Respiration Physiology (1983) 51,167-181 167

Elsevier Biomedical Press

QUANTITATIVE DESCRIPTION OF WHOLE BLOOD

CO~ D I S S O C I A T I O N C U R V E A N D H A L D A N E E F F E C T

J . A . L O E P P K Y , U . C . L U F T a n d E. R. F L E T C H E R

Research Division and Biodynamics Laboratory, Lovelace Medical Foundation, Albuquerque, N M 87108,
U.S.A.

Abstract. A simple procedure is presented to describe accurately the whole blood CO2 dissociation
curve on linear content (Cco2) and pressure (Pco2) coordinates with an exponential equation (Cco2 ---
K. Pco2b). A single coordinate and the hemoglobin concentration, Hb, are required. Whole blood
Cco 2 can be calculated from values for pH, Pco2, Hb and 02 saturation by empirically accurate
equations. The mathematical description of the CO2 curve was employed to quantitate the in vivo
Haldane factor (fH) from simultaneous arterial and mixed venous blood samples in 20 healthy exercising
subjects. The mean + SE was 0.28 + 0.03 (vol. % ACco/vol. % AHbO2). In 20 patients with severe
obstructive lung disease fH was 0.29 + 0.08 when calculated from arterial samples while breathing air
and 100% 02. Values for fH were not related significantly to acid-base status or Hb as suggested
by previous workers. By assuming these or other values for fH, the in vivo change in blood Pco2
resulting from a given change in oxygenation can be predicted.

Arterial CO2 curve Mixed venous CO2 curve

CO2 curve equation Obstructive lung disease
Exercise Oxygenated CO2 curve
Haldane factor

B l o o d a n d p u l m o n a r y gas exchange studies f r e q u e n t l y r e q u i r e a precise m a t h e -

m a t i c a l o r g r a p h i c a l d e s c r i p t i o n o f the shape a n d p o s i t i o n o f the CO2 d i s s o c i a t i o n
curve. T h e m a g n i t u d e o f the c h a n g e in Pco2 o r CO2 c o n t e n t w h e n h e m o g l o b i n is
o x y g e n a t e d o r reduced, k n o w n as the C - D - H o r H a l d a n e effect ( C h r i s t i a n s e n et al.,
1914), is also m o r e easily visualized with the help o f such curves. These precise
d e s c r i p t i o n s a r e p a r a m o u n t in r e b r e a t h i n g studies w h e r e 02 a n d CO2 e x c h a n g e a r e
a l t e r e d i n d e p e n d e n t l y a n d the b l o o d - t o - g a s CO2 g r a d i e n t s a r e often o f p r i m a r y
interest. C l e a r l y defined curves a r e also helpful for in vitro b l o o d studies d e a l i n g
with the p r o p e r t i e s o f gas exchange a n d r e a c t i o n rates o f h e m o g l o b i n w i t h 02
a n d CO2.

Accepted for publication 10 November 1982

0034-5687/83/0000-0000/$03.00 1983 Elsevier Biomedical Press

168 J.A. LOEPPKYet al.

Techniques to determine cardiac output, such as the CO2 rebreathing procedure

of Farhi et al. (1976) and the single breath method of Kim et al. (1966), ideally
require the CO2 dissociation curve for each subject under the conditions (e.g., work
load) that pertain during the breathing maneuver. The ultimate values of cardiac
output calculated from these procedures are extremely sensitive to the slope and
curvilinear characteristics of the dissociation curve. In studies involving these tech-
niques (Luft, 1973) and in others related to blood-to-gas gradients (Luft et al., 1981),
we have developed a relatively simple and useful mathematical description of the
CO2 dissociation curve of whole blood. The curve's description and assumptions
are based primarily on the classical biochemical studies of blood which took place
some 50 to 60 years ago and are well summarized by Henderson (1928) and Peters
and Van Slyke (1931). This presentation serves to consolidate some aspects of the
empirical nomograms that were derived in that era and offers some calculations
and deductions pertaining to the Haldane effect. Our characterization of the CO2
curve is simpler to calculate or program than some currently in vogue (Kelman,
1967; Olszowka and Farhi, 1968; Thomas, 1972) and does not require the use of
nomograms. It is empirically accurate in its derivation and application. We trust
that this simplified approach will be of interest to other laboratories in similar or
new applications dealing with CO2 transport.

