EuropeanJournal of
Eur J Appl Physiol (1987) 56:253-259
Breakdown of high-energy phosphate compounds and lactate
accumulation during short supramaximal exercise
J. Hirvonen 1, S. Rehunen 2, H. Rusko 1, and M. Hiirk6nen 2
Department of Biology of Physical Activity, University of Jyv~iskylii, SF-40100 Jyv~iskyl~i
Department of Clinical Chemistry, University of Helsinki, SF-00290 Helsinki, Finland
Summary. Muscle ATP, creatine phosphate and
lactate, and blood pH and lactate were measured
in 7 male sprinters before and after running 40,
60, 80 and 100 m at maximal speed. The sprinters
were divided into two groups, group 1 being
sprinters who achieved a higher maximal speed
(10.07+0.13 m - s -1) than group 2 (9.75_+0.10
m 9s - 1), and who also maintained the speed for a
longer time. The breakdown of high-energy phos-
phate stores was significantly greater for group 1
than for group 2 for all distances other than
100 m; the breakdown of creatine phosphate for
group 1 was almost the same for 4 0 m as for
100 m. Muscle and blood lactate began to accu-
mulate during the 40 m exercise. The accumula-
tion of blood lactate was linear (0.55_+0.02
m m o l . s - 1.1 - 1) for all distances, and there were
no differences between the groups. With 100 m
sprints the end-levels of blood and muscle lactate
were not high enough and the change in blood p H
was not great enough for one to accept that lac-
tate accumulation is responsible for the decrease
in running speed over this distance.
We concluded that 1) in short-term maximal
exercise, performance depends on the capacity
for using high-energy phosphates at the beginning
of the exercise, and 2) the decrease in running
speed begins when the high-energy phosphate
stores are depleted and most of the energy must
then be produced by glycolysis.
Key words: Adenosine triphosphate -- Lactate --
Muscle metabolism -- pH -- Phosphocreatine --
Short-term exercise
Of]print requests to: J. Hirvonen, Finnish Sports Institute, SF-
19120 Vierum~iki, Finland
Applied
Physiology
and Occupationa[ Physiology
9 Springer Verlag 1987
Introduction
In energy metabolism the converter of chemical
energy to mechanical work in the muscles is ATP,
but this is in such limited supply that during mus-
cular work it needs continuously to be resynthe-
sized. During heavy work the need for energy ex-
ceeds the supply from aerobic sources and energy
must then be produced from anaerobic sources by
the breakdown of creatine phosphate and anae-
robic glycolysis. In muscular work the breakdown
of creatine phosphate is a very rapid process, and
any A D P produced is at once rephosphorylated to
ATP.
Previous studies have shown that the speed of
creatine phosphate breakdown depends on the in-
tensity of the muscle work involved (Bergstr6m
1967). Karlsson (1971) reported that in muscular
work at 100% Vo . . . . in which exhaustion oc-
curred in 2 to 9 minutes, creatine phosphate stores
were reduced to very low levels in 2 to 3 min.
Other studies have shown that during maximal
voluntary dynamic or static muscular effort, crea-
tine phosphate stores are depleted in 20 s (Berg-
str6m et al. 1971; Keul et al. 1972). According to
the theoretical calculations of Margaria et al.
(1966); Newsholme (1980), and Mader et al.
(1983), creatine phosphate stores could be de-
pleted in maximal sprinting in 5--7 s. It seems
therefore that there is a difference between theory
and practice. This is possibly due to the fact that
in earlier studies the exercises were too light to
cause maximal breakdown of creatine phos-
phate.
The aim of this study was to investigate the
breakdown of ATP and creatine phosphate during
short-term maximal exercise in vivo and to try to
find a metabolic explanation for muscular fatigue
in supramaximal muscle work.
254 J. Hirvonen et al.: High-energy phosphates and lactate during short-term exercises

Table 1. Anthropometrical data and records of the test subjects

Subject Age Height Weight Fat Fast-twitch Record for 100 m

no. a (years) (era) (kg) (%) (%) (year made)

1. 26 179 69 8.3 52 10.68 (1982)

2. 25 186 76 11.3 59 10.97 (1983)
3. 23 179 78 11.2 56 10.79 (1983)
4. 26 179 72 10.1 66 10.94 (1982)
5. 39 178 74 12.3 58 10.99 (1983)
6. 33 187 80 7.6 65 10.90 (1982)
7. 20 172 72 10.5 64 10.94 (1981)

