616 Am J C /in Nuir l997;66:616-2l. Printed in USA.

1997 American Society for Clinical Nutrition

Nickel metabolism in humans investigated with an oral
stable isotope1
M arina Patriarca, Thomas David B Lyon, and Gordon S Fell
ABSTRACT W e report the results of the first complete study
of nickel metabolism in human subjects using a stable nickel
isotope (62Ni) as tracer. Four healthy adult subjects (two women
and two men) fasted overnight before ingesting 10 g 62Ni/kg
body wt. Blood samples were drawn after fixed intervals of time
and the total daily output of urine and feces was collected for the
first 5 d after dose ingestion. 62Ni in plasma, urine, and feces was
determined by isotope-dilution inductively coupled plasma-mass
spectrometry with 61Ni. The direct measurement of the fecal
excretion of the tracer allowed a reliable assessment of nickel
absorption from the gastrointestinal tract and we found no evi-
dence of the excretion of absorbed nickel via the gut. The percent-
age absorption calculated from the amount of 62Ni excreted in the
feces ranged from 29% to 40% . Urinary excretion over 5 d ranged
from 5 1 % to 82% of the absorbed dose. Plasma 62Ni peaked
between I .5 and 2.5 h after ingestion and decreased by a factor of
> 10 over the next few days. W e observed low between-subject
variability of nickel absorption and excretion. Confounding factors
such as contamination and dietary intake of nickel, which ham-
pered earlier measurements in subjects dosed with naturally abun-
dant nickel, were eliminated by using the tracer isotope 62Ni.
Am J Clin Nutr 1997:66:616-21.
KEY W ORDS Nickel, stable isotope, absorption, excre-
tion, inductively coupled plasma-mass spectrometry, humans
INTRODUCTION
Nickel plays an important role in human health as a toxin, an
allergen, and as a candidate essential element ( 1 ). Nevertheless,
information on nickel metabolism in humans is limited. Radio-
active tracers have been used to investigate nickel metabolism
in different animal species under various experimental condi-
tions (2-5). However, their use cannot be extended to human
subjects because of safety concerns. Direct investigations using
naturally abundant nickel were carried out in human volunteers
(6-10). Spruits and Bongaarts (6) showed that nickel was
rapidly absorbed from the gastrointestinal tract and then elim-
mated in urine in a fasting volunteer given 5 mg NiSO4.
Solomons et al (8) reported reduced gastrointestinal absorption
when nickel was ingested with food and studied the different
effects of food components. Hypersensitive subjects were
given 0.4-5.6 mg Ni504 orally to investigate whether oral
intake of nickel exacerbated their symptoms (7, 9). Although
Cronin et al (7) observed a dose-related worsening of symp-
toms, Gawkrodger et al (9), in a randomized double-blind
crossover study, did not confirm these findings but considered
that the occurrence of adverse effects due to nickel ingestion in
nickel-sensitive patients could not be ruled out because the
amount and the rate of nickel absorption and excretion varied
considerably among individuals. Sunderman et al (10) carried
out a complete study of nickel absorption and kinetics in 10
human volunteers given 12, 18, and 50 g Ni/kg body wt
orally, either after an overnight fast or with breakfast. Nickel
concentrations were then measured sequentially in serum,
urine, and feces for up to 4 d after ingestion. Nickel absorption
from the gastrointestinal tract was estimated as 27 17% in
fasting subjects but as only 0.7 0.4% when nickel was
ingested with food. Nickel elimination in both feces and urine
after 4 d was 102 8% and 104 21% in fasting and fed
subjects, respectively. The results of this study fit a two-
compartment mathematical model similar to that developed for
rats and rabbits by Onkelinx et al (2).
These studies provided fundamental and invaluable knowl-
edge about nickel handling by human beings. However, they
were limited because they required high doses, greater than
physiologic intakes, of naturally abundant nickel to produce a
measurable change in nickel concentrations in biological fluids
and excreta. Both contamination during sample collections and
dietary intakes contributed to the variability of the results.
