CHROMATOGRAPHIC DETERMINATION OF AMINO DICARBOXYLIC ACIDS IN PLANTS

• Document Type: CREST
• Collection: General CIA Records
• Document Number (FOIA) /ESDN (CREST): CIA-RDP82-00039R000100080033-7
• Release Decision: RIPPUB
• Original Classification: C
• Document Page Count: 9
• Document Creation Date: December 22, 2016
• Document Release Date: May 30, 2012
• Sequence Number: 33
• Publication Date: September 8, 1950
• Content Type: REPORT

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Title: C1fltOATOCRAPIIIC C'iTU1 INATION Off' AMINO OICA OX1LIC AC1DS
IN PLANTS by V. L. Krotovich and A. A. I3undc1' (USSR)
Source; Oo~clady Akad?mii Nauk SSSR LX:, 5, U61-~, 11 Auk; 1918
eriodical~
Thrice monthly
(
CONFIDENTIAL
50X1 -HUM
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CONFIDENTIAL.
G o ~ATOG iA :i J T MINATIQN Q1 AMINO
DIcAf 13oxYL:w AQII N PLANT8
V. L. Kr@tavlah anci A. , t1u tel
Tho relo oi, dicarb?xylc arn:Lna acids in plants ie v@ry gr?at.
From thorn aapara rro w c1 1utw r,re ?o'rn i; they ?ori a ~z aooaary
lint; in t1 on~ynatio tranow~rinatior{ roa~{tion; anc they urc of
aiT4 ular rAportww iil the ayntho is curioidg j!,7. 'or
thooo racoono, tau quuntitiativo dote aination is of vory ~raat
of nifioanco in invoati Ltinp tho rctaboliQ:n o1: the plint c1 1.
loYravar1 ?tho Ioror~n r thou It~rrOI~tly in uot; ?or this ictdrmination
is vary dii'f icult rind vary inaocurato.
The claosiccil invooti~aticno of M. Tovot Lor tho f iziet tiro
initiated tha wido-sprread applicrr,tin oi' the chronta o~raphic n thod
in or&unic and biolo ical chanistry. V ior~d then EJ establiehod
the pop ibi1ity oi' uoparatin icarboxylic amino acids ?runr other
arnir-o aacids by oxchan ;a adoorpUon on altualrrum oxide. Lator on,
ho u~od this 1~rincip1o to invoatiatO protoin lrydrolyzatea~~?
Krutovich :n.1 uuniol' uocd tho chromatotruphic mot' ~d with
plants to dots iuLnu w~tounts of amino dicarbo. rlic aciEIc. ThJir
pros duro convicted ?n passisrx an uquanuc axtrct w plant material
(tactivatod with boiling oth4 alcohol) through a column of
which had boon treatod with weak hydrochloric acid, end the subsoquont
elution of the adsorb?d dicarboyl~G Wino acids with a solution df
alkali.
The a1uuninurn oxido, standarizod according to Brokmann (11>re of glutamio and aspartio aoida oontaining 0.66
added to
mg of nitrogen; they found 0.68 mg of nitrogen of amino dioarbox~rlid
aoida.
In another teat, they wachod the adsorbed amino dioarbox rlic aside
which had ttot been added but had been preaent in the plant extract in
(
the natural state) with a hydrogen au1fido solution. Lupine sprouts were
used and the adsorbed e, traot washed the first time with 100 m1 of a
saturated HS solution, arid then with 50 ml of distilled water. In both
2
inatanoes, 0.37 mg of nitrogen of dicarboxylio acids was detected per 2
!121 oil ecti'aot.
Foregoing experiments wore instrumenta1 in establishing the foUow-
iang procedure in thic reaearohs
Fresh dried plant material (2 g) is inactivated by boiling with 96%
ethYl alcohol in a porcelain dish for 5 minutes, during which time part
of the alcohol evaporates. The remainder of the alcohol is removed with
benzene , and the dried material then pulvorized in a mortar, after which
,
rcr;'1t1#t /\L
Declassified in Part - Sanitized Copy Approved for Release 2012/05/30 : CIA-RDP82-00039R000100080033-7
 Declassified in Part - Sanitized Copy Approved for Release 2012/05/30 : CIA-RDP82-00039R000100080033-7
h
into a poraolain dish aovorod with
it is tranaforrod quantitativo~.y
30 n3. of distilled water, The maerial to woU mixed with the water and
a
11owd to eta for a ha at 20?, before being passed through the
a~,f hour
ol. To a fivon quantity of the ?iltrato
paper filter of a Bueohner funn
(usu drop of an alcohol solution of phenolphtha-
a1ly 2-~ m1,) im added a
with 0.05 N KON to a faint pink coloration
1ein~ It is then neutralised
and oorpLion tuba dontaining A2.203, whioh io pro-
introduced into the ad
pared in the f o11owtn mariner:
Hrokmann ~Rusaian epelling],
4 g of Al etandardiZed aooording to
2O3,
h 12 ml N UC1 for 5 minutec during which tim? it
are treated is aon-
mixtur'? is percnitted to stand for 1-'15
wit 0tinuoualy agitated . Then the
minutea, aft?r whioh rho cloudy liquid is poured off and 40-50 of die-
tilled water added. This mintt'r? is well ohakQn, and then daoanted after
as pr?oipitut? sottlos. The precipitat? i~ gashed by decantation until
there With 1~itm~as. The adsorhent prepared in this
ie no said roaotion wit
way is kept in th? same flask in whioh it wan treated, vnd?r water. bo 50 am in length and 'l-g mm in internal di-
A pleas adsorption to ~
0l~ 12 cm from the '.over end, is used for ohroma-
ameter wish a o?nstrioti
tog'raphic doterm A small piece of adsorbent cotton is placed
~.nation ~
in the oon~triated paxt, The tube itself is fastened with a rubber atop-
per into a Hansen flask and clamped to a rook The adsorbent is poured
into the tube, and thn water in which it was 0enta3=ned passes off entirely
(either because of the !' .oras of gravity or aided by a small amovuit of
be tooted is than ati1oked through the adsorbent
suat3.on) . The solution to
at all timea covered by the solution)
(taking oars that the adsorbent is
in 2 aeaonds. Next, the column is washed with
at the rate of one drop
As a resat, all of the neutral amino acids
50 ml of diat3.lled water.