Mean 27 180 74 10.2 60 10.89

+ SEM 3 2 1 0.6 2 0.94

a Subjects nos. 1--3 formed group 1 and nos. 4--7 group 2

Materials and methods
Subjects
Seven male sprinters, average age 27 years, volunteered for the
study. All were at the national top level of sprinters in Finland
and specialized in the 100 m distance (Table 1). They were div-
ided into two groups on the basis of their running results dur-
ing the study. The three fastest formed group 1 and the other
four, group 2.
Test protocol
All the sprinters ran distances of 40, 60, 80 and 100 m, two
runs being made on one day and two the following day. Before
each run they had a thorough warm-up for nearly one hour.
After five min rest and sample-taking, they then ran one of the
four distances selected at random. There was a rest of 2 h be-
fore the warm-up preceding the second run. The same proto-
col was repeated on the following day (Fig. 1).
All four runs were with maximal effort as in competitions.
The speed of the sprinters was determined by photocells
placed at ten-metre intervals. The speed curve was calculated
from the mean times. Conditions were constant throughout the
study and all runs were indoors. A mattress was placed at the
end of the track for the sprinters to fall onto at full speed so
that blood and muscle samples could be taken immediately
after each run.
Sample-takin9 and analyses
Venous blood samples for the measurement of blood lactate
were taken from the cubital vein with the subjects recumbent.
Samples were taken at rest (on the first day only), 5 min after
the warm-up about 30 s before the run and then immediately
after the run, and 2, 4, 6, 8 and 10 rain later to find the highest
level of lactate (Fig. 1). The blood was collected into EDTA-
containing tubes (1.4 rag. ml-~), frozen immediately and
stored at - 75 ~ C until analysed.
Capillary blood for the measurement of acid-base balance
was collected at rest, 5 rain after the warm-up about 30 s be-
fore the run and then immediately after each run. The capil-
lary tubes were placed immediately into an ice-bath, and the
determinations were made within 1 h by a P H M 72 Mk 2 Digi-
tal Acid-base Analyzer (Radiometer, Copenhagen, Denmark).
Muscle biopsies were taken after local anaesthesia (lido-
caine) of the skin, through a small incision (1--2 mm) with a
biopsy needle (Tru-Cut | Travenol Laboratories Inc., Illinois,
USA). They were taken from the lateral portion of the quadri-
ceps femoris at rest (on the first day only), 5 rain after the
warm-up bout 30 s before the run and then immediately after
each run. The latter samples were frozen in liquid nitrogen
within 10 s after the run. The weight of the biopsy samples
varied from 3 to 10 mg. Staining for the identification of slow-
twitch and fast-twitch muscle fibres was as described by Re-
h u n e n and H~rkOnen (1980).
Perchloric acid extracts were obtained for the measure-
ment of muscle and blood lactate and for the determination of
muscle ATP, creatine phosphate and creatine; the parameters
were determined fluorometrically with a Transcon 102 FN (El-
omit, Transcon Instruments, Ltd, Helsinki, Finland) by nico-
tinamide adenine dinucleotide-linked enzymatic methods, all
according to Nfiveri et al. (1978).
The substrates and enzymes were purchased from Boeh-
ringer M a n n h e i m G m b H (Munich, FRG) and the Sigma
Chemical Company (St. Louis, Missouri, USA), and dithio-
threitol (DTT) came from Calbiochem (Lucerne, Switzerland).
All other reagents were of analytical grade and were obtained

VENOUS
BLOOD ~ ; I~;;II i liilll
MUSCLE
BIOPSY; ;l II
1 ST RUN OF I 2 NO RUN OF

I
THE DAY I THE DAY
WARM-UP REST REST 120 MIN WARM-UP REST
5 MIN 5 MIN
; m m Fig. 1. Test protocol followed during two consecu-
~o 1~o ~;o ~o
TIME (MIN) tive days
 O

Table 2. Selected m e t a b o l i c p a r a m e t e r s in b l o o d a n d m u s c l e o f t h e s p r i n t e r s b e f o r e a n d after different r u n n i n g distances. A b b r e v i a t i o n s : B-lactate (blood-lactate); c B - p H

t<
(capillary b l o o d p H ) ; M - A T P ( m u s c l e A T P ) ; M - c r e a t i n e p h o s p h a t e ( m u s c l e creatine p h o s p h a t e ) . Values e x p r e s s e d as m e a n s +- S E M . * p < 0.05, ** p < 0.02; *** p < 0.01 for
significance o f d i f f e r e n c e s f r o m t h e p r e - e x e r c i s e v a l u e s or b e t w e e n g r o u p s 1 a n d 2 (z)