These limitations can be circumvented by the use of stable
isotopes, which are safe and convenient. There are five stable
nickel isotopes, of which two (61Ni and 62Ni) can be used for
tracer studies. The use of stable isotopes in metabolic studies
has been unpopular because of the need for extensive sample
pretreatment and expensive instrumentation. Inductively cou-
pled plasma-mass spectrometry (ICP-M S) now allows the de-
termination of the isotopic composition of biological materials
with less trouble. Stable isotopes determined by ICP-MS have
I From the Clinical Biochemistry Department, Istituto Superiore di
Sanita , Rome, and the Trace M etal Unit, Institute of Biochemistry, Royal
Infirmary, Glasgow, Scotland, United Kingdom.
2 Presented in part at the 4th International Congress on Plasma Spectro-
chemistry, Durham, United Kingdom, September 1994.
3 Supported by a grant from the European Science Foundation, Stras-
bourg, France, awarded to M P (ESF Program of Research Fellowship in
Toxicology, grant no. RF/93/4l/T).
4 Address reprint requests to M Patriarca, Laboratorio di Biochimica
Clinica, Istituto Superiore di Sanita, viale Regina Elena 299-00161 Roma,
Italy. E-mail: m.PATRIA@ NET.iss.it.
Received November 8, 1996.
Accepted for publication April 15, 1997.

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NICKEL M ETABOLISM IN HUM ANS 617
b e e n u s e d to s tu d y iro n a b s o rp tio n in in fa n ts (1 1 , 1 2 ), iro n
a b s o rp tio n in w o m e n b e fo re a n d d u rin g p re g n a n c y (1 3 -1 5 ),
z in c a b s o rp tio n in n e w b o rn s (1 6 -1 8 ), a n d c o p p e r m e ta b o lis m
in h e a lth y s u b je c ts a n d p a tie n ts w ith W ils o n d is e a s e (1 9 , 2 0 ).
T h e fe a s ib ility o f s tu d ie s o f n ic k e l m e ta b o lis m u s in g s ta b le
is o to p e s a n d IC P -M S h a s b e e n d o c u m e n te d b y T e m p le to n e t a l
(2 1 ), w h o m e a s u re d s e q u e n tia l s e ru m and urine c o n c e n tra tio n s
of 6 tN i in a h e a lth y subject given 20 pg 6tNi/kg body wt.
He re w e re p o rt th e re s u lts of an investigation o f th e a b s o rp -
tio n , d is trib u tio n , a n d e x c re tio n o f n ic k e l in fo u r h u m a n v o l-
unteers in w h o m 6 2 N i w a s used as tracer. S o m e o f th e s e
fin d in g s w e re p re s e n te d p re v io u s ly (2 2 ).
SUBJECT S AND M ET H ODS
Subj ects
T h e study was approved b y the Research Ethics Committee
at the Glasgow Royal Infirmary and volunteers gave their
w ritte n , in fo rm e d c o n s e n t to th e e x p e rim e n t. T h e v o lu n te e rs ,
two w o m e n (W l a n d W 2) and two m e n (M l a n d M 2) a g e d
21-30 y, w e re h e a lth y subjects re c ru ite d fro m th e h o s p ita l s ta ff
w h o had not suffered from any serious illness during the p a s t
5 y , h a d re g u la r b o w e l h a b its , a n d h a d n o h is to ry o f h y p e rs e n -
sitivity to nickel. All subjects were nonsmokers. Subject W 2
w a s ta k in g c o n tra c e p tiv e p ills a n d a n o ra l a n tih is ta m in e a n d
w a s a v e g e ta ria n . Die t w a s u n re s tric te d , a lth o u g h g u id a n c e w a s
p ro v id e d a b o u t s ta n d a rd fib e r a n d n u trie n t re q u ire m e n ts .
Or ganization of the study
Stable n ic k e l is o to p e s , 6 1 N i (9 3 . 6 1 % , a s N iO ) a n d 62Ni
(98.83% , as metal), were o b ta in e d fro m A E A T e c h n o lo g y ,
Oxford, United Kingdom. The study was carried out on two
s e p a ra te o c c a s io n s o n tw o s u b je c ts a t a tim e (W i a n d M 1 ; W 2
and M 2) a n d lasted 5 d (M onday to Friday). A 24-h urine
s a m p le and a fe c a l s a m p le w e re o b ta in e d fro m e a c h s u b je c t
b e fo re th e e x p e rim e n t.
