Eur J Appl Physiol
DOI 10.1007/s00421-011-2090-1
ORIGINAL ARTICLE
Blood lactate and sEMG at different knee angles during fatiguing
leg press exercise
Esteban M. Gorostiaga • Ion Navarro-Amézqueta • Miriam González-Izal
Armando Malanda • Cristina Granados • Javier Ibáñez • Igor Setuain •
Mikel Izquierdo
Received: 11 March 2011 / Accepted: 12 July 2011
Ó Springer-Verlag 2011
Abstract The purpose of this study was to examine the
changes in peak power output, blood lactate concentrations
and surface electromyographic activity (sEMG) of the
agonist [vastus lateralis (VL) and vastus medialis (VM)]
and the antagonist [biceps femoris (BF)] muscles at two
angular positions intervals (90–67° and 23–0° of knee
flexion), during a set of 10 repetitions leading to failure of
bilateral leg press exercise. Fatiguing exercise resulted in
increased blood lactate concentrations, the agonist mean
rectified voltage (MRV) at 90–67° of flexion, the antago-
nist average MRV at 23–0° of flexion and the spectral
parameter proposed by Dimitrov (FInsm5) (P \ 0.01–0.05).
Significant decreases (P \ 0.01–0.05) were observed in
power output, median frequency (Fmed) of the agonist
muscles at both angular position intervals and of the
antagonist muscle at 90–67° of flexion. No changes were
observed in the antagonist/agonist MRV activation ratio.
The present data suggest that the shift of frequency spec-
trum to lower frequencies and the accumulation of lactate
and/or H?, but not the antagonist/agonist MRV activation
Communicated by Toshio Moritani.
E. M. Gorostiaga (&)  I. Navarro-Amézqueta 
M. González-Izal  C. Granados  J. Ibáñez  I. Setuain 
M. Izquierdo
Studies, Research and Sport Medicine Centre,
Government of Navarre, Calle Sangüesa 34,
31005 Pamplona, Navarra, Spain
e-mail: egorosta@cfnavarra.es
M. González-Izal  A. Malanda
Department of Electric and Electronic Engineering,
Public University of Navarre, Campus de Arrosadia,
31006 Pamplona, Spain
•
ratio, may be relevant independent factors associated with
fatigue.
Keywords Leg press  Coactivation  Lactate  Fatigue
Introduction
Bilateral leg press exercise is one of the most common core
resistance exercises used to enhance performance in sports
and in knee rehabilitation processes as it develops powerful
muscles of the body that are activated during many func-
tional movements, such as running, jumping, squatting,
stooping and ascending or descending stairs (Escamilla
et al. 2001). A common repetition scheme of bilateral leg
press exercise adhered by trained athletes or knee injured
people includes one or more sets of 10 repetitions maxi-
mum (10RM) with a resistance equivalent to approxi-
mately 75–85% of their maximal dynamic strength (1RM).
During a set of 10RM, the subjects are unable to complete
the next repetition after 10 repetitions and the velocity and
power output of the repetitions slow naturally as fatigue
increases (Izquierdo et al. 2006).
The ultimate cause of the reduction in the maximal
capacity to generate force or power output during maximal,
short-term exercise remains obscure. In most cases, it has
been associated with impairments of various metabolic
events that provide energy for the contractile machinery
(Bogdanis et al. 1995; Hultman and Greenhaff 1991;
Karatzaferi et al. 2003; Westerblad et al. 1998) and/or with
rapid electrophysiological changes within the nervous
system or in the excitation–contraction coupling (Gandevia
2001). A non-invasive method for measuring electrophys-
iological changes and to assess neuromuscular function is
surface electromyography (sEMG). EMG amplitude (mean
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or root mean square) and spectral parameters (mean and
median frequency of EMG power spectrum) have been
traditionally used to evaluate the pattern of motor unit
activity as well as skeletal muscle fatigue. Thus, during
fatiguing exercise, muscle fatigue development is associ-
ated with changes in the surface EMG amplitude and EMG
power spectrum (Gerdle et al. 2000). However, the mean
and median frequency have relatively low sensitivities
because the established criteria for an objective selection of
boundary frequency and/or high and low-frequency bands
are subjective and depend not only on the development of
muscle fatigue but also on the muscle fibre-to-electrode
distances, electrode type, electrode longitudinal position,
muscle fibre length and volume conductor properties
(Dimitrov et al. 2006). To overcome the relatively low
sensitivity or low correlation of changes in median fre-
quency and muscle fatigue, a new spectral index was
proposed by Dimitrov (FInsm5) (Dimitrov et al. 2006). The
index is calculated as the ratio between the signal spectral
moment of order (-1), which emphasises the increase in
low and ultra-low frequencies in the sEMG spectrum and
normalising spectral moment of order 5, which emphasises
the effect of decreases in the high frequencies (Dimitrov
et al. 2006), and it has been shown to offer a powerful tool
for the assessment of muscle fatigability (Dimitrov et al.
2006; Gonzalez-Izal et al. 2010). To our knowledge, there
have been no investigations thoroughly describing the
metabolic and surface electromyographic activity (sEMG)
patterns using this new spectral index proposed by Dimit-
rov (FInsm5) (Dimitrov et al. 2006) across a single set of
bilateral leg press exercise leading to failure and the way
they are related to muscle fatigue. Thus, the first purpose of
the present study was to examine changes in blood lactate,
sEMG and power output during a set of 10 repetitions
leading to failure of bilateral leg press exercise and to
evaluate in what way lactate and sEMG changes could
contribute to the decline in muscle power production.
