From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
Radioactive
Direct
Vitamin
Measurement
By
ALFRE11
DOSCHERHOLMEX
lVitIi
B
ECAISE
pose,
to the
utilizing
(‘ol)alt
is determined
of vitamimi
method2
is l)a.sed
parenteraily
the isext
results
24 hours.
imi the
If
a
detected
surface
imi normals,
has
been
imi the blood
This
report
aft.er
gives
au
be excreted
iii
the
coumitimig
miot in patients
with
and
the
B12
vitamin
is easily
et al.3
vitamimi
liver
detected
is based
B,2
permiicious
anemia
simultaneously
with
the test dose.
evidence
of the ability
to absorb
made46
t.o the
measurement
by
omi the
omice
radioactivity
is given
indirect
use of direct.
amid
Schillimig
vitamin
in the urine
in
dose of orally
adminis-
urine
the
with
pur-
muethod’
dose
The
body
this
this
Heinle
tracer
by Glass
orgami for
over
absorb
for
deductiomi.
the
described
storage
to
In the
oral
simple
overloading
The method
is the miiain
but.
B12.’3
after
in megaloblastic
ability
of almiiost
all of the
able to absorb
a tracer
is
will
results
beemi described
vit.aniin
at. by
HAGEN
the
have
of
LIU
measure
feces
that
scintiilatioti
of intnimisic
factor
are all based
upon
referemsce
fact.
patient
too
imistrument.s.
that
the liver
By
labeled
excretion
MARGARET
techmiics
is arrived
the
UpOmi
to
in the
absorbed
B12, that
vitamiiin
absorbed.
three
of
Results
the Blood
S.
PAUL
B12 in humans
sought.
time
radioactive
anioumit
appropriate
knowledge
assistance
beeti
present
radioactivity
tered
have
Studies:
in
AND
of vitarnimi
methods
Jip
all
the
the teehnieot
malabsorpt.ion
anemiiia,
vitamuts.
B,2 Absorption
of Radioactivity
it is
is readily
unless
These
vitamin
a source
three
B,2.
of the
tests
Brief
radioactivity
the admimiistrat.ion
of oral tracer
doses
of labeled
vitamin
details
amid sunimarizes
the m’esults of furt.her
studies
by
B12.
this
method.
MATERIALS
The
mg.9
labeled
Co60
The
oral
the
pat ieuut
the
test.
vials
dose
after
(lose
was
givems.
tube.
diameter
by
pamuy. The
an additional
1.5
Per
ig
inches
wa-s
The
From
the
Minneapolis,
deep,
imuvestigatioms
Umiiversity
of
inches
of
in
C000
part.
in
two
a
Mc/
to
houmrs
which
after
20 ml.
elsewhere,7
photo-
6292
hole
Harshaw
sources
1082
disposable
detail
a Dumont
with
the
inches
Chemical
in
Com-
surrounded
was
were
by
24 ±
calibrated
source.
Veterans
of Medici
beast
bead.
The
crystal
was
(counts
per disintegration)
reference
Service,
with
thick
by
on secon(barv
Medical
at
of
administered
in mnore
coupled
efficiency
primary
for
coumutedt
2 imiches
unit
was
use,
by a grant
Admimuistration
of Mi nnesot
University
from
Eli
Lilly
amid Company
Hospital,
a Medical
to
the
Minnuesota.
June
Obtaimued
25, 1956;
from
t Facilities
were
supported
were
by
activity
(1.0sc)
(lescribed
in a sealed
tuud t.he Department.
was
plasma
crystal
by 1/
absolute
a sPecific
withheld
coumnter,
in diameter
determined
Sectiomi,
or
This
of Standards
Hematology
a,
blood
stmppbied
shielded
The
was
Bureaum
Mi munesot
Submitted
was
collar.
efficiency
School.
This
*
2 inches
or0.O2g
was
activated)
The crystal,
a National
of
hiad
(0.5/2c)
b.reakfast
counter.
(thallium
METHODS
imivestigatioms
B,
amid
ml.
ion
iodide
mu this
vitamin
Twenty
emit ire counter
2 inchi bead
cent.
against
used
fast
scimstillat
of a so(bium
multiplier
Ba
of O.46g
auu overmuight
itu a well-type
coussist.ed
pital
vitaniims
test
ANI)
of
umsed
Merck
the
for
accepted
and
Radioisotope
radioactivity
for
publicatiomi
Auig.
Rahway,
N. J.
Company,
Unit
of
the
measurements.
336
Minmseapolis
12,
1956.
Veterans
Administratiomu
Hos-
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
ALFRED
All
radioactivity
count
measurements
of 12,800.
changer.
modified
These
and
scaler,
a Simplex
described
in
the
cam
to
facilitate
shaft
Tvemutv
ml.
was
During
this
grou,mid
In 63 er
there
If, its a single
rums,
minute
amsy
were
count
on
equal
tests
week
pven
Heitule
the
t.o three
this
have
been
carried
anemia
after
the
(lose
active
in
patients
the
Schuilling
amid
was
(lay
was
out
WS
the
in the
counter.
