Colloid Osmometry
Elke Rudloff, DVM, DACVECC, and Rebecca Kirby, DVM, DACVIM, DACVECC
Complications related to intravascular fluid resuscitation and
maintenance can become life-threatening. Overhydration and
underhydration can lead to significant perfusion abnormalities and
can delay or prevent recovery. A working knowledge of
transcapillary fluid dynamics gives the veterinarian the basis for
evaluating the cause of alterations that make up Starling's forces.
By combining this knowledge with patient assessment and
laboratory diagnostics, the veterinarian can make a logical
decision about the best treatment to correct the Starling's force
alteration. Colloid osmometry is a particularly useful tool for
assessing a patient's colloid osmotic pressure. It allows the
veterinarian to distinguish between reduced oncotic pressure and
increased hydrostatic pressure as a cause for intravascular fluid
loss or edema formation.
Copyright © 2000 by W.B. Saunders Company
mber is a 6-year-old spayed female golden retriever that
A was being treated for peritonitis due to a ruptured pyome-
tra. On day 3 of hospitalization, she developed respiratory
distress. Auscultation revealed moist lung sounds throughout
her lung field, and she was hypoxemic. She had an albumin
level of 2.64 g/dL (normal range, 3.5 to 4.0 g/dL). An effective
treatment plan for her can only be determined after answering
several important questions pertaining to the cause of the edema
formation. Is the lung fluid due to overzealous fluid therapy or
heart failure? Is it due to a decrease in colloid osmotic pressure
associated with the hypoalbuminemia? Or is it due to acute
respiratory distress syndrome caused by an increase in pulmonary
capillary permeability secondary to sepsis? Measuring the colloid
osmotic pressure (COP) of the plasma or serum can provide
valuable information when the most likely cause of edema
formation in critical animal patients is undetermined.
Causes of Edema Formation
Sixty per cent of canine body weight is water, which is
distributed as shown in Figure 1. The fluid compartments are
separated by membranes that are freely permeable to water but
not to solutes. However, the capillary membrane is different. It
is permeable to all the ions in the plasma except for the plasma
protein anions. 1 The distribution of fluids from the capillary
into the interstitial space depends on Starling's forces (Fig 1).
The water moves under the force of hydrostatic pressure and
colloid osmotic forces. The hydrostatic pressure within the
capillary is generated by the blood pressure (ie, cardiac output
and systemic vascular resistance). The osmotic forces are
From the Animal EmergencyCenter, Glendale, Wl.
No reprints available.
Copyright © 2000 by W.B. Saunders Company
1096-2867/00/1503-0003510.00/0
doi:10.1053/svms.2000.18297
Clinical Techniques in Small Animal Practice, Vol 15, No 3 (August), 2000: pp 119-125
determined by the charges and concentrations of impermeable
ions on each side of the membrane (Fig 2).
Cations and anions must be present in equal concentration
across the membrane to maintain electrochemical neutrality. In
the plasma, the proteins constitute an important fraction of the
plasma anions. Because proteins are impermeable and nega-
tively charged, they are responsible for the retention of water in
the capillary; this is called the Gibbs-Donnan effect (Fig 1).
This force opposes capillary hydrostatic pressure (which favors
movement out of a fluid compartment) and is called COP, or
oncotic pressure.
When capillary hydrostatic pressure is greater than capillary
oncotic pressure or when capillary pore size is significantly
increased, fluids move from the capillary into the interstitium
(Fig 3). Excess quantities of interstitial fluid are normally
removed from the interstitial space by the lymphatic vessels.
This fluid volume is returned to the circulation and is
eventually excreted by the kidneys. When the quantity of fluid
moving into the interstitium surpasses the transport capabili-
ties of the lymphatic vessels, fluid accumulates in the intersti-
tium, causing edema. It is then necessary to determine if there
is a reduction in COP that has contributed to the pathophysiol-
ogy of the edema.
Clinical Consequences of Edema
Edema in the subcutaneous tissues usually causes only mild
clinical consequences and serves primarily as a diagnostic
indicator of fluid overload, COP imbalance, or increased
capillary permeability. Interstitial edema in the bowel promotes
mucosal fluid flow from the interstitial space into the intestinal
lumen. 2 This can result in vomiting or diarrhea. When
interstitial edema occurs in the lung, brain, or heart, the
function of that vital organ can be compromised, and the
consequences can be catastrophic.
It has been shown that when plasma COP is reduced, there is
an increased incidence of pulmonary edema and a decreased
rate of survival in humans) In a study of human head-trauma
patients, hypoalbuminemia contributed to gastrointestinal in-
tolerance of enteral feedings, and it was suggested that a low
COP plays an important role in this problem. 4 It is critical to
determine the contribution of decreased COP to the pathophysi-
ology of the edema formation process; the optimal goal is to
prevent edema formation from occurring. By assessing plasma
COP, it can be determined if colloid replacement is therapeuti-
cally necessary for achieving a proper fluid balance.
Monitoring clinical and laboratory parameters provides a
foundation for determining crystalloid and colloid fluid admin-
istration and preventing edema formation. Body weight, physi-
cal perfusion and hydration parameters, central venous pres-
sure, blood pressure, and total protein are commonly used to
guide the effectiveness of fluid therapy. As an adjunct to these
119
 4~ H20molecule
Colloid
molecule