On the other hand, examining the antagonist activity of
the hamstring muscles during knee extension is worthwhile
because the hamstring plays a relevant role in limiting the
resultant joint moment and stabilising the knee (Psek and
Cafarelli 1993). Many studies using sEMG signal ampli-
tude parameters have reported changes in the antagonist
activity pattern of hamstrings during non-fatiguing maxi-
mum (Aagaard et al. 2000; Kellis and Baltzopoulos 1996;
Osternig et al. 1995) and submaximal isokinetic dynamic
(Draganich et al. 1989) efforts of the knee extensors.
However, their results cannot be applied to dynamic leg
press exercise with either a resistance variable or constant
loading (e.g. isoinertial) approach. Escamilla et al. (1998)
have found during one set of 4 repetitions of non-fatiguing
bilateral leg press exercise with an intensity corresponding
to a load leading to failure in 12 reps that biceps femoris
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Eur J Appl Physiol
(BF) neural activity during knee extension is low and
constant at different knee angle positions. However, when
acute exercise is performed under fatiguing conditions until
failure, it has been found that changes in the antagonist
sEMG muscle activity (Gonzalez-Izal et al. 2010; Hassani
et al. 2006; Psek and Cafarelli 1993) and/or in the balance
of sEMG activity between agonists and antagonists (Kellis
1999; Psek and Cafarelli 1993) may take place. To the
author’s knowledge, there have been no studies that have
examined the coactivation levels at different knee angle
positions and have quantified the agonist/antagonist neural
activity ratio during a fatiguing set of leg press exercise.
Therefore, the second purpose of this study was to examine
the fatigue-induced antagonist muscle activity changes at
two angular positions during a fatiguing set of leg press
exercise.
Materials and methods
Subjects
Thirteen healthy male volunteers participated in this study.
Their mean (± SD) age, height, body mass and maximal
dynamic strength (1RM) during leg press exercise were
34 ± 5 years, 178 ± 6 cm, 74.1 ± 6.3 kg and 190 ±
28 kg, respectively. All where habitually recreational ath-
letes, mainly in endurance events, but none trained for
competition. The subjects were thoroughly informed of the
purpose, nature, practical details and possible risks associ-
ated with the experiment as well as the right to terminate
participation at will, before they gave their voluntary con-
sent to participate. A medical examination was also com-
pleted by a physician, and subjects with medical problems
were excluded. The present work was approved by the
institutional review committee of the Instituto Navarro del
Deporte, according to the Declaration of Helsinki.
Preliminary testing
Several pretest sessions were carried out during the
3 weeks before the experiment took place. First, the sub-
jects were familiarised with the experimental testing pro-
cedures about 3 week before. Second, two weeks before,
the first experiment subjects participated in a control test-
ing day where resistance-load verifications for one repeti-
tion maximum strength (1RM: the heaviest load that could
be correctly pressed only once using the correct technique)
and for 10 repetition maximum strength (10RM: the indi-
vidual maximum load that produced 10 repetitions to
failure) were determined in the leg press exercise machine.
After measuring 1RM, we estimated the absolute load that
Eur J Appl Physiol
should produce 10 repetitions to fatigue (10RM). Then,
after at least 10 min rest, the subjects performed a maximal
repetitive set until failure with the load that theoretically
should produce 10 repetitions to fatigue (*85% of 1RM).
If the number of repetitions until failure was equal to 10,
the load was defined as a 10RM and used during the
experimental main tests. If the number of repetitions until
failure was different than 10, several trials of a maximal
repetitive set until failure were performed on different days
with lower or higher loads during subsequent tests sessions
to determine the load leading to failure in exactly 10
repetitions.
Leg extensor power testing
Each subject randomly participated in two experiments
performed on separate days at the same time of the day,
between one and two months apart. In one experimental
day, the subject performed a set of 5 repetitions of leg press
exercise, and on the other day, a set of 10 repetitions was
performed. Both experiments were performed with the
same absolute load and averaged 158 ± 25 kg (*83% of
the individual 1RM). Subjects were requested to repeat
their pre-recorded normal diet and refrain from any form of
intense physical exercise for 48 h prior each test. On the
morning of the experiment, the subject arrived after a light
breakfast and a fast of at least 2 h. On arrival at the lab-
oratory, the subjects rested on bed for 20 min on which the
EMG electrodes were located. The subjects then completed
a period of warm-up consisting of a set of 5 repetitions at
50%, of three to four repetitions at 75% and 1 repetition at
90% of maximal bilateral leg press strength (1RM). Three
to four subsequent attempts were made to determine the
1RM. The rest between maximal attempts was always
2 min. After at least 10 min rest, arterialised blood was
drawn from the earlobe. Then, an intense bilateral leg press
exercise, 5 or 10 repetitions with the maximum load pos-
sible to achieve 10 repetitions (10RM), was performed. In
each repetition, the subject was instructed to move the
weight as fast as possible. The motion ranged from
approximately 90–0° of knee flexion (0° = full knee
extension). The time for each repetition and the concentric
and eccentric components were recorded. There was about
1-s pause between repetitions to ensure that stretch–short-
ening cycle did not enhance performance of the subsequent
concentric action. The power output of each repetition was
monitored continuously and measured during the concen-
tric phase of leg press action. Immediately after the last
repetition (within 5–10 s), an additional arterialised blood
sample was taken. All subjects were highly motivated, and
strong verbal encouragement was given to all subjects to
motivate them to perform each repetition maximally and as
rapidly as possible.