From
from
little
Nuclear
be
the
cent
more
7 cpm
of
to ninse backDuring
the
to 299 coumuts
per
±4
count
deviatiomi
daily
ruin.
cpm
from
thami
±7
from
the
was
the
cpm.
average
accepted
average.
the
of
(lose.
in each
no
the daily
amount
test
two
than
outside
cpni from
standard
oral
193
more
in 85 per
to
on
exCrete(l
variatiomu
no experimental
±7
until
Sevems
average
counts
backgroumud,
sigmiificaist.
omi 36 luomi-pernicious
Blood
administered.
with
161
microswitches
given
varied
found
consi(bered
patients.
more
varied
and
and
the
a preset
tinier.
malinerand
was
count
t.hami
times
figure
permuicious
less
withs
a model
of 3.8 determinations.
counts
repeated
with
same
there
given
count
rumi was
two
the
automatic
However,
the
of
of
tus average
backgrouisd
on
varied
above
mime
simugle
backgroumud
(lay,
counts
Forty-one
ouse
any
the
backgroumid
determimied
background
and
no
made
imucluded
radioactivity
with
ruin
of 242.
The
mu the
rummus on
were
model
1248 B electrical
reset
use of this timer,
instead
of the one
resetting
of the
each
study
ruins
of t.lsat
per
this
used
a(ldit.iomi
counted
cent
142
average
of the
background
backgroumrsd
were
during
of
an
per
counts
by
CPR.
the
337
HA(.EN
Company
the
and
were
in
ma(le
with
cent
average
of urinue
expressed
period
(cpm)
model
required
printing
S.
the use of au automatic
vial sample
described.8
However,
the chamuger was
facilitated
Instrument
timer,
report,
the
were
month
minute
Atomic
printing
study
counts
seven
to
PAUL
background
were
an
original
ahiquots
radioactivity
had
including
determinations
Company
register
test.s
AND
This device was essentially
as previously
to accept 20 ml. vials.
Commercial
equipmemit
Instrument
all
I)OSCHERHOLMEN
was
The
Addisoniamu
anemia
drawn
at
imstrinsic
factor
pernicious
comitrol
differemut.
subjects
time
concemitrate
amueniia
as
and
up
(IFC)
teste(l
17
intervals
with
umse(1
*
both
the
metho(ls.
RESULTS
Tables
1 and
2 include
the
results
of measurements
of radioactivity
and plasma
at different
time
imitervals
imi 28 comit.rol subjects
anemia
patients
examined
with
the smualler
test dose.
Figure
curve
of the plasma
radioactivity
values
in control
subjects
Each
point
on the curve
represemits
the average
of from 5 to
In t.he control
subjects
hours
of the test. Out
only
four
contained
appeared
imi all five
in radioactivity
curve
reveals
tests
the
it was difficult
to detect
activity
during
the first three
of 28 blood
or plasma
samples
counted
from this interval,
more
in 11 out
present
than
of 19 blood
cases
examined
sevemi
out
value
was
found
of five
instamices.
at that
plasma
patients
added
counts
with
definite
were
permiicious
radioactivity
to the
* Kindl
four
hours
samples.
time.
measurable
At. five
From
t.o be just
activity
hours
themi omi there
test
activity
was
supplied
as the
amoumit
in radioactivity
was still presemit
with
permiicious
was
au increase
composite
imidividual
at eight
hours.
over the ensuimig
in the plasma
in
amieniia
(E.L.)
was
fol-
every
two hours
for ten hours
after
t.he imigestion
showing
amiy definite
radioactivity.
When
the same
niomit.h later with t.he addition
of 100 mg. IFC,
l)lood
w-ithimi
amiemia
or
as high
decline
activity
Omie patient
low-ed with blood
measurement
of the oral test. dose without
procedure
was repeated
one
and
At.
amid plasma
Follow-ing
the peak
there
was a slowdays.
Evemi after
one week significamit.
three
cpmn.
until a peak was reached
imi the 8 t.o 12 hour period.
The
an average
peak
of 38 cpmu at eight. hours,
but in some
12 hour
blood
iii
amid 8 pernicious
1 is a composite
from
these
tables.
17 determimiations.
omily
the
were
traces
range
foumid
examined
w-ere
foumid
in control
with
at
subjects.
similar
ten
test
hours
Sevemi
doses
unless
atid
more
no
IFC
dose.
by
Dr.
R.
\V. Heinle,
The
Upjohn
Company,
Kalamazoo,
Michigan.
was
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
338
RADIOACTIVE
VITAMIN
B12
ABSORPTION
STUDIES
I-.
:7
i-
‘-
-
----
-
00
00
-
-
i
0I
‘1
:‘7
9
01
00
-------
-
-
Lt’)
0
#{149}1’
‘O
N.
-
00
N.
-
----CO
n-.
0
0
.0
cu
0.
CO
I-
0
r-
0
.0
-
-
-
m0-)
-
‘7
-
01
0
C’7
C’)
C’)
N.
C’7
-
-
7’7
07
‘7040’s
r-
-
C’S
-
-
C’7
C’S
N.
0
0
0
CO
-‘S
l
E
N.
-
-
0’7 07
9
z
-
-1’
0
00
-07
0
L’7k’7
-
tm’
07
C’S
07
C’S
N.
00
-
07
07
C’S
C’)
C’)
07
00
0707
©
N.
1’
07
-
07
‘1
C’)
c’o
-
-
N.
-
C’S
C’S
C’S
C’)
1’
C’)
i’
C
-
C’S
.
00
C4
07
C’S
C’S
00
C’S
00
-
0
07
C’S
07
00
C’S
00
07
C’S
01
07
C’S
CC0’-4C7N.
C’s
00
07
N.
‘)
e1
00
1’
07
07
00
0
L’7
N.