Ii' ol Q- 0
Solute
molecule

Colloid osmotic
pressure

. ,D ~ ,,,, I~" h ~l' Hydrostatic

t / • pressure
. Na+\ - / 1
" ~" "~ | ] ~, Gibbs-Donnan
" i ~ '--'~"~, - ~ 0 ) ~" effec //N~a+
U Na+ -~'--~..-- "--_ j/

" O-Na:

O Na+ -. Na+ -,<

Intravascular Interstitial Intracellular Lymphatic

compartment compartment compartment compartment
8% TBW 25% TBW 66% TBW
25% ECF 75% ECF
Fig 1. Fluid dynamics: 66% of total body water (TBW) exists intracellularly, and 34% exists extracellulary. Of the extracellular
fluid (ECF), 75% exists in the interstitial compartment and 25% in the intravascular compartment. Fluid movement across the
capillary membrane is under the influence of Starling's forces:

v =/<(Pc - P~) - f (1% - I~i) - Q

where v = volume, k = filtration coefficient, P = hydrostatic pressure, c = capillary, i = interstitial fluid, f = membrane pore
size, IX = oncotic pressure, and Q = lymph flow. Within normal tissues, there is a constant dynamic flow of fluid across the
capillary membrane with net movement of fluid, colloid molecules, and solutes across the interstitium into the lymphatic
vessels, where it is returned to the cranial vena cava.

parameters, it can be beneficial to determine plasma COP
through colloid osmometry. This should be done during fluid
resuscitation of patients with compromised cardiovascular
states in which the hydrostatic pressure is altered, in patients
with hypoproteinemia in which the COP is altered, or in
patients with systemic inflammatory response syndrome dis-
eases in which capillary pore size is enlarged.
Colloid osmotic pressure can be estimated from calculating
the plasma total protein (TP) by refractometry using the
Landis-Pappenheimer equationS:
COP = 2.1(TP) + 0.16(TP z) + 0.009(TP 3)
This equation, however, becomes inaccurate with alterations in
plasma pH, alterations in ion binding capacity by the proteins,
and deviation of the albumin-to-globulin ratio from normal
limits. Also, a poor correlation exists between calculated and
measured COP in the critically ill, making direct measurement
the more accurate method of determining COE 6-8 Finally,
synthetic colloids are not reflected when the plasma total
protein is measured by refractometry, making the Landis-
Pappenheimer equation invalid for determining the COP in
animals receiving synthetic colloids. 9 However, the COP effect
of the administered synthetic colloid is determined by direct
measurement of COP. Therefore, direct measurement of COP
with colloid osmometry is the method of choice.
Colloid Osmometry
Human hospital laboratories commonly run tests for COP
levels. It is important to provide them with a set of normal dog
and cat samples to determine the normal range for each animal
on that machine. There are also a variety of osmometry

120 RUDLOFFAND KIRBY

 s,~,~ H~O molecule

~ Colloid
O molecule

Solute
O
molecule
O

A Colloid osmotic
~"" 0 Na+ pressure

lID
O
Hydrostatic
pressure
~.: .....N a +

13 4

1
.Dffa.
• :. N a +
o
O O Na+ --
i ~- r s "1"
" "

Na+ ,::
:~ .._..:;~i"
m

Fig 2. Example of osmotic forces. Water and solutes that are freely permeable across the capillary membrane will distribute
themselves in equal concentration across the capillary membrane, When nonpermeable colloids are placed into the capillary,
there exists a greater concentration of solute within the capillary. Because the concentration of water molecules is then higher
outside the capillary, more water molecules hit the capillary membrane and pass through into the capillary. The result is
osmosis: a net migration of water from the interstitium into the capillary. The pressure exerted by the presence of colloid
molecules within the capillary membrane opposes the movement of water from the capillary into the interstitium and is called
colloid osmotic pressure.