40 m 60 m 80 m 100 m ~z

Before After Before After Before After Before After

B-lactate
(mmol l)
Group 1 1.5 _ 0 . 1 4.5 +-0.2** 1.7 +-0.1 5.9 -t-0.4"** 1.5 +-0.2 6.8~+-0.4 *** 1.6 +-0.3 8.3 + 0 . 6 * * *
O-,
Group 2 1.6 +-0.2 4.4 +-0.4*** 2.0 +-0.3 5.5 +-0.7*** 1.8 +-0.5 7.8 +-0.7*** 2.4 +-0.8 8.0 ___0.8***
cB-pH
Group 1 7.44+0.02 7.36+-0.02* 7.42-t-0.02 7.31+-0.01"* 7.41__.0.01 7.28+-0.01"** 7.42+-0.01 7.24+0.01"**
Group 2 7.42+0.01 7.33+-0.01"* 7.41+0.01 7.32+-0.02** 7.41___0.01 7.26+-0.01"** 7.41+-0.01 7.24___0.01"** O
Lactate a c c u m u l a t i o n
( m m o l s - 1 x I - 1)
Group 1 0.56 +- 0,02 0.57 +- 0,05 0.56 + 0.05 0.59+_0.03
Group 2 0.49 +- 0.08 0.52 _ 0.03 0.62 + 0.05 0.52 +- 0.07 cp
M - A T P (retool x kg - i;
wet weight)
Group 1 5.4 + 0 . 3 3.5 _+0,6 5.5 +-0,2 3.2 + 0 . 0 " * * 5.4 + 0 . 1 3.3 +-0.2*** 5.2 +-0.2 3.7 + 0 . 4
Group 2 5.3 +-0.3 5.0 +-0,5 4.9 +-0,5 4.2 5.8 +-0.3 5.0 _+0.4 z 5.1 3.7 _ 0 . 1 " *
M-creatine phosphate
( m m o l x kg - J ;
wet weight)
Group 1 10.3 + 0 . 1 3.8 ___0.4* 10.8 + 0 , 6 4.1 + 0 . 8 * * * 10.3 ___0.4 2.5 +-0.3*** 9.1 +-0.6 2.6 _+0.4**
Group2 12.0 +-1.7 6.5 +-0.9** 10.3 +-1,0 5 3 +-0.6*** 12.3 + 1 . 1 5.4 _+0.5 *** Z~Z 11.8 +-1.8 4.4 __+0.1" ~
256 J. Hirvonen et al.: High-energy phosphates and lactate during short-term exercises

from E. Merck (Darmstadt, FRG) or BDH Chemicals Ltd 40 m and 100 m was not significant. Muscle total
(Poole, GB). creatine was 29.1+2.4 m m o l . kg -~ on the first
day at rest, and the mean after all warm-up peri-
Statisticalanalyses ods was 24.5+2.1 mmol 9 kg -1. After 40 m the
creatine concentration was 23.3 _ 1.9
Statistical analyses were by the matched-pair t test (Richterich m m o l - k g -~, after 60m, 22.8+2.1 m m o l . kg -~,
1968) or the Mann-Whitney U test (Siegel 1956).
80m, 23.1+1.4 m m o l . k g -~ and after 100m
22.4 + 2.2 mmol 9 k g - ~.
ATP did not decrease significantly after the
Results
runs (Fig. 2).
Changes in runnin9 speed, blood and muscle
lactate, blood pH, and muscle A TP, creatine Differences in metabolism with reference to
phosphate and creatine in the different runnin 9 performance capacity
distances
The maximal running speed was 10.07+0.13
Running speed was highest from 40 to 60 m m . s -~ for group 1 and 9.75+0.10 m - s -~ for
(Fig. 2), the highest value being 9.89 +0.22 m- s --1 group 2. After 100 m the speed for group 1 was
( m e a n + S E M ) at 50m. At 8 0 m speed had de- 5.7% lower than maximal, and that for group 2,
creased significantly from that at 50 m (p < 0.001). 10.8% slower (Fig. 4). The speed of lactate accu-
Muscle and blood lactate levels began to increase mulation did not differ between the groups (Table
as soon as running began (Fig. 2). The correlation 2).
coefficient between the highest recovery of blood For the 40 m and 60 m distances postexercise
and muscle lactate was 0.60 ( n = 2 6 ; p<0.001). ATP and creatine phosphate concentrations
The speed of accumulation was constant for all tended to be lower for group 1 than for group 2.
distances (0.55 + 0.02 mmol 9s-~, Table 2), as cal- After 80 m the difference between the groups was
culated from peak blood lactate values (Fig. 3). statistically significant for both concentrations
Blood p H decreased as lactate increased (Fig. 2), (p<0.05 and <0.01) and after 100 m for creatine
and the decrease was also linear from the start of phosphate (p < 0.02, Table 2).
running. Muscle creatine phosphate was 21.7 + 0.8 Significant differences between the groups
mmol 9k g - ' on the first day at rest, and the mean were found in the use of high-energy phosphate
after all warm-up periods was 10.9+0.4 (2 x AATP + Acreatine phosphate). After 40 m
mmol- kg -~ (p<0.001) (Fig. 2). After 40 m the group 1 sprinters had only 50.9+7.8% of their
creatine phosphate concentration was 5.6+0.8 phosphate pool left, whereas group 2 still had
retool, kg -~, significantly lower than the prerun 73.2+4.8% (p<0.05) (Fig. 4). At 60 m and 80 m
value (p<0.001). The decrease tended to be the differences between the groups were still sig-
greater with distance, but the difference between nificant (p < 0.02 for both distances), but at 100 m