N.
N.
N.
L’7
N.
3
4’
07
07
00
0
00007
0000a0a
0-
fi-o
0-
fi
0
‘S
c
2072073020
S
.0)
.i
0)
0)5)
0
-:
.1
;
0
0
.
-:
C
:
-
.
.0
-
.
C..
‘
-
-
-
-
-
:::
-
‘
‘
0)
-
.
.0Q
-
.2
-a
J’o
Q
CO
Z
0)03
00
0)
.fi fi
“0
C’
S
0’.
.
L’7
S
-
C’s
07
.
00
-1
C’)
C’S
C’S
-
-
-
-tm’
-
-
)
CO
z
N.
-
07
C’S
L’7
C’S
C’S
-
-
-
07
C’S
07
0
07
C’.
-4’
C’S
-4’
0
C’S
-
-
-
-
-
-
-
-
-
Z
C’S
C’S
-
-i
07
B
:i
-
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
.‘uLFlIEI)
2.-(’mnparison
TABLE
Excretion
m)0SCHERH0LMEN
the Radioactivity
bet iveen
of Radioactivity
nimiscimlarly
after
24 hours
Name
after
Sex
J.A.L.
ML.
PAUL
AXI)
in 20 ml.
the
ldministration
the Oral
Test
Age
(if Plas
of 1,000
Dose
HAGEN
S.
of 0.46
q
ma
and
339
24 hour
the
Vitamin
g
B,
Co6#{176}
Vitamin
B,2
(0.5
Net
cpm at different
hours
after
the orat
test dose
Diagnosis
24
54
50
39
30
48
39
37
31
40
17
26
20
IntraLc)
Urinary
excretion
per cent
orat test
36
Bronchial
23
56
Healthy
Parkimisomiism
S.S.
F.J.O.
H AG.
F
\I
M
M
M
63
Arteriosclerosis
39
Bromucisiecta.sis
63
Coromsary
E.A.
M
67
Lumug
Tumor
27
D.C.
M
57
Optic
Atrophy
25
I).M.
F
27
Healthy
21
I.W.B.
S.F.P.
P.!4.
P.L.*
M
59
Psychoneurosis
20
9.3
M
33
Cholelithia.sis
19
12.7
M
67
PA.
Disease
64
M
10.9
11.8
10.4
8.2
23
15
7.0
4.9
0
10
2
22
F.B.
0
5.3
6
PA.
1.3
8.6
27
HAG.
H.A.G.*
M
F.C.
M
62
PA.
0.6
12.4
0.6
11 .0
8
32
59
PA.
8
42
M
O.F.K.
O.F.K.*
14.L.
14.E.
AM.
*
M
1(X) mg.
obtained
alteration
ordimsary
two
85
imutrimusic
A rompam’ison
the
plasma
vitamin
(lose.
In six of the
of IFC to the test
and imi 11 instances
latter
(fig.
The
2). This
tested
larger
was
especially
with
dose
of 0.92
1.6
PA.
12.5
0.9
of the
al)sorptiomi
B,2 was
dose.
also
well
direct
test.
vitamin
B,2,
the
same
avoid
with
demomi.strated
in the
repeated
six patients
dose
imistead
was carried
the smaller
of
of
out. in
tracer
after
done
at ten
correlation
amid
possible
paremiteral
oral dose
study
was
technic
2. To
amid the activity
in the plasma
w-as increased
urinary
excretion
the
hours
was
ad-
its all
foumid
or blood,
i.e.
amid vice versa
with
pernicious
IFC.
Mg. vitamin
was
procedure
The blood
samplimig
was
at 24 hours.
A reasonable
amid omie patient
gemieral
fimidings,
there
measurement
in table
curve
the
“flushing”
givemi 24 hours
after
the
B12 comitainimig
a five-fold
1.0 Me. of Co’#{176}
was
givemi
to
with pernicious
anemia.
The results,
table
except
that
higher
count.s
were
obtaimied.
The patiemit
w-it.h permiicious
amiemia
showed
plasma
at four amid eight
hours,
but
whemi
labeled
blood
is found
original
method.2
The
with permiicious
anemia
patiemits,
amid without
three
(‘ontrol
subjects
3, revealed
the same
8
44
5
Schilling
between
the um’imiary radioactivity
w-ith imicreased
blood
values
there
anemia
0.8
14.1
PA.
results
for imi Schilling’s
amid 7 patiemits
(lition
(‘ases
7
40
concemutrate.
a modified
as called
subjects
hours
factor
between
with
of the
(‘rystalhmie
11 (‘ont.rol
PA.
81
M
With
those
56
in
of
dose
18.2
17.7
17.4
I\I
HEM.
Asthma
10
(‘rotary
Given
traces
of radioactivity
IFC
was added
to
imicrease
in his inaxiniuni
the
in blood
amid
test dose of
plasma
counts.
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
RAI)IoA(TIVE
VITAMIN
B0
AI3SOIIPTION
STUI)I
ES
55
50
45
E
In
40
-
E
30
-
25
1
tO)
zo
3In
-IC
0
tO)
5
246
12
I0
8
24
72
120
168
Hours
Fuu.
(t).5
c )
auseuuuia
imi 20 ml.
I -Radioactivity
.
Each
poi
subjects.
Although
on
nit
The
shsaded
whole
fowsd
early
that
the radioactivity
t he cu rye
after
is anu average
ims(hcates
area
blood
plasnutu
om’ l)lasma
oral
test.
