final-y of nethylglyonl u an important inton-
”ditto in 513001;“. that me into being 321m!!! 1912. The clan
Iranian]. minimum” batman lactic and pyravic acid- Ind methyl-
glyonl contributed math to the “trauma: of the 1d“, ad 1%
In mun: “hauntintod by identification of. W, which
catalyze- tho conversion of uthylglyonl to lactic told, u tell u
by inflation. of the «stud oocummo of uthylglyml in a variety
or biological systems. than, u ”:1: u 1913 Dunn nnd many (1)
menumoormonhuinbbodmdmudm tin-nu
(11m, hart, m1... basin. and mm”) o: - umber or animal upon.-
(an, dog, ox, cheap, rabbn, “foul“, and oodfish). More fluently,
Hopkins and Harm (2) found us amending: ads distribution or
W mung Wombat”, nan, unwed, fungi, Ind higher
p10“. An early 6181! to the identiticntion of mthylglyonl in bio-
legion mm occur- in the report by Toenail-on md Fish" (3) who
hoisted uthylglmnl p-nitrophomrlouuom Iran an incubation nixtnro
consisting of {NJ-awn and much and panama Mam.
Anya. (h) upon-d this work in! mm mm tor the pmnoo o:
nnglyonl 1n the color {crud on auction with mcmphoophotmmia
«1:: ma ama- 10a. Bomber: and Kabul (5) reported 1.01m» o:
lathylglyml n it! Zfi-dinitrvplnnylomono full bacterial extract.

ta which mun-1,6-diphoaphlu lad been um. 511113117 , tubal
and m (6) mum mammal 2,h-d1n1tmpm1mmm m-
m» And (no: do: ”cl. when thou tutorials were incubated with
hem-0 Womb-N. loodhu ma Lou-am: (7) an own-ad identifi-
mm: o: uthylglyonl by tho pmoduro or flouburg ma lob-1 (5) 1n
ohioknn embryo um. Cu. and Cook (a) reported an Mhflglyoxal
occur: a la Mk 1:: lactic said tuition.

Reuben and m (9), Lame and Hey“- (10), Duds! and Dudluy (11),
Mimunndlobol (12) mmpaodplmnihhcohman for 31:-
only.“ thich inwlnd nthylglyonl. Probably the no“ inns-nuns
mum of Lmnomxmrmchhflzmbalu.

9m 930 cm gm (soon

on can |:o o quon

HEB «H . qr?! g—a an. L CH,

CH .____) Q8 0

on upon—a 4mm no cm
3.03 3:08 (13.03 on on on 0
(28.03 énzon —’2* u "’3

CH,

The. culmination of then ”mm. m mashed in that. auggutod in 1929
by ”saber-g and Kobe). Mob 1. arm in the following autumn

m —-> human“. phosphwyhtion

l

cn,cocno
glyonluo l

exposes-H -—> cacao + co.

011,090me / enigma

8*

 

Tbs work of Royal-In: shsflorsd thus hypotheses, so that than
ideas which an run: nave:- buflt upon fix-A sxpsrimntsl grmd
hats sinus fallsn Mo durum.

sou-1mm “is sad tho not that its tn:- rols X's-sins s
sushi-y, Abs name“. of uthylglyonl 1:: 510103100. systems
cannot in disregarded.

Boysr (13) am that Isthylglyonl rmuen is possible rm
WuMdshyds snd ours:- ”ions phosphstss. This tor-sues is s
shaded num- tum sn may-sue rescues, bocsnss scan mols
sum-sci did not fluid um ssthylglyoml than boiled lunch sxtrsot.

