Acta physiol. scand. 1960.48.

454-460
From the Department of Physiology, Kungl. Gymnastiska Centralinstitutet,
Stockholm 0, Sweden

Myohemoglobin as an Oxygen-Store in Man

BY
ASTRAND,
IRMA PER-OLOFASTRAND,
ERIKHOHWUCHRISTENSEN
and RUNEHEDMAN
Received 23 September 1959

Abstract
ASTRAND,
I., P.-0. ASTRAND,
E. H. CHRISTENSEN
and R. HEDMAN.
Myohemoglobin as an oxygen-store in man. Acta physiol. scand. 1960. 48.
454-460. - The aim of the present research was to investigate further
the possible r81e of myohemoglobin as an oxygen-store during the initial
stage of muscular work. One subject worked intermittently with a work
load of 2,520 kpm/min with varied duration of work and rest pauses on
a bicycle ergometer. A highly significant difference in the blood lactic
acid concentration during the experimental time of 30 min was found,
at work with short work periods (10 sec, lactic acid concentration about
10-20 mg per 100 ml) compared with relative long ones (60 sec, lactic
acid concentration 1 10-140 mg per 100 ml). The conclusion was drawn
that the first type of work is performed aerobically. The calculated
oxygen demand, during the work period of 10 sec, however, does not
correspond to the measured oxygen intake. A deficit of about 0.43 1 0,
for each period of work will occur. It was suggested that this amount
of 0.43 1 0,is supplied to the working muscle mainlyfrom oxymyo-
hemoglobin. This store function of myohemoglobin is discussed in rela-
tion to the present findings and to the results mentioned in the literature.

In an earlier investigation concerning intermittent heavy work (2,160 kpm/

min) it was shown that blood lactic acid concentration remains low if work
alternates with rest pauses every half minute. If work and rest periods are
increased to 3 min the lactic acid concentration will reach high values and
the total work time will be limited due to exhaustion (ASTRAND et al. 1960).
A possible explanation for the aerobic work metabolism when 0.5 min
periods are used was given by the assumption that the oxygen bound to the
myohemoglobin plays an important r61e in the supply of oxygen to the working
muscles in the initial stage of work. With increasing duration of the work

454
 MYOHEMOGLOBIN AS AN OXYGEN-STORE 455
period the relative importance of this oxygen fraction diminishes, and if the
transport of oxygen is insufficient for the local demand, anaerobic processes
have to cover a certain fraction of work metabolism, and lactic acid accumu-
lates in the muscles and in the blood.
The present work was planned to further elucidate the r61e of myohemoglobin
in this respect. To obtain more conclusive results the work load was increased
to 2,520 kpm/min. Furthermore, the periods of work and rest were varied
independently of each other so that more decisive results as to the relative
importance of the rest pauses could be obtained.

Methods
Work was performed by one male, physically well trained subject, R. H., on a Krogh
bicycle ergometer. The subject and the methods used were the same as in the earlier
experiments (ASTRAND et al. 1960). The work of 2,520 kpm/min corresponds to a
load of 6 kg with 70 pedal revolutions per minute. The total experimental time was,
if possible, 30 min. The work periods were always of the same duration throughout one
experiment. They lasted for 10, 15, 30 or 60 sec. The rest pauses lasted from 20 up to
240 sec (compare Table I ) . Due to the different duration of rest pauses the total quanti-
ties of work produced on the different experimental days ranged between about 6,000
and 38,000 kpm, and the average work load for the 30 min varied between some 200
kpm/min and 1,260 kpm/min.