Derivation of equation for the CO2 dissociation curve

Peters and coworkers (Peters, 1923; Peters et al., 1924a,b) were apparently the
first to note that the CO2 dissociation curve of whole blood was linear when plotted
on logarithmic coordinates. This can readily be verified in the physiological range
(Pco~ values from 20 to 80 mm Hg) by plotting the values for Pco~ and CO2 content
(Cco2) of Dittmer and Grebe (1958), compiled from numerous sources, and those
of Henderson et al. (1924). Thus, the first assumption is established:

ln~(Cco~) = a + b. llh(Pco2) (1)

where Cco~ and Pco~ are expressed in vol. ~o and mm Hg, respectively.
It is also necessary to know the coordinates for a single point on the curve
(C~o~ and P~o~) so as to establish the position or height of the curve (Peters et al.,
1924b). Equation 1 can be written in terms of P~o2 and C~o~ and subtracted from
the general form of eq. 1 giving
ln~(CcoJC~o~) = b. lne(Pco2/P~o:) (2)
alternatively, eq. 2 can be written as
Cco~ = K . Pco2b (3)
where
K = C~o,/P~o~ ~ (4)
 CO 2 DISSOCIATION CURVE AND HALDANE EFFECT 169

Peters et al. (1924a,b) clearly demonstrated that the slope of the curve is linearly
related to the hemoglobin concentration (Hb) or 02 capacity (O2cap). The following
equation was presented (Peters et al., 1924b) which related the Cco2 difference (A)
between Pco2 values of 60 and 30 mm Hg to O2cap:

ACco2(6(~30) = 0.334 (O2cap) + 6.3 (5)

where O2cap is expressed in vol. ~ . Equation 5 can be converted to be a function
of Hb (eq. 6), knowing that 1.36 ml of 02 combine with 1.0 g of hemoglobin
(Lenfant, 1974) :

ACco2(60_30 ) = 0.4542 (Hb) + 6.3 (6)

where Hb is expressed in g ~ . . For example, this means that if Hb is 15.0 g ~ ,
the curve will pass through the points at Pco2 of 30 and 60 mm Hg to result in a
Cco2 difference of 13.113 vol. ~ . Although it is possible (knowing Hb, C~o2 and P~o2)
to construct the curve on logarithmic paper (Peters et al., 1924b), it is more
expedient and precise to arrive at an exponential equation of the form shown in eq. 3.
Using eqs. 3 and 4 to determine Cco~ at Pco2 values of 60 and 30 mm Hg and
substituting in eq. 6 gives
C~o2(60/P~o) b - C~o~(30/P~o~)b = 0.4542 (Hb) + 6.3 (7)
Equation 7 cannot be solved algebraically for b; however, solutions can be obtained
by iteration using the following equation:
(60/P~o) b - (30/P~o) b - (0.4542 (Hb) + 6.3)/C~o~ -- Z (8)
and systematically substituting numerical values for b until Z is equal to zero and
thus eqs. 7 and 8 are equivalent. It is apparent from eq. 3 and the constraints of
the Pco: versus Cco2 relationship that b must always lie between values of zero
and 1.0. An example follows for the solution of b. Assuming that fully oxygenated
blood has P~o2 = 40 mm Hg, Cbo: = 50 vol. ~ and Hb = 15.0 g ~ , eq. 8 becomes
1.5b - 0.75 b - 0.26226 = Z (9)
Zero and 1.0 can be taken as initial lower (bL) and upper (bu) estimates of b. By
substituting bL and bu into eq. 9 we obtain ZL = --0.26226 and Zu = 0.48774. In
order to arrive systematically at a value for b the following equation is used:

b' - bLZu -- buZL (10)

ZU - ZL

This computed value for b' is 0.34968 and is substituted in eq. 9 and the computed Z'
is then -0.01423. Because Z' is less than zero, b' and Z' became the new values
for bL and ZL in eq. 10 and the cycle is repeated, retaining the original values for
bu and Zu. If Z' is greater than zero, b' and Z ~ become the new values for bu
and Zu, and the original values for bL a n d ZL are retained. Sufficient accuracy is
achieved by repeating this cycle until I Z'[ is less than 0.00005. In this specific
170 J.A. LOEPPKY et al.

example, onlythree trial-and-error cycles are required, and the final value for b
is 0.3692 (rounded). The other coefficient, K, is then obtained from eq. 4.
The coefficients for eq. 3 have now been determined and this whole blood curve
is defined as follows:
Cco2 = 12.8105 Pco20'3692 (11)

and is shown as curve B in fig, 1. As further examples, two other curves with
equations are shown in fig. 1 having Hb values of 20.0 (curve A) and 10.0 g~o
(curve C) and passing through the same point. Curve D was computed for blood
having Hb = 14.7 g ~ , P~o: = 40 mm Hg and Cbo: = 54.3 vol. ~ . This was a single
point value noted by Henderson et al. (1924) for reduced blood. The other 8 points
shown were also directly determined from their blood after equilibration. It is
apparent that the mathematical curve from the single point is an excellent approxi-
mation of the measured points between Pco2 values of 20 and 80 mm Hg. The mean
absolute difference between estimated and measured values for these 7 points is
only 0.27 vol. ~ .
Although eq. 11 can be obtained with simple hand calculators, it is much more
practical to utilize a programmable calculator with decision-making (Z ~ 0 ?) capa-
bilities. We have used the Hewlett-Packard model 97 desk calculator for these
procedures where the above operations can be carried out in about 90 programmed
steps. The previous iteration technique will not always arrive at appropriate values
for b when using extreme values of Hb, P~o: or Cbo~ which are well beyond the