5 to
17 dci ermi
ramige
of values
of
the
could
doses
be used
plasma
gave
higher
coumits.
was so (‘lose to the background
for
of 0.46
ag.
uuat iomis
in 25 muon jwrmuicioums
for
each
poimut
coumiting
Except
that
Co’#{176}vitamimm
Bm.7
-
purposes,
it was
for a few imist.amiees
where
it. was difficult
t.o evaluate,
plasmna
coumits
were comisistently
higher
thami whole
blood
counts.
borne
out by the experiment.s
(‘ited imi table 4. [nwashed
red blood
This was w-ell
cells contained
sonic
have
m’adioact.ivity,
to the
salimse,
The
order
but
less
always
trapping
of plasma.
When
they
(‘omstained
mit) definite
question
arose
to explore
this
whether
thami
smaller
possibility
whole
1)100(1.
This
were
washed
red 1)100(1 cells
radioactivity.
the
scintillatiomi
same
samiiple
counters
of is-hole
is-as detected
in volumes
of 20 amid 8 ml.,
in 4 miil. However,
when
plasma
even
with the 4 ml. well (‘ounter.
The
the
reproducibility
urinary
secomsd
from
draw-n
injected
activity
excretion
study
was
made
plasma.
The
0.46
t.emi hours
later.
paremit.erally
mneasurememits.
was
of the
plasma
method
by
in each
g.
(0.5
coumited,
coumiting
duplicate
but
mio definite
te(’hmiic
was
on
after
the
radioactivity
dose
was
given
amid the urine
The results
g.
normiial
used.
In
counted
activity
was
could
control
and
of nomiradioact.ive
collected
for the
are summiiarized
be
was
compam’e(I
five
instance
1,000
could
activity
Mc.) oral
Thereupon,
with
in
volumes
of 20, 8 amid 4
blood
definite
radioac-
significant.
tests
due
beets
twice
blood
thm’ee differemit
sized well-type
scintillat.iomi
counters
usimig
miil., m’espect.ively.
Table
5 shows
that. imi the case of whole
tivity
could
emisuimig
in table
foumid
be found
with
that
subjects.
had
1)lood
of
The
disappeared
samples
vitamin
Bm2
24 hours
for
6. Iii getseral
were
were
radiothere
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
\LFIIEI)
I)OSCHEIIHOLMEX
Co rretation
60
between
A
Gontro[
o
PA.
ANt)
S.
HAGEN
341
Pt asma and Urina r Rad ioactivit
Subjects
without
with
#{149}
P.A.
PAUL
IFC
IFC
#{163}
A
A
S
40
A
A
.
#{163}
30
A
A
20
A
j50
10
I
m
2
0
4
i
I
6
Percent
FI(;.
2.-Correlatioms
activity.
between
A “flushing”
oral
test
(lose
(lose
of 0.46
g.
3.-Radioat-tivity
T.Bm,F
1112
(1.0
Mc)
in Blood
Om-all:,
to
Three
I
10
I
10 hour
(0.5
Sex
Age
M
61
or Plasma
after
the
Subjects
Diagnosis
M
24
One
M
F.McA.
M
.
F.McA.
M
with
mug.
t
B,2
Urinary
obt
=
iuuse(l
excretioms
itst.ramuiusculmtrly
I FC
75
2(K)
IFC
Plasma
*
75
4
hours
--
-
12Ol44i1b8
14
2233
56*
38
fIt)
Permuicious
Blood
Amieniia
Plasma
Permuicious
A,semi a
Blood
Plasma
S amid
of
radioactivity
after
oral
comuceust
rate.
2l
31
Plasma
Blood
pooled
Patient
after oral test dose
-
Plasnua
froni
Vitamin
(‘)6O
g
.1 nemia
___________
-
senla
or
u-ions
108*
Blood
Psychoneu-
24 huours
I mit ri music fact
at different
________
Plasma
Eniphy-
62
Pei-n
Blood
rosis
CC.
of 0.92
Administration
and
I
Bronchial
20
Mc).
(.‘ontrol
Ast.hnia
F.!!.
18
and 24 hour urimsary radiogivems paremuterally
24 huours after
the
was
0
MilK.
16
14
radioactivity
B,2
Cpm
Name
t
Padioactivittj
plasma
vitamin
t
12
Urinarq
of l,00Og
Co6#{176}
B,2
I
8
910
1110i
6
5
7 029
953
44H
blood
0.8
8.4
30
4
12 hour
15.6
specimuuemss.
its per cemut. of oral
test.
dose.
tracer
(lose.
1 ,000-
Mg vitamuuits
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
342
RADIOACTIVE
4.-Radioactivity
TABI.E
VITAMIN
20
of
nil,
of
B,
ABSORPTION
Whole
Blood,
Blood
Cells
STUDIES
Plasma,
Net
Unwashed
and
ttashed
Red
cpm
Name
Plasma
Whole
Blood
RBC
Unwashed
EL.
42
28
F.H.
56
32
22
55
38
20
M.R.K.
108
CC.
60
33
EL.
5.-Radioactivity
TABLE
of
______-
Blood
hour
Sample
17
6
of
Scintillation
or Plasma
Blood
Countei-s,
Efficiency
or
Plasma
in
20,
8 and
4 ml.
of counter,
Volume
counter,
24
Blood
Blood
1
0
Different
ml.
