BRIEF REPORT

Steady-State Acid-Base Response at Exercise Levels Close to Maximum Lactate Steady State
Pedro J.B. Peinado, MD,* Valter Di Salvo, MD,w Fabio Pigozzi, MD,w Ana I.P. Bermudez, MD,* Ana B. Peinado Lozano, MD,* Francisco J. Calderon Montero, MD,* and Nicola Maulli, MD, MS, PhDz

Objective: To study acid-base status during a constant-load treadmill test at exercise levels close to the maximum lactate steady-state. Design: Two tests were performed: one maximal and one steadystate at a load corresponding to the mean of the two ventilatory thresholds observed in the rst test. Setting: University Sports Science Laboratory. Participants: Twenty-three male Sports Science students, aged 26.7 ( 4.9 SD) years, 176.1 ( 6.3) cm in height, and weighing 72.8 ( 6.7) kg. Main Outcome Measures: Capillary blood gases, metabolites, electrolytes. Results: Acid-base status variables declined signicantly during the maximal test. Lactate concentrations were above the values observed at onset of blood lactate accumulation. All the blood variables did not vary signicantly at the various experimental times, except pH values and PCO2 values. Bicarbonate concentration remained constant. Plasma potassium, chlorine and sodium concentration did not increase. Conclusion: During a steady-state test at a load corresponding to approximately 80% of VO2max, the acid-base status in capillary blood remained constant even though the lactate concentration was over 4 mmol/L. However, despite the maintenance of a constant acid-base status, other physiological variables did not behave in the same fashion. Key Words: acid-base balance, exercise, steady state, hydrogenion concentration (Clin J Sport Med 2006;16:244246)

etabolic acidosis during maximal exercise is attributed to an increase in lactic acid concentration in the serum.13 Plasma and muscle are major buers, and help maintaining a constant pH. Little attention has been paid to the acid-base status at a constant load.2,47 Maximal Lactate Steady State (MLSS) is a good indicator of training status in endurance events,2,4,5 but its assessment can be extensive and complex.2,5 We tested the null hypothesis that the mean of the ventilatory threshold 1 and 2 is not an acid-base steady state. A secondary aim was to determine whether there were dierences in the acid-base status during a graded maximal test, and a constant-load steady-state test at intensities close to the MLSS.

SUBJECTS AND METHODS Subjects

Twenty-three (n = 23) male Sports Science students participated in the study. All subjects: 1) engaged in intense physical activity at least three times per week; 2) did not consume substances that might have interfered with the test results (e.g., alcohol, tobacco, anabolic steroids); and 3) did not have metabolic or other health problems, as assessed by pre-participation medical examination. Ethics committee permission was granted. All subjects gave written informed consent.

Tests
Expired air volume and composition was measured with a Jaeger Oxicon Pro (Erich Jaeger, Germany). Electrocardiography was performed using a 3-channel Hellige Cardiotest EK 53 (Hellige, Freiburg, Germany). The tests were conducted on a H/P/COSMOS PULSAR 3P 4.0 treadmill (h/p/cosmos sports & medical, NussdorfTraunstein, Germany). Capillary blood lactate concentration was measured with a YSI 1500 (YSI, Yellow Springs, USA). Peripheral blood gas concentrations were assessed using an ABL 77 (Radiometer, Copenhagen, Denmark).

Received for publication July 23, 2005; accepted January 10, 2006. From the *Facultad de Ciencias de la Actividad Fisica y el Deporte INEF, Universidad Politecnica de Madrid, Spain; wIstituto Universitario di Scienze Motorie, Dipartimento di Scienze della Salute, Roma, Italy; and zDepartment of Trauma and Orthopaedic Surgery, Kelle University School of Medicine, North Staordshire Hospital, Stoke on Trent, Staordshire, England. Reprints: Pedro J.B. Peinado, MD, Facultad de Ciencias de la Actividad Fisica y el Deporte-INEF (UPM), C/Mart n Fierro, S/N, Madrid 28040, Spain (e-mail: pedroj.benito@upm.es). Copyright r 2006 by Lippincott Williams & Wilkins

Research protocol
The maximal treadmill test used speed increments of 0.2 km/h every 12 seconds and a xed slope of 1%.8 Two independent observers detected the aerobic-anaerobic transition using the ventilatory method.9 The steady-state
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Steady-State Acid-Base Response

6,00 5,00 Lactato [mmoll-1] 4,00 3,00

Lactate [mmolL-1]

Bicarbonate (mmolL-1)

25,0 24,0 23,0 22,0 21,0 20,0 19,0 Bicarbonato (mmoll-1)

TABLE 1. Characteristics of the Subjects

Subject Characteristics Age (years) Height (cm) Body mass (kg) Duration of test (min) VO2 peak (l/min) VO2 /kg (ml/min/kg) VE (l/min) HRmax (beats/min) Speed (km/h) Pet O2 (kP) Pet CO2 (kP) Steady State 29.12 (0.5) 3.886 (0.447) 54.26 (5.7) 134 (17.7) 176 (11) 14.2 (1.3) 13.76 (0.4) 4.33 (0.4) 26.7 ( 4.9) 176.1 ( 6.3) 72.8 ( 6.7) Maximal Test 15.45 (5.2) O 4.813 (0.514) O 66.47 (7.49) O 166 (21.2) O 191 (7) O 17.4 (1.3) O 15.57 (0.6) O 5.35 (0.5) O

2,00 1,00 0,00 Rest 10 min. 20 min. 30 min.

