H.

Kuipers
G. Rietjens
F. Verstappen
Effects of Stage Duration in Incremental Running Tests H. Schoenmakers
on Physiological Variables G. Hofman
Training & Testing

Abstract with different stage duration. The mean V4 was significantly

lower in the 6 min per stage protocol compared with the 3 min
To study the effect of stage duration on some physiological vari- per stage protocol (12.9 vs. 14.4 km/h). Mean ventilatory
ables in an incremental running test, 8 well-trained runners threshold was not different between the 1, 3 and 6 min per stage
underwent 3 running tests, with stage durations of 1, 3 and protocols. No threshold behaviour was found in respiratory rate.

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6 min. To study maximal lactate steady state (maxLASS) and its
corresponding speed, every subject underwent a 4th test with
three 15-min stages at three speeds, based on the running speed
at 4 mmol/l blood lactate (V4) in the 6 min per stage protocol.
The first load in the 15 min per stage test was V4 ± 0.5 km/h, the
second at V4, and the third V4 + 0.5 km/h. To compare the max-
LASS speed with outdoor performance, the subjects also ran 5 km
at this speed on an outdoor track. Mean maximal running speed
(Vmax) in the incremental test was significantly lower in the 6-
min (15.1 km/h) and 3-min stage protocol (17.1 km/h), compared
with the 1-min stage protocol (18.3 km/h). Mean peak VÇO2 and
486 mean peak heart rate were not different between the protocols
MaxLASS can be estimated from V4 in the 6 min per stage proto-
col, and verified by three 15-min intensities being V4 ± 0.5 km/h,
at V4, and V4 + 0.5 km/h. The mean blood lactate concentration
at the maxLASS speed was not different between treadmill run-
ning and outdoor running on a track. In conclusion, for measur-
ing peak values of physiological variables in an incremental run-
ning test, the duration per stage is of less importance, however,
when measuring blood lactate concentration as a function of
running speed, the duration per stage should be at least 6 min.
Key words
Lactate steady state ´ ventilatory threshold ´ breathing frequency

Introduction ious studies because of differences in variables used, definitions,

A high maximal oxygen uptake (VÇO2max) is an important pre-
requisite for endurance performance. However, VÇO2max intensi-
ty can only be sustained for approximately 7 ± 10 min [2, 6,10].
Several endurance events last beyond 10 min and are performed
at an intensity below the maximal oxygen uptake [2, 9,10, 21].
Studies in which the relationship between plasma lactate
response, VÇO2max, and endurance performance were investigat-
ed showed a higher correlation between endurance performance
and lactate variables such as anaerobic threshold, than with
VÇO2max [7,18, 23]. During the last decade several papers have
been published on lactate variables as indicators of training
adaptations and predictors of exercise performance. In using lac-
tate variables and thresholds, it is difficult to compare the var-
methodology and measuring techniques.
Threshold indices are usually assessed in incremental exercise
tests, often using the blood lactate response as a function of exer-
cise intensity. A potential drawback of lactate measurements in
incremental exercise tests is that it may take 3 min or more to
establish steady plasma lactate concentrations at a given exer-
cise intensity [3,11,19, 20, 27]. This would imply that studies on
lactate measurements in incremental exercise tests in which
stages of less than 3 min were used, may have failed to assess
the adequate plasma lactate concentrations belonging to a given
exercise intensity, and consequently assessed improper thresh-
old values. The duration per stage and the ramp slope appears
to be related to the blood lactate response, as has been shown in

Affiliation
Department of Movement Sciences, University Maastricht, The Netherlands

Correspondence
H. Kuipers, M.D., Ph.D ´ Department of Movement Sciences ´ University Maastricht ´ PO Box 616 ´
6200 MD Maastricht ´ The Netherlands ´ E-Mail: Harm.kuipers@bw.unimaas.nl