Equipment
The study was performed on a modified horizontal bilateral
leg press variable resistance machine (Technogym,
Gambettola, Italy). The sitting was individually adjusted to
minimise displacement between the lower back and the
backrest during muscular force exertion and, therefore, to
avoid posture change. Each subject was instructed to fix
their feet in the same position on the force platform to ensure
a constant foot position. The exercise machine incorporated
four force transducers on a foot platform located below the
subject’s feet. The strain gauges recorded the applied force
(N) within an accuracy of 1 Newton at 1,000 Hz. The force
platform and leg press plate all remained stationary
throughout the lift, while the body moved away from the
feet. In addition, a rotational encoder (Computer Optical
Products Inc, California, USA) was set to weight plates to
record the position and direction of the displacement within
an accuracy of 0.2 mm at 1,000 Hz. Customised software
was used to calculate the power (instantaneous product of
displacement velocity and applied force) and work output
(vertical displacement of the weight plates times applied
force) per repetition. After the end of the exercise, results
were integrated over 1-ms time intervals. The maximum
10-ms integral of applied load and displacement velocity,
during each repetition, is referred to as the ‘peak power
output’. The average 10-ms integral of applied force and
displacement velocity, over the total concentric contraction
time period of each repetition, is referred to as ‘mean power
output’.
Surface electromyography
Surface electromyography (sEMG) during the extension
and flexion actions of the leg muscles was recorded from
the vastus lateralis (VL), vastus medialis (VM) and the
short head of the bicep femoris (BF) of the right leg by a
pair of bipolar surface electrodes (Blue Sensor N-00-S,
Medicotest) with a distance between the electrode’s centres
of 22 mm. The agonists vastus lateralis (VL) and vastus
medialis (VM) muscles were averaged because they have
demonstrated similar patterns of activation during bilateral
leg press exercise (Escamilla et al. 1998). The antagonist
biceps femoris (BF) muscle was chosen because during
maximal dynamic knee extension contractions the level of
coactivation is 3-fold higher for the BF compared to
semitendinosus muscles (Aagaard et al. 2000). After
careful preparation of the skin (shaving, abrasion and
cleaning with alcohol), the electrode pairs were placed
longitudinally on the medial portion of the muscles, fol-
lowing the recommendations of the SENIAM (Surface
ElectroMyoGraphy for the Non-Invasive Assessment of
Muscles). The positions of the electrodes were marked on
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the skin by small ink tattoos. The dots ensured the same
electrode positioning in each test over all experimental
period. EMG signals were recorded at a 1 kHz sampling
rate, with Muscle Tester ME3000 (Mega Electronics Ltd)
(bandwidth of 8–500 Hz/3 dB and a common mode
rejection ratio [100 dB). To facilitate and normalise the
analysis, the concentric phase of the leg press was divided
into 4 intervals of 22.5° of knee angle. The parameters
analysed in the present study corresponded to the first
(from 90° to 67°) and the fourth (from 23° to 0°) interval of
the knee extension actions for the VL, VM and BF
muscles.
Data analysis was performed offline using the MATLAB
2006a software environment (The MathWorks Inc., Natick,
Massachusetts, USA). The following sEMG parameters
were calculated:
(a) mean rectified voltage (MRV), calculated after a full-
wave rectification and filtered by a moving root-
mean-squared filter with a time constant of 50 ms, as
the integrated sEMG divided by the time.
(b) median frequency (Fmed), calculated numerically
from the following equation:
FZmed Zf2
PSð f Þ  df ¼ PSð f Þ  df
f1 Fmed
where PS(f) is the sEMG signal power spectrum calculated
using Fourier transform, f1 = 8 Hz and f2 = 500 Hz
(determined for the bandwidth of the surface
electromyography).
(c) The spectral parameter proposed by Dimitrov (FInsm5)
(Dimitrov et al. 2006) was also calculated. This spectral
index (FI nsm5) has been proposed to overcome the
relatively low sensitivity of mean and median fre-
quency (how good they are at correctly identifying
fatigue or power output changes) (Dimitrov et al. 2006)
because it predicts changes in muscle power most
accurately than other frequency or amplitude sEMG
parameters (Gonzalez-Izal et al. 2010).
The index is calculated as the ratio between the signal
spectral moment of order (-1), which emphasises the
increase in low and ultra-low frequencies in the sEMG
spectrum and normalising spectral moment of order 5,
which emphasises the effect of decreases in the high fre-
quencies (Dimitrov et al. 2006).
R f2
f f 1  PSðf Þ  df
FInsm5 ¼ R1f2
f 5  PSðf Þ  df
f1
where PS(f) was the sEMG power spectrum calculated
using Fourier transform and f1 = 8 Hz and f2 = 500 Hz.
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Eur J Appl Physiol
(d) the coactivation ratio was calculated by dividing the
MRV of the antagonist muscle (BF) by the MRV of
the agonist muscles (average of VL and VM) for a
common period of time and a given range of motion
of knee movement during each repetition. An increase
in this coactivation ratio may be indicative of a
greater level of muscular coactivation.
After calculating all of these parameters, an arithmetical
mean was calculated between the parameters of VM and
VL to obtain an index that included the effects of the
fatigue of the two muscles.
Blood samples
In both experimental days, capillary blood samples were
obtained from a hyperemic earlobe prior exercise and
immediately after the last repetition (5 and 10 repetitions).
Blood lactate concentrations were used because blood lac-
tate gives valuable information of muscle lactate accumu-
lation during one set of 10 repetitions to failure of leg press
exercise (Gorostiaga et al. 2010). After cleaning and
puncturing, a 5-ll sample of whole blood was automatically
aspirated into a single use, enzyme-coated electrode test
strip. Lactate concentration was determined via ampero-
metric measurement and the result displayed in 60 s
(Lactate Pro LT-1710; Arkray KDK Corporation, Shiga,
Japan). In terms of reliability, the manufacturers report a
coefficient of variation (CV) of 3.2% and 2.6% with lactate
standards of 2 and 11 mmoll-1, respectively. The Lactate
Pro was checked for accuracy according to manufacturer’s
instructions before every test.