Results
Table I summarizes the experiments done and gives the maximal values for
blood lactic acid concentration, the total quantity of work performed in the
30 min experiment and other calculated values of importance for the dis-
cussion. The total quantity of 25,200 kpm of work will be represented in all
series, I-IV arranged according to the duration of the work period in the
table, and the quantity of 15,120 kpm will be found in three of the sections.
This makes a direct comparison possible. If the work period lasts for 10 sec,
420 kpm will be produced at every work occasion; with longer duration this
quantity increases and reaches, at 60 sec, 2,520 kpm. The calculated oxygen
demand for these different work quantities are given in the table. These values
are, of course, approximate but still they give a good illustration of the varying
demands, which are 0.9 1 of 0, for the 10 sec periods and 5.6 1 for the 60 sec
periods. The calculated values are based upon a mechanical efficiency of
23.0 per cent and a caloric coefficient for oxygen of 4.85.
Fig. 1 and 2 illustrate the changes in blood lactic acid concentration during
work with a total quantity of 25,200 and 15,120 kpm respectively. The relation-
ship between work and rest time is always 1 to 2 in Fig. 1, and 1 to 4 in Fig. 2.
With the short work periods of 10 sec followed by pauses of 20 sec, the blood
lactic acid concentration was about 20 mg per 100 ml, while in the experiment
with 40 sec pauses it was approximately 15 mg per 100 ml; that is, in both cases
31-593790. Acta physiol. scand. Vol. 48
ak blood lactic acid concentration in exfieriments with intermitterrt work of various duration of work and pause perio
tion, ejective work time and oxygen demand have been calculated and tabulated
I I1 IV
, sec ............ 10 15 30 60
s, sec.. . . . . . . . . . . 20 40 30 180 30 60 120 180 240 120 180
ed in each work
................. 420 630 1260 2520
ion in 30 min, kpm 25,200 15,120 25,200 5,800 37,800 25,200 15,120 10,840 8,320 25,200 18,900
rk time in 30 min
t, min ........... 10.0 6.0 10.0 2.3 15.0 10.0 6.0 4.3 3.3 10.0 7.5
in each workperi-
................. 0.9 1.4 2.7 5.6
lues for blood lactic
. mg per 100 m l . . 23 17 31 29 78 70 56 58 41 142 114
 MYOHEMOGLOBIN AS AN OXYGEN-STORE 457
mg % Producfion 25.200 kpm In 30mn
150- work:r& 1r2
rho-
& 60sec work
f20sec ,muses
130-
mg % Producfion f.5120 kpm In 30m;n
120- I201 work:res/ t4
6Osec uork
110- If0 - 24Oscc pauses

-
f00 too.
90- 90.
80 -

70 -

60. Joscc work

60 sec p a u s e
SO.
30sec work
LO. f20 set pauses

30-
f O s e c work
20. 20scc peu.%=s 20
10 10
0 0
0 io 20 3Ominufe.5

Fig. 1 and 2. The blood lactic acid concentration at a total work production of 1) 25,200 kpm
and 2) 15,120 kpm during a n experimental time of 30 min. The work is performed with a load
of 2,520 kpmlmin. The work periods last for 10, 30 and 60 sec and the corresponding rest peri-
ods for l ) 20, 60 and 120 sec and for 2) 40, 120 and 240 sec.

very close to the normal rest value of about 10 mg per 100ml. If the work periods
are lengthened to 30 sec and the pauses to 60 sec, one finds a significant increase
in the blood lactic acid concentration; after about 9 min a maximal value of
70 mg per 100 ml is reached (see Fig. 1). After that there is a small reduction
and the values remain approximately at 60 mg per 100 ml until the end of the
experiment. At the longer pauses of 120 sec, and consequently a smaller amount
of work performed (Fig. 2), a maximal value is also obtained after about 9 min,
now at 56 mg per 100 ml, and thereafter the values are stabilized between 40
and 50 mg per 100 ml. Therefore, even with work periods of 30 sec a certain
equilibrium is reached between the production and the elimination of lactic
acid. With work periods of 60 sec, however, a corresponding balance is never
reached; the lactic acid concentration increases until the end of the experiment.
With the relatively short pauses of 120 sec (Fig. l ) , the blood lactic acid con-
centration reached a maximal value of 142 mg per 100 ml after 22 min. The ex-
periment was then interrupted because the subject was no longer able to continue.
With pauses of 240 sec (Fig. 2) the task could be fulfilled for 30 min and the
blood lactic acid concentration reached a value of 1 14 mg per 100 ml a t the end.
-158 I. ASTRAND, P.-0.ASTRAND, E. H . CHRISTENSEN AND R. HEDMAN