Accepted after revision: February 15, 2003

Bibliography
Int J Sports Med 2003; 24: 486±491  Georg Thieme Verlag Stuttgart ´ New York ´ ISSN 0172-4622
cycle ergometer exercise [24, 26]. Stockhausen et al. [24] pro-
posed for cycling a duration of 3 min when using a 20 W incre-
ment and a stage duration of 4 min when using a 30 W incre-
ment. A potential drawback of incremental exercise protocols
with relatively long duration of each stage is that peak values of
VÇO2 and heart rate may not be reached, because of premature
fatigue, while for full exercise diagnostics maximal values may
be relevant as well. In contrast to cycling, only few studies have
been conducted on running exercise to assess the most suitable
protocols for measuring peak values of VÇO2, heart rate and also
for measuring lactate response to incremental exercise. One of
the few studies conducted in this respect on running is from
Foxdal et al. [11] who investigated lactate variables with incre-
mental protocols with different stage duration. This study focus-
sed on lactate and it was not searched for peak values. They
found that lactate equilibrium between muscle and blood
requires at least 8 min in running exercise.
To confirm as well as extend the data obtained by Foxdal et al.
[11] and to study peak values as well, the present study was con-
ducted in which our first aim was to compare the response of
oxygen uptake, heart rate, and blood lactate concentration as
well as their peak values in incremental running protocols in
which the duration per stage was different (1 min, 3 min, and
6 min). A second aim was to relate the blood lactate response
and ventilation rate obtained in the different protocols with the
maximal lactate steady state (maxLASS) and to compare the
blood lactate concentration at the maxLASS speed on the tread-
mill with that during outdoor track running at the same speed. A
third aim was to shed light on conflicting data about a deflection
in ventilatory rate as intensity indicator reported by some inves-
tigators [8,12], but not confirmed by others [15].
Material and Methods
Subjects
After approval from the local ethical committee 6 male and 4
female well-trained competitive middle distance runners of
regional and club level entered the study after written informed
consent. All subjects trained at least 4 times per week for at least
one hour per training session, and all had been in training for at
least 2 seasons prior to the study. The subjects ran between 60 ±
150 km per week, and the training included interval training at or
slightly above the individual race pace. Because of injuries two
male subjects were unable to complete participation in the
study. The mean age of the remaining 4 male subjects was
32  12 y, mean weight 72  6 kg, mean height 186  10 cm. For
females the mean age was 35  15 y, mean weight 57.5  6 kg,
and mean height 167  6 cm.
The study took place during the summer (May ± June), and all
subjects had been training before the start of the study for at
least three months, in preparation for the race season.
Procedures
The subjects refrained from strenuous exercise 24 h before each
exercise test. After the subjects reported to the lab, a venous
catheter was inserted into a forearm vein. A stopcock was con-
nected to the catheter, to enable collection of blood. The subject
Kuipers H et al. Stage Duration in Incremental Running Tests ¼ Int J Sports Med 2003; 24: 486 ± 491
was connected to a heart rate monitor (Polar, Kempele, Finland),
and wore a face mask, connected to a portable metabolic system
(Cosmed K4). Subsequently the subjects stepped on the tread-
mill (Medifit, The Netherlands) after which the test was started.
A 1 % incline of the treadmill was maintained, in order to match
energetic cost of outdoor running at the same speed [14].
Exercise protocols
To study the physiological response in an incremental running
test with different duration per stage, 3 incremental running
protocols were performed on a treadmill. The order of the first
three protocols was random, and at least one week separated
two consecutive tests.
Training & Testing
Protocol A started with a warming up of 5 min at 8 km/h in
women and 10 km/h in men, and thereafter the exercise intensity
was increased by 1 km/h every minute until the subject was
unable to continue.
Protocol B also started with a 5 min warming up at 8 km/h for
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women and 10 km/h for men, and thereafter the running speed
was increased by 2 km/h every 3 min, however, from a heart
rate of 85 % of the individual maximal heart rate the speed incre-
ment was 1 km/h every three min.
Protocol C was similar to protocol B with the only difference that
every stage lasted 6 min.
If in protocols B and C a stage could not be completed to the full
length the maximal running speed (Vmax) was calculated accord-
ing to the equation:
487
Vmax = Vcompleted + t/T ” speed increment
in which Vcompleted is the running speed of the last stage that was
completed, t the number of seconds that the uncompleted run-
ning speed could be sustained, T the number of seconds required
to complete the stage, and speed increment is the speed load
increment.
To assess maxLASS, a fourth protocol D was conducted, that con-
sisted of 3 incremental workloads each lasting 15 min, preceded
by a 5 min warm up at 50 % of the previously assessed maximal
running speed attained in the 6 min per stage protocol. The three
intensities of protocol D were based on the running speed from
protocol C at which the blood lactate concentration was
4 mmol/l (V4). The first speed was 0.5 km/h below V4, the second
at V4, and the third 0.5 km/h above V4. Maximal lactate steady
state (maxLASS) is defined as the running speed during constant
running at which the blood lactate concentration between 10
and 15 min of running exercise varied less than 0.5 mmol/l. For
drawing blood the subject interrupted running for 15 ± 20 s by
stepping next to the belt.
To compare the blood lactate concentration at the speed of max-
LASS measured on the treadmill and running outdoor, the sub-
jects also ran 5 km without interruption on a 400-m tartan track
at the maxLASS speed as assessed on the treadmill. In order to
maintain a steady speed a supervisor on a bicycle remained in
 Table 1 Values (mean  SD) of maximal running velocity (Vmax), peak VÇO2, peak heart rate (PeakHR), peak blood lactate concentration (Peak-
La), the running speed at 4 mmol/l blood lactate (V4), and the running speed at breakpoint VE/V ÇO2 (Break VE/V ÇO2) in incremental
protocols with stage length of 1 min (protocol A), 3 min (protocol B), and 6 min (protocol C)

