Full text

NOTES
341
CNROm.3648 .,:
;,. ,( ,:, .‘, .’ .,‘. ,‘.
.., :
.’
,.;
.:,
..
., .I
,,,,
.,.,,’
,‘,,
,:
of secondary amines on paper.with
I..
.”
go.me~critical:remarks’on.the;colours
sodium nitroprusside reagent
,‘,: ,,; :‘:
,’
‘,
<.
(’ .
‘,’
: ; In,. paper elktrophoretic
and. chromatographic
: studies, on ihe : occurrence
of
biogenic ~amines. and basic .amino acids in the, %aecesl-3 a rumen’ fluiS and urine6 of
dairy cows, we used .ninhydrin as a colour reagent, though in some cases diazotized
sulfanilic acid :.(Pauly.‘s reagent) was used for .the final detectioix ,of aromatic compOunds.;,.Generally,good: results had been found; with the ninhydrin r&gent, but. ,the
colouris withthe se’condary amines, dimethylamine and diethylamine, :were’often faint.
.:.
;
TABLEI.
.’
;
*
,:;‘_.
;
‘,‘,,
RELLTIVE
INTRNSITY.'OF
NITROPRUSSIDE
REAGENT;'
Compoicnd
,,:.I
.,
”
!’
,.’
.,
,,.:
hmh&iline
Dinkthylrimine
Ethylamin6
Diethylamin&
Propylamine
l3utylamine
Amylaminc
Phenylcthyhimine
Tyramine
Diaminopropanc
Putrescine
Ctidavefine‘
Spermine
Spermidine
Ethanolamine
3-Aminopropane
4-Aminobutane
Agmatine
Lysine
Arginine
Histidinc
Ornithine
P-Al anine
y-Aminobutyric
acid
s-Aminocaproic
acid
Taurine
Tryptamine
Histamine
Carnosine
Norephedrine
Kynurenine
Glutamine
Citrulline
~10OENlc
,,,,
Cbnlen+ation
‘,
i
(
AMINII~AND
,,
,‘I
,,”
,,.
i3AS1c
AMINO
ACXDS
,.
in’ pg
.(
.,
,.’
COLOURIZD'WITH
.,
.:
‘Papt?v: Si?di?iciber& S#iU
Untreated
S%ICJM
’
2643
b
T,veated with *
A
B ,.
i
c,
2.8
1.7
-
acid-water
*A== Phenol saturated with water; l3 - ro-butanol-acetic
acid-water
(70 : 15 : rg), saturated
glycol
monomethyl
ethk’-propionic
-t_ P Strong; (+) 3 faint; (( -1_)) = very faint; - = negative.
(4 : r :5) ; C = ethylene
with sodium citrate.,
J. Chromatog.,
37 (rgG8) 34x-341
3pz
NOTES
..
.
,/,., Inlater ,studies we therefore decided to make use of the calours produced, by
secondary. amines with the sodium nitroprusside reagent recommended by STEIN VOX
I(AMIENsKr~:;i-AcCording'to~this :author,...this reagent:should,.be:highly
:sensitive.,and
..l),,, ‘..‘;.,‘:.‘,Z, ,”I .:.*:, ;:
specific’for. secondary. amines:
On spraying an electrochromatogram of a faeces ‘extract, in which dimethylamine,‘:and,..diethylamine Iwere present;, however, we found .with this reagent ‘a blue
spotin Rhe:position: of spermidine. .This .unexpected result decided ,us on a closer study
_.,l:,’
.‘j,
‘,.,,
. .
:
df:thisicolour.reagent;
‘.: : ,: ‘. ,.r: >:.‘::,,‘,‘.: A-: great,. number. of .:,biogenic amines: and basic amino ,acids were treated on
chromatographic paper’ (Schleicher & Schiill; “zoq3b) ,with .,the ‘sodium., nitroprusside
reagentAll compounds tested irrthis way gave a blue-purple spot after:$o-6o.minutes.
The experiment ‘.was repeated with chromatographic paper treated beforehand with
:
the solvents:used2# 3. The results of these experiments are given in Table I.
.‘.
‘. .,:.Alfhough STEIN EON KAMIENSKIO stated that primary amines give pale red-blue
spots &th this reagent above IO pg, we found, however, that this also happened
below fo+g, .and.that;: moreover; the, colour was the same as with secondary amines.
~.,‘. Furthe&~ore it appears that the sensitivity depends ‘strongly on the solvent used.
Thus we .were unable :fd ‘confirm‘ the ‘&eat sensitivity and specificity of this reagent.
: .:,,’
“,’
.of
,
L&oratory
~?ete&&y
Bioche&ry,
Stats ~k&wsi~y C$ Utreckt, Biltstraat ~72, Utrecht
(Tee Nefherlands)
I/.,
,‘.’
_’
:’
,,’
.,
‘,
r D. L, SAN-RHEEN~N, .iVat&rd, rg3 (&62) 170.
2 D. L.,.vANRHEENEN, Rec. Trav. Ckim.; Ba‘(rg63)22.5.
3 D. L.VAN RHEENEN, Tibesis,
Utrecht, 1964.
4,I-L J. H~ENDRIICS,R, L. PRESTON, R. A. PR~NS AND D. L.VAN RNEENEN,
-91
5 M.
91
‘6 E.
D. L. VAN, I$I-TEEN~?
Miss C. B. J.! SIPMAN
,.
!
',,
T~jclschv.
Dier&eneesk.;
(rg66) ‘369.
:’ ’
J. H. GEELEN, D. L.VAN R~IEENEN, H;J. HENDRIICS AND L.SEEICLXS, TijdscAr.Di~~geneesk.,
(1966) ~577. ‘..
STEIN VON .KAMIENSKI, ‘PZanta, 50 (x957) zgr.
.:
:
Received: June zzth, 1968
J. CiWO~a~~g., 37 (rg68)
“.
.
34x-342

Download Report