URINARY
EXCRETION
OF
AMINES
IN NORMAL
CHILDREN
Thomas L. Perry, M.D., Kenneth N. F. Shaw, Ph.D., Dorothy Walker,
and Dorothy Redlich, Ph.D.
Division of Chemistry and Chemical Engineering (Contribution No. 2815),
California Institute of Technology, Pasadena, California
Supported by grants from the Ford Foundation and the National Institutes of Health.
ADDRESS: (T.L.P.) Department of Pharmacology, University of British Columbia, Vancouver 8, Canada.
PEDIATRICS, October 1962
576
MINES ARE EXCRETED in human urine in
minute amounts as compared to
amino acids. Certain amines have marked
pharmacologic activity, and therefore their
low concentration in urine does not
neces-sarily reflect their physiologic importance.
A better knowledge of the normal pattern
of urinary amines, and their biochemical
origin, is needed to provide a firm basis for
exploring their physiologic role and
pos-sible utility in diagnosis of certain
meta-bolic disorders, particularly those which
in-volve malfunctioning of the central nervous
system.
A number of investigators have studied
the excretion of one or more amines in
human unine.’13 As a result of their studies,
the following amines have been reported to
occur in the urine of normal individuals:
methylamine, dimethylamine, ethylamine,
ethanolamine, pyrrolidine, piperidine,
epi-nephrine, norepinephrine, dopamine
(3,4-dihydroxyphenylethylamine),
normeta-nephrine, metanephrine, p-tyramine,
tryp-tamine, serotonin (5-hydroxytryptamine),
histamine, 1-N-acetylhistamine, and
1-methylhistamine. Recently techniques have
been developed that make possible the
re-covery of more than a single or limited
group of amines from human urine. Jepson
et al.14 found phenylethylanline, o-tyramine,
and m-tyramine, as well as other previously
reported amines, in urine from normal
sub-jects. Kakimoto and Armstrong15 have
shown that normal human urine also
con-tains octopamine
(p-hydroxyphenyletha-nolamine), synephrine
(N-methyloctopa-mine), and 3-methoxytyramine.
The purpose of this paper is to report a
method by which amines can be separated
from other urinary constituents and subse..
quently identified by paper
chromatog-raphy, and also to report the pattern of
amines present in the urine of normal
cilil-dren.
MATERIAL AND METHODS
Urine was collected for 24 to 36 ilours
from 20 normal children; 13 were boys and
7 girls, and their ages ranged from 23 to 14
years. Each subject was in good health and
well nourished, lived at home, and
con-sumed a normal diet. None received any
medicament except as indicated herein. In
six of these subjects a second specimen of
urine was obtained between the fifth and
seventh days of continuous administration
of a monoamine oxidase (MAO) inhibitor.
Three children were given pheniprazine
(Catron, JB-516,
-phenylisopropylhydra-zine) in a daily dosage of 12 mg, and the
remaining three were given nialamide
(Niamid,
N-isonicotinoy1-N’-[3-(N-benzyl-carboxamido) ethyl] hydrazine) in a daily
dosage of 100 mg. Additional urine
collec-tions were also made on three children
while on a plant-free diet and during the
oral administration of neomycin
(Mycifra-din N, 0.2 gm four times daily) to reduce
the intestinal bacterial flora.
Urine specimens were stored at - 20#{176}C
during collection and until processed. After
determination of the creatinine
concentra-tion by the Jaffe method, each urine
speci-men was treated with the resin Amberlite
CG-50, type 2, in the hydrogen form, to
ad-sorb amines and other urinary bases, by a
modification of the method of Awapara et
al.16 The resin had been purified initially
ac-cording to the procedure described by Hirs
et
ARTICLES 577
of creatinine was first adjusted to pH 6.5
by addition of hydrochloric acid or sodium
hydroxide, and then stirred for one hour
with 50 gm of Amberlite CG-50. The
suspen-sion of resin and urine was then poured
into a glass column (3.3 X 50 cm) and the
urine was allowed to drain from the resin.
The resin was washed in the column with
50 ml of water to displace residual urine,
and this was added to the effluent urine.
Free amines and basic amino acids were
held by the resin, while the neutral and
acidic amino acids and conjugated amines
remained in the effluent urine. The effluent
urine was reserved for later hydrolysis. The
resin was then washed with 2,000 ml of
water to remove traces of neutral and
acidic urinary constituents, and the bases
were finally eluted with 400 ml of 4 N
acetic acid.