On each o c c a s io n , M o n d a y m o rn in g , a basal b lo o d s a m p le
(10 mL) was o b ta in e d b y v e n ip u n c tu re fro m e a c h o f th e tw o
subjects a fte r a n o v e rn ig h t fa s t. W e ig h t a n d h e ig h t o f e a c h
subject w e re re c o rd e d . S u b je c ts w e re a s k e d to e m p ty th e ir
b la d d e rs b e fo re d rin k in g 1 0 g 6 2 N iJ k g b o d y w t d ilu te d in
water. Together with th e is o to p e d o s e , e a c h s u b je c t in g e s te d
t w o gelatin capsules, each containing 10 radioopaque barium
s u lfa te -im p re g n a te d p o ly e th y le n e p e lle ts to a c t a s a fe c a l
m a rk e r (23). The subjects fasted for 2.5 h after the isotope
ingestion a n d th e n w e re a llo w e d to re s u m e th e ir u s u a l e a tin g
habits. Additional blood samples (10 mL) w e re o b ta in e d 0 . 5 ,
1 . 5 , 2.0, 2.5, 3.0, 3.5, 4.5, 6.0 (for subjects W l and M l), 6.5
(for subjects W 2 and M 2), 24, 48, 72, and 96 h after isotope
a d m in is tra tio n . Urin e w a s v o id e d in to s e p a ra te a c id -w a s h e d
p la s tic bottles at the fo llo w in g tim e in te rv a ls : 0-3, 3-6, 6-12,
12-24, 24-48, 48-72, 72-96, and 96-120 h. Fecal output on
e a c h o f th e 4 d a fte r is o to p e in g e s tio n w a s c o lle c te d d ire c tly
in to a c id -w a s h e d p la s tic c o n ta in e rs .
Sample collection and stor age
Blood samples w e re c o lle c te d in to l0 -m L a c id -w a s h e d p la s -
tic tubes containing 1 .5 g K,EDTAIL (shown to be free fro m
nickel as assayed by graphite-furnace atomic-absorption s p e c -
trometry) a n d g e n tly m ix e d s e v e ra l tim e s b y in v e rs io n . P a c k e d
c e ll v o lu m e w a s m e a s u re d o n each sample b y using a micro-
h e m a to c rit c e n trifu g e . Plasma was separated by centrifugation
at 500 X g for 10 mm at 15 #{176}C and stored at -20 #{176}C. Red cells
w e re w a s h e d th re e tim e s w ith s a lin e s o lu tio n (0 . 9 % N a C 1 ,
Aristar grade, 99.99% ), then diluted to the original volume of
th e b lo o d s a m p le w ith d e io n iz e d w a te r a n d s to re d a t - 2 0 # {1 7 6 }C .
Urin e volumes were determined by multiplying th e w e ig h t o f
th e s a m p le s b y a n a v e ra g e u rin e d e n s ity o f 1 . 0 2 kg/L. After th e
u rin e w a s m ix e d th o ro u g h ly , a l0 0 -m L s a m p le w a s d ra w n a n d
stor ed a t -20 #{176}C.
Fe c e s w e re w e ig h e d a n d s to re d at 4 #{176}C until treated. The
number o f ra d io o p a q u e p e lle ts e x c re te d in e a c h s a m p le w a s
counted b y v is u a liz a tio n w ith a P h illip s B V 2 1 5 Im a g e In te n -
s ifie r (P h illip s M e d ic a l S y s te m s , Lo n d o n , Un ite d Kin g d o m ).
P e lle ts w e re counted tw ic e w ith th e c o n ta in e r in tw o d iffe re n t
p la n e s . E a c h s to o l w a s h o m o g e n iz e d w ith a m o d e l L2R ho-
mogenizer (Silver son M a c h in e s Ltd, Buckinghamshire, Un ite d
Kin g d o m ) a fte r a d d itio n o f a k n o w n a m o u n t o f w a te r; a 5 0 -m L
sample w a s fre e z e -d rie d fo r 2 4 h o r u n til a constant weight was
reached. T h e ly o p h iliz e d s a m p le s w e re th e n g ro u n d in a m o rta r
to a fine powder a n d s to re d in p la s tic c o n ta in e rs a t ro o m
te m p e ra tu re .