Discussion
The present results confirm earlier findings by ASTRAND et al. (1960).
They show conclusively that a principal difference exists in man’s reaction a t
intermittent work to short and relatively long work periods, even if the total
amount of work in a certain time is the same. Furthermore, the results answer
the question, whether the length of the work period or the length of the pause
is the deciding factor for the blood lactic acid concentration. I n the earlier
series of experiments, as stated above, the length of the work periods and
pauses were always equal in the same experiment, which brought about
difficulties for making a conclusive interpretation in this respect. I t can be
seen from the results in Table I that the most decisive factor is the length
of the work period (compare series I-IV). From the results in series I11 it is
most apparent that even the length of the pause has a certain significance. In
this series the work period was 30 sec and the pauses on the different experi-
mental days varied between 30 and 240 sec. With a pause of 30 sec there were
30 work occasions during the half hour, with a pause duration of 240 sec there
\\.ere only 7 work occasions. Fewer work periods naturally decrease the possi-
bilities to produce lactic acid, and longer pauses provides greater possibilities
for the elimination of lactic acid. That explains the decrease in blood lactic
acid concentration from 78 mg per 100 ml, which was the maximal value with
30 sec pauses, to 41 mg per 100 ml with 240 sec pauses. Accordingly, the duration
of the pauses has a secondary importance in comparison to the duration of
the work periods. This is also illustrated by the results given in Fig. 1 and 2.
It is of great interest whether or not the present results are in agreement
\\-ith the hypothesis mentioned above that the difference in the reaction at
short (10 and 15 sec) and long (30 and 60 sec) work periods can be explained
b?- the r6le of myohemoglobin as a n oxygen-store.
One knows that the oxygen transport by the blood to the working muscles
increases with the duration of the work period, and that equalization between
oxygen need and supply can take several minutes to occur. In the series of
experiments performed with 10 sec of work and 20 sec of rest, the oxygen
intake was determined during the work period, and corresponds to 2.80 l/min.
During the 10 sec of work the actual oxygen intake was 1/6 of this, or 0.47 1.
The oxygen intake during work was 10 times greater than a corresponding
rest value of 0.043 1. The oxygen intake during the last 30 sec of a 60 sec work
period with 120 sec pauses gave a maximal value of 4.08 1 O,/min, or 0.68 1 0,
for 10 sec. I n the latter case the oxygen intake was 16 times greater than the
rest value. To be adequate for a load of 2,520 kpm/min a n oxygen supply of
5.6 I/min or 0.9 1 per 10 sec is required according to Table I. During the
experiment with work for 10 sec and pauses for 20 sec we must calculate with
a deficit of 0.90 -0.47 = 0.43 1 0,. With work for 60 sec and pauses for
120 sec, the oxygen intake during the whole work period was maximally 3.26 1
 MYOHEMOGLOBIN AS AN OXYGEN-STORE 459

09

Oa

0 7

06
1 ;,~flcl+

02 &und fo
myohemog/ Q63/
05

04
02 transp.
03 3 26/

02

01

0 BMR 0 2 6 1
IOSar I-0 rec
Fig. 3. The oxygen demand for 10 and 60 sec work with a load of 2,520 kpmlmin. An attempt is
made to illustrate the fraction of 0, that is a) bound to myohemoglobin, b) transported by the
blood and c) 0, deficit.

