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J. clin. Path. (1954), 7, 66.
THE SERUM VISCOSITY AND THE SERUM PROTEINS
BY
ROBERT D. EASTHAM
From the Department of Clinical Pathology, University College Hospital, London
(RECEIVED
FOR PUBLICATION JULY
Equal weights of the various protein fractions in
plasma affect the plasma specific gravity equally,
whereas weight for weight the fractions exert
differing influences on the plasma viscosity. Thus
fibrinogen has a greater effect on the plasma viscosity than does globulin, which, in turn, has a
greater effect than albumin (Bircher, 1921 ; Naegeli,
1931; Oberst, 1939; Cowan and Harkness, 1947).
Working with mixtures of albumin and globulin,
Naegeli (1931) and Nugent and Towle (1935) have
shown that at a constant total protein content the
viscosity of the mixture falls as the ratio of
albumin to globulin rises, and vice versa.
It was considered that perhaps, given the total
protein content and the viscosity of a serum, it
might be possible to calculate the albumin: globulin
ratio. Thus, using the copper sulphate specific
gravity method for estimation of the total serum
protein level (Harrison, 1947) and a modified
Ostwald viscometer for the estimation of the serum
viscosity (Lawrence, 1950), the estimation might be
completed in under 15 minutes per serum, a much
more rapid test than either the Kjeldahl or the
Biuret methods. The specific gravity of a seruim
can be estimated to ± 0.0005 (± 0.18 g. %), and the
serum viscosity ratio is accurate to ±0.2% (Lawrence, 1950). In fact, the estimation of the
albumin:globulin ratio by this proposed method
was not achieved.
Method
Blood, 7-10 ml., was taken by venepuncture from
various patients and normal controls, allowed to clot,
and the serum then removed. The total serum protein
level and the albumin:globulin level were estimated
by the micro-Kjeldahl method described by Harrison
(1947). The serum viscosity was estimated by the
method described by Lawrence (1950). Each serum
was progressively diluted with physiological saline, and
the viscosity of each dilution was estimated. From the
curve obtained by plotting the total serum protein
levels against their respective viscosities it was possible to calculate the viscosity of each serum at a
constant total protein level. For convenience, this
arbitrary level was taken as 6.5 g.%. Saline was used
7, 1953)
as diluent so that the various protein fractions should
remain in solution (Bingham, 1945).
Also, a normal serum and serum from a case of
advanced cirrhosis of the liver were mixed in varying
proportions, and the viscosities of the various dilutions
of the sera and the mixtures were estimated. Thus
both the albumin:globulin ratios and the proportions
of the various globulin fractions were varied in the
mixtures.
Resuls
In Fig. 1 the viscosities of the two sera and their
mixtures are shown at various dilutions. The
S.iS
7
4J~~~~~~~~~~~~~~~~4
0~~~~~~
0~~~~~~~~~~~~~~N
12
14
16
1.8
24
22
Serum Viscosity Ratio (Water= 1.0)
FIG. 1.-The serum viscosity ratios of two sera and mixtures, plotted
against the serum protein concentration.
results resemble the graphs of Naegeli (1931) and
Nugent and Towle (1935), in that as the albumin:
globulin ratio rises so the serum viscosity falls.
Thirty-five results are shown in Fig. 2, in which
the serum viscosity of each serum at a total pro-
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SERUM VISCOSITY AND SERUM PROTEINS
tein content of 6.5 g. % is plotted against its corresponding albumin: globulin ratio. The sera were
taken from two normals and from a number of
patients with rheumatoid arthritis (nine), congestive
cardiac failure (seven), multiple myelomatosis
(three), carcinomatosis (two), unexplained anaemia
in old age (two), advanced hepatic cirrhosis (one),
lymphoma (one), diabetes with nephrosclerosis
(one), acute rheumatic fever (one), dermatomyositis
3-01-.0
0
*0
C~
._
-
0 2.011.0
0
0
10
0
0
0
0*
0
0
0
0
10/1.0.
0
a
0
0
14
1.5
1.6
.~~~~~~1.8
1.7
-
1.9
Serum Viscosity Ratio (Water=l.O)
FIG. 2.-The albumin: globulin ratios of sera plotted against their
serum viscosity ratios at a total protein concentration of 6.5 g.%.