Statistical analysis
Standard statistical methods were used for the calculation
of the mean and standard deviation (SD). Results in the
figures are given as mean and standard deviation (SD)
(Fig. 1) or standard error (SE) (Figs. 2, 3 and 4) values.
Student’s paired t test was used for comparisons of values
between the two different exercise models in this study,
whereas one-way analysis of variance for repeated mea-
sures was used to examine the differences in performance
indexes and metabolite concentrations over time. Were
significant F value was observed (P \ 0.05), the means
were compared using a LSD post hoc test to locate the
pairwise differences between the means. For the purposes
of comparison, the power output, blood lactate and sEMG
changes for the second five repetitions were compared with
the first five repetitions by means of Student’s paired t test.
Pearson product-moment correlation and second-degree
polynomial relationships tests were performed to examine
the relationship between variables.
Eur J Appl Physiol
Fig. 1 Peak power output
profiles (average for n = 13
subjects) for each exercise
during the two experimental
conditions: when exercise was 5
repetitions (open circles) and
when exercise was 10
repetitions (filled circles). Boxes
represent mean of the peak
power output for the first and
the second half of a set of 10
repetitions. #Significant
difference (P \ 0.01) between
the first and the second 5
repetitions. Values are
means ± SD
Results
Leg extensor power
The peak power output profiles during each repetition in
the two experimental conditions (5 or 10 repetitions) are
shown in Fig. 1. Average peak power output during the
first 5 repetitions was maintained similarly in both exper-
imental periods. After that, however, average peak power
declined progressively (P \ 0.05), reaching a value of
61 ± 15% of the highest value after 10 repetitions. Aver-
age peak power decreased 13 ± 14% (P \ 0.05) from the
first (861 ± 230 W) to the second (718 ± 237 W) 5 rep-
etitions of the exercise (Fig. 1).The magnitude of the
decline in average mean power output between the first and
the second 5 repetitions was 22 ± 11% (from 369 ± 94 to
287 ± 86 W; P \ 0.05). The total duration of the exercise
was 33.6 ± 5.8 s and was longer (P \ 0.01) for the second
(19.3 ± 5.6 s) than for first five repetitions (14.3 ± 2.3 s).
Surface electromyographic activity
The pattern of activation of the average mean rectified
voltage (MRV) during exercise was similar for VM and VL
muscles. Thus, the average MRV activation of VM muscle
increased progressively (P \ 0.01) from the first (490 ±
248 lvms-1) to the last repetition (568 ± 303 lvms-1)
at 90–67° of flexion, whereas similar increase (P \ 0.05)
was observed in VL muscle at the same angular position
(473 ± 302 lvms-1 and 505 ± 249 lvms-1 for the first
and the last repetition, respectively). The increase
(P \ 0.01) in the average MRV activation from the first to
the second 5 repetitions of the exercise was similar in VM
(9 ± 5%) than in VL (8 ± 7%) muscles. Similarly, the
changes in the average median frequency (Fmed) during
exercise were similar for VM and VL muscles. Thus, the
average (Fmed) of VM muscle decreased progressively
(P \ 0.05) from the first (67.4 ± 1.4 Hz) to the last repe-
tition (58.2 ± 9 Hz) at 90–67° of flexion, whereas similar
decrease (P \ 0.01) was observed in VL muscle at the
same angular position (72.2 ± 21 Hz and 65.8 ± 18 Hz
for the first and the last repetition, respectively). The
decrease (P \ 0.01) in average (Fmed) from the first to the
second 5 repetitions of the exercise was similar in VM
(11 ± 8%) than in VL (8 ± 6%) muscles. Since the vastus
lateralis (VL) and vastus medialis (VM) muscles presented
similar patterns of activation during leg press exercise,
their average (MRV) activation and average (Fmed) were
averaged.
Mean rectified voltage (MRV) and median frequency
(Fmed) values of the quadriceps (averaged for VM and VL
muscles) at 90–67° of flexion were similar in both exper-
imental days during the first 5 repetitions. Figure 2 shows
the average MRV of the quadriceps (averaged for VM and
VL muscles) (Fig. 2a) and BF muscle (Fig. 2b) over the
repetitions at two different angular position intervals
(90–67°of flexion and 23–0° of flexion). A significant
effect of angular position was found on MRV activation
characteristics during exercise. Thereby, during exercise,
the agonist (averaged for VM and VL muscles) average
MRV activation was *3 times higher (P \ 0.001) at
90–67° of flexion than at 23–0° of flexion (0° = full
extension), while the antagonist (BF) average MRV acti-
vation was *2 times higher (P \ 0.001) at 23–0° of
flexion than at 90–67° of flexion. For each angular position
interval, there were significant differences between repe-
titions. Thereby, at 90–67° of flexion, the agonist average
MRV reached the maximum levels obtained during 1RM
already at the second repetition and increased 7%
(P \ 0.01) from the first to the second 5 repetitions of the
exercise. No change was observed at 23–0° of flexion in the
agonist MRV activation. On the contrary, at 23–0° of
flexion, the antagonist average MRV increased 49%
(P \ 0.01) from the first to the second 5 repetitions of the
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 Eur J Appl Physiol

Fig. 2 Changes in mean

rectified voltage (MRV)
(averaged for vastus lateralis
and medialis muscles) (a) and
for biceps femoris (b) over the
repetitions at two angular
position intervals: 90–67° (filled
cicles) and 23–0° (open circles)
of knee flexion. Boxes represent
mean of the antagonist/agonist
MRV activation ratio during the
whole exercise at 90–67° (filled
histogram) and 23–0° (open
histogram) of knee flexion.
*
Significant differences
(P \ 0.01–0.05) between the
knee flexion intervals.