The effluent urine was adjusted to pH
0.5 by the addition of concentrated sulfuric
acid, and was hydrolyzed at 120#{176}Cfor one
hour in a pressure cooker. The hydrolyzed
urine was filtered, adjusted to pH 6.5 with
sodium hydroxide, and stirred with 50 gm
of resin to adsorb the amines freed by
hy-drolysis. The resin was washed on a
col-umn with 3,000 ml of water, and the amines
were then eluted with acetic acid as
de-scribed above.
The acetic acid eluates from the
tinily-drolyzed and the hydrolyzed urines were
then lyophilized. The lyophilized residues
contained besides amines large amounts of
basic amino acids and salts. The amines
were separated from the latter by
extrac-tion with ethanol and acetone as described
by Rodnight.3 Each eluate residue was
dissolved in 1 ml of water; then 4 ml of
absolute ethanol and 50 ml of acetone were
added. The suspension was filtered, and
the filtrate was concentrated to dryness
under reduced pressure on a rotating
evaporator, using a water batil with
tern-perature not exceeding 50#{176}C.The resulting
dry material was dissolved in 0.5 ml of
ethanol, and 50 ml of acetone was added to
tilis. The suspension was again filtered,
and the filtrate was concentrated to dryness
as just described. The final material,
con-taming amines and small amounts of other
urinary bases, was dissolved in 1.0 ml of
methanol, and was stored at - 20#{176}Cuntil
chromatographed.
The methanol extracts from the
unhy-drolyzed and hydrolyzed urines were
chromatographed two-dimensionally on
Whatman No. 1 paper, using two pairs of
solvent systems. Phenolic and indole amines
were most effectively separated by using
n-butanol-acetic acid-water (12:3:5)
(BuAc)
as the first solvent, and isopropanol-amrnonium hydroxide-water (8:1:1)
(IpAm) as the second solvent. Aliphatic
amines were better separated by using
2-methyl-3-butyn-2-ol--.formic acid-water
(75:5:20) (MBF) as the first solvent, and
butanol-acetic acid-water (12:3:5) as the
second solvent. Phenolic amines were
vis-ualized on the chromatograms by spraying
them with diazotized p-nitroanilin&8
(DPNA), diazotized sulfanilic aci&8 (DSA),
and 2,6-dichloroquinone chloroirnide
(N,2,6trichloropbenzoquinoneimine)l8
fob-lowed by 0.1 M borate buffer, pH 10
(DQCI). Indole amines were demonstrated
by spraying chromatograms with a 0.4
solution of dimethylaminocinnamaldehyde
in methanol-fl N hydrochloric acid (9:1)
(
DMCA). Primary and some secondaryaliphatic amines and some of the aromatic
amines were visualized by spraying
chroma-tograms with a 0.2% solution of ninhydrin
in 95% ethanol containing 1.0%
2,4-luti-dine.bu The ninilydrin-sprayed
chromato-grams were allowed to develop at room
temperature for 3 to 4 hours, tilen were
heated at 70#{176}Cfor 5 minutes, and finally
were sprayed with a 6.5% aqueous nickel
sulfate solution to preserve the ninhydrmn
colors. Quantities of amines ranging from
0.1 to 1 lL were detectable with these
re-agents; the amounts varied with the spray
reagent and the individual amine.