Nickel analysis
P la s m a , red c e ll, u rin e , a n d fecal samples were analyzed
by IC P -M S a fte r is o to p ic d ilu tio n w ith 6 tN i (s p ik e ), a c id -
d ig e s tio n , a n d s o lv e n t e x tra c tio n w ith a m m o n iu m te tra m e th y l-
enedithiocarbamate and 4-methyl-2-pentanone (24) with a
PlasmaQuad SX300 (VG Elemental, W insford, United King-
dom) lo c a te d a t th e R o w e tt R e s e a rc h In s titu te (A b e rd e e n , S c o t-
land, United Kingdom). Between-day p re c is io n w a s 0 . 8 % fo r
s e ru m and 2.4% for urine samples. R e c o v e ry o f 6 2 N i added to
s a m p le s o f s e ru m , re d b lo o d c e lls , fe c e s , a n d u rin e w a s 9 9 . 8
3.2% (24). M inor modifications of th e m e th o d in c lu d e d re d u c -
tio n o f th e c o n te n t o f sulfuric a c id in the mixture for acid
digestion to 1 0 % , to m in im iz e th e b la n k . A ls o , u rin e s a m p le s
(10 mL) w e re e x tra c te d w ith o u t a c id d ig e s tio n . B e fo re e x tra c -
tion, u rin e s a m p le s w e re a c id ifie d to a fin a l c o n c e n tra tio n o f
1% HNO3 and centrifuged at 500 X g for 10 mm at 15 #{176}C to
remove a n y s e d im e n t. T h e 6 tN i-s p ik e d s o lu tio n w a s c a lib ra te d
against a standard nickel solution (BDH, P o o le , Un ite d Kin g -
dom). Further details of the method and calculations in v o lv e d
are g iv e n e ls e w h e re (2 4 ).
B o th 6 # {1 7 6 }N i and 62Ni w e re d e te rm in e d in e a c h fe c a l s a m p le to
assess both dietary input ( #{176}Ni) and tracer elimination (62Ni).
Although b o th is o to p e s could be determined simultaneously,
their concentration w a s e x p e c te d to d iffe r b y m o re th a n o n e
o rd e r o f m a g n itu d e . B e c a u s e is o to p ic d ilu tio n m e th o d s p ro v id e
more accurate re s u lts w h e n th e a m o u n t o f th e is o to p e s p ik e is
similar to th a t o f th e is o to p e s e le c te d fo r a n a ly s is , th e tw o
nickel is o to p e s in e a c h fe c a l s a m p le w e re d e te rm in e d fro m tw o
separate aliquots (0. 1-0.2 g), which allowed different amounts
of 6tNi to be added. The amount o f is o to p e a d d e d in each case
w a s adjusted to match the expected concentrations of either
60Ni o r 6 2 N i to g iv e ra tio s o f # {1 7 6 }N i to 6 tN i a n d 6 2 N i to 6tNi
of 1.
Each set of samples, for a given subject and sample ty p e
(plasma, re d c e lls , u rin e , a n d fe c e s ), w a s a n a ly z e d o n th e s a m e
d a y to re d u c e a n a ly tic v a ria b ility . C o n tro l s a m p le s w e re p re -
pared fro m p o o le d s e ru m a n d u rin e to w h ic h k n o w n a m o u n ts o f

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618 PAT RI ARCA ET AL
I W I and W 2 are women and M 1 and M 2 are men.
T ABL E 1
General data on volunteers taking part in the 62Ni study
Subject
Wi Ml W2 M2
Age (y) 30 28 21 30
W eight (kg) 66.2 77.5 59.2 86.5
Height (cm) 162 186 169 179
Body surface area (m2) I .70 2.02 1 .68 2.06
Creatinine clearance (niL . min 1.73 m2) 89 1 12 92 22 84 6 100 I l
Total 62Ni dose (pg) 670 760 57 1 879
I W I and W 2 are women and M 1 and M 2 are men.
2 t SD.
62Ni were added and analyzed together with the plasma and
urine samples obtained from the subjects.
Ad d it ion a l a n a lysis
Serum and urine creatinine concentrations were determined
by the Jaff#{ 233} (25) method on an automated analyzer. Creatinine
clearance was normalized to body surface area, according to
Gowan and Fraser (26).
R E SUL T S
Details on each subject taking part in the study-including
sex, age, weight, height, body surface area, average creatinine
clearance, and 62Ni dose-are given in Table 1 . The daily fecal
output of the tracer (62Ni) and the excretion of the fecal marker
(radioopaque pellets) for each volunteer are compared in Table
2. The plasma concentrations of 62Ni observed in each of the
four volunteers at various time intervals are shown in Figure 1 .