(3,nolud the 1S to aysteine) pass through the ad-
wash water is then poured out of the receptacle
sorption column ? The
which is rjnsed. per the elution of the adsorbed diaarboxlia amino
aside, 3 ral 3 N KaN and then 0 ml o?= 0 ~ 05 N ItCH are introduced into the
3
-w 4 -
,CONFIDENTIAL
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F i )4P Al
R~
auakod through into the roooptao1o. Tho liquid #rom the ool-
tube a
until tho' reoiduo is drys ao that the salts of dicer-
'unn is drawn of
o amino aoido are tranafarrod to tho rooeptaole, The oolution
bo~li
quantitatiVOly tranafer'61 into a K j?1deh flank, aM the d trogen
A
i
,~uaoian apeiiing7 mioro epparatua.
dotormired with a Parnaa-VaSner RD
Rea~11ta aro oxprosaed in nilliSrama of nitrogen of amino dicar-
bo io aoida per gram of th7 substance. A oontro1 solution is pro-
aubatitutinS an OqV 1 quantity of pure wator for the oxtraot.
dared by
Krotovioh and nndol' attemptod the dotormination o f froo amino
dioarboxYlio aotde ooourrinS in thn natural state in plants, and pre-
pared tho data shown in Table 3.
Thou' method iQ dosoribad as aoourate, quiok, and not roquiring
Ppstatus. They nay it can bo usod, too, for detormina-
oomplioat?d a
tion of the amino dioarbo'rlie acids in hydro1zateO of proteins.
The authors of this paper, which waA au1 fitted 15 June 1948 and
?nted the name day by Aoadomioian A. I. Oparin, aro affiliated
prey
the Inctitute of Biochemistry imeni A. N. Balch, Aoademy of Soii. c
with
enoee'USSR.
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 Declassified in Part - Sanitized Copy Approved for Release 2012/05/30 : CIA-RDP82-00039R000100080033-7
,v9eae,iz
Table l
S.ialb? is the D.terminatian o!' it*rbos93i
Aoi6e which wero 6ddod to Zctrao4? o~
Various !1,.nt 8peoimeas
Aaount added to the
e:treat
Sabha 2
Adsorption of Oyetine by 11203 under Different
-
With water
with drogenatt3.fide
eolntion
-
Oonditioa$ ~
CONFIDENTIAL
Declassified in Part - Sanitized Copy Approved for Release 2012/05/30 : CIA-RDP82-00039R000100080033-7
 Declassified in Part - Sanitized Copy Approved for Release 2012/05/30 : CIA-RDP82-00039R000100080033-7
Wi1uN I
?ob:o
of pros ~aarbo l~ ~~ ~oida t plo*t
'V.. f r
Katorts1o (t1 tor~a o! 1 ~ of
th dsto4 apooi)
faros beets ~ Mhsst ipro.ti1 Lupine sprouts I eisu i?aves~ MilloK 1?V~e
1
0,69 I 2.76
- s.s I 7.gs
7-
(ONFIDENTIAI
0.75
Declassified in Part - Sanitized Copy Approved for Release 2012/05/30 : CIA-RDP82-00039R000100080033-7
 Declassified in Part - Sanitized Copy Approved for Release 2012/05/30 : CIA-RDP82-00039R000100080033-7
?1
RKNCLS
1. A. fl'ft,tnDhteyfl, Uepokhi eovremonnoy hiokhimi, I, 40, 1947.
2, Kretovioh a 4 Duide1', Dok1ady Akadomii Nark CSC, LIX, 9, 1947.
3. H. Tpvot, Chronophyllae in tho Flant and Animexi WAr1d, WArw,
1910.
4, T. Wioland, Uoppa-Saylor' a Z, CA'III, 24, 1942.
. Wieland, Naturwiooeneohatten, XXX, 333, 1942.
6. Wieland, Box'., L C V, 1001, 1942.
7. Wieland and L. Wirth, Eer., LXXVI, 823, 1943
8. 0. Sohraim and J. Primoeiah, Box'., L~CCVI, 373, 1943.
9. S. Darlin, Aota phyoiolo ioa eoandinavioa, X, faoo. 1, 91, 1945.
10, C. Dent, W. Stepka, and F. Steward, N, CLX, 652, 1947.
.END-
CONFIDENTIAL
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