Amines were identified by
chromato-graphing equivalent amounts of urine
ex-tracts on duplicate sheets, to one of which
was added a small amount of an authentic
amine. By chromatographing the extracts
both from unhydrolyzed and from
Form
No. Compound
Present
RF Colors Giren with Dereloping Reagents
Bu.4c IpArn DPNA DS.4 DQCI !)3fC.4
Purple
I Tryptamine F 0.89 0.77
Q Serotonin F and C 0.53 0.63 Red-brown Orange-brown Grey Blue
3 Bufotenin F and C 0.53 0.81 Red-brown Orange-brown Blue
4t Kynuramine F 0.70 0.66 Red-purple
.5 UnidentifIed F 0.71 0.55 Red-purple Purple-’red
I; Unidentified F 0.75 0.84 l’urple
7 histamine F 0.44 0.56 Ileak orange Orange-red
S $-N-acetylhistamine F 0.34 0.73 Weak orange Orange-red
9 Unidentified F 0.52 0.46 Weak orange Orange-ret
10 Unidentified F O.6 0.81 Weak orange Orange-red
11 p-Tyramine F and C 0. 64 0. 75 Light purple Orange Grey
I m-Tyramine F and C 0.66 0.73 Pink Yellow Blue
1St p-hlydroxybenzylamine F and C 0.61 0.63 Pink Yellow Blue
14 (ktopamine F and C 0.55 0.54 Pink \ellow Blue
15 Synephrine F and C 0.38 0.72 Pink Yellow Blue
16 3-Methoxytyramine C 0.67 0.63 Grey-blue Orange-brown
17t S-Metlioxy-4-hydroxy- C 0.37 0.56 Purple Weak orange Blue
henzylamine
is Normetanephrine F and C 0.54 0.49 Purple Weak orangs Blue
19 Metanephrine F and C O57 0.66 Purple Weak orange Blue
o Unidentified F 0.58 0.60 Yellow Blue-grey (fades)
21 Unidentified F 0.74 0.84 light purple Orange
Unidentified F 0.56 0.41 Grey-brown
23 Unidentified F and C 0.68 0.87 Green
4 Unidentified C 0.53 & O.7S 0.30 lIed-purple Brown Purple (fades)
5 Unidentified C 0.72 0.59 Light purple Orange Weak green
211 Unidentified C 0.69 0.69 Purple Grey-blue
7 Unidentified C 0.77 0.83 Grey-brown
88 Unidentified F 0.71 0.49 Purple-pink Yellow Blue
9 Unidentified F 0.76 0.59 Light purple Yellow
the free as well as the conjugated amine
content of urines. The concentration of
cer-tam amines present in urine was estimated
semiquantitatively by visual comparison of
the color intensity of spots on
chromato-grams prepared from a graded series of
aliquots of urine extract and a similar series
of standard authentic compound.
Most of the authentic amines used in this
study were obtained from commercial
sources. Octopamine was generously
pro-vided by Dr. Sydney Archer,
-N-acetyl-histamine by Dr. Richard W. Schayer, and
-N-methylilistamine by Dr. Herbert Tabor.
o-Tyramine was prepared enzymatically
from o-tyrosine by incubation with
Strepto-coccus fecalis decarboxylase, after which
tile amine was recovered by the use of
Am-berlite CG-50 as previously described.
p-hydroxybenzylamine was prepared from
p-methoxybenzylamine by demethylation
with hydriodic acid.
-N,N-dimethylhis-tarnine was obtained from histamine via
-hydroxyethylimidazole by a modification
of the method of Wrede and Holtz,20 and
tile procedure of Garforth and Pyman.21
RESULTS
Tile pattern of amines observed in the
urine extracts of 20 normal cilildren proved
to be quite uniform, except for a few
varia-tions dependent on diet or on tile
admin-istration of an MAO inhibitor. Table I
lists the aromatic and heterocyclic amines
which were detected in most or all of the
urine extracts by chromatography in the
solvent system BuAc-IpAm. The Rf values
listed were measured following
cochromat-TABLE I
AnoIATIc AMINES IN URiNE OF NORMAL ChILDREN5
SAbbreviations used in this table: F =free amine; C =amine r&eased from conjugated form by hydrolysis; BuAc =butanol-aetic
aci(l-water (l :3 : 3); IpAm =isopropanol-ammoniu.m hydroxide-water (8: 1 : I); DPNA =diazotized p-nitroaniline; DSA diazotized sulfanilic
acid; I)QCI =dichloroquinone chloroimide followed by borate buffer; DMCA =dimethylaminocinnamaldehyde.
t Amines not previously reported to be normal constituents of urine.
SOLVENT FRONT
a
I
0 0
ciD
5,
2 - ISOPROPANOL -ANNONIIJN HYDROXIDE -WATER 181:1)
‘FABLE It
!