#
Significant differences
(P \ 0.01) between the first and
the second half of a set of 10
repetitions for a given knee
flexion interval, n = 13. Values
are mean ± SE

Fig. 3 Changes in average median frequency (Fmed) (averaged for

vastus lateralis and medialis muscles) (a) and for biceps femoris
(b) over the repetitions at two angular position intervals: 90–67°
(filled circles) and 23–0° (open circles) of knee flexion. *Significant
differences (P \ 0.01–0.05) between the knee flexion intervals.
#
Significant differences (P \ 0.01) between the first and the second
half of a set of 10 repetitions for a given knee flexion interval, n = 13.
Values are mean ± SE
Fig. 4 Changes in average fatigue index (FInsn5) averaged for vastus
lateralis and medialis muscles (a) and for biceps femoris (b) over the
repetitions at two angular position intervals (90–67° and 23–0° of
knee flexion). #Significant differences (P \ 0.01) between the first
and the second half of a set of 10 repetitions for a given knee flexion
interval, n = 13. Values are mean ± SE

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Eur J Appl Physiol

exercise while the lower (12%) MRV increase observed in

the same period at 90–67° of flexion approached statistical
significance (P = 0.07). The ratio of antagonist/agonist
MRV activation was *4 times greater at short muscle
lengths (0–23° of flexion) than at longer muscle lengths
(90–67° of flexion) (Fig. 2b). However, this antagonist/
agonist ratio did not change significantly over the repeti-
tions at both angular position intervals.
The average median frequency (Fmed) of the agonist
(averaged for VM and VL muscles) and BF muscles over
the repetitions at two different angular position intervals
are showed in Fig. 3. From the first to the second 5 repe-
titions, the agonist average Fmed decreased 11–15%
(P \ 0.01) at both angular positions intervals (Fig. 3a). In
BF muscle, average Fmed decreased 8% (P \ 0.01) from
the first to the second 5 repetitions at 90–67° of flexion
while no changes in average Fmed were observed at 23–0°
of flexion (Fig. 3b).
Figure 4 shows that the decreases in Fmed in the agonists
and antagonist muscles during exercise were mirrored by
concomitant increases in the spectral parameter proposed by
Dimitrov (FInsm5). Thus, from the first to the second 5 rep-
etitions, the agonist average FInsm5 increased 79–92% (P \
0.01–0.05) at both angular positions intervals (Fig. 4a). In
BF muscle, average FInsm5 increased 63% (P = 0.05) from
the first to the second 5 repetitions at 90–67° of flexion, while
no changes in average FInsm5 were observed at 23–0° of
flexion (Fig. 4b).

Blood Lactate
Blood lactate concentrations increased significantly
(P \ 0.05) after 5 (2.5 ± 0.6 mmoll-1) and 10 repetitions
(4.8 ± 0.9 mmoll-1) from rest (1.1 ± 0.2 mmoll-1)
values.
Relationship between sEMG parameters, muscle power
output and blood lactate changes
Significant second-degree polynomial relationships were
observed between changes in power output, sEMG
parameters and blood lactate. Thus, changes in absolute
mean power output during exercise were negatively cor-
related with percentage changes in FInsm5 (R2 = 0.56,
P \ 0.05) (Fig. 5a) in VL and VM muscles at 90–67° of
flexion as well as with relative changes in blood lactate
(R2 = 0.68, P \ 0.01) (Fig. 5b). The relationships
observed between changes in absolute power output and
changes in median frequency (Fmed) as well as changes in
MRV activation of the agonist (averaged for VM and VL
muscles) were lower (R2 = 0.34–40, P [ 0.05) than those
found between changes in power and changes in FInsm5.
Fig. 5 Relationship between the absolute changes in mean (a) and
peak (b) power output with (a) the relative changes in agonist muscles
FInsm5, at 90–67° of knee flexion, during the second 5 repetitions
(expressed as a percentage of the first 5 repetitions) or with (b) the
relative changes of blood lactate concentration during the whole
exercise (expressed as a percentage of the average rest values)
Discussion
The results of the present study showed that a set of 10
repetitions leading to failure of bilateral leg press exercise
resulted in increases in blood lactate concentrations, ago-
nist MRV activation at 90–67° of flexion and the antagonist
MRV at 23–0° of flexion, as well as in the spectral
parameter proposed by Dimitrov (FInsm5). In contrast,
10RM led to significant decreases in mean and peak power
output, average median frequency (Fmed) in the agonist
muscles at both angular position intervals and in the
antagonist muscle at 90–67° of flexion. No changes were
observed in the antagonist/agonist MRV activation ratio.
The decrease in absolute power output was negatively
correlated with changes of the spectral analysis proposed
by Dimitrov (FInsn5) in the agonist muscles and with
changes in blood lactate concentrations.

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Myoelectrical and metabolic manifestations of fatigue
One of the findings of the present study was that the
changes in power output were negatively correlated with
changes in the spectral analysis proposed by Dimitrov
(FInsn5) in the vastus lateralis and vastus medialis muscles
as well as with changes in blood lactate. No relationships
were observed, however, between the changes in power
output and the changes in median frequency (Fmed) or in
MRV activation. The finding that the application of FInsm5
on surface sEMG provided greater accuracy to map losses
in muscle power output than the other sEMG-based
parameters studied agrees with previous findings obtained
during unilateral knee extension protocols (Dimitrov et al.
2006) and during five sets of 10 repetitions leading to
failure of bilateral leg press exercise (Gonzalez-Izal et al.