Stohr (1h) man an Isthylglynnl u toads a run. In (15)
presented strong evidences shoving tbs Inhibitory sfrsct of Isthylc
monl on succinic dam-dragons”. Appsrontly mthylglyonl commas
um: ~53 smynss to tors s stabls condensation canpound. Other .53
sums, a110, azlntuic, Mouuhydrogames, hamkinssss,
sdsnosinstdplnsphatsses srs she Mdbitad sinnsrly two succinio doc
hydrogsnsss. Sale! (16) shwsd thst muxylglyonl 1s slanted 1n tbs
mam—dancim rsts. Thusppesrs tabs omssdbys
defloisncy 1n muslin activity in tho 1.1m. Thurston, tbs symptoms
at thinnins Gauchos: can bs sttributod to mug/1mm intonation.

Tbs physiologist]. sols of glyonlsss in tissue as suggested by
Knox-hot (17) when hs pointed out thst this sum my slain“. tbs
spontsnsausly found uthylalyonl.

balm (18) shout! tbs: reduced slutsthiom is we mm a:
mom. Imoys (19), Jowtt sud “Qua-u: (2e), and kasr (21)

rspsrtsd thstssthyldyonlsndzluuudons fetuses-panama»;-
gsstsd thst this sight bs sn mtsnsdh’as in the ghouls» rescues.
Jon-u sud mam-1 (so) shssmd um. tbs .53 groups mum. by
iodinssssrssssd m 3W us sddsd is s slutsthisus (031i)
' schism. Thsy propsssd tbs follsdng mohai- fsr 31mins scum
QR
gm . can :0 3,0 $333 + 0311
CH. —_) Iis -—-? '3;

Flatt. and Schroeder (22) noticed that iodoacetis acid inhibits
glysnlsss scuba by ass tram elutstbisns , the sum itself not be.
in; has-nod, which us show by the fact that addition of glutatldsns
ssuld rsstsnrs the ssuvity.

1mm (19) slaved was the We! rm by momma.
sad glutsttdoas in the prosonss of 11m: mm was differs“ fro-
tbssfonsdinitssbssnss. Homsblstoisolstsboth. Tbs mound
fmsdtnthssbssnsssttbssmmnryhbflssndbshsndnhs
m of mamas sud ”mam. m macaw" mm
mumprsssnsssfthssnzysssbmpflnslsflmeksmu
tam glumm and mu: said. Espldns sad mm (2) demon-1mm
“ass as bass-t aim was side up of two entrapment-s. cm- of than
is Abs 'snsyas' slush converts mthylglysnl Ind phenylglyonl to the
mspudin: hydroxyssids st «and rates. Tbs other obnponent, which
thus authors sailed 'fsctor', when sddod to tho systems mom“! tbs
rsts of conversion of mthylglyonl, hm bad no «If 'sct m plmnylglyonl.

In the presence of the 'onzyms', the 'faotor' was separated by ad-
sonption of the letter on calciu- phoSphste 301 st pH 5.6. Becker (21)
deserts-std tbs pmsencs of so intemedists by its sbsorption st

2&6 a , since neither glutsthione , nor mthylglyonl show Appreciabls
sbsorption st that wave length. Using enzyme prepsrstions fmn yeast,
Escher slso studied the function sod properties of ghouls» I and II,
which be chimed corn-pond to not-gun's ham“ and 'fsctor', so! be
proposed the following nscknnisn for glyoxflssesction.

a
gun I Hogso so 11 con

0 0-) OH our) 08 __.) 0. no.) OH + 63H ‘
CH, a, (:31! ca, on, 3,0 ca,

Using sn snows prepsrstion fros ox hurt, and different tectmiques,
Crook sud Lat (23) arrived essentially to the some conclusions ss
Becker.

Although sethylglyonl is no longer ooneidored an intsmedistc
in ghoolysis, there still remain sue‘possibility that it sight be
present in biological systems. This work presents sn sminsticn of
”NM biological systms using nethods which no specific for methyl-
glyonl end per-it so investigation for this substance under slnost
physiological sonditions.

buxom-mu. mimosa

mm'm. HS‘I‘HODS

Hatcrisls

Ethenol, nickelous sulfate, sodiun soetete, snhydrous ethyl ether ,
disodim hydrogen phosphste end sodixn chloride were ell 0.1”. reagents.
Hetheson chrolotropic scid uns recrystellised full 50 per cent stlanol.
Sulfuric said as Herc}: resgent (reds. Iodosoetic scid, n-heptenol,
hydroxylsnine hydrochloride end soaiosrbsside hydrochloride uere
Esstnen Kodak Cowper: best creole. Fructose-1,6-diphosphste use ob-
teined fro- flutritionel Biochemicals Corporstion. The yeast as s
Fleischen product. Wreldehyde us obtained through the cour-
tesy of Dr. John c. Speck, Jr.