.‘%O. (OfliJX)Ufld
‘
1orm Present
llb
---
---
-,
.1/LIP BU.1C
(I..50 1).43
, . ,. ( olor tilt/i .\ 1
It-. . .
h,,vlrin-Iut,dine
l5khije
‘
( olor after
,. .\ ickel eu/fate
Piiik
3() Ethianolaniine F
31 I.thiylatnine F I).SI) (1..56 Purple Pink
3’2t #{216}-I1ydroxypro)ylanhine F 1)..53 0.Si Purple Pink
33 Pyrrohidine F’ 1).64 (1.59 \ellow \elli)w
1 1’ryptamine F’ I).85 I).76 Purple Pink-hroevi
11 p-1’yramine F and C 0.61 0.7() Purple Pink
ISt p-llydroxybenzylamine F aiid C 1)59 0.66 Yehtow-spurple Pitik
17t S-Ietlioxy-4-hiydroxvheiizylainine C 0 .59 (1 .66 Yehlow-spurple Pink
34 Benzylarnine 0.6K 0.79 Purple Pink
55 Unidentified F I).17 0.13 Yellow Yellow
36 Unidentified F’ (I.44 0.58 Purple Pink
37 tJnidentified F 0.51 0.35 PurpLe Piiik
38 Unidentified F 0.55 0.43 Purple 13i,ik
Si) Unidentified F’ t).48 0.‘35 Purple Brown
40 Unidentified F’ 0.5 (I.46 Yellow \ehlov
41 Unidentified F’ 0.4.5 I).36 Pink Pink
4 Unidentified C 1)46 0.4R Pink Orange-p;ik
43 Unidentified F and C 0.77 0.78 Yellow Yellow-brown
44 Unidentified F 0-79 0.90 Purple Grey-pink
ARTICLES
ograpily of each compound with urine
ex-tract. The volume of extract applied to tile
ciiromatogram usually was equivalent to
25 mg of creatinine in the original urine, or
15 mg of creatinine in hydrolyzed urine.
Rf values for some amines varied
consid-erably from tilose listed if the cOnlpOunds
were chromatographed alone, or in a much
smaller volume of urine extract. Colors
given by developing reagents are listed for
DPNA, DSA, DQCI, and DMCA. Free
amines are indicated by an F and those
re-leased from conjugated form and detected
only after hydrolysis by a C. Figure 1 is a
schematic diagram illustrating the
posi-tions taken by these aromatic amines \vllen
a urine extract is sul)jected to
two-dimen-sional chromatography in tile solvent
sys-tern BuAc-IpAm.
Table II lists tile ninhydrin-reactive
amines WiliCil were detected in most or all
urine extracts of normal cilildren. Tile Hf
values are those found for each compound
FIG. 1. Aromatic amines in urine of normal
chil-dren. The numbers refer to the compounds listed
in Table I.
wilen cocilromatographed
two-dimension-ally in a 15 to 25 mg creatinine equivalent
of urine extract in tile solvent system
Nisin-DoIN R:kcTIvF: IN tIIJNE OF Nouiz (‘hhhI,nuP:N5
S Abbreviations used in this table: F=free amine; C=amine released from conjugated form by hydrolysis MBF = 2-methiyl-3-butynol-formic acid-water (75 : .5 : O) ; BuAc = butanol-acetie acid-water (H : S : .5).
t Amines not previously reported to be normal constituents of urine.
_____
SOLVENT FRONTMBF-BuAc.
Colors
listed are those given: by each compound when developed with
ninhydrin-lutidine, and when
counter-,-
0
sprayed with nickel sulfate. Figure 2 is aI z poundsurinaryschematictaken byoncompoundsthethesemapchromatogram.rnnhydrin-positiveshowinglisted in theTableManypositionsI reactofcorn-the
-I poorly or not at all with ninhydrm. These
I substances were not detected in the
MBF-BuAc solvent system, and therefore are not
recorded in Table II or depicted in Figure
2.
I
The urinary concentrations of 1 1aroma-tic amines which were estimated
semiquan-2 - BUTANOL-ACETIG ACID-WATER (12:3:5) titatively are recorded in Table III. The
lowest and highest urinary concentrations
FIG. 2. Ninhydrin reactive I)ases in urine of
nor-mail children. The numbers refer to the compounds encountered in 20 normal children are
listed in Table II. listed. In addition, Table III gives values
TABLE III
CONCENTRATION OF CERTAIN AMINES IN URINE OF NoIL%IAL CHILDREN5
Subjectt
Serotonin
C MAO
Bufotenin
C MAO
Tryptarnine
C MAO
Normet
C anephrine
MAO
p- Tyrarnine
-
--C MAO
1 1.3 4.6 0 5.1 50 2.4 l3.’2 2t4 77
1.9 5. 0 0.5 10.7 91 0.8 6.3 16 60
3 4.4 9.1 0 0 10.4 10’ 2.2 10.8 29 85
4 7.7 10.6 0 0 10.4 55 2 I). 5 81
5 4.4 7.7 0 0.5 11.6 88 3.3 13.8 49 115
6 4. 6.9 0 1. 8.1 6 So 9.2 7 59
o subjects:
Minimum 1.S 0 3.9 0.8 10
Maximum 7.7 0.3 .0 3.3 5’2
.