80-- Hb(9/o)
.14.7 (D)
j o - 2;-2Q0 (A)
~o...I"7" .. ~5.0(B)
60, C c 6 2 ( 5 4 . 3 ) ~ ~ : . . - ~ o (c)

~"// B Y= 12.8105 Xo3692

~/// C Y =16.1357x 0"3066

O" ,

Pco2(mm Hg)

Fig. ]. CO 2 dissociation curves defined by simple exponential equations as derived from H b and a
single coordinate as described in text. Curves A, B and C show effect of Hb. Curve D is for reduced
blood from one measured coordinate reported by Henderson et al. (1924) with other measured
coordinates (circles) also shown.
 COz DISSOCIATIONCURVEAND HALDANEEFFECT 171

ranges reported here. The following tests will determine when solutions for b are
possible. A solution for b will always be found when Z is positive for b = 1.0
from eq. 8 ( ~ = ,). If Zb = l is negative and P~o~ < 120 mm Hg, no solution exists.
If Zb = ~ is negative and Pco2 > 120 mm Hg, there may be none or two solutions.
This decision is made by computing a b value from the following equation:

bp = lne[lne(30/P~2)/ln~,(60/P~)] (12)
lne2
A Z value is computed from eq. 8 utilizing bp (Z b= bp)" There is no solution when
Z b = bp is negative and two solutions when it is positive. If there are two solutions,

one value for b can be solved by substituting bL = 0 and bu = bp and corresponding

Z values in eq. 10. The other solution is found using bL = bp and bu = 1.0 and
appropriate Z values. Additional information is required (e.g., another coordinate)
to determine which value for b is most appropriate.

The estimation of the single point (P~o2, C~o~)

The analysis of whole blood C O 2 content, as was customary by the method of

Van Slyke and Neill (1924), is today no longer a routine measurement. It is now
more convenient to calculate CO2 content of plasma (COEp) from pH and Pco2 of
the blood sample via the Henderson-Hasselbalch equation:
CO2p = 0.0301 Pco2(1 + 10pn-6I) (13)
where CO2p, expressed in mEq/L, constitutes HCO3 plus the dissolved CO2 in
the plasma. It is then necessary to convert CO2P to Cco~ of whole blood. This
conversion is dependent on the hematocrit, pH and 02 saturation (So) in a complex
fashion (Van Slyke et al., 1923), but Van Slyke and Sendroy (1928) devised an
empirical nomogram which facilitated this conversion knowing these variables.
A subsequent nomogram was later constructed which required Pco: instead of pH
(Van Slyke et al., 1932). Fortunately, Visser (1961) has converted the former
nomogram into an empirical equation which was subsequently modified by McHardy
(1967) and is shown below in a form utilizing Hb instead of O2cap to calculate
whole blood Cco: :
[ 0.02924(Hb) ]
Cco2 = 1.0 (2.244 -0.422 So)(8.740 - p H ) CO2p-2.226 (14)

where So2 is expressed as a fraction, Hb in g%, COzp in mEq/L and Cco~ in vol.%.
In order to test the validity of Cco: as predicted by pH, Pco:, Hb and So_. from
eqs. 13 and 14, we compared the latter with values obtained manometrically
(Van Slyke and Neill, 1924) from whole blood obtained during an exercise study
at a barometric pressure of 630 mm Hg (Luft, 1973). In that study, 20 arterial and
20 mixed venous samples were meticulously analyzed for pH, Hb, and 02 and CO2
172 J.A. LOEPPKY et al.

55-
- - Identity Line J/
~
--- Regression Line .,"~/F..