Protocol Vmax PeakVÇO2 PeakHR PeakLa V4 BreakVE/ VÇO2

(km/h) (l/min) (beats/min) (mmol/l) (km/h) (km/h)

A 18.3  1.9 4.48  1.13 183  6 14.0  1.8
B 17.1  1.6# 4.46  1.04 187  5 11.0  2.5 14.4  1.4 13.3  1.6
C 15.1  1.5## 4.41  1.01 189  5 10.8  2.6 12.9  1.5# 13.4  1.4

# p < 0.05 between C and B.

## p < 0.01 C compared with A.
Training & Testing

close proximity of the running athlete and clocked the split time
every 200 m, and gave feedback to the athletes for maintaining
the proper velocity. Immediately after completing the 5-km run,
a venous blood sample was taken for lactate measurement.

Variables
Respiratory variables and heart rate were measured continuous-

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ly, and displayed every 30 s on a portable computer that was
connected to the receiving unit of the K4 metabolic system.

The ventilatory threshold, based on VE/VÇO2 was determined

from the respiratory variables by the software provided with
the Cosmed K4 measuring system using linear regression, and
defined as the running speed at which the VE/VÇO2 started to Fig. 1 Example in one male subject of blood lactate concentration
depart from linear increase [25]. In addition a break point in the (mmol/l) as function of treadmill speed in two incremental exercise
linear increase of the ventilation rate was searched for by visual protocols with stages lasting 3 min (squares), and 6 min (circles).
inspection and with linear regression analysis.