Sized
Net
cpm
29
20
8
10
30
8
8
42
4
5
42
4
0
24
20
19
S.P.
S.P.
Plasma
30
8
S.P.
Plasma
42
4
J
‘I’hree
30
Blood
Blood
Plasma
it)
D.M.
Plasma
24
2()
54
D.M.
Plasma
30
S
25
D.M.
Plasma
42
4
19
6.-Comparison
TABLE
Same
the
Blood
F.H.
F.H.
F.H.
F.H.
F.H.
Washed
.
Well-Type
Name
RBC
Urinary
Between
Excretion
after
the
the
Test.
Oral
Reproducibility
1,000
g
Administration
Cpm
in
20 ml.
-
of
Vitamin
B,2
of 0.46
of plasma
at
Mg
the
Plasma
Was
Given
(‘p60
Vitamin
10 hr.
Urinary
Radioactivity
and
Intramuscularly
B,2
excretion
(0.5
the
24
10 hours
Mc).
of radioactivity
of oral
test
dose
in per
cent
Name
Test
E.B.
Test
B
WI?.
31
34
33
51
23
39
38
49
L.K.
17
28
P.13.
E.H.
was
A
good
reproducibility
the repeated
plasma
excretiomi
percentages.
1)ut w’ith
values
some
Test
25.8
28.5
B
14.4
27.4
20.6
18.2
1
variations.
correlated
Test
A
a little
34.9
26.8
9.9
20.7
How-ever,
better
than
in t.his
small
t.he duplicate
series
urimiary
DISCUSSION
After
physiologic
peak of the
Imisignificant
foumid
consistently
of labeled
amounts
radioactivity
activity
in the
was
not
cious anemia
when
IFC
the delay
its appearance
blood
present
only
during
in control
was given
of blood
vitamin
B12
w-ere
administered
orally
amid plasma
the
subjects
simultaneously
radioactivity
was found
8 to 12 hours
first
three
hours.
This
trend
but
also
in patients
with the test dose.
could
not be attributed
with
the
later.
was
perni-
Therefore,
to lack
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
ALFRED
of readily
available
activity
above
started
intrinsic
three
to appear
after
the
DOSCHERHOLMEX
start
factor
times
at four
of the
the
it was
This
Our
absorption
curves
different
from
of vitaniin
B12 w-ith
concemitrations
closely
physiologic
anioumits
massive
In
i’as
with
deviation
of
muot always
physiologic
observed
bioassay
betw-een
absorptiomi
after
one
S.
and
resemblimig
of vitamin
hours
Schillimig’s
origimial
obser-
b)egami four
t.o six hours
vitamin
determined
massive
oral
doses.
They
hours
after
the
ingestiomi
B12 were
usually
with
of labeled
glucose.
Although
umitil five
ct al.9 who
of
B12.
background
by Ross
two
that
vit.amimi
the
in plasma
imi line
doses
343
HAGEN
found
in the urimie usually
of vitamin
B12.2
after
those
PAUL
imit.eract.iomi
fimidimig was
vatiomi that significamit.
excretion
the pam’ent.eral
“flushing”
dose
imigly
for
standard
hours,
test.
AND
These
B12 wem’e strikserum
levels
peak
serum
found
of
the
differemices
absorbed
after
in a different
test
dose,
indicate
that
maminer
from
doses.
studies,
a comparatively
after
our
rather
slow
the peak values
slow amid gradual
disappearamice
parenteral
microbiological
was
admimiistratiomi
method,
of gamma
m’adiat.iomi
declimie
of the blood
imi marked
previously
were unable
contrast
to
wem’e attaimied
radioactivity.
the
rapid
there
This
clearamice
after
reported.
(1omiley amid associates’#{176} using a
to demonstrate
imicreased
serum
vitamimi
B12
concemitrations
six hours
after
the imitravenous
vit.aniimi
B,2. Mollin
et al.” injected
imitravenously
vitamiiimi
12 and
foumid a rapid clearamice
of plasma
administration
of I ,000 ,g. of
1 .5 g. doses
of Co55-labeled
radioactivity.
Imideed, 50 per cent
had
oiily
disappeared
activity
in 0.3 to 0.9 hours.
remained,
clearance
w’ith
curves
If we
different..
14 hours
later
after
accept
from
peak radioactivity
Similar
figures
samples
F.McA.,
In
43.6
counted,
to the
table
32 control
By ten
average
to 92.0
hours
per
cent
w-ith
in the plasma
with t.he larger
test
3).
subjects
dose
givemi
per
an
average
ceiit
cemit.
of the
Our
peak
plasma
doses
highest
were strikimigly
comicentratiomi,
of 68.1
per
cent
of the
in 13 comitrol
subjects
(tables
1 amid 2).
test dose (table
3) w-hemi w’hole blood
of vitamin
the
17.5
of physiologic
as close to the
amid in t.w-o patients
oral
5 to 29 per
of approximately
oral admimiistrat.iomi
the ten hour
value
persisted
were found
were
w-as added
an
with
B,2
smaller
test
permiicious
(pat.ient
dose
anemia
w-hemi IFC
P.L.,
table
2 amid patient
w’e found
from
17 to 54 cpm,
with an average
of 36 cpni imi the 8 to 12 hour period,
which
represented
the peak
of the radioactivity.