The concentrations of 62Ni in plasma and red cell samples
taken at the same time intervals for one of the subjects are
compared in Table 3. Cumulative urinary excretion as a per-
centage of the absorbed dose is plotted against time in Figure
2. The absorbed, excreted, and retained fractions of the dose of
62Ni for ea ch su b j ect a r e sh own in T a b le 4. T h e d a t a ob t a in ed
in this study are compared with data from the study by Sun-
derman et al (10) in T a b le 5.
DI SC USSI O N
The 62Ni dose given in this study (Table 1 ) was relatively
small and comparable with the amount of nickel that can be
ingested from diets rich in certain types of food, such as dark
chocolate, nuts, and soy products, although several studies
showed that nickel absorption is greatly reduced when ingested
with food (8, 10). The dose (0.6-0.9 mg), although not phys-
iologic in form, was the closest in size to a typical physiologic
dose used to date. Smaller doses (eg, 0.4 and 0.6 mg Ni) have
been administered during studies of clinical responses of nick-
el-sensitive patients to oral nickel intake (7, 9), but these
studies had only a limited number of nickel determinations.
Gastrointestinal absorption of 62Ni was estimated as the differ-
ence between ingested nickel and nickel excreted in feces. The
completeness of fecal excretion was confirmed by the complete
recovery of all radioopaque pellets in the feces within 5 d of
ingestion, although patterns of fecal excretion were different from
subject to subject (Table 2). One of the volunteers, who consumed
a vegetarian diet, excreted 18 of the 20 pellets within 24 h,
whereas the others required a longer time. In all volunteers, the
pattern of excretion of the tracer closely followed that of the fecal
marker and there was no evidence of secondary peaks. Therefore,
we are confident that the recovery of the dose of 62Ni not absorbed
from the intestine was also complete and that reexcretion of
absorbed nickel via the gut was n egligib le. The average transit
time (AU) ofthe digesta through the gut was 47.4 h (SD: 18.3 h;
SE M : 9.1 h ) a n d , a ft er exclu sion of t h e vegetarian subject, 54.4 h
(SD: 14.4 h; SEM : 8.3 h). Cummings et al (23) reported an AU
of 54.2 h (SEM : 2.6 h) in 50 subjects given a single dose of pellets
and a shorter AU was observed when additional fiber was in-
cluded in the diet.
The mean concentration of dietary nickel observed in the
feces of all subjects (except in one basal sample, which was
considered incomplete) was 2.2 1. 1 p.g/g wet wt (range:
0.9-5.0 ; .L g/g wet wt; n = 19) and 7.9 3.3 g/g dry wt
(range: 3.5-15 p g/g dry wt; n = 19). Daily fecal excretion of
T ABL E 2
Fecal marker and tracer excreted per day by each subject
W I M 1 W 2 M 2
Fecal Fecal Fecal Fecal
Day marker Tracer marker Tracer marker Tracer marker Tracer
2 4 5 5 2 55
% of tota/ excr eted
6 9 9 0 7 0 2 5 2 2
3 4 0 3 6 2 5 1 9 1 0 2 9 1 5 1 7
4 1 0 1 0 2 0 1 1 0 2 3 5 3 8
5 5 2 0 1 0 0 2 5 2 3
r 0.982 0.965 0.953 0.947

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0 10 20 30 40 50 60 70 80 90
100
4 5
NICKEL M ETABOLISM IN HUM ANS 619
_1
0
E
C
z
04
(0
E
(I)
0
Time (h)
FI GURE 1. Pl asma concentrations of 62Ni after its oral administration in four subjects.
naturally occurring nickel averaged 255 92 j.tg/d (range:
74-388 p.g/d; n = 18; one outlier excluded). M ean individual
nickel excretion, computed from the mean value of daily nickel
excretion over 5 d for the four subjects, was 288 60 p.g/d
(range: 199-326 .tg/d; n 4). The daily variation for individ-
ual subjects ranged from 14% to 51% of the mean. The daily
fecal output of nickel agrees with that of previous studies (10,
27, 28) and is consistent with dietary intakes in the United
Kingdom, which have been estimated as 140-150 p.g Ni/d
from food plus 100 g Nild from cooking utensils (29).