~,= 43.12 ~ "

45 S.D.=5.58 ../" .

(Cm~ed) 3~ Y'=D'47";'7~/ ""

f n=40
,~" " -y-O.61+1.01x
,~/" r= *0.99
" ~EJE.=1.1vol.%

25
25 3~5 4'5 ~
CcD2(Van Slyke)

Fig. 2. Comparison of 20 arterial and 20 mixed venous whole blood CO2 contents (Cco2) measured
manometrically (Van Slyke) and computed by eqs. 13 and 14 from pH, Pco2, Hb and So2 values given in
table 1.

contents and pressures in 5 men during rest and steady state exerdse up to a X[o:
of 2.3 L/min (table 1). The comparison of measured and computed Cco: is shown
in fig. 2. The mean values differed by less than 0.1 vol. ~ and the regression line
is almost indistinguishable from the line of identity. The scatter in the two methods
was nearly identical. This comparison adequately justifies the prediction of Cco2
by eqs. 13 and 14 and allows one to compute the dissociation curve as previously
outlined without a direct determination of whole blood Cco2.

Determination of effect of in vivo oxygenation or reduction of hemoglobin on the

CO 2 dissociation curve

If the values for So2 and Hb are known for the blood sample from which Pbo2 and
C~o2 are obtained, then it is possible to predict changes in Pco2 or Cco~ that will
result for a known change in Sos from the Haldane factor (frO. The value for fH is
best expressed as the change in CO2 content in vol. ~ per vol. ~ change in Oz
content (ACcoJAHbOE), i.e., Cco2 will increase when HbO2 is reduced and will
fall when HbO2 increases at a constant Pco2-
The standard COE curves under consideration here (fig. 1) were by necessity
obtained by in vitro equilibrations in the older studies. Theoretical considerations
of the differences between in vivo and in vitro dissociation curves indicate that the
Haldane factor is less in circulating blood, but during respiratory exchange in the
lungs or tissues the in vitro conditions prevail (Michel, 1968). With this in mind
 CO2 D I S S O C I A T I O N CURVE A N D H A L D A N E E F F E C T 173

we have analyzed the 02 and COz pressures and contents of whole blood obtained
directly in an earlier study mentioned above in which single breath and direct
Fick cardiac outputs were compared (Luft, 1973). With the graphical considerations
suggested by other authors (Mithoefer, 1959; Kim et al., 1966) and procedures
outlined above we have computed in vivo values for f. for five human subjects
during rest and exercise whose CO2 contents were shown in fig. 2. All variables
necessary to compute fH are presented in table 1. The methods employed to obtain
fH are depicted in fig. 3 with an example for subject 4 (table 1) working at 1.53 L/min.
His Hb at this work load was 18.3 g ~ , with a mixed venous (point V) Pco2 and Cco2
(computed) of 43.6 mm Hg and 48.38 vol. ~ , respectively. Corresponding values for
arterial blood (point a) were 18.3 g~o, 31.8 mm Hg and 39.39 vol. ~ . The equations
for the two CO2 curves (A and B) satisfying these conditions were obtained as
described previously and are shown in fig. 3. The vertical displacement of the two
curves must be entirely due to the Haldane effect and fH was computed at PVco2
and Paco~. In this case ACco: at PVco~ is 2.76 (V - V l ) , obtained by solving curve B
for .Cco: at a Pco: of 43.6 mm Hg and subtracting from Cco~ at v. Since AHbO2
(02 content minus dissolved 02) was 10.68 v o l . ~ , f. is 2.76/10.68 -- 0.258. At the
arterial end Cco~ at al is computed from curve A at 31.8 mm Hg and Cco~ at a is
subtracted, giving a ACco2 of 2.73 and fn ----0.256. The value for fH will be slightly
lower at lower Pco~ levels because the curves ultimately converge when Pco~ is zero.
The greatest difference noted in arterial and mixed venous fn values was 0.007.

45- J"~'~' 1
Cco2 Gc..,O~o # ' c ~ ~ '

40- ' 7 ~ ~ ~ 1if'" -

a - x : 1.61 mm Hg
I V l - X l : 1"85 rnm Hg
I
I
I

%o2(mrnHgl
Fig. 3. CO2 dissociation curves for subject 4 exercising at 1.53 L/rain (table 1). Equations are shown
for curves passing through arterial (a) and mixed venous (V) points. The difference in Cco 2 between
curves A and B divided by AHbO 2 (10.68) is the Haldane factor (0.26) which varies slighdy with Pco:.
Curve C is computed for this blood if it were fully oxygenated and horizontal distances a - x and
V1 - x I show the increases in 1;co z at a and V that would result if So2 rose from 0.86 to 1.0 and Cco 2
did not change.
174 J.A. LOEPPKY et al.

