Studies on trimethylamine oxide

78 Pages·09.03 MB·English

by  Benoit, George Julien, Jr.

9TODXB8 OR OXIDE
w
GioRGi benoit -mum
m,im
A tbesia submitted for the degree of
DOCTOR Of PHILOSOPHt
IMIVERSITT Of WASHIHGTGH
.XH*.
I 'P.. .
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4S.|*WI*lIlilBiS
An autiiGr wishes to express Ms
. t® wmmrnm® s«ri &» Wmm&m for tl* «#-
gestioG of the problem and for his interest, eaeoor*
«gm®at, a*4 *#*£«*} to -ffow H*-
.M y M A itM w r
«*M» Me* SojmM. X» iQntifc&m* Godrio I*
«ai rMlMMt x*nm i t* fMa***** nil, of t&# intror--
* ' ,.
*ftgr of' ftMfelAtto** fair tlioSr ooofoamttoii out <Mt»
M$o«t*13y Is the tmwmmm o# aoae of' tfc* ajarino ®r«*
ganisms studied; and to Professor Donaldson and Mr.
Jttoiwirt f» of tte tAufciait«* glut® StfftrtoMiat
Of vioterlott far' pmmmw&m. aoae of ti» aoootiaow!
ttOOft*
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#
the feedings continue, ite ab ility o f tte animal to
WBWft the ooapettai into the Iumwmwo UiMtfcv&wlM oxide
jseeift MLOIeMft ■hfcefc.nfcJiMgM-jn; *> SEmWSSh UtoBKSMMOSaift'a Vk< i%
OSd*UtS Ok SP^gfiu ^8 JiipUvUf8st8m—ifcs*n8 to iw-'Ot'
Mtlter(SYf offars a different oxplaiiaiieii* Enlarging
©a tte finding: ®f HtetetefSBf tte t intestinal teeterts in the
«Bt and «ut oMld apXit o ff targMlte&satn* fte» telMthjrteiiiJt#
oxide and choline, but only In troees from betaiae, he ascribes
aid. tef ^thyi-ate »«* and/or tyiteteftete.te oxide found in urine
o f animals to which choline or tetftin* tea. teen fad to tte
above bacterial action rather than to a conversion by the ani­
mal. He found the urinary iriisebftyliiiateS. oxide output of dogs
to te ’imiepem^Mt of in jested choline or be tain©, hut tee tr i-
mo thylnmine output was increased{37,39}.
Mdller (3?) found that when large amounts {Z * 3 gms
nitrogen per' animal) of -la&MtteteMMftiiMi chloride were fed to
tely half Bf • the is jested material teu il te yeeotered as
amine « oxide with m eoasldateftte ftttt&M uu«te»ied|. the *•»
MiBdip teeiBE fe*«" found in the manm*tm fraotion* terse
amounts of tiiiiiterteiitee oxide (2.6 » » nitrogen per animal)
'UHiUPeFewHNp'nesst ,w-.»i» w e<*» .wswwf imn^mMMwiar w.«mim* "* m*r • kww ' ^gyw,ty — W!Swl; — ... •■ . jp
©are ateut 1/8 recovery as triM thylaiiine exids * teteeihyi-
amine, the Vfttto of tte tee teteg #*!». tte wmriMa* w » te.
the ammonia end urea ImetieB*
FuehsUO) seald detest m more ttem traces of IrS**
metfeylamiae or tvlM tejlftaiM oxide te the urine of dogs to
which ted teM fed Smrgs aasntabs (20 pm) of eteltee, m itten#
it ess nearly completely decomposed.
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10
m € Liatsol<33) J^sojiMgato#: th® noraal
Mm® w.lifitttS' output of trims thy Isuftln® oxid© on a pay*!