(one) and pooled sera (five). Although, in general,
the serum viscosity increases as the albumin:
globulin ratio falls, there is a wide scatter in the
readings of the ratio at any given serum viscosity
value. Thus the estimation of this ratio from the
serum viscosity and the total serum protein concentration is not feasible.
Discussion
Although the molecular weight of globulin is
given as 155,250 by Harkness and Whittington
(1947), they state that this is an average estimation.
Globulin consists of many fractions, and Wuhrmann and Wunderly (1947) give the molecular
weights of alpha-globulin as 500,000, beta-globulin
as 2,000,000, and gamma-globulin as 150,000. It
67
is reasonable to suppose that these fractions will
have different effects, weight for weight, on the
serum viscosity. Harkness and Whittington (1947)
showed that a mixture of albumin and globulin also
contained a combined (albumin-globulin) complex
with a molecular weight somewhere between those
of its components, and hence with an intermediate
effect on the viscosity of the mixture. This being
so, it is obvious that variations in the proportions
of the globulin fractions, quite apart from any variations in the albumin: globulin ratio, will affect the
serum viscosity. This would account for the wide
scatter apparent in Fig. 2, and the resulting gross
inaccuracy were the serum viscosity taken as a
measure of the albumin: globulin ratio: it has a
direct bearing on the erythrocyte sedimentation
rate. It is known that the erythrocyte sedimentation rate depends on the aggregating action of
the plasma, which is a complex function of the
plasma viscosity (Whittington, 1942). Also globulin is known to have a direct effect on the sedimentation rate, although not as great as the effect of
fibrinogen (Ley, 1922 ; Linzenmeier, 1923 ; Naegeli,
1931; Bendien and Snapper, 1931; Vogt, 1941).
Aldred-Brown and Munro (1934) could find no
close correlation between the sedimentation rate
and the plasma globulin content or the albumin:
globulin ratio, and from Fig. 2 it may be concluded
that sera with the same viscosity will probably not
influence the erythrocyte sedimentation rate to the
same degree, since the viscosity may be the result
of varying proportions of protein molecules of
various sizes.
Ehrstrom (1939) suggests that euglobulin accelerates the sedimentation rate; also it has been
shown that euglobulin has a greater effect on the
erythrocyte sedimentation rate than has pseudoglobulin, and that albumin positively depresses the
sedimentation rate (Gray and Mitchell, 1942;
Gordon and Wardley, 1943). Shedlovsky and
Scudder (1942) found a direct qualitative relation
to exist between the plasma alpha-globulin and the
sedimentation rate, but not between either the betaglobulin or gamma-globulin fractions and the sedimentation rate, while Hardwicke and Squire (1952)
found that alpha-globulin and gamma-globulin
fractions were the most active of the globulin
fractions in increasing both the sedimentation rate
and the plasma viscosity. A further complication
is suggested by the work of Kunkel and Tiselius
(1951); using a two-way electrophoresis technique,
they found that some plasma protein fractions, in
particular the gamma-globulins, may often be
heterogeneous. Thus, while the serum viscosity in
conjunction with the total protein content will indi-
Downloaded from http://jcp.bmj.com/ on June 17, 2017 - Published by group.bmj.com
68
ROBERT D. EASTHAM
cate a qualitative change in the serum proteins, an
accurate assessment of the nature of the globulins
present or the albumin: globulin ratio is not
possible.
Summary
Although sera with a high albumin: globulin
ratio are less viscous than those sera in which the
ratio is low, no simple relation has been found to
exist between these two variables. Thus it is not
possible to calculate the albumin: globulin ratio
accurately from the total serum concentration
and the serum viscosity.
This lack of correlation is probably explained by
the different effects of the various globulin fractions on the serum viscosity, and by the possibility
that some of these fractions may be heterogeneous.
The significance of these findings is discussed
with relation to the effect of the plasma globulin
on the sedimentation rate of erythrocytes.
I am indebted to Professor M. Maizels for encouragement and helpful criticism, and to Dr. J. S. Law-
rence for the loan of one of his viscometers. The
substance of this paper formed part of a thesis submitted for the degree of M.D. (Cambridge).
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The Serum Viscosity and the
Serum Proteins
Robert D. Eastham
J Clin Pathol 1954 7: 66-68
doi: 10.1136/jcp.7.1.66
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