P" _.111 (...........

1~ 2~ ", ~ _ .~ ~_ ~:s speed

g 9t-~8 I-
Z35- L
"~ 71-'~ 14 P

' ~ //' l a c t a t ~
E -o
"~4~ 8 F Fig. 2. Changes in running speed, blood lactate,
I-~ 3 6 ~- . . . . . . . . . . . 75~
capillary blood pH and muscle ATP and crea-
tine phosphate (CP) for the different running
g 2F~ 41- distances (n=7). The values are expressed as
Z20 ~ 1L:~ 2L it
/
means and the vertical bars denote SEM. The
ol o 210 I I 810 i
warm-up 0 40 60 100 m
SEMs for pH are so small that they lie inside the
distance of running symbols
J. Hirvonen el: al.: High-energy phosphates and lactate during short-term exercises 257

8 -
B-- Before running T
A - After running
7-

6 -

5-

~_ 4
---6
-~E3

,'r 2

~-- 1 Fig. 3. Accumulation of blood lactate

compared with muscle lactate (columns)
for the different running distances (n = 7).
Time 0 2 4 6 8 1 0 (rain) 0246810 0 2 4 6 810 0246810 The values are expressed as means and
40 rn 60 m 80 m 100 m the vertical bars denote SEM

~o muscle work lasting for 11 s was depleted after

,.6 . ~ 10.50 5.5 s. We also found that the sprinters of higher
~ 10.00 ~ i ~ performance capacity used about 100% of the
cr
o~ 9.50[ _re_. . . . -~2~_~
creatine phosphate in this same time. Relatively
ff~a 9.o0ii "Z
small changes have been reported in the ATP con-
T
100
centration in fatigued muscle (Wilkie 1981) and
~
X2~
80 this was also seen in our study; ATP never fell
o~
o._ 50 below about 60% of the prerun level. It seems
therefore that in supramaximal sprinting most of
,c_ 40
c_ "6 the creatine phosphate store is used during the
o~
E Om 20 *-- g r o u p 1
first few seconds of exercise in the acceleration
c.- g r o u p 2
o
phase, and that the capacity to do muscle work
40 m 60 rn 80 m 100 m is related to the ability to use the high-energy
Fig. 4. The use of high-energy phosphates in running different phosphate pool; the sprinters of higher running
distances. Group 1 (n = 3) consists of the three fastest runners ability depleted their high-energy phosphates
and group 2 (n =4) the others. The 100% values are calculated more effectively in the beginning of the 100 m run
from the values of ATP multiplied by two and from the values than those of lower running ability. It can be sug-
of creatine phosphate (see Table 2). The values are expressed
as means and the vertical bars denote SEM
gested that, with the help of more chemical energy
from the high-energy phosphate pool, the better
sprinters were able to produce more "mechanical
work" and thus run faster. Better running ability
no differences were seen (group 1 52.4_+4.7%, could also be due in part to greater efficiency in
group 2 54.0+4.5%). the working muscles, i. e. better running technique
and better utilization of the elastic properties of
the muscles.
Discussion The great reduction in the concentration of
creatine phosphate seen after the warm-up period
The use of high-energy phosphates during is confusing. It could partly be due to the physical
maximal sprinting exercise involved, since the warm-up was thor-
ough and included sprints from 20 to 60 m. How-
The main purpose of the present study was to ex- ever, the time from the end of warm-up to sample-
amine the utilization of high-energy phosphates taking was only 4--5 min, which is too short for
during supramaximal exercise. We found that the complete breakdown of creatine phosphate
about 88% of the creatine phosphate expended in after maximal sprinting. This is supported by the
258 J. Hirvonen et al.: High-energy phosphates and lactate during short-term exercises
results of our previous study (Rehunen et al.
1982), where breakdown was still incomplete
5 min after maximal exhaustive exercise. Earlier
Harris et al. (1976) have reported the half-time of
the fast component of creatine phosphate recov-
ery to be 21--22 s, the half-time for the slow com-
ponent being over 170 s. Our exercise was, howev-
er, maximal, which was not the case in their study