488 From the venous catheter a 2 ml blood sample was drawn during
the final 30 s of every stage, except for protocol A (increments
every minute) in which no blood samples were taken, because
of too short time available for blood sampling. From all samples
the first 0.5 ml of the sample was discarded because of dilution
with saline. After sampling the catheter was flushed with saline.
To obtain a blood sample the subject grasped the railing and
stepped next to the running belt. Blood sampling required 15 to
20 s during which the running speed for the next stage was
adjusted. Immediately after sampling the subject stepped back
on the running belt and continued running. In protocol D (15-
min stages) for assessing maxLASS, blood was sampled every
5 min. The blood lactate concentration in whole blood was meas-
ured immediately after sampling (EML-105, Radiometer, Copen-
hagen, Denmark).
V4 was determined by straight line interpolation of the blood
lactate curve.
Data handling and statistical analysis
The data obtained during the last 30 s of the various protocols
were used for analysis. To define a breakpoint in VE/VÇO2 and
respiratory rate as function of running speed, these data were
analysed with regression analysis.
The other data were analyzed with a SPSS software packet ana-
lysed using a repeated measure ANOVA. Depending on the
ªMauchly`s test of sphericityº an uni- or multi-variate test was
employed. Differences between the protocols were analyzed
with a paired t-test. In case of univariate testing a Bonferroni cor-
rection was applied. Differences were considered statistically
significant at p < 0.05.
Results
The mean peak running speed during the incremental protocols
was inversely related to the duration of the stages (Table 1). The
highest mean peak running speed was attained in protocol A in
which the load was increased by 1 km/h every minute (mean
18.3  1.9 km/h), which was significantly different from the peak
running velocities in the protocol with 3-min stages
(17.1  1.6 km/h; p < 0.05), and with 6-min stages (15.1  1.5 km/
h; p < 0.01).
Mean peak VÇO2 (Table 1) was slightly higher in the protocol with
1-min stages, compared with the protocol with 6-min stages,
although the difference did not reach statistical significance.
From Table 1 it can be seen that the mean peak heart rate was not
significantly different among the incremental protocols.
The blood lactate responses were different between the 3- and 6-
min protocols (p < 0.025), i. e. in all subjects the curve obtained
from the 3 min per stage protocol was shifted to the right com-
pared with the 6-min protocol (Fig. 1). The mean peak blood

Kuipers H et al. Stage Duration in Incremental Running Tests ¼ Int J Sports Med 2003; 24: 486 ± 491