These data confirm that the concentration of naturally abun-
dant nickel in feces is quite high and rather variable on a
day-to-day basis. The problem of variable endogenous nickel
was overcome by using an enriched 62Ni tracer (98.83% ). The
tracer 62Ni was obtained by measuring the total output of 62Ni
and 6#{176}Ni. The measurement of 60Ni allowed a correction for
endogenous 62Ni to be made. The size of the correction ranged
between 5.1% and 13.2% of the total amount of 62Ni excreted
in feces. Fecal excretion of the tracer by the four volunteers
averaged 66.7 4.2% of the dose given and the absorbed
fraction was therefore 33.3 4.8% .
T A BL E 3
Kinetics of 62Ni appearance in plasma and red cells in one subject (W 2)
Time (h) 62Ni in plasma 62Ni in red cells
nmoL/L
0 0 0
0.5 68 2.1
1.5 310 2.1
2.0 318 2.6
2.5 335 4.4
3.0 324 3.4
3.5 318 3.9
4.5 279 4.7
6.5 200 5.5
2 4 6 1 1 5 . 2
48 26 17.7
7 2 1 3 1 6 . 6
96 8 1 2 . 7
The plasma concentrations of 62Ni observed in each of the
four volunteers at various time intervals are shown in Figure 1.
Plasma 62Ni rose rapidly after ingestion and reached a peak
between 1 .5 and 2.5 h, with concentrations ranging between
269 and 344 nmol/L; 62Ni was rapidly cleared from plasma but
was still detectable, at concentrations < 32 nmolIL, until 96 h
after ingestion.
Few data are available on the distribution of nickel between
plasma and red cells. Hopfer et al (30) found no significant
difference between the concentrations of nickel in serum and
whole blood from the same unexposed subjects. However, in
a )
Cl)
0
#{149}0
a)
0
Cl)
.0
C
9-
0
a)
C)
C
C
a)
e
a)
0
0 1 2 3
Time (d)
FI GURE 2. Cumulative urinary excretion of 62Ni as a percentage of the
absorbed dose after its oral administration in four subjects.

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620 PATRIARCA ET AL
T A B L E 4
Absorption, excretion, and retention of the 62Ni dose in each of the four subjects, expressed as a percentage of the dose and a percentage of the
absorbed dose
WI Ml W2 M2 i SD
Percentage of dose (% )
Total 62Ni dose 100.0 100.0 100.0 100.0 -
Total fecal excretion 71.3 67.6 59.9 68.7 66.9 4.9
Absorbed dose 28.7 32.4 40.1 31.3 33.1 4.9
Total urinary excretion 14.5 22.3 32.9 18.9 22.1 7.8
Retained amount 14.2 10.1 7.2 12.4 1 1.0 3.0
Percentage of absorbed dose (% )
Absorbed dose 100.0 100.0 100.0 100.0 -
Total urinary excretion 50.4 68.5 82.0 59.8 65.2 13.4
Retained amount 49.6 31.5 18.0 40.2 34.8 13.4
vitro s tudie s o f cellular uptake of nic ke l indic a te d tha t hig h
c o nc e ntra tio ns o f a lbum in a nd a m ino a c ids m a y pre ve nt nic ke l
accumulation in red cells (3 1, 32). In this study we observed in
vivo the dis tributio n o f nic ke l be twe e n re d c e lls a nd pla s m a .
The c o nc e ntra tio n o f 6 2 Ni wa s de te rm ine d in re d c e ll s a m ple s
taken from two of the volunteers (W 2 a nd M2 ) o ve r a period of
tim e a fte r ing e s tio n o f the is o to pe . Fo r bo th s ubje c ts , the
c o nc e ntra tio n o f 6 2 Ni in re d c e lls wa s < 1 6 nm o l/L, ie , m uc h
lower than that in the corresponding pla s m a s a m ple s . Be c a us e
the concentrations o bs e rve d we re c lo s e to the de te c tio n lim it o f
the m e tho d, the re s ults c o uld be qua ntifie d o nly fo r o ne s ubje c t
(W 2; Table 3). The upta ke o f 6 2 Ni in re d c e lls wa s s m a ll,
de s pite the hig h 6 2 Ni c o nc e ntra tio n in the s urro unding pla s m a .
The peak wa s reached 48 h after ingestion. The de c re a s e in
6 2 Ni c o nc e ntra tio n started when the c o rre s po nding pla s m a c o n-
c e ntra tio ns we re lo we r tha n tho s e ins ide the re d c e lls .