instruments available for purchase that are relatively simple to
operate and maintain.
Instruments
Direct assay of plasma COP is possible with several instru-
ments. Figure 4 shows the Wescor 4420 colloid osmometer
(Wescor Inc, Logan, UT). This instrument measures COP
when a 0.325-mL sample of heparinized whole blood, plasma,
or serum is injected into a test chamber. The test chamber is
separated by a semipermeable membrane from a reference
chamber filled with normal saline. The saline-filled reference
chamber mimics the Gibbs-Donnan effect of interstitial fluid.
Water migrates from the reference chamber into the test
chamber under the influence of osmolality changes, causing a
negative pressure gradient in the reference chamber. After
equilibration, the negative pressure gradient is measured by
the sensing diaphragm of a pressure transducer. As the
pressure changes, electrical impedance is altered and the
output signal is amplified, then converted to a readout in mm
Hg.
When necessary (as with small mammals, cats, or small
dogs), smaller sample volumes can be used for measurement.
Once the osmometer is calibrated to zero, 0.3 mL of normal
pooled plasma/serum is introduced, followed immediately by a
0.125-mL of sample plasma/whole blood. With this procedure,
the error due to contamination in the sample chamber is
reduced to an acceptable level.
Maintenance of the Wescor 4420 is relatively simple but is
required to ensure the accuracy of the results. Both chambers
are rinsed daily with normal saline. The instrument is cali-
brated daily with a test solution of albumin of a known COP

COLLOID OSMOMETRY 121

 :+,++ H~O molecule

D molecule

Solute
molecule

,-,,'~ . - ~ Colloidosmotic
,, pressure
Na+ " * " N ~ Hydrostatic
• o " pressure
7 • Na+
,1+
Na+ o
-- ., O ,.

.~=~ • Na+
B o "~* •
-+ --" o o . Na+ "•

+
a+ I

" "- Na+_

• O O O'-Y 0 D - - m :

.. -t- •

,,+ 0~- a 4- (t -- • "" N i

. Q.a+ Ha+ +.

la+ oNa+e _.
e

Intravascular Interstitial Intracellular Lymphatic

compartment compartment compartment compartment

Fig 3. Causes of increased interstitial water. The top figure depicts the result of isotonic crystalloid administration. When the
hydrostatic force of the additional fluid increases the force of capillary hydrostatic pressure over colloid osmotic pressure,
permeable solutes, sodium (Na+), and water are forced into the interstitium. In addition, when more crystalloid is added, there is
a decrease in the amount of colloid per unit volume which decreases the colloidal effect that also promotes water flow into the
interstitium. The additional movement of fluid will expand the interstitium and can cause edema if the lymphatic compartment
is overwhelmed. The bottom figure depicts the result of increased capillary pore size, or loss of capillary integrity (change in f).
Colloid molecules are distributed into the interstitium and exert an attraction force promoting sodium and water movement into
the interstitium. The additional movement of fluid will expand the interstitium and can cause edema if the lymphatic
compartment is overwhelmed.

(commercially available through the instrument manufac-
turer). The instrument is calibrated to zero with saline before
and after each use. All solutions must be free of air bubbles
before injection.
An instruction book and service manual are available for
detailed troubleshooting. The most common problem recog-
nized with use of the instrument is failure to reach a plateau
pressure. This may indicate an improper seal on the test
chamber or drying of the semipermeable membrane. It is
usually corrected by tightening the chamber or replacing the
membrane.
Other osmometers have been used to determine of COE The
IL 186 Weil Oncometer (Instrumentation Laboratory Inc,
Lexington, MA) operates under the same principles as Wescor's
4420 Colloid Osmometer. However, it differs significantly in
the test cell design. The reference compartment flow-through
capability, facility of membrane replacement, and fluid-to-
membrane-surface contact make the IL 186 more complicated
to maintain.
A needle-tTpe colloid osmometer consists of a sensing probe,
a manometer system, and a sample bath. 1° This system uses a
hollow fiber rather than a semipermeable membrane to filter
the sample. This removes any problems associated with
pressure leakage or membrane damage during installation. In
addition, continuous measurements can be made, However,
the needle system can be slightly altered with hydrostatic
pressure increases that can develop during flow by the sensing
probe. This system has not found favor with those performing
point-of-care diagnostics.
Normal Values
Normal canine plasma COP (samples collected with lyophi-
lized heparin) measured with the Wescor 4400 for the authors'
lab is 21 to 25 mm Hg, and normal feline plasma COP is 23 to
25 mm Hg. The COP values and their clinical significance are
listed in Table 1. Reported normal mean canine whole blood
COP values (samples collected with lyophilized heparin)

122 RUDLOFF AND KIRBY

Fig 4. T h e W e s c o r 4420
colloid osmometer.