2010). This suggests that the spectral index (FInsn5) may
confer greater sensitivity for assessing peripheral muscle
fatigue during dynamic resistance exercise than the tradi-
tionally used amplitude or spectral sEMG parameters. In
addition, one may suggest that the association observed in
the present study between fatigue-induced changes in peak
power and in blood lactate concentration seems likely to
reflect that during exercise the inability to maintain a
required or expected power appears to be closely linked to
some biochemical changes such as an abrupt accumulation
in muscle lactate and/or hydrogen ions (Hultman and
Greenhaff 1991).
An interesting finding of the present study was that over
the course of repetitions, a clear dissociation between
surface MRV amplitude and median frequency took place
because the increase in MRV activity was accompanied by
a progressive decrease in median frequency. Increases in
sEMG activity during exercise in parallel with decline in
median frequency have been observed during sustained
submaximal isometric (Fuglevand et al. 1993) and dynamic
one-leg knee extension exercise (Masuda et al. 1999)
contractions. It has been suggested that the decline in
median frequency with fatigue seems to be primarily due to
decreases in muscle fibre conduction velocity associated
with decreases in intra-muscular pH (Brody et al. 1991),
lengthening of the intra-cellular action potential (IAP) and
decreases in firing rate of fatigued fast motor units. These
changes in median frequency are probably associated with
the recruitment of additional motor units over the course of
repetitions to partially compensate for the loss of force of
the motor units that are already active (Bigland-Ritchie
et al. 1986; Fuglevand et al. 1993; Kellis 1999). Although
different hypothesis have been advanced (Basmajian and
DeLuca 1985; Gandevia 2001), several authors suggest that
these changes in median frequency and in the recruitment
of motor units are causes, at least in part, of the observed
increase in the surface MRV amplitude (Arabadzhiev et al.
123
Eur J Appl Physiol
2010; De-Luca 1984; Gandevia 2001). The fatigue of the
fast motor unit associated with the decrease in intra-mus-
cular pH and lactate accumulation is in agreement with
previous biochemical studies using high-intensity cycling
(Casey et al. 1996; McCartney et al. 1983) sprinting in a
non-motorised treadmill (Greenhaff et al. 1994), knee
extension exercise (Gorostiaga et al. 2010) or electrically
evoked isometric contractions (Soderlund et al. 1992)
showing that the development of fatigue observed under
these conditions may be attributable in part to accumula-
tion of muscle lactate and H? and to the inability of the
initial predominantly recruited high firing type II fibres to
maintain the required very high rate of ATP resynthesis
(Gollnick et al. 1973; Koopman et al. 2006; Soderlund
et al. 1992). Taking these data and the present data toge-
ther, it is not unreasonable to suggest, therefore, that events
occurring locally within the muscle such as an abrupt
increase in muscle lactate or by-products of ATP hydro-
lysis (hydrogen ions), activity-induced changes in the fibre
action potentials and/or impaired ability in the number of
contracting fast twitch (FT) fibres to regenerate ATP at
high rates make a significant contribution to the reduction
in median frequency and power output and to the increase
in MRV amplitude observed during the final repetitions in
the present study.
Agonist and antagonist coactivation
An additional purpose of the study was to examine the
agonist and antagonist electromyographic activity levels at
different angular positions during leg press exercise. A
significant effect of angular position on mean rectified
voltage (MRV) activation characteristics was found. The
main findings were as follows: 1) the agonist (average of
VM and VL muscles) MRV activation was significantly
higher at 90–67° of flexion than at 23–0° (0° = full
extension), whereas it was the opposite with the antagonist
(BF muscle) coactivation pattern, 2) significant increases in
neural agonist and antagonist activation levels were
observed over the repetitions mainly at the angular posi-
tions of higher neural activation and 3) the ratio of average
hamstring/quadriceps MRV activation was 4 times higher
at 23–0° than at 90–67° of knee flexion and was maintained
during progressive fatigue at both angular positions. The
higher activations in VL and VM in the mid range of joint
movement (90–67°) and in BF towards full knee extension
(23–0°) are in agreement with other findings observed on
non-fatiguing submaximal leg press exercise (Escamilla
et al. 1998) and during maximal isokinetic fatiguing (Kellis
1999) or non-fatiguing (Aagaard et al. 2000) knee exten-
sion contractions. The increase observed over the repeti-
tions in the agonist activity level at the angular positions of
higher neural activation (90–67°) has been observed in VM
Eur J Appl Physiol
during isometric fatigue test (Weir et al. 1996) and during
maximal isokinetic concentric efforts of the knee extensors
(Kellis 1999). As explained above, this increased agonist
activity has been related to an attempt of the central ner-
vous system to compensate for the failing muscle by
increasing the motor units available (De-Luca 1984; Enoka
1995; Kellis 1999) and/or to a shift towards lower fre-
quencies due to reduction in the muscle fibre conduction
velocity (Aagaard et al. 2000; Basmajian and DeLuca
1985) and to lengthening of the intra-cellular action
potential (Arabadzhiev et al. 2010; De-Luca 1984; Gandevia
2001). The considerable increase in the antagonist BF
muscle activity observed during exercise in the present study
has been also found during intermittent submaximal iso-
metric (Psek and Cafarelli 1993) or isokinetic (Hassani et al.
2006) knee extension contractions suggesting that the
decline in the mechanical output of the knee extensors
immediately after fatigue may be due, in part, to increased
force exerted by the antagonist muscle (Bentley et al. 2000).