As indicated by the da ta in Fig ure 2 , the re wa s a la rg e be twe e n-
s ubje c t va ria bility in urina ry nic ke l e xc re tio n, whic h wa s o nly
pa rtia lly e xpla ine d by the diffe re nc e s in urina ry vo lum e s a nd
flows. Average da ily urina ry vo lum e fo r a n individua l ra ng e d
from 1015 to 1755 m L and the a ve ra g e urina ry flo w wa s be twe e n
0.71 and 1.22 mlJmin. Values for creatinine clearance and intra-
in d iv id u al v ar iab ilit y we re within the e xpe c te d re fe re nc e ra ng e fo r
he a lthy s ubje c ts (2 6 ) a nd indic a te d no rm a l re na l func tio n (Ta ble
1 ). A lo we r da y-to -da y va ria bility wa s o bs e rve d fo r the ve g e ta ria n
s ubje c t (W 2), according to the la c k o f die ta ry c re a tinine inta ke .
Five days after ing e s tio n, the e lim ina tio n o f nic ke l wa s no t c o m -
ple te in a ny o f the s ubje c ts . The a ve ra g e a m o unt e xc re te d wa s
65.0 1 3.5% of the absorbed do s e .
T A B L E S
Estimates of nickel absorption, excretion, and retention determined by
using naturally occurring nickel or a stable isotope (62Ni)
Sunderman Ct al (
nickel2
10), This study,
62Ni3
% of dose
Fecal excretion 76 19 66.9 4.9
Urinary excretion 26 14 22.1 7.8
Total excretion 102 8 89.0 3.1
Absorbed fraction 27 17 33.1 4.9
Retained fraction 0 1 1.0 3.0
1 SD.
2 Length of study: 4 d.
3 Length of study: 5 d.
In this s tudy, the pe rc e nta g e o f the o ra l nic ke l do s e a bs o rbe d
from the gut wa s fa irly s im ila r in thre e o f the vo lunte e rs ,
whe re a s the fo urth subject (W 2) had higher gastrointestinal
a bs o rptio n (25%). S he is a yo ung ve g e ta ria n wo m a n, who m a y
ha ve ha d ina de qua te iro n inta ke . It ha s be e n re po rte d tha t
nic ke l a bs o rptio n inc re a s e d in iro n-de fic ie nt ra ts (3 3 ). This
suggests that iro n s ta tus s ho uld be assessed in volunteers cho-
s e n for studies o f nic ke l a bs o rptio n. The pe rc e nta g e o f the
absorbed do s e e xc re te d in urine s ho we d a wide r va ria bility
fro m s ubje c t to s ubje c t, whic h wa s pa rtia lly e xpla ine d in te rm s
of the to ta l a m o unt o f urine pa s s e d. The pe rc e nta g e o f the do s e
re ta ine d in the bo dy a t the e nd o f the e xpe rim e nt ra ng e d fro m
1 8% to 49.6% of the absorbed dose (Table 4).
Biliary excretion has be e n s ug g e s te d as a ro ute o f nic ke l
e lim ina tio n o n the ba s is o f e vide nc e o f bilia ry e xc re tio n in ra ts
a nd rabbits (2) a nd the m e a s ure m e nt o f nic ke l in hum a n bile
s pe c im e ns (3 4 ). Ho we ve r, we o bs e rve d no e vide nc e o f e nte ro -
pathic circulation of nic ke l, a s judg e d by the a ppe a ra nc e o f
s e c o nda ry pe a ks o f pla s m a o r urinary nic ke l c o nc e ntra tio ns .
The pattern o f fe c a l e xc re tio n o f nic ke l ha s be e n a s s e s s e d fo r
the firs t tim e be c a us e the tra c e r could be distinguished from
na tura lly a bunda nt nic ke l ing e s te d with fo o d. The e lim ina tio n
of 62Ni in fe c e s fo llo we d the same pattern observed for the
fecal marker (Table 2). This suggests that biliary excretion of
nic ke l is ve ry s m a ll o r a bs e nt in hum a ns .