TABLE 1
Simultaneous mixed venous (V) and arterial (a) blood variables utilized to compute the Haldane
factor (fH) in 5 human subjects during supine rest and submaximal steady-state ergometer exercise
(data from Luft, 1973)

Subject Vo2 B.E. Hb So2 AHbO z

(L/min) (mEq/L) (g ~ ) (vol. ~o)
a

1 0.31 + 1.0 16.5 0.930 0.738 4.31

0.96 -0.7 18.1 0.849 0.494 8.74
1.54 -4.4 18.6 0.853 0.389 11.74
1.95 -6.1 19.1 0.840 0.335 13.12
2 0.33 + 1.1 16.1 0.979 0.806 3.79
0.86 -1.0 17.5 0.893 0.569 7.71
1.44 -2.7 18.2 0.889 0.440 11.11
3 0.26 + 1.9 15.6 0.949 0.735 4.53
0.75 + 1.0 16.2 0.881 0.551 7.27
1.40 + 0.9 16.5 0.870 0.467 9.04
1.97 - 1.5 16.9 0.881 0.433 10.30
2.32 -2.7 16.9 0.889 0.366 12.02
4 0.33 + 1.4 16.8 0.933 0.714 5.00
0.93 + 0.5 17.9 0.867 0.576 7.08
1.53 -0.9 18.3 0.858 0.429 10.68
1.79 - 1.2 17.8 0.903 0.407 12.01
5 0.28 +0.7 16.1 0.883 0.737 3.20
0.88 -0.1 16.4 0.888 0.519 8.23
1.55 -3.8 17.1 0.899 0.395 11.72
2.12 -8.4 18.0 0.899 0.317 14.25
17.2 0.892 0.521 8.79

Base excess (B.E.): computed for oxygenated blood by equation of Siggaard-Andersen (1974), p.51.
Mean of a and V (no significant difference). Hemoglobin (Hb): from O2ca p (determined manometrically
after equilibration of arterial blood to Po2 = 122 mm Hg) divided by 1.36 and confirmed by oximeter.
02 saturation (So2): from 02 content (determined manometrically) minus dissolved 02 and divided by

These computations were repeated in identical fashion for the other 19 cases. The
two values for fR (arterial and mixed venous, using computed Cco2) were averaged
and the values for each subject at rest and during exercise are shown in table 1.
The mean value for fH was 0.28 (SE = 0.03), indicating that there is a 9 5 ~
probability that the true population mean lies between 0.22 and 0.34 in 95~o of
cases. The fH values in table 1 range from 0.10 to 0.64. Undoubtedly, some or all
of this variation is due to random errors in the measurement of variables utilized
to compute Cco2 by eqs. 13 and 14. With the mean values from table 1 and assuming
no errors in the arterial value for Cco2, it can be shown by calculations that a 1.0~
overestimation assumed individually for mixed venous So2, Hb, Pco2 and p H will
result in respective errors in fH o f - 1 , + 1, + 19 and" + 3 0 0 ~ . Even a 0 . 1 ~
overestimation in mixed venous pH ( + 0.007 unit) will overestimate fH by 2 8 ~ . In
 CO 2 DISSOCIATION CURVE AND HALDANE EFFECT 175

pH Pco2 ( m m Hg) C c o 2 (vol. %) frt f~l

a V a V a v

7.510 7.493 28.6 31.4 40.33 43.27 0.31 0.15

7.471 7.421 31.0 38.6 39.63 45.56 0.20 0.25
7.433 7.351 27.3 42.1 32.11 43.10 0.24 0.25
7.411 7.321 26.4 43.0 29.52 41.30 0.19 0.29
7.500 7.469 29.8 33.8 41.34 44.49 0.26 0.07
7.455 7.415 31.7 38.6 39.51 45.21 0.26 0.29
~7.432 7.352 31.1 45.2 36.64 46.49 0.22 0.27
7.474 7.435 34.8 40.0 46.26 49.68 0.19 0.03
7.471 7.413 34.0 42.3 44.68 50.26 0.20 0.13
7.447 7.383 36.1 48.3 45.02 53.98 0.36 0.17
7.436 7.351 32.6 48.7 39.51 50.82 0.34 0.20
7.437 7.345 30.2 47.1 36.65 48.69 0.30 0.26
7.485 7.450 31.1 38.7 41.51 48.76 0.64 - 0.01
7.467 7.421 33.9 39.8 43.08 46.91 0.10 0.35
7.457 7.390 31.8 43.6 39.39 48.38 0.26 0.29
7.469 7.382 30.0 43.6 38.25 47.96 0.21 0.30
7.473 7.462 32.3 34.8 42.68 45.32 0.41 0.24
7.442 7.403 33.5 43.9 41.38 51.05 0.55 0.15
7.394 7.308 33.7 49.6 37.39 47.40 0.20 0.30
7.366 7.245 26.5 49.5 27.44 41.35 0.14 0.24
7.452 7.391 31.3 42.1 39.12 47.00 0.28 0.21