122mm.
1 Model 12-8 Celenen plutonuoronstor, equipped with s B 2 prinsry

filter end s P04 secondary filter, end the standard curettes supplied

for this instrunent were used in seeking sll fluorescence morensnts.
l lyopbilieing * apperstus ves nede from s 0 tube fitted with

ground gloss Joints. it one end was sttscbed s SO-ul flesh in shicb

use pieced the ssterisl to be dried. The other end use comected to

s tube which use plsced into s dry ioe-ethsnol bsth. The sppsretus use

enousted with en oil pump.

P ti end Stsndsrdisstion of he 1 Stock Solutions

Those stock solutions were prepared end standardised sooording to
the procedure described by 2mm end Speck (2h).

One can of DL-glyoersldehyde was dieeolved in 25 ll. of l H
sulfuric said, end this solution use distilled. Fifteen milliliters
of the anemia or. collected sud diluted to mo :1. volule with
distilled teeter. The resulting solutions were stored st 5°.

The nethylglroxel solutions were standerdised by preoipitetion of
the disesiosrbssons. To s 1041. sliquot of the solutim to be
stendsrdisod use added e filtered solutim of l g. of senioerbseide
hydrochloride end 1.5 g. of sodiuu soetsts in 15 ll. of distilled
nter. The We use sllolsed to stead folIi 2h hours. The preoipitste
use collected in s filter oruoihle end seabed with s for nilliliters of
amt. It as then dried in so oven st 105° end weighed.

Quoroastgo humus; of Hetlglgmgl

These datomimtions were denied out screaming to the procedure
of Thornton sud Speak (21.) else.

Stsndsrd solutions of sethylglyonl were node up freshly every day
fro-\the stock solution. One milliliter of the steward solution end
1 ll. of freshly propsrsd 2 per oent ohroaotropio said were plsoed in
s 25-:1. volumetric (leek. Seperste solutions oonteining the unknon
end the bleak were prepsred in the sane my. To each of these solutions
sore them gradual; sdded lo :1. of ice-cold concentrated sulfuric said.
The flssks were oooled during the oddition by esirling in so ice-water

mixture. After oddition of the sulfuric scid, each flask use stoppemd
end plsced in s ehellou tutor both containing inc-set” mixture. The
fluke vere then trsnsferrsd together to s tater bsth saintsined st

50° shore they sore Mrlod for five ninutes. At the end of this tine,
the flash sore trsnsferred together to s cool aster bsth more they
11ers quickly brought to rcoe “aperture. Each flesh use diluted to
the Isrk with concentrated sulfuric said, end the fluoresccms of the
new use compared with the etsndsrd, using s blue print] tum,
sndsyellesesoendsxyfilterfortheeeesurenente.

The sword-cuts described belcv involve the eminsticn of seversl
biologicsl systems shioh it us thought sight centsin ssthylslyousl.
The asteriel stained use lyophili'sed' end l-Il slicuote of the dis-
tillste sore trestsd Iith chromtropic scid, scccniing to the pro-
cedure described shove. The fluorescence as compared eith thst of
s knoll stsndsrd. ‘ I

Emulation Iese
Five grsns of hsker's soft yeut me pleoed in the lyophilitr
in; sppsrstus, sndfthe distillste use collected.

st Est
1 Isle ret use sserifioed. Five eillilitere of blood were is-
ssdistely collected fru the eninsl end placed in the lyophillting

Iowans.