Assuming
that the Co6#{176}
remained
with the vitamimi,
this corresponded
to from two to five MMg. vitamin
13,2 per ml. of plasma,
with an average
of 3.5. This
B,, present
was equal
to about
1.0 per cent of the total
concentration
in serum
which
averaged
337 MMg. per ml. imi 48 tests imi our
Assuming
determimied
an average
from tables
two
per
cent
the
average
day
of the
of the
of 11.7
studies.
plasma
volume
1 and 2 that the
amount
per
cent
It is also
given
(230,000
we were
plasma
24 hours
a fraction
of the
blood
at a given
The relatively
moment.
late accumulation
cpm).
able
of interest
activity
in the
then,
that
only
of approximately
plasma
radioactivity
to “flush
that
This
figure
out”
imi the
we were
able
peak
of
t.he
radioactivity,
was
of radioactivity
imi the
and
the
finding
niore
of definite
it can be
was only
in contrast
urimie on the
to detect
after
such a flushing
procedure.
absorbed
vitamimi
w’as presemit
ance from the plasma,
the fact that
five to six times
creted
in the urine
omi the second
day of the experiments
the
2800 nil.,”
at its peak
of vitamin
laboratory.
plasma,
defimiite
itt
to
second
radio-
It is apparent,
the circulatimig
the
slow’ clear-
radioactivity
was
thami was circulating
radioactivity
in
exat
the
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
:14-i
IIAm)I0AcTIvE
even
plasma
prolonged
of
after
such
iliflux
of
physiologic
VITAMIN
a flushimig
of
ABSORPTION
procedure
radioactivity
amounts
B12
vitamin
may
the
into
B,2.
STUDIES
all
be taken
bloodstream
slow
A
as evidence
after
oral
of the
transport
vitamimi
the imitestinal
wall w-it.h gradual
release
into the blood
could
explain
phemioniemia,
although
alternative
explanations
are possible.
Nine
patiemits
with
permiicious
anemia
were examined
amid in all
w’ere able
to separate
them
from
the control
subjects.
The plasma
tamed
seven
itt
given
the
given
a similar
was added
to the
IFC
radioactivity
snialler
test
dose
rangimig
from
22 to 44 cpni.
The
cpm.
three
control
subjects
the
slightly
rest had
absorptiomi
above
three
less activity.
of vitaniimi
B,,.
given
radioactive
vitamimi
B,,
years it has become
absorb
smnall but
Ami additional
agreememit
Smaller
w-ith this
scintillation
therefore
given
expected
that
they
after
evident
amounts
opinion.
coumiters
have
also
test
makes
obtained
niargimi
of radioactivity.
contrast
to
5.3
have
able
beemi
oral
t.est
0.5
c.
Since
betweemi
certain
years
to obtain
of
Co6#{176}
labeled
the direct
miornial
advantages
requires
Schillimig
interference
is given
less
availability
for
2
c.
amid the
the
had
but
the
of the
be used
B,,)*
g.
subjects
and
had activity
Kimudly
supplied
labeled
of radioactivity
from
Dr.
the
is less
N.
pain
start
opinion
is the
test.
absolute
Over
efficiency
for counting
whole
blood
fact
the
amid
purposes.
past
anemia
are in
it
Our
was
fail-
blood was used
(table
cells and the fact that
of whole
blood.
magmiitude,
thus
small
quantities
vitamin
B,,
in the
plasma
as with
20 ml.
has
differentiated
in
we
an
after
B,,.’4
amid t.he result
the bone marrow
by
ex-
subnormal
amount
of
dose
of 1,000 g. non-
patient
pernicious
disease.
imi comparison
anemia,
The test
it appears
to have
is easy to perform,
with
the
Heinle
can be obtaimied
within
12 hours.
morphology,
since no parenteral
than
the
daily
intake
of vitamin
B,,
diet..
*
the
plasma
that patients
with pernicious
of vitamin
B,,.”
Our findings
subjects
and patiemits
with
in the diagnosis
of this
dose
peak
of Co58 and Co56 with half lives of 72 days
permit
the use of larger
test doses.
Indeed,
same
average
activity
in 4 ml. of plasma
after
(2.5
vitamin
measurement
test.
was
Co6#{176}will
Co58
cooperation
methods,
with
while
blood
it. difficult
to ensure
a homogeneous
mixture
with
the smaller
counters
is of a lower
of accuracy
when
work
is done with
such
The
dose
dose
of this
ure to detect
radioactivity
imi the 4 ml. counter
when
5) must
be ascribed
to the diluent
effect
of the red
their presemice
The activity
reducing
the
whole
a definite
parent.eral
a higher
could
we
oh-
deviatiomi
of the background,
finding
as evidemice
for slight
confirmation
hours
24
instances
counts
17 to 54 cpm. When
anemia
had plasma
betw’een
control
latter,
mievertheless,
times
the standard
We interpret
this
increasimigly
insufficient
through
all of these
2 to 8 cpm,
case
a similar
that in all but one case w’e were able to detect
radioactivity
imi the urimie after
a “flushing”
few
can
eighth
a
from
normals
was obtained.
The
dose amid revealed
a maximum
plasma
of IFC the radioactivity
increased
to
activity
ranging
from
56 to 108 cpm.
Although
w-e had mio difficulty
in separating
tients
w-ith pernicious
anemia,
four of the nine
the plasma
while
the
from
ranging
from
with pernicious
In
amimied,
but also here a good
differentiation
nimith case was tested
with the larger
test
radioactivity
of 1 1 cpm. With
the additiomi
53
ranged
test dose had activity
test dose the patiemits
miormals
for
administration
S. Ritter,
Merck
and
Co.,
Rahway,
N.