Subject
Octopamine
C MAO
Synephrine
-C MAO
. .
Iltstamzne N-Acetyi-. .
istamsne
p-Il yd
rosy-.
enzy amine
3-Methoxy-4-H yd rosy-benzylainine
1 0 64 5 690
2 <1 <1 8 75
S 0 0 4 16
4 3 0 56 7
5 0 <1 <1 10
6 0 5 6 115
20 subjects:
Minimum 0 0 0.4 1 0 0
Maximum 5 56 .4 108 14 19
aAll values are expressed as .ig free base/100 mg of creatinine; C = control, MAO during MAO blockade. t Subjects 1, 2, and 8 were given pheniprazine as an MAO inhibitor, while Subjects 4, iS, and 6 were given
ARTICLES 581
for seven amines which are metabolized by
monoamine oxidase, both before and
dur-ing blockade of this enzyme in six of the
cilildren. Tile amine content of urine is
expressed as .g free base/100 mg of
creatinine, and values are not corrected for
losses in recovery.
The actual concentration of many of
these amines in urine may have been two
Or tilree times greater than tile values
found. In several experiments in which
authentic amines were added to tile
orig-inal urine, over-all recovery averaged 20%
for octopamine, 30% for p-tyramine and
normetanephrine, 35% for synephrine,
his-tamine and N-acetylhistamine, 50% for
sero-tonin, 70% for tryptamine, and 85% for
bufotenin.
COMMENT
Fifteen aromatic and heterocyclic amines
have been identified chromatographically
in the urine of normal children. An
addi-tional 14 unidentified aromatic bases were
commonly found, many of which also may
be amines. Further studies are being
di-rected toward identification of these bases.
Three of the arnines, kynuramine,
p-hydroxybenzylamine, and
3-methoxy-4-hy-droxybenzylamine, have not been reported
)reviOusly as constituents of human urine.
Tentative identification is based on
mi-gration of the urinary compound and
au-tilentic amine at tile same rate in the
sol-vents BuAc, IpAm, MBF, and also in a
fourth solvent, acetonitrile-formic
acid-water (80-2-18). Authentic kynuramine and the urinary compound gave identical colors
with DMCA, and with tile Ekman
re-8 Likewise, urinary
p-hydroxyben-zyiamine and
3-methoxy-4-hydroxybenzyia-mine gave the same colors as the respective
authentic amines with DPNA, DSA, DQCI,
and ninhydrin-lutidine.
Kynuramine was detected only in urine of
subjects on MAO blockade, in a
concentra-tion of 0.5-1.5 g/100 mg. of creatinine.
It may be formed in vivo by the action of
aromatic L-amino acid decarboxylase22 on
kynurenine, a tryptophan metabolite, but
further work is required to exclude
exog-enous sources such as intestinal bacteria
or diet. This amine is an excellent substrate
for monoamine oxidase in vitro.23
p-Hydroxybenzylamine and
3-methoxy-4-ilydroxybenzylamine are of exogenous
on-gin, since they were not found in urine of
cilildren on a plant-free diet. These
corn-pounds may be present in foods or may be
formed metabolically by amination of
p-hydroxybenzaldehyde and vanillin,
deniva-tives of which are widely distributed in
plant foods.
The occurrence in normal urine of
m-ty ramine,’4 octopamirie, synephnine, and
3-methoxytyramine’ was confirmed.
Bufo-tenin, a psychotomimetic amine reported to
occur in normal adult urine,2 was found in
small amounts in unines of 4 children given
an MAO inhibitor and one child not on
MAO blockade.
As shown in Table III, the administration
of pheniprazine or nialamide led to an
in-creased excretion of serotonin, bufotenin,
tryptamine, normetanephrine, p-tyramine,
octopamine, and synephrine. MAO
block-ade also increased excretion of
metaneph-rine, 3-methoxytyramine, and m-tyramine,
but these substances could not be
quanti-tated due to insufficient separation from
neighboring spots on the chromatogram.