During the whole exercise, the average MRV hamstring/
MRV quadriceps ratio was 4 times higher at short muscle
length (23–0° of knee flexion) than at long muscle length
(90–67° of knee flexion). These ratios were maintained at
both angular positions during progressive fatigue. Higher
MRV hamstring/MRV quadriceps ratio towards full knee
extension may serve to prevent possible damage caused by
the uneven pressure distribution on the articular surface of
the joint created by agonist contraction at these knee angles
(Basmajian and DeLuca 1985; Psek and Cafarelli 1993)
and may aid the knee’s ligaments in maintaining joint
stability at these knee angles (Psek and Cafarelli 1993;
Solomonow et al. 1987). Maintained ratio over the repeti-
tions between the degree of activation of quadriceps and
hamstring has been found during isometric fatiguing
extensions of the knee (Carolan and Cafarelli 1992; Psek
and Cafarelli 1993) suggesting that the behaviour of
coactive muscles persists during progressive fatigue (Psek
and Cafarelli 1993).
The present study has several limitations that need to be
mentioned. First, the accuracy and interpretation of sEMG
data are the subject of debate, as the surface EMG signal can
be influenced by several ‘non-physiological’ factors that
affect the volume conductor (Farina et al. 2004; Gandevia
2001). These include, among others, the ‘crosstalk’ phe-
nomenon (the fact that a signal that is recorded by the
electrode placed over one muscle is actually generated by a
nearby muscle), factors of anatomical nature (e.g. thickness
of the subcutaneous tissue layers, distribution and size of
motor units and fibres, geometrical changes in the position
of the detection system with respect to the muscle) or fac-
tors related to fibre membrane and motor unit properties
(e.g. conduction velocities, motor unit synchronisation).
These factors can, however, be minimised by appropriate
electrode placement, which we believe was the case in this
study. And secondly, the conclusions of this study are
generalised only to subjects with characteristics similar to
the present study’s participants and to studies that examine
knee extension responses in a similar fashion to the current
study. We do not believe these limitations jeopardise our
conclusions.
In summary, this study showed that one bout of 10
repetitions of bilateral leg press exercise leading to failure
resulted in increased agonist average MRV activation at
90–67° of knee flexion and in blood lactate concentrations.
The decrease in power output was negatively correlated
with changes of the spectral analysis proposed by Dimitrov
(FInsn5) in the agonist muscles and with relative changes in
blood lactate but was not correlated with the other sEMG-
based parameters studied, suggesting that (FInsn5) may
confer greater sensitivity for assessing muscle fatigue.
Moreover, the antagonist (BF) activation levels increased
over the repetitions at 23–0° of knee flexion, whereas the
ratio of hamstring/quadriceps MRV activation was constant
towards full knee extension and higher than in the mid
range of joint movement. These results suggest that the
shift of frequency spectrum to lower frequencies and the
accumulation in muscle lactate and/or hydrogen ions are
relevant factors in the fatigue process and support the
concept that the behaviour of coactive muscles persists
during progressive fatigue.
Acknowledgments This work has been done with the support of the
Spanish Ministry of Education and Science (Plan Nacional I?D?i
2004–2007 Strategic Action: ‘Sport and Physical Activity’. Ref:
DEP2006-56076 and SAF2007-65383) and the Public University of
Navarre. Current address of Professor Dr. Mikel Izquierdo is Public
University of Navarra, Department of Health Sciences, Navarra,
Spain. Current address of Dr. Cristina Granados is University of the
Basque Country, Physical Education Department, Alav, Spain.
Conflict of interest No known conflicts of interest associated with
this publication, and there has been no significant financial support for
this work that could have influenced its outcome.
References
Aagaard P, Simonsen EB, Andersen JL, Magnusson SP, Bojsen-
Moller F, Dyhre-Poulsen P (2000) Antagonist muscle coactiva-
tion during isokinetic knee extension. Scand J Med Sci Sports
10:58–67
Arabadzhiev TI, Dimitrov VG, Dimitrova NA, Dimitrov GV (2010)
Interpretation of EMG integral or RMS and estimates of
‘‘neuromuscular efficiency’’ can be misleading in fatiguing
contraction. J Electromyogr Kinesiol 20:223–232
Basmajian JV, DeLuca CJ (1985) Muscles alive. Their functions
revealed by electromyography. Williams and Wilkins, Baltimore
Bentley DJ, Smith PA, Davie AJ, Zhou S (2000) Muscle activation of
the knee extensors following high intensity endurance exercise
in cyclists. Eur J Appl Physiol 81:297–302
123

Bigland-Ritchie B, Cafarelli E, Vollestad NK (1986) Fatigue of
submaximal static contractions. Acta Physiol Scand Suppl
556:137–148
Bogdanis GC, Nevill ME, Boobis LH, Lakomy HK, Nevill AM
(1995) Recovery of power output and muscle metabolites
following 30 s of maximal sprint cycling in man. J Physiol
482(Pt 2):467–480
Brody LR, Pollock MT, Roy SH, De Luca CJ, Celli B (1991) pH-
induced effects on median frequency and conduction velocity of
the myoelectric signal. J Appl Physiol 71:1878–1885
Carolan B, Cafarelli E (1992) Adaptations in coactivation after
isometric resistance training. J Appl Physiol 73:911–917