EnniEtion of get Liver
i fensle rot, eight nonths old, use sacrificed. The liver was

reacted inedietely, plsoed in s fleck, frozen end lyophilized;

Eminsticvn of least remntstiog Mixtume
The fereentetion mixture use prepsred in the folloedng more

To 10 ll. of e 10 per cent solution of glucose end 10 ll. of s 0.1 I!
disodiul hydrogen phosphste solutia were sdded 5 g. of yeast. This
nixture us slloved to stood for too hours st recs teqereture. l'ive
nilliliters of this suspension were lqophilieed. .

Exemdnstion of Liver root Inmbatod with Fructose-1 6-D h

     

he Isle rsts em sacrificed sni their livers (totsl night
18.} I.) were moved. These were then ground in e sorter with clesn
send snd suspended in 50 ll. of berets buffer st pl! 7.0. The borste
buffer use prepsred by titrsting 200 ll. of h ‘por cent borio scid solu-
tionsith 0.11 sodiu hydroxide,ueing epnum u. check the pa.
'ren nilliliters of this suspension sure sdded to 25 al. of 1.29s! cent
fructose-1,6-diphosphsts solution. This ninture sec incubated for
an hours u 37". run use lsbellsd mixture 1. the reminder of the
suspension on slsc mouse a 37° for an hours. he Iilliliters of
thisnixturelere thenedded toZSnl.efsl.2per oentsolutionof
fructose-1,6-diphosptnte end incuhsted further for s 214-th period.
This sss labelled sixture II.

it the end of the incubstion period, 5-nl. sliquots were removed

from each nixture end lyophillzed. .

 

10

Eminstion of Yeast ‘fiothzlglg on]. Added)

To 3 g. or baker's sort yeast see added 1 ml. or 0.1 H iodoocotio
soid solution. The mixture use slloued to stand for one hour st room
tomsnture. One Iflliliter of s solution continuing 0.00251 leg. per

ll. of nethylglyonl use sdded to this. The nixture ‘ns inedistely
phoed in the lyophflising ' sppsrstus sad the distillste collected
(h IL).

 

i fermentation limn'e use prepsrod in the following mm Ten
milliliters of s 0.1 H gluooee solution, 10 ll. of 0.1 H dieodiu
hydrogen phosphate solution end 5 g. or yeast were mixed. After stand-
ing for two hours is m mnzm, 1 :1. or 0.1 a woman said
modded. Tinnixtm'eussthonsnomdtostandtormhslthour
longer. Four nillilitere of this meponeion were removed end 1 ml. of
sethylglyonl solution oontsining 0.0385 mg. of this substance use
sdded. This portion was then lyoptdl‘ited‘.

Waco t Blood Math 1 Added

i not use snoritioed end the blood inedistely collected. Five
-Iillilitereo£thehloodssreplscedinsnssktowiohssssddsd
1 .1. ed’ e solution of snowman omtnining 0.0021 lg. per .1.
The flesh use connected tothe lyophill‘zm~ eppsrstue end the die-
tillete use collected.

m tion o; fits for Heth‘glglzgg

The leaves o! the follow; pleats were investipted for methyl-
ghouls Haple (5221; W), lulborry mg), white osk
(% rcue _s_1_b_s_), shite fir (£33.22 gmolos), blue gross, ivy
Qanthonoogeue tubuggidsts , end tobsooo (gm m). In ed-
dition,theroelssndstmottobsocoplentsuereslsoeanined.

Mention or Log Extrect
Fresh lessee were collected, dthout the steels, fished end placed

in e deep treese mrnisht. When roedy for use, they sere the-ed,
plsced in s Inelin cloth, eud‘pressed in the hzedrsulio press. The Juice
see inedistsly {men eith dry-ice end lyophilizedy

Paar aluminum:

Ltteqts were sede to identity sethylglyonl is ssple lessee by
tomstieuet thesetlvlglndseendsopu-stiensnd identitioetionot
thissubetsncehypsperchro-etomlysocordingtothspfloedmot
Spock (25).