and in other earlier studies.
The influence of warm-up bouts on the low
values of creatine phosphate may not be the only
explanation. The reduction could also be partly
explained by the fact that the blood flow is higher
in working muscle, thus making fresh muscle
samples heavier than those taken at rest (Rehunen
et al. 1982; Sabina et al. 1983). In this study the
influence of increased blood flow was calculated
by measurement of total creatine; this showed
that the effect of increased blood flow was about
15% due to the warm-up and about 20% due to the
runs.
Glycolytic energy production
The rate of lactate production was about the same
for all running distances, suggesting that the
power of glycolytic energy production is constant
from the start to the end of an exercise, at least in
one of up to 11 s in duration. Lactate production
rate does not seem to depend on performance ca-
pacity in trained athletes, and thus glycolytic
power can be exluded from the metabolic factors
explaining differences in performance capacity.
The results suggest that in the beginning of a
maximal 100m run (acceleration phase) both
high-energy phosphate stores and glycolytic en-
ergy production are used maximally to produce
energy in the form of ATP for the contractile ma-
chinery. During the middle part of the run (maxi-
mal speed phase) the contribution of the high en-
ergy phosphate stores is reduced and at the end of
the run (deceleration phase) glycolysis is the main
anaerobic energy source.
Muscle fatigue
In maximal bicycle ergometer tests and when run-
ning up stairs a reduction in power output occurs
about six to seven seconds after the start (Mar-
garia et al. 1966; Ikuta and Ikai 1972). In 100 m
sprinting a similar reduction has been seen in the
form of a decrease in maximal sprinting speed
(Murase et al. 1976). It has been suggested that
muscle fatigue during anaerobic muscle work is
caused by the accumulation of hydrogen ions
(H+), which inhibit glycolytic energy production
at the level of phosphofructokinase (Sahlin et al.
1981; Nassar-Gentina et al. 1981; Sahlin et al.
1983). In athletic performance it takes at least 40 s
of heavy work to reach maximal lactic acidosis
(Kindermann and Keul 1977) and because of the
low level of lactic acidosis seen in the 100 m run it
is obvious that this cannot be the primary reason
for muscle fatigue and the decrease in running
speed.
The energy production rate from high-energy
phosphate stores is much higher than that from
glycolysis (Mar6chal 1981). Therefore, when crea-
fine phosphate stores are depleted to certain level
the total energy production in the form of ATP is
noticeably reduced (Hultman et al. 1981). The
critical level of creatine phosphate is reached
when the enzyme creatine kinase is no longer sa-
turated with the substrate, creatine phosphate.
According to the results of this study the critical
level seems to be reached after 50 m running.
After this, ATP production does not cover the de-
mand for muscle contraction, thereby causing a
decrease in running speed.
Conclusions
1. In supramaximal voluntary muscular work,
creatine phosphate stores are depleted after 5
to 7 s. If the work continues, the role of glyco-
lysis becomes increasingly more important in
energy production.
2. The metabolic explanation of the fatigue which
appears after 5 to 7 s in maximal muscle work
may be that the energy production rate is re-
duced due to lack of energy supply from the
high-energy phosphate stores.
3. Sprinting performance correlates with the de-
pletion of high-energy phosphate stores in
muscle.
4. Lactate accumulation is constant throughout
the work, at least up to 11 s, reflecting glyco-
lyric energy production power in supramaxi-
real exercise.
Acknowledgements. This study was supported by grants from
the Finnish Olympic Committee.
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Accepted October 29, 1986