Benzylamine was found in relatively
large amounts in hydrolyzed urine of
chil-dren given nialamide; the amine is derived
from nialamide, part of which is excreted
unchanged. Benzylamine is produced when
nialamide itself is hydrolyzed under the
same conditions as urine. Children given
nialamide also excreted much larger
amounts of p-hydroxybenzylamine than
those not receiving this drug, and therefore
it appears that the benzylamine moiety of
part of the administered nialamide may be
hydroxylated in vivo.
When plant foods were excluded from
the diet, p-hydroxybenzylamine,
3-methoxy-4-hydroxybenzylamine, synephnine, and
un-identified compounds 26, 27, 28, and 29
disappeared from urine, but they were
582 AMINE EXCRETION
normal diet. It is concluded that these
seven amines are of dietary origin, and
probably not metabolites of intestinal
bac-tenia. Tile excretion of serotonin did not
diminish on a plant-free diet, even though
appreciable amounts of this amine are
pres-ent in certain fruits such as banana and
pineapple. Urinary excretion of senotonin
is probably a reliable indicator of its
en-dogenous production, at least in the kidney.
More N-acetylhistamine was excreted by
children on a plant-free diet, because of a
compensatory increase in meat
consump-tion and acetylation in the liver of
hista-mine present in meat.24
The daily urinary excretion of amines by
normal adults, which has been measured
by other investigators, and by normal
chil-dren may be compared if values found in
tile present study are multiplied by a
fac-ton proportionate to the expected daily
crea-tinine excretion of an adult of average size.
Assuming a daily creatinine output of 1,700
mg for a 70-kg adult, the excretion ranges
of six amines listed in Table III can be
mul-tiplied by 17. The calculated serotonin
ex-cretion of 22 to 130 &g for children
corn-pares well with 60 to 120 tg found for
adults.3 Tile calculated tnyptamine
excre-tion of 66 to 370 tg for children is higher
than reported adult ranges of 25 to 130 &g,25
45 to l20ig,3 and 50 to 101 tg.7 Calculated
p-tyrarnine excretion of 170 to 890 g for
children compares with 190 to 430 tg
re-ported for adults. Calculated histamine
ex-cretion of 7 to 41 lL for children is similar
to 6 to 19 reported for adults,26 but
ex-cretion of 17 to 1,840 hA of
N-acetylhista-mine is much higher than 2 to 56 g cited
for adults.26 Only calculated
normetaneph-nine excretion of 14 to 56 pg for children is
lower than 100 to 300 tg reported for
adults.13 Since recovery of these amines
from children’s urine was not complete, it
may be concluded that children excrete
proportionately larger amounts than adults,
or that values previously reported for adults
may be low.
Ethanolamine was the most prominent
ninhydrin-reactive amine found in all urines
in the range 150 to 250 .g/100 mg of
crea-tinine. -Hydroxypropylamine has not been
reported previously as a constitueilt of
nor-mal urine, and its identification on the basis
of similar RF in four solvents is considered
tentative. Methylamine, dirnethylamine,
and pipenidine, which llave been found by
others in human urine,1’ 12 were not
sep-arated adequately from other compounds
by the solvents used, but may have been
present in the unines studied. Besides tile
10 unidentified ninhydnin-positive bases
found regularly in urine (Table II), 10 other
ninhydnin-neactive unknowns were
en-countered occasionally. i’t’lany of these
com-pounds probably are aliphatic amines.
Other amines which were not detected ill
tile present study have been reported in
human urine and include norepinephrine,
epinephnine, and dopamine.6 These
cate-cholamines are oxidized and destroyed
when sheets are chromatograpiled ill tile
solvent IpAm, and their separation from
other compounds is inadequate in the
solvents BuAc and \‘IBF. Norepinephnine
and epinephnine could not be detected at
all, and dopamine was detected in only
a few unines. Five additional amines
re-ported in normal adult urine by other
in-vestigators could not be detected in any
child’s urine studied, even though the
solvent system BuAc-IpAm allowed their
ready identification \‘ilen the autileiltic
amines were aded to urine extracts. These
were phenylethylamine and o-tyramine,
found in the urine of normal adults on
MAO blockade,’ iNmethy1serotonin,2
c
N-methylhistamine, and
-N,N-dimethylhis-tarnine.4
More complete information needs to be
obtained about tile nature of tile many
unidentified amines present in human urme,
as well as about the physiological function
of those of known identity. Knowledge of
the normal pattern of amino acid excretion
in urine preceded the discovery of tile
aminoacidunias, many of which are
as-sociated with forms of mental defect. It
seems reasonable to expect that the finding
ARTICLES 583
amine excretion in the urine of children
Wilo are mentally defective or mentally
iii may provide clues as to the mechanism
of the abnormality in brain function. A
pre-liminary study27 has shown, for instance,
tilat phenylketonunics excrete much less
serotonin, and less of several otiler amines,
than do normal children. This lends support
to the suggestion that underproduction in
the brain of amines vital to normal cerebral
function may account for the mental defect
characteristic of untreated phenylketonunia.