02 cap. AHbO2: computed as (Sao2-SVo2 ) . H b - 1 . 3 6 . pH and Pco:: from Corning (model 16)
blood-gas analyzer. Cco2: computed for whole blood from eqs. 13 and I4 (see text) and confirmed
manometrically (fig. 2). fh: calculated similar to fH, but using Cco 2 measured manometrically.

order to determine whether any of the variation in fH could be accounted for by

the variations in Vo2, base excess, Hb, pH or Pco~ we computed correlation coeffi-
cients by linear regression for each with fn. Arterial and mixed venous values for
pH, Pco2 and base excess were averaged. The mean difference between correspond-
ing arterial and mixed venous values in base excess was + 0.2 mEq/L (arterial minus
mixed venous). The differences ranged from - 1.2 to + 1.2 mEq/L. These differences
were directly and significantly related to fn values because HCO3 is the main
determinant of calculated base excess and Cco2 and the latter determined the
calculated values for fn. The r value closest to statistical significance was - 0 . 4 2
with Hb (0.05 < P < 0.10); however, Hb would have been expected to correlate
positively according to von Mengden et al. (1969).
Also included in table 1 are the fn values computed as above but with the Cco2
176 J.A. LOEPPKY et al.

values determined manometrically (fH). The mean in this case was 0.21 (SE = 0.02).
This mean is not statistically lower than the 0.28 fH value. The relatively low
(although statistically significant) correlation between fH and f~ (r = -0.54) indi-
cates that the difference is due to the small random differences in Cco2 determi-
nations by both techniques (fig. 2). It is the venous - arterial Cco2 difference which
directly determines fH and f'H. The 'true' values for this difference should be the
arteriovenous 02 content difference x respiratory exchange ratio, if these are
perfectly measured, since cardiac output determines both differences. This mean
was 8.01 vol. ~ , that from the Cco2 calculated by eq. 14 was 7.88 and the mean
from Van Slyke determination of Cco~ was 7.64. Since the latter is farther from the
'true' value, it would suggest that fH is a less accurate estimate of the Haldane
factor than fH" The mean f~ was lower than f. because the Van Slyke values for
Cco2 were 0.19 vol. ~o higher on the average for arterial samples and 0.05 vol. ~o lower
for venous samples than Cco2 calculated by eqs. 13 and 14. However, the mean
difference was insignificant (fig. 2). The differences in fH and fH happen to be
consistent in all subjects at rest and give the impression that f~ correlates directly
with Vo2. However, if resting values are omitted, then this correlation is again
statistically insignificant as it was with fH. This would not occur if there was a true
relationship because V'o~ shows a greater change from the first to the last level of
exercise (table 1) than it does from rest to the first level. We conclude that the
variability in our fH and f{x values, and the difference between the two, is caused by
small random errors in the determination of Cco, and this variability cannot be
attributed to any systematic physiological influences resulting from exercise.
An opportunity to compare our mean value of 0.28 for fH obtained from the
exercise study on healthy subjects (table 1) with that of individuals with impairment
of pulmonary gas exchange was available from a previous study (Luft et al., 1981)
on 20 severely hypoxic patients with chronic obstructive lung disease. Comprehensive
gas exchange and acid-base analyses on these patients while breathing air (mean
So: = 0.73) and then 100~ 02 resulted in two C O 2 coordinates for which CO2
curves were calculated. The fH values were computed similarly to those in the
exercise study. The mean value for fH in these 20 patients was 0.29, essentially
identical to the exercise study, although the variation around this mean was
considerably greater (SE = 0.08) with a 95~ confidence interval between 0.13 and
0.45. Manometric measurements of Cco: were not available in these patients. Some
of the additional variations in the patients could have arisen from the fact that
the reduced and oxygenated points on the CO2 coordinates were not simultaneously
obtained, as were the mixed venous and arterial points in the exercise study, so
that changes in Cco~ or Pco~ may not have completely compensated for alterations
in ventilation, ~'/t) distribution or pulmonary shunting induced by 100~o 02. Also,
AHbO2 was considerably smaller (5.7 vol. ~ ) and this would tend to magnify varia-
tions in fH due to random errors of the measurements required to compute Cco:.
We correlated individual fH values with the same variables as in the exercise study
and again noted no statistical significance. The highest correlation was again with
Hb (+ 0.41), but opposite in sign to that noted in the exercise study.
 CO2 DISSOCIATION CURVE AND HALDANE EFFECT 177