J.
and
There
vitamin
the
is no
B,,
omi a normal
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
ALFREI)
Heimirich
imi the
amid Heimburg’5
blood
intrinsic
iii
They
w-ith
comitrol
g.
test
dose.
but
in figure
How-ever,
1 ,000
oral
subject,
depicted
recently
after
the
umiable
the
of mionlabeled
findimig
our
test
345
HAGEN
patients
to
to
of
after
radioactivity
the
additiomi
observations,
dose
they
amid found
explain
these
vitamin
12
of
found
no activity
differences
re-
from
parent.erally
carried
out. a similar
curve
miot materially
our
simultamie-
procedure
in omie
differemit.
from that
1.
SUII
miew method
A
S.
reported
amiemia
We have
an absorptiomi
foumid
PAUL
in comstrast
first three
hours
hours.
We are
gave
the
ANt)
in permiicious
amid
material.
the activity
in the
mainimig
after
four
ously
have
normals
factor
results.
1)OSCHERHOLMEN
for
the
described
usimig
ingestion
of physiologic
RY
determinat.iomi
measurement
of the
absorptiomi
of radioactivity
test
doses
of vitaniimi
in the
of Co6#{176}
labeled
Although
doses
of 1.0 Mc. (0.92
g. vitamin
reliable
results
were obtaimied
with 0.5 Mc. (0.46
blood
vitamin
B,,)
g.
gave
vitaniin
B12 has
been
after
the
or plasma
B,2.
higher
counts,
equally
The radioactivity
B,,).
was
found
in the plasma
port.iomi
of the blood.
With
t.his method
it was possible
to differentiate
betw-een
all
with permiicious
aneniia
amid 36 comitrol
subjects.
Iii miomi-pernicious
anemia
subjects
and in pernicious
anemia
intrinsic
factor,
activity
there
until
w’as a relatively
a peak
of the
test
dose.
served
by
others
w-as reached
This
mode
of absorptiomi.
Following
the peak
and
the
small
amounts
and
B,,
oral
blood
states,
Illo
consiste
de
que
nove
doses
physiologic
of vitamin
del
doses
de 1,0 Mc (0,92
the
in the
the
the
ingestion
early
indicating
rise
quite
declined
different
of intrinsic
factor
of vitamimi
ob-
a differemit
gradually
week,
evaluation
from
other
activity,
B,2.
INTERLINGUA
determimiar
radioactivitate
de
B,,,
as one
given
radio-
after
from
radioactivity
metabolism
IN
pro
interval
patiemits
or plasma
administration
previously
reported.
diagnosis
of pernicious
anemia
and
of the
methodo
imi le mesuratiomi
le imigestion
Ben
un
blood
different
for as long
SUMMARIO
Es describite
(luite
concemit.ration
aspects
in the
8 to 12 hour
doses
persisted
of various
rise
was
after
parenteral
valuable
in the
malabsorptive
in studies
curve
massive
usually
rapid
disappearamice
This
niethod
appears
vitamimi
imi the
absorptiomi
after
delayed
of mime patients
proba
Mg de vitarnimia
de
le absorption
de vitamina
B,,.
in le sanguimie
o le plasma
post
vitamina
B,,)
B,,
marcate
resultava
imi plus
comi Co60.
alte
contos,
le resultatos
obtenite
con 0,5 c (0,46
Mg de vitamina
B,,)
non esseva
minus
fidel. Le radioactivitate
esseva
trovate
in le portion
plasmatic
del sanguine.
Le methodo
permitteva
le differentiation
inter
omne
membros
de un serie de
nove
patiemites
subjectos
habeva
recipite
In
del
radioactivitate
intra
curva
terisate
doses
tiomi.
un
del
intervallo
de anemia
perniciose
e umi serie de 36 subject.os
de comitrolo.
sin anemia
perniciose
e imi subjectos
con anemia
perniciose
qui
factor
intrinsec,
ii occurreva
un relativemente
retardate
elevation
del
sanguine
de inter
o del
8 e 12 horas
plasma
post
absorption
de vitamina
B,, esseva
per un pronipte
elevation
que altere
oral de vitamina
B,,. Le differemitia
pare
usque
umi culmimie
le imigestion
del
dose
esseva
attimigite
de proba.
Iste
satis differente
ab le curva
characautores
ha observate
post massive
indicar
differemite
modos
de absorp-
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
346
1tAIOACTIVE
Post
le culmine
del
radioactivit.at.e
se
durante
de
statos
factor
imit.rinsec,
a
devenir
radioactivitate
e
de valor
de malabsorption
del
parve
samiguine,
residuos
de vane
B,,,
de
le nivello
de
niulto
post
in le diagmiose
de vit.amina
e in studios
STUDIES
septimana.
Isto esseva
rapide
del radioactivitate
liii
disparitiomi
promitte
altere
de
ABSORPTION
gradualmente,
usque
reportate
parenteral.
Le niethodo
B12
comicentration
abassava
periodos
previemnente
VITAMIN
illo
differente
ab
administrationes
anemia
perniciose
in le evalutation
aspectos
del
del
persisteva
de
metabolismo
le
e de
activitate
de
de vitamina
B,,.