" .... : -" . . ~. ' k. 2 & . ae ~. .e'r. ~e">:

. . . . . ,:~. >. " #':'2".. .
• z, ~..'-2. g" -:-': (-7"~- ~..,)" .. , ¢" '~ g~-~.~e>,?{}~-" C ~'o'-<I . . . . ~-

measured with the Wescor 4400 fall around 19.95 __ 2.1 mm standardized. Samples should only be compared with the
Hg, and feline whole blood COP values fall around 24.7 __ 3.7 reference values for the type of sample collected. During
mm Hg. 11 Compared with whole blood, serum and plasma point-of-care monitoring, whole blood may be the more
COP values were higher in this study. Additional canine studies efficient way to sample. When samples cannot be immediately
reported other resuhs, 1°,12,13but they generally were within the run, plasma or serum can be frozen and thawed with minimal
same range. effect on COE 14
For more accurate comparisons, normal values for COP Circumstances not related to protein concentration that can
should be established by the reference laboratory performing increase COP measurements include excessive hemolysis and
the measurement. Samples from clinically normal dogs and synthetic colloid administration. Falsely low COP readings can
cats are used to determine the reference range, and collection
result when liquid anticoagulants (eg, ethylenediamine tetra-
and sample type (either whole blood, plasma, or serum) is
acetic acid, citrate, or liquid heparin) are mixed with the blood
sample, producing a dilutional effect. Dry anticoagulant (lyophi-
lized heparin, commonly found in green-top blood collection
T A B L E 1. M e a s u r e d P l a s m a C O P V a l u e s a n d T h e i r Clinical tubes) produces a clinically insignificant change in COP in the
Significance
sample. 1<15 This is due to its relatively high molecular weight
COP Result Clinical and low concentration.
(mm Hg) Significance
Physiologically, severe acidemia and alkalemia and electro-
>25 Pathologic hyperproteinemia or significant hemoconcen- lyte changes alter the net negative charge on the plasma ion
tration
18-25 Normal values in dogs and cats
and affect the Gibbs-Donnan effect on measured COE When
14-18 End-point goal of colloid resuscitation; edema not likely the blood pH decreases or the sodium concentration decreases
caused by reduced COP significantly, the measured COP is lowered, and vice versa.
11-14 At risk for edema formation; colloid therapy beneficial
<11 Edema likely due to decreased COP; colloid therapy When alterations in sodium concentration exist, a reference
highly recommended sample having the same sodium concentration as the sample is
NOTE. Clinical significance based on author's experience with the
used. This can be made by mixing normal saline with
Wescor 4400 device. hypertonic saline or water to a concentration that approxi-