(during the first half min 2.43 l/min and during the second half 4.08 l/min).
This means a deficit of 5.60 - 3.26 = 2.34 1 0,. The reason that the oxygen
intake during the first half minute of a 1 min work period is relatively smaller
than during the 10 sec periods (2.43 and 2.80 l/min respectively) is that during
the short pauses of 20 sec the circulation and respiration never decline severely
before the work is begun again. If the pauses are lengthened to 2 min, the time
for adjustment becomes significantly increased.
With work for 10 sec and pauses for 20 sec we must assume practically
aerobic conditions in the working muscles. If such were not the case, the
60 work occasions should have brought about a successive accumulation of
lactic acid as a consequence, compare Fig. 1. We believe that the conclusion
can be drawn that approximately 0.43 1 0, have been available in the working
muscles at the beginning of each new work period, naturally even a t the 60 sec
periods. Quantitatively this means, that a supply of oxygen for the 10 sec
periods is assured by that amount, which is already in the muscles and by
the amount which can be transported by the blood during the work itself.
For 60 sec work, a deficit of 1.91 1 arises. This must be covered by anaerobic
processes, which results in an increase of the lactic acid concentration in
the blood.
Fig. 3 illustrates schematically the relative importance of the postulated
amount of oxygen in the muscles at 10 sec and 60 sec work periods respectively.
Naturally, the oxygen supply to the muscles becomes smaller at a single
work occasion, and the anaerobic factor is of greater importance quanti-
460 I. ASTRAND, P.-0.ASTRAND, E. H. CHRISTENSEN AND R. HEDMAN

tatively. If the work time is sufficiently short the amount of oxygen bound
to myohemoglobin should, however, be able to play a decisive r61e for the
muscle metabolism even in those cases. A revaluation of the so-called Simonsen-
effect must be the consequence. According to MULLERand HETTINCER ( 1 957)
this effect is important during the first 10 sec of work and results in a n extra
oxygen consumption as a result of the anaerobic conditions in the muscles.
On the basis of the results given here the amount of the oxygen which is
available a t the beginning of the work can not, of course, be determined. If
one attempts to calculate the amount of oxygen bound to myohemoglobin,
one is immediately confronted by a whole series of more or less unknown
factors. One is not familiar with the size of the active muscle mass, the myo-
hemoglobin concentration of the musculature or the degree of reduction of
oxymyohemoglobin. If one uses the values given in the literature for myo-
hemoglobin for example b y BIORCK(1949, p. 131), one finds that each gram
of muscle can bind about 10 mm3 0,. If one assumes 20 kg of active muscles
for the subject in the work mentioned here, one arrives a t a value of 200 ml 0,.
There is still a deficiency of about 230 ml, according to the above. The values
given by BIORCKare in this case too low, since it is generally accepted that
the amount of myohemoglobin increases with training, and the values given
above are not derived from specially well trained individuals.
It is quite evident from the investigations of SCHOLANDER, IRVING and
GRINNELL (1942) on diving seals that myohemoglobin can constitute a n im-
portant factor for oxygen supply to the musculature. According to our concep-
tion, the experimental results laid forth here indicate that myohemoglobin
has a n important function as an oxygen-store even in man. The reader is
referred to BIORCK(1949, p. 42), regarding references for and against such a
conception based on earlier findings.
Before a definite answer can be given to the question of the quantitative
r61e which myohemoglobin plays in this respect, further investigations are
required on myohemoglobin concentration in trained individuals. I t is our
hope that the experimental results related here will help to create greater
interest into this field of research.

References
.)\SIR+ND, I., P.-0. .&STRAND, E. H. CHRISTENSEN and R. HEDUAN, Intermittent muscular work.
.kta physiol. scand. 1960. 48. 448-453.
BIORCK, G., On myoglobin and its occurrence in man. Acta med. scand. 1949. Suppl. 226. 42. 131.
~ I U L L EE.
R ,A. and TH.HETTINGER, Der Energimehrbedarf bei Arbeitsbeginn. Arbeitsp/ysiologie.
1957. 16. 480-499.
SCHOLANDER, P. F., L. IRVING and S. W. GRINNELL, Aerobic and anaerobic changes in seal
muscles during diving. 3. biol. Chem. 1942. 142. 431-440.