Tmtygrusttnshlesvesmcellsoted,‘shedendpoendio
ewmum. Totldssizturesere eddod 100-1. of distilled
ester, 1 a. of hydrewl-iae hydrochloride end 1.5 s. e: sodiu acetate
. (trihydnte). This We use «We eith sodiul mm, m
extanted'flth an 504-1. serum at snhydrous ethyl ether. The
mined extrsots were dried overnight over anhydrous sodium enlists.
The ether use distilled off, the lost portion being removed under vecuun
st the nter m in order to «Bid overheeting of the residus.

The residue use then dissolved in 0.5 ml. or ulster end e for drape

of the solution plsced on e sheet of Whetmn No. 1 filter paper.

The solvent used for paper ohronetog'sphy as enter-sstunsted n-hsptsnol.
After ellouing the solvent to rise for 20 hours, the sheets were dried
in the sir end sprsyed sith s 3 per cent solution of niokelous sultete
ts idol) had been edded 0.1 ml. of 28 per cent monie per 100 nl. The
chronstogrm' indiosted sover‘sl throw mote end considerable 'tsiling'.

In soother ettmpt to identify sethylglyoxsl in lseves, 35 g. of
freshly collected ssple leeves vers mum in s nortsr with said-flushed
lssni end pressed with 15 ml. or distilled aster. To one solution use
edded 1 c. of lwdroxylssine‘hydrochloride end 1 g. of sodiun ecetste.
The solution see filtered end trested for pspor chrometogrsphy scoot-ding
to the procedure described m. The Mum obtsined in this
ssnnsr sleo shoved oonsidershls 'tsiling' end severel brown spots.

In order to definitive]: prove the presence or meme of methyl-
glyonl in pleats, neple leeves sore collected, vsshed end pleced in
the deep treeee overnight. They were then themed, placed in e suelin
cloth end pressed in the hydrsulio press. The eetrsct was inedietely
trcesn sud lyophillieed. To the distillsts (epproadnstely 50 :1.) was
edded l s. of hydrcxylnine hydrochloride end 2 g. of sodim ecotate.
This solution see treeted for peper chronfogreplv eccording to the
procedure previously described . The results sre indiosted in Tehle IV.
it reuts'. Id. ,. - Heth~l-._-xel h _‘ or Chrometogaue .'- : t Liver

i ssle ret us ssorilioed end the liver imedietoly removed.

1)

limplscedinewsringhlendersithsscluticactlg. othydronle
sumhydrwhleridesndlé s.e£sodimsoetstein50sl.eddistilled
enter. This suspension use treated for peper ohmstogrsphy seem-din;
tetheprocedu-epnvieulydeeoribed.

it t to Identit Roth 6 t h P or anatomy;
Twat: grass of hsker's so.“ yeast were plsced in s Wsring blender

Iith 50 I1. ct distilled nter outlining l g. of hydrcxylsnine hydro-

chloride snd 1.5 s. 01' sodiul scetete. This suspension sse trested

for pspel' chrmstogmphy seem-em to the procedure previously described.

 

 

 

 

 

 

 

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Laws hm 2:331: 2:125

 

$1313.78 .1231) EISCUSSIGR

Eo mthylglyonl could be round in mt, yeast “mutation
Wurst blood or rst liver. The methods employed for this
investigsticn offer several advantages over the previous techniques
used to:- detersining sethylglyonl. They ere quite sensitive sod
specific end they persit on examination at hiclogiosl syetons under
slnost physiologicel conditions.

lstudyussede inerdertodeternine whether itiepoeeible
to recover sny sothylgb'onl when it is added st low concentrstioo to
these cystezne. It was found the“ rim setln'lglyml. see added st
constitutions of 0.0013 mg. per 31.550 yeast, yeast {orientation
sutures «- rst blood none could he recovered. These results on he
sttributed to the imodiste {ammo or s complex between sethyl-
glyoxsl end auutrdone. (Severn mammal-e (16.19.20) have
shown thst glutethicue ects es s ooensyme in the trsnel‘omtien of
uttvlglyonl te lactic said.) Since this reaction can be inhibited
by edding icdescetete (22) these experiments were repeated with inco-
scetete present. The results, which sore not sleeps reproducible,
ere reported in Table I.