l3jtt«fe»firo#) dlot* Mm tm m m r ftsfttag .11#' • *■§§ s§s par 14
tours; the latter, 31 - 11#* farridl), aoafiraiag LIntzel’s
work (32), showed that mm is also oapahle of transforialag in-
jested trimethylamine quantitatively into trio® thylaaia® oxide,
white o*»i t.hytataj«w oxide when fed is auoaMts&ivoly excreted
In view of the jjofw.l excretion o f trimftthyiaiaaltw* ox­
ide in mammalian mrlne, It would he interesting to see It there
is # ocourreac© of this compound in the fM##ls tlesue ot
these anfaals. Kapellar-Mler and >2rael (42) claimed ■%&■ M m
shown by indirect aeons that hoof muscle Mesne contained Ml*
aothylaaine oxide to lie extent of 8.0 «gt.$rt»eMyi«itgiao oxide
ner lOO ooa fresh muscle. do« m scle contained 8*4 au&s
per H# gms fresh muscle. However their experimental method#
are open to mpIm* oytMolsa. »#y roftiixe#. « wmmM extra**#
in what we# essentially 15% sodium hydroxide* This drastic
treatment hy iholr mm ®#»l##toa saponified m.MflmtmA m m
pounds, giving w$.m to mono* aal tsiaalliylaaSiaa ®v«a whoa none
a < jttMMBt la the prefer*®# condition* Mtmt they reported Mai
I## gas of «N»dflMi .wool® co»#ol»i ©»i#l#. ga #f ppo-fonwd trl-
aethylamime and 0.0167 gn of trimethylamine oxide. Since it it*#
Mm, shown repeatedly that fro sh fish auecle tissue, such as
thoy ololm to tor®' used, is very low la fro# Marti|rl*atft#«
tfeoir result# are doflaltoly qmesMeaafeie*
M m tm W S} found m w&0t$SUmmM iaeyea#® 1» tr i-
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u
mtkfUa&m or ©xiis mammUm tr©m tmmm sub­
jects fbllM&tg 'Vigorous ■ mmmtm*
m?mrnmimt9 using m m%m tm t& Sm * sr tlt
gotoot no trass of trimethylamne in frasji,. noriaal, M m urine,
from M s m m It® ana the Stullar cMM'tt? fa&sinfMt, It Is
probeblo that the triaethylaraine reported in iunam urine from
fclaa to tin*® |41f 4%til#4?f jronN&teft s ithmr tmm MsMrial mm
otisr raduotion ©I part of tfis oxide prassnt or
fr©j» the splitting off' daring tfcs laaiyiM al -proosiiirs of trl* -
w«-tkyianm from ©losely related oonpounds ff#snsr anwgfl^iy
la arias» SvMsass fsr th@ rebusiton mm found bjr m m m l worts-*
era(34,41,44).
If «*. «re justified in anamlng that 'IrlMtiiflaiBims
fsan# In urine of .all asaawal# resu lts iw » o®*
14s, sag is not aeSglaftUff prsssttl as «»«&* then th* sttttx*
known pis'tar® of ustntw llm of trinethylaaln® onMs and related
. soapowiis in smmsM o«n "is swBoarisod as follows*
JU • feiaslfeyiiwilns ©*Ms is mm«roM rapidly «•§ near­
ly quantitatively without fitriliar sfraftfl** .
$m . when lajootsff is aonvsrtsd at
least largely Into trims tl^laada* mmM# «wt ip an tllalivsly sap® .
ovotoft as sack. .
S* .. Swire. Is a definite,, ansll sssrstlon of triiaethirl-
snls* srtde by Imnmmlob a atxot diet* %here is a
ooaparahle onsvwtlMi i f stk r wwwals* .
4* . » s injestion of sons ooapMds .U»*nS**t eholiae,
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12
m tk fl ««tcv a t diwteylateteaoeti® m«M| timfamXt? *te!2ftr to
trlaeteylaalit® oxide gt»#s rlee te an elimination a t
.totm tkyhudm oxide* tka mmmmt a t tte m m m m mhloh mm bo
maamata* .tow am tte teals at tte telteteylam ia# oxide exoreiei.
defends on tte coiapound, on tte dosage, and possibly oa tte' diet
meed* Iter* la evtdem* teat tte triiaetbylamiae ®xi«® eliminate
ed M e t tm » tte trimethylamino sp lit o ff ttese eompoaads by
Intestinal bacterial action.
5. latestiaal bacteria can re&tsee tei mfttbyisaln* ax**
ide to te* .m*** aeyi *»itey If tte aiaount of iajeeted triwteylamlme
oxidO' I« large,. tte. *ovy*apo*aiBglf> Iar®@ bmmit of trlmateyte
amine formed nay be excreted in part as sate instead of ia tte
form of triaathylaminG oxide*. By tte same token, i f a large
amount of trlmsteylanlae t e . tajest*#, part of it may te excrete'
e i ia tte free few*
aMte®o»el#i| elelme to tee* stem tte*. tte urinary
elimination of teJ^teytealne from elfelae rets Increased merted*
ly daring starvation* A* soneindes teat telwteylai& lse tee m
endogenous origin* la tte author** opinion, tee yetelts aad
•owKtaioa are grotem y of fft*stlan*fc!« w J*»# teeamee of tee
feU eeliig reasons!
1* He aeei « poorly em tw lled matted of analysis
tmt M w teylm dM # 'fain®* efe&efc te ■ obtained do not, in a ll
'fgotetelity, represent teiaeteylesiiae at ell* -
g* i t ie entirely peeeible teat te® new el deviation
of M e titration vale® .ie smf fie lent te aoooaat for te® it* ■
@r«aa® efeleli te. reported*
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m
II* i m W&BKt ftt BftlftteXKASSflR i f m im mrr.«m«f mrim
JMf9#mW&
fo r' tea puxpmmmm o f this m u i t m * tea^imSte to « n
a method of setemI.teSft teteetete wjii«i|i. waMia x* bo
spaelfio, not being influenced by otter naturally occurring oo»~
pounds o f tte^ totals* and olioline types; 2. hav® good sm sttta*
Ity* because the concentration of trftmtfcf&MMfa* oxide In most
organisms in which i t oocurs is rather low (order of magnitude
of *«#£ trtathySttriUMi oxiis ftiteitato* (0%) 3$$h* GBUHg©, on tte
M int weight basis); . 3* have good eccuraey; and, 4*. eoasuae te
lit t le tfm as possible* ft*# i#n#mma tm teStss end ©resist te*
tion methods uaed ly early workers te white m m arystelltiia te*
vtmrtl*** 'Msnslly tte m rtattnrtte* 'm s obtained end meljwat*.,
m m teflalaat te tte test term etem -asntitesi erlteri® « i
« n tterofteo raJested* tte mttoft o f W ilflrfS?), which In*
Totees Isolating tte tea® as tee reteeteate m lt fWUtemft by
TOlwsetrt® analysis of te® «br®§itwijt. present, aptmmft' slase
this work m s sterteS, tet it , too, is «f#n te tee. sr ltte im
o f undue time consumption.