COLLOID OSMOMETRY 123

mates that of the sample. Should a clinician believe this is a
common consideration in their practice, he or she may elect to
make stock reference solutions of varying sodium concentra-
tions for ease of application during the sampling process.
Osmolality changes caused by solutes other than sodium (eg,
abnormal amounts of glucose, urea, or mannitol) do not cause
an error in osmometry.
In humans, it is also reported that upright patients have an
approximately 15% higher COR compared with patients in a
prone position. 16 In the adult human, normal upright COP is
22 to 29 m m Hg, arid supine COP is 17 to 24 m m Hg. In the
supine position there is a mobilization of protein-free fluid into
the central circulation. This process can take up to 4 hours. It is
not known at this time if postural changes cause a significant
change in COP values in animals.
Clinical Application of Colloid Osmometry
Colloid osmometry is useful when diagnosing the origin of
edema, preventing edema during aggressive fluid therapy, and
administering synthetic colloids to animals with increased
capillary pore size from systemic inflammatory response syn-
drome diseases. When used in combination with other physi-
cal and monitored parameters, COP values can aid in determin-
ing the end point of fluid resuscitation, guide fluid selection,
and direct other therapeutic modalities. The following cases
will illustrate these clinical applications. (See Table 1 for COP
values and their significance).
Increased Capillary Permeability
Amber the 6-year-old, 30-kg spayed female golden retriever
mentioned at the beginning of this article, was being treated for
peritonitis due to a ruptured pyometra. She had an open
abdomen, and she had been resuscitated and maintained with a
combination of isotonic crystalloid, the synthetic colloid
hetastarch, and plasma transfusions. On day 3 of hospitaliza-
tion, she became hypoxemic. Auscultation revealed moist lung
sounds throughout her lung field. She had an albumin of 2.64
g/dL (normal range, 3.5 to 4.0 g/dL). A thoracic radiograph
showed a diffuse interstitial and alveolar lung pattern. The
heart and the vena cava size were within normal limits. The
blood pressure was normal and central venous pressure was
5.0 cm H20 (normal range, - 1 to 5 cm H20; above 10 cm H20
suggestive of vascular volume overload, increased intratho-
racic pressure, or right heart failure). The COP was 16 m m Hg
(see Table 1). An echocardiogram showed a mild decrease in
contractility but no evidence of heart failure.
It was important to determine the cause of the lung edema.
First, an increase in hydrostatic pressure from volume overload
or heart failure needed to be ruled out. The central venous
pressure and blood pressure were within normal limits for
resuscitation of an animal with systemic inflammatory re-
sponse syndrome (SIRS) from sepsis, making volume overload
unlikely. Next, because the dog had hypoalbuminemia and had
been on synthetic colloids, it was important to determine if the
colloid therapy had been adequate to maintain the COP. The
COP of 16 m m Hg should have provided an adequate intravas-
cular COP to maintain the fluid within the blood vessel as long
as the hydrostatic pressure was not elevated. By elimination,
the most likely cause was an increase in capillary pore size
associated with SIRS. This was supported by finding an
albumin level of 2.84 g/dL in the lung fluid aspirated from the
124
trachea. The dog required 5 days of ventilator therapy for
stabilization.
Ashley was a 12-year-old, l l-kg sheltie dog that presented
with a thoracic mass. The mass was removed, and a histopatho-
logic diagnosis of round cell sarcoma was made. Ashley
developed evidence of SIRS and disseminated intravascular
coagulation. She had an albumin level of 1.79 g/dL in spite of
plasma administration. She had been resuscitated and main-
tained with a combination of isotonic crystafloids and heta-
starch. Despite what was believed to be aggressive colloid
replacement, Ashley developed peripheral edema of the distal
forelimbs. An echocardiogram did not show evidence of
myocardial dysfunction. The central venous pressure was 8.4
cm H20, and systolic arterial blood pressure was 110 m m Hg.
The COP was 16.2 m m Hg.
The normal echocardiogram, central venous pressure, and
arterial blood pressure readings made an increase in hydro-
static pressure due to volume overload or heart failure unlikely.
The normal COP indicated that we were successful in our
colloid replacement in spite of the low albumin level. The limb
edema was probably due to an increased hydrostatic pressure
(approximated by measuring the central venous pressure) as
well as an increase in capillary permeability due to the SIRS.
Decrease in COP
Marley was a 6-year-old, 25-kg, mixed-breed dog that pre-
sented with an intestinal foreign body and infectious peritoni-
tis. An intestinal resection and open abdominal drainage were
required. Marley developed SIRS and had an albumin level of
I. 17 g/dL despite large volume plasma administration. He was
fluid resuscitated and maintained with a combination of
isotonic crystalloids and hetastarch. Despite what was believed
to be aggressive colloid replacement, Marley developed periph-
eral edema of the distal rear limbs. The central venous pressure
was 5.0 cm H20, heart rate was 120 bpm, and blood pressure
was 135/82 m m Hg. The COP was 12.8 m m Hg.
The normal central venous pressure, heart rate, and blood
pressure made an increase in hydrostatic pressure due to
volume overload or heart failure unlikely. The low COP
indicated that we needed to intensify our colloid replacement
to prevent worsening of edema. There was probably an element
of increased capillary permeability due to the SIRS that
accounted for our difficulty maintaining COP in the face of
plasma and hetastarch administration.
Preventing Edema During Aggressive Fluid Resuscitation
Max was a 5-year-old, 30-kg, mixed-breed dog that presented
with an intestinal linear foreign body and hypotension. The
heart rate was 138 bpm, and the blood pressure was too low to
detect by Doppler on the limb. The prefluid plasma COP was
22.9 m m Hg, a result of increased total protein and significant
intravascular fluid deficit. Resuscitation was accomplished
with a combination of isotonic crystalloids (600 mL) and
hetastarch (500 mL). This combination brought the heart rate
down arid the blood pressure up, while only slightly decreasing
the COP to 19.4 m m Hg. Continued resuscitation with 1400
mL of crystalloid alone diluted the impermeable ions and
dropped the COP to 12.9 m m Hg. This left the dog at risk for
developing edema. Hetastarch was added to the maintenance
fluid therapy plan to restore the COP.
RUDLOFF AND KIRBY
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