There is some disagreement in values reported for f. and about whether or not
it varies significantly under physiological conditions which alter pH, Pco~ or Hb.
In the original description of this effect on the blood of J. S. Haldane (Christiansen
et al., 1914) the value for fH was not precisely determined. However, it was clearly
demonstrated that ACco2 increased linearly in relation to the reduction in HbO2
and not Po2. The statement was made that when oxygenated blood was completely
reduced, "At the physiologically important part of the curve, between 35 and 60 mm
pressure of CO2, the extra amount of CO2 taken up is about 5 to 6 vol. per 100 vol. of
blood." For blood having an Hb of 15.0 g ~ this translates to a value for fn some-
where between 0.25 and 0.29. However, these observations were made from
generalized curves and their tabled values (n = 13) give a mean of 0.40, ranging
from 0.06 to 0.62 with a trend toward higher values at greater Pco2. A more
systematic analysis of the CO2 curve and the effect of reduction of HbO2 was
performed some 10 years later on the blood of A.V. Bock by Henderson and
associates (Henderson et al., 1924; Bock et al., 1924). They reported an increase
of 6.3 vol. ~ in Cco~ for blood having an O2cap of 20 vol. ~ when it was taken from
the oxygenated to the reduced state (f. = 0.32). However, actual data points pre-
sented gave a mean f. of 0.30 for values of Pco2 between 3 and 80 mm Hg, with 0.28
at the lowest Pco~, a peak of 0.32 at 50 mm Hg and then a decline at higher Pco~
values (Henderson et aL, 1924). This diminution of f. at high and low Pco2 has
also been reported for dog blood, but a higher peak value of 0.4 was noted
(Mithoefer et al., 1963). More recently Mithoefer and coworkers (1969), using dog
blood, concluded that f. declined in linear fashion with increasing H , irrespective
of Pco2. Their regression equation indicated that f. would be 0.35, 0.38 and 0.41
at pH values of 7.3, 7.4 and 7.5, respectively. Von Mengden et al. (1969), have
reported results from 150 human subjects where Hb and base excess were manipu-
lated in vitro. They described the Haldane effect as the increase in pH when
oxygenated blood was reduced and presented an equation which demonstrated
that the factor was directly related to Hb and inversely to Pco~. By utilizing eqs. 13
and 14 their values can be converted to vol. ~ . For Oxygenated blood having a pH
of 7.4 and Hb of 15.0 g~o the values of f. at Pco~ values of 20, 40 and 60 mm Hg
are 0.19, 0.27 and 0.31, respectively. Their mean value is therefore about 30~o
lower than that reported by Mithoefer et al. (1969). A value of 0.32 is often utilized
(Kim et al., 1966). However, this has arisen mainly from theoretical considerations
during breathholding (Mithoefer, 1959) or has been obtained indirectly (McHardy,
1967) by analyzing nomograms such as those derived by Dill et al. (1937) during
the 'classical period' of blood analyses. One popular computer program (Olszowka
and Farhi, 1968) utilizes the values from Dill et al. (1937) where AHCOa-/AHbO2
was shown to fall at high and low pH values, but remained fairly constant at 0.43
between pH values of 7.3 and 7.5. Assumptions about base excess and Pco~ must
be made to convert these values to vol. ~ .
A review and discussion of the Haldane effect and its values is provided by
Siggaard-Andersen (1974). Theoretically, f. should be defined as the change in
178 J.A. LOEPPKYet al.

hemoglobin-bound H resulting from AHbO2. If given in these terms, the conver-

sion to fn in vo1. ~o again requires the knowledge of the other variables in eqs. 13
and 14. In vitro studies with erythrocyte fluids have shown that this Haldane value
is especially sensitive to Pco2 and pH and also affected by the range over which So~
or HbO2 is changing, i.e., the binding of H is somewhat greater if So~ changes
from 0.50 to 0, than from 0.75 to 0.25. The Haldane factor is also dependent on
the concentration of 2,3-DPG (Siggaard-Andersen, 1974) which may account for
some of the variations in values previously cited. The relative contribution of
HCO3 and carbamino compounds in determining fH also depends on pH and
2,3-DPG (Klocke, 1980). These complex interrelationships indicate that the value
for fn may well be different in various blood sources even when the common indices
of acid-base balance have been corrected, e.g., by eqs. 13 and 14. Also, small
systematic errors in measuring Hb, So:, pH and Pco:, as we have shown, may
account for relatively large differences in reported values for fn-
From a practical standpoint the important question is whether the in vivo fn in
whole blood varies significantly under physiological conditions to introduce appre-
ciable errors in computations of Pco~ and Cco: from CO2 dissociation curves if fH
is assumed a constant. Our results indicate that variations in fH cannot be accounted
for by changes in other commonly measured indices of gas exchange or acid-base
status. Therefore, a case may be made for considering it as a constant within the
physiological range of contents and pressures encountered in healthy men at rest
and in exercise up to 70~ of Vo2max and in resting patients with obstructive lung
disease. Our mean value is somewhat lower than the often-assumed 0.32, but is
relatively close to that noted from in vitro studies of erythrocyte fluid (Siggaard-
Andersen, 1974; Klocke, 1980). During exercise it is almost identical to the value
of 0.27 calculated by the equation of von Mengden et al. (1969) when utilizing our
mean values for pH, Pco2, So2 and Hb. From the mean of our patient data their
equation resulted in a value of 0.30 for fH. The value of 0.28 is also within the
range stated in the original description of the Haldane effect (Christiansen et al.,
1914). In the following computations our mean value of 0.28 for fH is assumed,
although other values may easily be substituted.