I&»t*sl(9S) a ite ms® of the fa st tte t trlm thyiaaiiie
oxide I® easily ratesoft te tetmtkqrtealte*. sftlte te teen anti*
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M
m M U Bb M il mwmrn*& alley (IQ jnrtn aiaaijuua, I farts
#Ofjp®r, 1 jpnft sine) in HG1 solution as the reducing agent, fit®
metliod M® found to be highly speeifie for *»®iM orttsnf ah®**
line t be tains, y-butyrohataines and carnitine «r@r# found m* to
I»tSpftMFS*
‘i’o 'test Ida tael* s ae thod, Syf .MMyVMim oxide was
■prepared by %be method of Smst®* mmI. flsn&tJ&cfl?}* lb® hytrats
melted at 9g« U yt».e gpye 'tbe fhw r ^ f.e*l peroeatage nitrogen
irite* analysed by t b# aethod. Galoulated for
%*m mm* m # » * u*9, is«e* m*s*
In cemflraatloa of Lintzel» s worlc, tbs author found
tb s. redaction of WlmmwXmAm- enlde la ***■*£ bfiroah^rio
said in %k m presence of :e varda* a alloy (o.5 gm/5 mis solution)
to be ©onplate la 30 atatsn at 10t® % IM minutes at if* %
and 3S nihates at ®§* 0# AbM* tihs rednebten m% Mf® « MS no-
companied by a ear tala amount of frothing, stoat of the reduc­
tions ware aade at 95* 0. ifce .period of heating should be as
short as possible, commensurate with complete reduction, so
that possible splitting off of trimothylamina from choline,
batalae, e tc ., be eliminated or at least reduced.. Under the
conditions of mxpmrtmmt used, ohollse-t hoMlm^ aieibylgnaiild*
ins* creatine* tyrosine, eystln®,, gSptiss* «M proteins Mrs
found not to interfere:' 'to' «n «pprectaMe m M I*
fasts for complete redaction sera made on 1. pure
aqueous solutions containing known amounts of triaethylamin©
oxide; a, bt»le§i#«I fin ite farise|f ant, S, SfMons fct*l*«r
lea l estreats ffrea psttatt) to Shiah. had been added t e m
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m
« w a t» of ttim tfcftettte* oxlte« fsiactioa ate rssowwy were
template to w itite tte U a lt of - sapiirtesatel » f f» fU l*
te* tettewttetteft of trijaethyXaiaiae «*SA*ff then, r*~
tette to the teterateatfoa of tte triteiaylaiitea fM te frwa
it* fh© text 9^ttM i was to offs ot a sepiiyattea of trlteihyl-
amine fmm otter steatite tasos* Stebtyftt) showed tint tte
amount of cliaathylamine present in fresh cod muscle press Jmis#
was m m .i®w* tel** te t «% a ll of tte same order of magnitude
m- tte trimethylaraine oxide present. Meager evidence, mostly
of a negative nature, indicates that tte mmm®thylaalae contest
of fish muscle is also much lows? than that of irlstethylaaiae
by tte bacterial mteotlon of tetasteyiaiatae axtte#
Since higher hoaologues have oof been reported from these Ma­
terials, amiaonia is the principal, if not indeed the only, vola­
tile base other tte*- trimethylamine present in reduced fish
atetee «xtm*t*»
The 'late: of oetem *** o f mono- and dimethylaainea in
fish ft*** aot M M M lteU their aba®®** te otter b iological ma­
ten®# a nebbed of aaaiyate ter tv&tetkytexlte
te r ia ls, a teo li
bate la te account itelr je«tete» tettN R H t
The fir st attteP* te saturate fiiaattttatirelf tarte
methylaaine from primary aai secondary amines and ammonia was
by te tlliffilte l* m s xteteft was ifctei oa tte- rosisteao# of'
tel»teyl«»ia« te. .MtUiiti** by stetw* fcffoteoialhe and tte vteft*
tlte case of' oxidation of ®ri»sry .ate secondary antes* ate aa-
aonia by It*
m is wetted has te rsssmt .ysars be®a nearly coapletely
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