ADDENDUM
Simice
the
l)r(’l)au’atiomi
ttIi(l
submnissiomi
Energy,
Lnited
Nations
2: 223, 1956)
have
published
their
sorption
crve
obtained
aftem- the
B,, are similar
to those
oi)SerVPd
of
10:
of Atomic
475,
this
l)aI)(-m,
1956)
and
experience
\Iolhin
and
Booth
w-ith
and
plasma
admiiinistration
of
Sniith
(Peaceful
Mollin
(Brit.
countimug.
Their
doses
physiologic
Uses
J.
Haernat.
plasmna
of
Co56
ab-
vitamin
us.
by
REFERENCES
HEINLE,
of
WEI.CH,
A. D.,
amid absorption
65:
Phys.
Am.
2
R. W.,
excretion
ScilmLLus(;,
R.
excretiomu
of
214,
F.:
\.,
SCHARF,
of
Intrinsic
factor
II.
studies
aft.er
oral
The
& Clin.
Med.
42: 860-866,
1953.
G. B. J., BOYD,
L. J., GELLIN,
G., AND
vitamimu
B,2 l)y the liver in huma,is:
Test for
vitamin
B,
and imutrinsic
factor
activity.
Arch.
A.,
AND
New
:
6
observations
Medical
Minnesota
juice
Studies
Tr.
27:
Bulletimu
on the
radioactive
A,
L.: Uptake
of intestinal
STEPHANSON,
measurement
urinary
B12. J.
vitamimu
Biochem.
& Biophys.
S.: Presented
in part to the Society
Section,
Jan. 25, 1956.
on vitamin
B,.
absorption,
Editorial,
Minnesota
Medicine,
and
of gastric
of
of radioactive
absorpt.ion
of
51: 251-257,
1954.
for Experimental
P.
HAGEN,
AND
effect.
administration
Lab.
G,ss,
Biology
\V. H.:
amuemia,
AND
B,
vitamin
1952.
radioactivity
DOSCHERHOLMEN,
PRUSOFF,
in pernicious
G. C.,
MEACHAM,
labeled
Co60
University
of
1956.
204-205,
vitamin
B,2 absorption
stt,dies:
Results
of direct
measurement
blood,
Forty-eighth
Annual
Meeting
Am. Soc. Cliii. I,ivest.,
p. 17,
1956,
amid J. Climi. Invest.
35: 699, 1956.
BASKIX,
R., DEMORE5T,
H. L., AND SANDHAtS,
S.: Gamma
counting
efficiency
of two
Na! crystals,
Nucleomuics
12: 46-48,
1954.
DEMOREST,
H. L., ERIK50N,
J. H.: Automatic
sample
changer
for
well-type
scintillation
Radioactive
AND
of
in the
radioactivity
coumuter.
Ross,
and
I. M.,
tions
“Mouix,
of
B,7 without
intrinsic
84-94, 1951.
D. L., PITNEY,
and
urinary
Cox, E. V.,
B,2 in serum
vitaniirs
C. L., KRE VANS,
on the absorpt.ion,
‘#{176}
CONLEY,
1954.
D. L.,
MOLLIN,
comscemutration
vitamin
38:
12: 68-69,
Nucleomsics
G.
J. R.,
W. R.,
of
and
C.,
BARROWS,
excretion
S. J.,
BAKER,
parenterally
C.:
and urine
following
9: 473-488,
1954.
B. F.,
CHOW,
utilization
excretion
Blood
factor.
C.
UNGLEY,
AND
BRADSLEY,
AND
administered
Co58
oral
AND
of vitamin
Hematologic
respomises
administration
J. E.:
vitamin
J.
A.:
C.
LANG,
B,,.
Lab.
B,1,
Observa-
& Clin.
The plasma
Blood
of
Med.
clearance
11: 31-43,
1956.
P.
12 M0LLISON,
Publicatiomis,
‘
MOLLIN,
gastric
‘
D.
L.,
L.: Blood
transfusion
1951, p. 47.
“intrinsic
in a young
A.,
DOSCHERHOLMEN,
“HEINRICH,
S. J.,
BAKER,
atrophuy
VON
factor.”
H.
AND
C.,
AND
man.
HAGEN,
AND
Ztschr.
in
medicine.
clinical
DoNmAcul,
L.:
Brit. J. Haemat.
P. S.: Observations
HEIMBURG,
f. Naturforschung
R.
G.
Oxford,
Addisonian
pernicious
1: 278,
to
V.:
lib:
Blackwell
Scientific
amiemia
without
1955.
be published.
Radio-vitamin
113-114,
1956.
B,,
resorption
and
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
1957 12: 336-346
Radioactive Vitamin B12 Absorption Studies: Results of Direct
Measurement of Radioactivity in the Blood
ALFRED DOSCHERHOLMEN, PAUL S. HAGEN and MARGARET LIU
Updated information and services can be found at:
http://www.bloodjournal.org/content/12/4/336.full.html
Articles on similar topics can be found in the following Blood collections
Information about reproducing this article in parts or in its entirety may be found online at:
http://www.bloodjournal.org/site/misc/rights.xhtml#repub_requests
Information about ordering reprints may be found online at:
http://www.bloodjournal.org/site/misc/rights.xhtml#reprints
Information about subscriptions and ASH membership may be found online at:
http://www.bloodjournal.org/site/subscriptions/index.xhtml
Blood (print ISSN 0006-4971, online ISSN 1528-0020), is published weekly by the American Society of
Hematology, 2021 L St, NW, Suite 900, Washington DC 20036.
Copyright 2011 by The American Society of Hematology; all rights reserved.