Fig. 2 Example of blood lactate concentrations (mmol/l) in one fe-
male subject during the maxLASS test at V4 ± 0.5 km/h (11.0 km/h),
at V4 (11.5 km/h), and at V4 + 0.5 km/h (12.0 km/h).
lactate concentration in the protocol with 3-min stages was not
different from the protocol with 6-min stages. Since the 4 mmol/l
intensity is often used as parameter for endurance capacity, the
running speed at which the blood lactate concentration was
4 mmol/l (V4) was assessed in both protocols with 3- and 6-min
stages. V4 was significantly higher in the 3 min per stage proto-
col (14.4  1.4 km/h) compared with the 6 min per stage protocol
(12.9  1.5 km/h; p < 0.05).
As can be seen from Table 1 the running velocity at which VE/VÇO2
departed from a linear increase was 14.0  1.8 km/h in the 1 min
per stage protocol, 13.3  1.6 km/h in the 3 min per stage protocol,
and 13.4  1.4 km/h in the 6 min per stage protocol (p > 0.05).
Using linear regression as well as visual inspection no threshold
behaviour was found in respiratory rate during incremental exer-
cise in any of the protocols with different stage duration.
The running speed of the 15-min protocol was based on the run-
ning speed at 4 mmol/l blood lactate concentration (V4) in the
6 min per stage protocol. The running speed at V4 in the incre-
mental 6-min protocol was 12.9  1.5 km/h, whereas the running
speed at which the maxLASS was reached in the 15 min per stage
protocol was 12.9  1.7 km/h (p > 0.05). The mean blood lactate
concentration reached at maxLASS was 6.03  1.47 mmol/l (range
between individuals 3.8 ± 7.2 mmol/l). All subjects completed the
15 min at V4, and were able to start with a running speed of V4
+ 0.5 km/h, however, 6 stopped prematurely because of fatigue.
Blood lactate concentrations at V4 + 0.5 km/h increased progres-
sively (varying between individuals from 3.2 to 10.9 mmol/l).
Only 2 subjects were able to finish the 15 min with difficulty,
and blood lactate levels rose to 8.5 and 10.2 mmol/l in these two
subjects, respectively (Fig. 2).
The mean blood lactate concentration after the 5-km run on the
outdoor track was 6.32  2.12 mmol/l and was not different from
the mean blood lactate concentration at the maxLASS speed dur-
ing the 15-min protocol. All subjects experienced the speed at
maxLASS as strenuous, however, all indicated that this was be-
low the competitive speed.
Kuipers H et al. Stage Duration in Incremental Running Tests ¼ Int J Sports Med 2003; 24: 486 ± 491
Discussion
The present study demonstrates that in a protocol in which the
running speed was increased by 1 km/h every min, the maximal
running speed is on the average 1.2 km/h higher compared to a
protocol in which the running speed was increased every 3 min,
and on the average 3.2 km/h higher compared with a protocol in
which the running speed was increased every 6 min. The higher
maximal running speed in a protocol with a short time per stage
can be explained by a greater contribution of anaerobic power at
the later load steps, due to the delayed lactate accumulation that
is associated with local muscle fatigue. Similar to cyclists where
maximal power output on a cycle ergometer can be used as a
parameter for endurance capacity [17, 22], also in runners the
Training & Testing
peak running velocity can be used as parameter of endurance
capacity. Therefore, when reporting maximal running speed in
an incremental protocol, the duration per stage has to be taken
into account.
Although the mean peak oxygen uptake in the 6 min per stage
protocol was slightly lower than the two other protocols, this dif-
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ference was not statistically significant. This implies that for
measuring VÇO2max and peak heart rate in an incremental proto-
col on a treadmill the duration per stage is of less importance and
may vary from 1 to 6 min.
The results of the present study show that when the duration per
stage in incremental protocols is shorter than approximately
6 min, blood lactate concentrations lag behind the corresponding
running velocity. Thus for measuring blood lactate concentra-
tions belonging to a given exercise intensity, a duration of 5 ±
6 min at a minimum per stage appears to be required. This is in
line with the findings of Foxdall et al. [11] who compared the 489
blood lactate response in incremental treadmill protocols with
4-, 6- and 8-min stages, and suggested that stages of at least
8 min are required for adequate lactate equilibrium between
muscle and blood. This would also imply that incremental run-
ning exercise protocols for measuring blood lactate parameters
such as the 4 mmol/l intensity, load steps less than approximate-
ly 5 min yield non-steady state values for blood lactate. The exer-
cise intensity at which blood lactate concentration equals
4 mmol/l is often used as a parameter for endurance capacity
[1, 7]. When looking at the incremental protocol with different
duration per stage at the running velocity at which blood lactate
concentration is 4 mmol/l (V4), we found that V4 was on the
average 1.5 km/h lower in the protocol with 6 min per stage,
compared with the protocol with 3 min per stage. This probably
reflects the insufficient time for lactate equilibration between
muscle and blood in the incremental protocol with 3-min stages.
This implies that also when using other lactate parameters (i. e. 1
or 2 mmol/l above resting levels), the time per stage has to be
taken into account.
An important finding in the present study was that maximal lac-
tate steady state (maxLASS) speed can be estimated from the
running speed at V4 in the incremental running test with 6-min
stages, and be measured by a second incremental protocol with
15 min per stage on a separate day. The mean running speed at
V4 in the incremental protocol with 6-min stages was not differ-
ent from the mean velocity at maxLass. However, on an individ-
 ual basis the running speed at maxLASS could be in the range of
plus or minus 1.0 km/h of V4. The mean blood lactate levels in
the 15-min protocol were significantly higher than the 4 mmol/l
at V4 in the 6-minute protocol. A relevant finding is that the
speed at maxLass can be assessed just by doing an incremental