thhcuda those recoveries ere tether low, they nevertheless one
be considemd significsot-opsrticulsrly in view or the instability of
sethylglyonl either st love concentrstions er in the presence of
biologic“ system. These results inflict“ the?! if nothylglrcnl

7.431433 I
nae-ma w nanmmoxn min; To saws cmzrmzmo macaw/:75

 

 

 

3;th Flethglglyonl E‘fi’gthflglyoxal Po:- Cont
:Ldied , Mg. I‘Lecove rad , Hg . fiacowry
Yam 0.0025]. 0.009 '51; 32 .3
least he mutation
matures O .0355 O .0531? S .0
Blood (Rat) OJE’A O O

 

”emulated appreciably at any stage of. gig-0013313 one limld be able
to damn-t it by the“ methods.

Ariyama (Is) and several other worknro reported tint 11m extracts,
when: incubated with hexane diphosphate yield nomylglyoxal. It was
decided to repent. this work in order to damn-nine whether mafiaylglyoxal
could be detected under those oirmmtancoo by motion with cam-amo-
tropio told end flumtry. The result» of thin experiment, which
are mportod 1n Tabla II M Ihich indicate the presence of. nethy -
glyoml, mpport Ariym'e work.

The original llvor homognnatec-borata ammo m nloo watod for
nothylglyoml. no mfliylglyonl coulfi be detected in this suspension
when no thief-owl MpEnopLato had boon added.

Thou maul“ elm 1n Tabla III indicate tm presence of 11321111-
oant mount: of mthylglyonl in plants, emxmy 1n the olzsor green

lemme. In ordain to detomim definitely 1’“ preaonco or obsonce,

 

15

mm; II

ammmma (1' ISLE 1372400 12' .1153 IfiiL‘Bl‘f 55;} WITH
:3me 2-1 ,6-EIPEEOEPKE'E3

“w

Blank Standard. Solution I Solution 11

F1 scram-tor

Bowling! 0 . 7L ‘ 100 37

 

‘ The mm contained 0.00095 mg. of uuzylgh-onl

u- “u. :

~'\ 4"

TLRBLE III
Similflfi PEG! THE} 513313310}! C? W33

“— _._ “ ‘M A M .J
._ 7 J

Plant Finer-ornate:- Roam
Emined Ufiom Em

A 7 Q fi' _

0 e" . -; “7.. .
i"

 

Kaple Lame 346 85
Gran O Eh
Ivy Leave: h 95
Kulbo my Lame 0 92
Oak Leave. 10 92
Mite Fir Lame 16 92
Tobacco lama 1h 69
Tobacco Stem 27 100
Tobacco Route 0 73

 

" The mm solution contain“! 0.00096 mg. of nothylgljonl

 

SO-nl. porno“ of flotilla“ from lyophu intion or naplo lea: Juice

were eminod by the prooedum involving paper dxmtogrmhy or :32.
ounce. The moun- of those oxperimonte era elm in Table IV.

 

17

TAfiLE- IV

ESLLTS Q9 P¢W:fi CIfiiCi‘fi’ECfluiPkfl G“ LBJ" EXIEL‘xCT (mmmv)

{H

 

 

 

R! Vain. futility of spa
Hemlzlyoxim 0.58 Bright. pink and fill”
Un’mmm 0.96 fellow, um: 'tailing'
Then result: indium that uuwlglyonl doe. not occur in the

have! of thin plmt . Instead tom other mum. compow having
the preparty of forming an crime which given t yullov oomph: with
mission- ion teen. to he want. This cub-m appear- to met
with chroma-epic acid undar the conditions 0! tbs procedure for
determination of wthylglyoul to yield n fluorescent W. The
moans undoubtedly sex-1t: mum- investigation beam» or no
probable dgrdticanco in ptmtosmthosia.