The results of this inve s tig a tio n s ho we d a much lower be twe e n-
s ubje c t va ria bility tha n that observed in e a rlie r s tudie s (8 -1 0 ),
which us e d na tura lly a bunda nt nic ke l. This is a ttribute d to the
improved ability to dis ting uis h the tra c e r fro m o the r s o urc e s o f
e nviro nm e nta l nic ke l. The e s tim a te s o f nic ke l a bs o rptio n, e xc re -
tio n, a nd re te ntio n o bta ine d in this s tudy a re c o m pa re d in Ta ble 5
with those reported by Sunderman e t a l (1 0 ), the o nly o the r
c o m ple te s tudy o n hum a n subjects. W e observed a higher gastro-
inte s tina l a bs o rptio n (6 %) a nd lo we r fe c a l (-9 %), urina ry (-4 %),
and total nickel excretion (- 1 1 %). The m o s t s triking re s ult ob-
s e rve d in this s tudy wa s tha t a c o ns ide ra ble fra c tio n o f the ab-
sorbed nic ke l do s e (34.8 13.4% ) was s till retained 5 d after
ing e s tio n. The s e re s ults fo r nic ke l re te ntio n a re c o ns is te nt with
tho s e o f re c e nt a nim a l s tudie s [m ic e fe d phys io lo g ic do s e s o f 5 7 Ni
(5 )1 a nd a study by Te m ple to n e t a l (2 1 ), who , in a s ing le -s ubje c t
study found urinary e xc re tio n o f 6 tNi persisting 96 h after a n o ra l
dose of the is o to pe .
The us e o f a s ta ble is o to pe to inve s tig a te nic ke l m e ta bo lis m
in hum a ns has several advantages; the doses can be reduced to
a m o unts c lo s e r to phys io lo g ic o ne s a nd c o nta m ina tio n is e f-

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NICKEL M ETABOLISM IN HUM ANS 621
fectively re d u c e d b y a fa c to r o f 1 8 . 5 , a llo w in g th e fa te o f th e
tra c e r to be followed for a long time.
On the assumption that nickel absorbed from aqueous solu-
tions during fasting behaves as does nickel absorbed from th e
d ie t, th e re s u lts o f th is s tu d y s h o w th a t e lim in a tio n o f n ic k e l
fro m th e b o d y ta k e s lo n g e r th a n re p o rte d p re v io u s ly a n d th a t
nickel excretion a n d in te rin d iv id u a l v a ria b ility m a y h a v e b e e n
o v e re s tim a te d . Estimates of long-term retention o f th e a b -
sorbed nickel fraction should b e c o n s id e re d w ith c a u tio n b e -
cause th e n o n e x c h a n g e a b le o r s lo w ly e x c h a n g e a b le fra c tio n s
can a p p a re n tly b e in c re a s e d b y a n y lo s s e s o f n ic k e l d u e to
in c o m p le te fe c a l and u rin e c o lle c tio n s . Ho w e v e r, in th is s tu d y
the fecal marker given with the nickel dose was completely
recovered from a ll subjects and the retention of nickel w a s
c o n firm e d b y its c o n c e n tra tio n in p la s m a a n d u rin e .
T h e in v e s tig a tio n o f th e s ite s w h e re n ic k e l is re ta in e d a n d th e
processes in v o lv e d in th e s lo w e x c h a n g e o f n ic k e l fro m tis s u e s
will re q u ire fu rth e r s tu d ie s . P re lim in a ry re s u lts o n th e k in e tic s
of nickel uptake fro m e ry th ro c y te s in v iv o h a v e b e e n re p o rte d .
A three-compartment mathematical model fo r n ic k e l m e ta b -
o lis m has b e e n d e v e lo p e d w ith d a ta fro m th is s tu d y a n d p re -
lim in a ry r esul t s w e re p re s e n te d p re v io u s ly a t th e Fifth C O M -
T OX Symposi um on T oxi col ogy a n d C lin ic a l C h e m is try o f
M etals (u n p u b lis h e d o b s e rv a tio n s , 1 9 9 5 ). A fu ll d e s c rip tio n o f
the model w ill b e th e s u b je c t o f a fo rth c o m in g p a p e r. E l
W e are grateful to Andrew Duncan, Dennis OReilly, and Naveed Sattar
from the Glasgow Royal Infirmary, who took part in the organization of the
metabolic study; to the participants of the experiment; and to Brian M cGaw
(Robert Gordons University, Aberdeen) for his advice on the isotopic-
dilution techniques. W e are indebted to Ian Bremner for the use of the
ICP-M S facility at the Rowett Research Institute, Aberdeen, and to M artin
Reid for his skillful assistance.
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