protocol with 6-min stages. However, for assessing the highest
blood lactate levels at which a steady state can be maintained,
exercise of longer duration is required. This supports the finding
that for attaining a true steady state in blood lactate concentra-
tion a duration of 6 min is still insufficient. The 15-min duration
at a given exercise intensity for assessing maximal lactate steady
state is shorter than used by other investigators who used 20 ±
30 min of continuous exercise [4,11,16]. Jones and Doust [16]
proposed to adopt 30 min constant exercise for assessing max-
Training & Testing
LASS in running. However, their results suggest that when no
steady state is reached in approximately 10 min the exercise is
not at maxLASS. Jenkins and Quigley [13] also observed that
blood lactate levels remained stable between 10 and 30 min of
continuous exercise. The 15-min protocol was based on a pilot
study conducted before the present investigation, in which in 5
subjects the blood lactate response during 30 min exercise per
stage was studied. It was found that when no steady state (in-
crease < 0.5 mmol/l) in blood lactate concentration was attained
between 10 and 15 min, no steady state was attained in 30 min.
Fluctuations in blood lactate concentrations beyond 20 ± 30 min
may be attributed to a shift in substrate utilization from carbo-
hydrate towards fat, inducing a different level of lactate steady
state. The validity of 15-min duration for assessing maxLASS
was supported by the outdoor 5-km run at the maxLASS velocity
that lasted 20 ± 25 min. The longer duration elicited the same
blood lactate concentration as the 15-min treadmill run in the
laboratory. Therefore, maxLASS speed can be approximated by
490 V4 obtained from an incremental protocol with 6-min stages,
whereas the accurate assessment of lactate levels requires a pro-
tocol consisting of 15-min stages. The mean maximal blood lac-
tate concentration at which a steady state could be maintained
was 6.02 mmol/l (range 3.8 ± 7.2 mmol/l). The considerable range
reflects the different response between individuals.
Ventilatory threshold is also used as parameter for endurance
capacity. In the present study ventilatory threshold was defined
as the running velocity at which VE/VÇO2 started to increase non-
linearly [25]. The results of the present study show that the run-
ning speed at which the non-linear increase in VE/VÇO2 started,
was not different between stage durations of 1 to 6 min (Table 1),
which is in line with the findings of Zhang et al. [28]. This sup-
ports the assumption that in the experiments conducted, venti-
latory response is not directly related to blood lactate response,
and is a variable which is hardly affected by stage duration in
incremental protocols.
Another aim of the present study was to investigate whether
breathing frequency shows threshold behaviour and can be
used as a parameter for exercise intensity and endurance per-
formance. The results show that the pattern of breathing fre-
quency is rather variable, and failed to show a stable and consis-
tent pattern. Therefore no clear and consistent threshold could
be detected in most subjects. This is in line with the study by
Jones and Doust [15], but appears to be in contrast to a study by
Cheng et al. [8], and James et al. [12]. Cheng et al. [8], however,
Kuipers H et al. Stage Duration in Incremental Running Tests ¼ Int J Sports Med 2003; 24: 486 ± 491
employed the Dmax method, which always yields some kind of
threshold as long as the variable increases non-linearly, irrespec-
tive of a real threshold behaviour. The study of James et al. [12]
indicates that breathing frequency can be used as an indicator
of anaerobic threshold, defined as the disproportionate increase
in breathing frequency. However, in the present study no consis-
tent disproportionate increase in breathing frequency could be
found and indicates that respiratory rate is not a useful variable
to be used as threshold variable, or marker of exercise intensity
in runners.
A question that is relevant for translating laboratory results into
sport practice is the relationship between laboratory tests and
running speeds in competition. A point that has received little
emphasis in recent sport scientific literature is the observation
that there is no such thing as one specific speed at which endur-
ance performances are done. When the average speeds of world
records at different distances in certain disciplines are compared
it appears that the average speed declines with increasing dis-
tance. This indicates that there is not one single critical speed,
but it decreases with increasing distance. This is supported by
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the results from the study of Jones and Doust [16] who studied
the relationship between 8 km running performance and lactate
parameters, and Bentley et al. [5] who studied the relationship
between lactate variables and 20-min and 90-min time trials.
Foster et al. [10] studied lactate levels during a simulated time
trial over 5 km in cyclists, and observed that plasma lactate levels
increased throughout the trial. From the present study it is
concluded that the running speed at 5 and 10 km are difficult to
predict from lactate variables obtained during running tests in
the laboratory. Therefore, lactate threshold variables may be
helpful for assessing endurance characteristics and training-in-
duced adaptations in athletes, however have limited value for
predicting endurance performances.
In conclusion, peak VÇO2 and peak heart rates can be obtained in
incremental protocols with increments varying from 1 to 6 min.
When the relationship between blood lactate levels and running
velocity are required, the duration per stage should be at least
6 min. An important finding is that the maxLASS running
velocity can be estimated from running speed at 4 mmol/l (V4)
in an incremental protocol with 6 min per stage. However, for
precise assessment and for corresponding blood lactate levels,
stages of 15-min duration at V4 ± 0.5 km/h, at V4, and 0.5 km/h
above V4 can be used.
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