«1
I1
.1 *4

53. ma .1172!

1. least, yeast tomontauon mixtures, r41". blood, rat liver and
plants tors tannin-d for hour-{151mm, Tho oathodo involved:

a) Mow-11133131011 sud fluororao’a-io dotomination of hotly]:-
£13031 in the distillate after reaction with chromatroplo
acidouulmrio told 311mm.

b) Lyophil’ization of tho fixture followed by conversion of
carbonyl compounds in the distillate to their om. one!
oopmuon of the claims by papa;- mumphy.

2. Ho mthylglyoml owls! ho dotoctod in yeast, smut {emanation
mixturoo, rot blood or rat liver. Hothylglyonl could be mavens!
“to? being added to the" Oyutmo‘, providing iodoaoetato m addod
also.

3. Evidence for noting-7131:0103. production as obtained with rat. liver
homo—gonna. 1noubotod with (mason-1 ,Miplmptato .

h. A volntilo mums m found in plants which gin- . nmooont
oompmmd on "action with olumotmpio cold under the oomutiona
of the pronoun for methylgmonl.

5. Thin whom: cannot be nothylglyoml “no: it could not be con-
noted to an om baring on R! value identical with that of
nothylglyoximo. .

117;}: EGRJ CIT EB

19

LIT—LB. iTURB CITED

(1) a. :3. mm m: a. w. Dudley, J. 3101. Choir... 35. 135:3, (1911‘).

(2) r. o. aopzm m: 5:. .8. Roman, mom. 3., )3, 320. (191.5).

(3) a. Toonniom and v. 7mm, 2. m:- Phyoiol. cm... 13, 257, (1926).
(L) 1:. Ariym, J. 3m. cm... 11. 395. (19253).

(S) c. venom ma 1!. label, Bioolm. 2.. 33:1, 236, (1929).

(5) 2.. Aux—in and 3. Simon, Comps. rent)” Soc. Biol. Pm... 119, 905.
1933 .

(1) J. 5mm and a. Lolmnn, Biochon. 4.. 31,. 1186, (1937).

(8) a. a. Can. we! a. r. Cook, Biochen. J.. g, 1319. (1931).

(9) c. Nouborg an a. mm, 2. mgcpxma" _1_. m. (1912).
(10) r. 1. 1mm and o. x. Meyer, J. 3101. cm... _1_1_. 351. (1912).
(11) B. 9. Dakin and R. W. Dudley. J. 8101. Chem. 33;. 555, (1913).
(12) c. amber; god a. no.1, Mocha. 2.. 391. 236, (1929).

(13) 1'. Mayor, Bull. Soc. cm. 3101., :5. 1395. (1953).

(1h) a. Star, 2. fur mum. cm... 19.5,, 15, (1932).

(15) x. m, J. Biol. Chem. m. 289. (1950).

(16) a. a. Salon, Bios-hen. 4.. 21, 227. (1951:).

(17) o. Eeyorhof, Elgob. Emflunoh” y. 208. (1935).

(13) 1. Lemma. Bioolm. 4., 2g. 32, (1932).

(19) s. Xmoyo, J. 31mm. (Juan). 3;. 319. (193i).

(20) x. Joann and J. a. M1, 31mm. 4., 31, has (1932).
(21) 3. Backer, J. Biol. Chem, $22, 683, (1951).

20

(22) H. 2.211;“ and a. 2. Schroodor, J. Biol. Chem... 192.. 2:31. (193k).
(23) :3. a. Crook and 1:. Law. Biochsm. J.. 2;, 1132. (1952).

(21) a. J. Thornton and J. c. Speck, Jr... 1511. Ciao... 33. £99, (1950).
(25) J. C. Speck, Jr., momma work.

(25) r. a. Hopidnl and a. J. sou-gm. Biochem. J., g2, 73, (1318 .

 

WTBY LIBRARY

Date Due

 

Demco-293

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III
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