Change in blood Pco2 resulting from Haldane effect

A corrollary to fH as depicted in fig. 3 is that the Haldane factor can be expressed

as APco2 if no change is assumed in Cco2 and the fH value is known in vol. ~ . This
is of considerable significance when blood-to-gas CO2 gradients are measured and
debated (Lenfant, 1966; Scheid and Piiper, 1980), i.e., what is the maximum amount
that pulmonary capillary or arterial Pco2 can change due to the Haldane effect
alone and thereby contribute to alveolar-arterial gradients otherwise altered by
shunting or V/t) distribution?
As an example of this computation, consider the subject in fig. 3 (subject 4 at
 co2 DISSOCIATIONCURVEAND HALDANEEFFECT 179

1.53 L/min in table 1). If he were to breathe 100Y/o O5 and increase his So2 to 1.0,
curve B would shift to curve C as a result of the increase in HbO2 and pass through
point a2 at Paco~. The value for Cco2 at this point can be computed as follows:
Cco2(a2) = Cco2(a) - 0.26(24.89 - 21.35) (15)
Retaining fH for this particular sample at 0.26, the value for Cco~ is 38.47 vol. ~ .
The distance from a to x (fig. 3) is the rise in Pco~ due to complete oxygenation
of Hb. This is now computed by deriving the equation passing through point a2
as described previously and solving it for Pco~ at the Cco2 calculated or measured
for point a. The Pco= at x calculated in this fashion from the equation shown in
fig. 3 for curve C is 33.41, thus Pco: has risen 1.61 mm Hg. At the venous end a higher
value of 1.85 mm Hg is obtained due to the lower slope of the parallel curves.
In practical applications only one blood point is known and if So2 and Hb are
known or assumed, as well as f . , then the above calculations are possible. For
example, an arterial blood sample from a patient breathing air is found to have
an Hb of 15.0~, So~ of 0.80, pH of 7.45 and Pco2 of 50 mm Hg. We wish to know
the rise in Paco2 that could result from the Haldane effect when,the patient breathes
pure 02. First CO2p is calculated by eq. 13 (35.20 mEq/L) and partially-reduced (r)
whole blood CO2 content by eq. 14 (64.38 vol. ~o)- The Cco~ resulting from full
oxygenation (o) of Hb is computed as follows:

Cco~(O) = Cco~(r) - 0.28[(I - So~) Hb. 1.36] (16)

and equals 63.24 vol. ~o. The coefficients for eq. 3, K (18.4894) and b (0.3143) are
computed for Hb, Pco2 and Cco2(O) and the increase in Pco2 (2.96 mm Hg) is
calculated from the following equation:
APco: = e[lne(Cco2 (r)/K)]/b -- Pco: (17)

If an oxygenated point is known and one wants to compute the decrease in Pco~
with Hb reduction then eq. 16 is solved for Cco~(r) and the constants for eq. 3 are
computed for the curve passing through Cco2(r) and Pco2, and then Cco2(O) is utilized
in eq. 17.
Values obtained for Pco~ by this mathematical model conform well to those
predicted from a nomogram constructed by Lenfant (1966) from the nomogram of
Dill et al. (1937), where the former shows APco~ as a function of ASc2 and Pco~.
Our model has the advantage that it takes into consideration Hb variations as well.
Utilizing these computations we demonstrated that the Haldane effect could account
for 78~o of the rise in Paco~ noted when the 20 pulmonary patients were given
100~o O2 (Luft et al., 1981).
Whether the Haldane effect produces an in vivo change in Cco~ or a change in
Pco2 or both (vertical or horizontal displacement of a coordinate in fig. 3) is not
easily determined, especially during rebreathing procedures. Altered gas exchange
will affect the direction of the shift of a given CO2 coordinate, but the magnitude
of the shift will be determined by the CO2 dissociation curves passing through the
180 J.A. LOEPPKY et al.

coordinates. The value of models such as those presented here lies in showing to
what maximum extent Pco2 and Cco2 can be altered by the Haldane effect. Com-
putations of pressure and content in both reduced and oxygenated blood samples
can be made with this model in order to differentiate the Haldane effect from
clinical impairments or experimental alterations in gas exchange.

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