ADDENDUM
Since the sul)mission of this article for publica-tion, the occurrence of p-hydroxybenzylamine and 3-methoxy-4-hydroxyhenzylamine in normal urine has been reported (Kakimoto, Y., and Armstrong, M.D. : Tue phenolic amines of human urine. 1. Biol.
Chem., 237:208, 1962).
SUMMARY
The urinary excretion of amines was
in-vestigated in 20 normal children by means
of two-dimensional paper chromatography,
following preliminary separation of amines
from other urinary constituents by the use
of a cation exchange resin. Normal children
were found to excrete the following 19
amines : tryptamine, serotonin, btifotenin,
kynuramine, histamine,
-N-acetylhista-mine, p-tyramine, m-tyramine,
p-ilydnoxy-benzylamine, octopamine, synepiirine,
3-methoxytyramine,
3-methoxy-4-hydroxy-henzylam me, normetanephrine,
metaneph-nine, etllanolamine, ethylamine,
-hydroxy-propylamine, and pyrrolidine. Of these,
kynuramine, p-hydroxybenzylamine,
3-methoxy-4-ilydroxybenzylamine, and
1-hy-droxypropylamine have not been previously
reported as constituents of iluman urine.
The methods used were not stisfactory for
tile detection of methylamine,
dimethyl-amine, pipenidine, nonepinephnine,
epineph-nine, and dopamine, but the presence of
these six amines in urine has been reported
by others.
An additional 24 unidentified
corn-pounds, probably amines, were found in
most or all of the unines studied, and some
of them appeared to be present in
rela-tively large amounts. Other unidentified
bases were encountered in a few urines.
The effects of monoamine oxidase blockade
on the urinary excretion of arnines have
been described, as well as the effects of
exclusion of plant foods from the diet, and
antibiotic restriction of intestinal flora.
Fig-tines are presented for the approximate
range of daily excretion of 12 amines. It
is suggested tilat study of the excretion of
amines in the urine of children who are
mentally defective or mentally ill, and
com-panison with the pattern of urinary amines
in normal children, may provide clues to
the mechanisms of certain forms of cerebral
dysfunction.
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Acknowledgment
The authors wish to express their appreciation
to Prof. Linus Pauimg for helpful discussions of
this work, and to Morris Gutenstein and E. Elmo
Jacobs for technical assistance. Thanks are also
given the children who enthusiastically co-operated
in this study.
DIAGNOSTIC Tisi’s IN INFANTS AND
Cmi.-DREN, Ed. 2, Hans Behrendt, M.D.,
Phila-delphia, Lea & Febiger, 1962, 617 pp.,
$15.00.
This book is advertised as being aimed at a
wide audience of students, pediatricians,
pa-thoiogists, and general practitioners. As a
ne-suit of this approach the book falls far short
of satisfying the needs of any one of these groups. It is a mixture of the old and new,
neither of which is critically appraised. Many
of the laboratory tests included are of purely
historic interest. Dr. Behrendt utilizes 39 pages
to discuss tests of basal metabolism rate, even
though he acknowledges that such procedures
have “outlived their usefulness.” In the
dis-cussion of hypoproteinemia no mention is
made of protein-losing enteropathy. Medical
students will be confused by the fact that on
one page hypernatremia is said to be “rare,”
and that on the following page it is said to
occur in “30% of infants with diarrhea.” A
petechiometer, reported in 1949, is described
as “new.” These are examples of small errors,
but the book contains many of them. This
tends to destroy your faith in other areas of
the book where you are not qualified to judge.
The author’s inability or unwillingness to
dis-card, select, and be critical, severely limits
the usefulness of this book. I would
specifi-cally not recommend it to pediatricians.