Casey A, Constantin-Teodosiu D, Howell S, Hultman E, Greenhaff
PL (1996) Metabolic response of type I and II muscle fibers
during repeated bouts of maximal exercise in humans. Am J
Physiol 271:E38–E43
De-Luca C (1984) Myoelectric manifestations of localised muscular
fatigue in humans. Crit Rev Biomed Eng 11:251–279
Dimitrov GV, Arabadzhiev TI, Mileva KN, Bowtell JL, Crichton N,
Dimitrova NA (2006) Muscle fatigue during dynamic contrac-
tions assessed by new spectral indices. Med Sci Sports Exerc
38:1971–1979
Draganich LF, Jaeger RJ, Kralj AR (1989) Coactivation of the
hamstrings and quadriceps during extension of the knee. J Bone
Joint Surg Am 71:1075–1081
Enoka R (1995) Mechanisms of muscle fatigue: central factors and
task dependency. J Electromyogr Kinesiol 5:141–149
Escamilla RF, Fleisig GS, Zheng N, Barrentine SW, Wilk KE,
Andrews JR (1998) Biomechanics of the knee during closed
kinetic chain and open kinetic chain exercises. Med Sci Sports
Exerc 30:556–569
Escamilla RF, Fleisig GS, Lowry TM, Barrentine SW, Andrews JR
(2001) A three-dimensional biomechanical analysis of the squat
during varying stance widths. Med Sci Sports Exerc 33:984–998
Farina D, Merletti R, Enoka RM (2004) The extraction of neural
strategies from the surface EMG. J Appl Physiol 96:1486–1495
Fuglevand AJ, Zackowski KM, Huey KA, Enoka RM (1993)
Impairment of neuromuscular propagation during human fatigu-
ing contractions at submaximal forces. J Physiol 460:549–572
Gandevia SC (2001) Spinal and supraspinal factors in human muscle
fatigue. Physiol Rev 81:1725–1789
Gerdle B, Larsson B, Karlsson S (2000) Criterion validation of
surface EMG variables as fatigue indicators using peak torque: a
study of repetitive maximum isokinetic knee extensions. J Elec-
tromyogr Kinesiol 10:225–232
Gollnick PD, Armstrong RB, Sembrowich WL, Shepherd RE, Saltin
B (1973) Glycogen depletion pattern in human skeletal muscle
fibers after heavy exercise. J Appl Physiol 34:615–618
Gonzalez-Izal M, Malanda A, Navarro-Amezqueta I, Gorostiaga EM,
Mallor F, Ibanez J, Izquierdo M (2010) EMG spectral indices
and muscle power fatigue during dynamic contractions. J Elec-
tromyogr Kinesiol 20:233–240
Gorostiaga EM, Navarro-Amezqueta I, Cusso R, Hellsten Y, Calbet
JA, Guerrero M, Granados C, Gonzalez-Izal M, Ibanez J,
Izquierdo M (2010) Anaerobic energy expenditure and mechan-
ical efficiency during exhaustive leg press exercise. PLoS One
5:e13486
123
Eur J Appl Physiol
Greenhaff PL, Nevill ME, Soderlund K, Bodin K, Boobis LH,
Williams C, Hultman E (1994) The metabolic responses of
human type I and II muscle fibres during maximal treadmill
sprinting. J Physiol 478(Pt 1):149–155
Hassani A, Patikas D, Bassa E, Hatzikotoulas K, Kellis E, Kotzama-
nidis C (2006) Agonist and antagonist muscle activation during
maximal and submaximal isokinetic fatigue tests of the knee
extensors. J Electromyogr Kinesiol 16:661–668
Hultman E, Greenhaff PL (1991) Skeletal muscle energy metabolism
and fatigue during intense exercise in man. Sci Prog 75:361–370
Izquierdo M, Gonzalez-Badillo JJ, Hakkinen K, Ibanez J, Kraemer
WJ, Altadill A, Eslava J, Gorostiaga EM (2006) Effect of
loading on unintentional lifting velocity declines during single
sets of repetitions to failure during upper and lower extremity
muscle actions. Int J Sports Med 27:718–724
Karatzaferi C, Myburgh KH, Chinn MK, Franks-Skiba K, Cooke R
(2003) Effect of an ADP analog on isometric force and ATPase
activity of active muscle fibers. Am J Physiol Cell Physiol
284:C816–C825
Kellis E (1999) The effects of fatigue on the resultant joint moment,
agonist and antagonist electromyographic activity at different
angles during dynamic knee extension efforts. J Electromyogr
Kinesiol 9:191–199
Kellis E, Baltzopoulos V (1996) The effects of normalization method
on antagonistic activity during concentric and eccentric isoki-
netic knee extension and flexion. J Electromyogr Kinesiol
6:235–245
Koopman R, Manders RJ, Jonkers RA, Hul GB, Kuipers H, van Loon
LJ (2006) Intramyocellular lipid and glycogen content are
reduced following resistance exercise in untrained healthy males.
Eur J Appl Physiol 96:525–534
Masuda K, Masuda T, Sadoyama T, Inaki M, Katsuta S (1999)
Changes in surface EMG parameters during static and dynamic
fatiguing contractions. J Electromyogr Kinesiol 9:39–46
McCartney N, Heigenhauser GJ, Jones NL (1983) Power output and
fatigue of human muscle in maximal cycling exercise. J Appl
Physiol 55:218–224
Osternig LR, Caster BL, James CR (1995) Contralateral hamstring
(biceps femoris) coactivation patterns and anterior cruciate
ligament dysfunction. Med Sci Sports Exerc 27:805–808
Psek JA, Cafarelli E (1993) Behavior of coactive muscles during
fatigue. J Appl Physiol 74:170–175
Soderlund K, Greenhaff PL, Hultman E (1992) Energy metabolism in
type I and type II human muscle fibres during short term
electrical stimulation at different frequencies. Acta Physiol
Scand 144:15–22
Solomonow M, Baratta R, Zhou BH, Shoji H, Bose W, Beck C,
D’Ambrosia R (1987) The synergistic action of the anterior
cruciate ligament and thigh muscles in maintaining joint
stability. Am J Sports Med 15:207–213
Weir JP, McDonough AL, Hill VJ (1996) The effects of joint angle on
electromyographic indices of fatigue. Eur J Appl Physiol Occup
Physiol 73:387–392
Westerblad H, Allen DG, Bruton JD, Andrade FH, Lannergren J
(1998) Mechanisms underlying the reduction of isometric force
in skeletal muscle fatigue. Acta Physiol Scand 162:253–260