18,0 17,0 16,0 15,0

FIGURE 1. Analysis of variance (ANOVA) results. Lactate and bicarbonate in [mmol/L] showing mean standard error. O = Differences between rest state and steady-state exercise (P < 0.01).

Mean SD (N = 23). VE indicates Ventilation; HRmax, Heart rate; Pet O2, End tidal O2 alveolar pressure; Pet CO2, End tidal CO2 alveolar pressure. O = Signicant dierences between the two tests with Pr0.01 (Students paired t-test).

test involved 30 minutes of running at a constant intensity between VT1 and VT2 (82.2 1.7% of VO2max). Fingertip capillary blood samples (at rest, every 10 minutes during steady-state test, and at the end of the test) were taken, and analyzed immediately for gas levels and lactate concentration.

RESULTS
Lactate and bicarbonate levels remained constants throughout the test. There were, however, signicant dierences with the values at rest (Fig. 1). There are signicant dierences (Fig. 2) between T10 and T30 for pH values and PCO2 values, which decrease signicantly (P < 0.05). There were signicant dierences between maximal and steady-state tests for all variables (Table 1 and 2). Apart from Na+ and PO2, all the other variables showed signicant dierences between steady state and maximal tests.

Statistical analysis
Descriptive statistics were calculated. We used the Students paired t-test to detect dierences between maximal and steady-state tests. Repeated measures analysis of variance (ANOVA) was used to detect dierences in the steady-state test results at 10, 20 and 30 minutes. Where dierences were observed, the post hoc Bonferroni test was applied to establish their direction. Statistical analysis was conducted using SPSS v11.0. Signicance was set at P = 0.01.

DISCUSSION
The work load at MLSS in this study was 82.2% (1.7) of VO2max.5,6 In other investigations, MLSS occurred at approximately 70% of VO2max.4 This dierence may result from the training status of the subjects, and the protocol and type of ergometer used in

7,65 7,6 7,55 7,5 7,45 7,4 7,35 7,3 7,25 7,2 7,15 Rest 10 min. 20 min. 30 min. 28 c 30 34 a 32 bc
pH

PCO

38

36

TABLE 2. Biochemical Variables in the Maximal and Steadystate Tests

Biochemical Variables LH (mmol/L) PH HCO3 (mmol/L) Na+ (mmol/L) K+ (mmol/L) Cl (mmol/L) Ca+2 (mmol/L) PO2 (mmHg) PCO2 (mmHg) Steady State 4.57 7.343 18.92 144.6 6.715 114.4 1.341 82.84 32.36 (1.84) (0.046) (2.33) (2.2) (1.036) (2.4) (0.087) (11.47) (2.34) Maximal Test 9.04 7.190 14.09 145.6 7.555 115.5 1.423 95.09 36.65 (2.24) O (0.045) O (1.71)O (3.3) (2.071)C (2.8)C (0.115)O (23.23) (4.57)O

FIGURE 2. Analysis of variance (ANOVA) results. pH, showing mean standard error, carbon dioxide pressure, showing mean standard error. A) Differences between rest state and steadystate exercise; B) Difference between 10th and 20th min of the test; C) Difference between 10th and 30th min of the test (P < 0.05).
r

Mean SD (N = 23). LH indicates Lactate concentration; PO2, Partial pressure of oxygen; PCO2, Carbon dioxide pressure. O = Signicant dierences between the two tests with P < 0.01 (Students paired t-test); C = Signicant dierences between the two tests with P < 0.05 (Students paired t-test).

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the test. The latter introduce the greatest variability into determining MLSS: MLSS ranges from 71.8 (5.9)% on a cycle ergometer to 78.1 (4.4)% of VO2max using a speedskating track.4 The load used in our study produced a steady-state test, as evidenced by the constant heart rates, VO2 levels and the lactate concentrations throughout the test. Also, all our subjects completed the tests. The acid-base state at a constant velocity does not stay stable in all variables. This is shown by the pH and the PCO2. Baron et al. work obtain a pH of 7.33 (0.03), 7.34 (0.04) and 7.35 (0.04) for 10, 20, and 30 minutes, respectively, of stable pH state in arterial blood, with signicant dierences (P < 0.05) between minutes 10 and 30.7 Our own pH data were 7.31 (0.06), 7.34 (0.06) and 7.35 (0.05) at 10, 20 and 30 minutes, with signicant dierences (P < 0.05) at the same time points in capillary blood. PCO2 data in Baron et al.s study were 40.8 (4.6), 39.4 (5.0) and 37.5 (5.0), with signicant dierences (P < 0.05) in all measurements.7 Our own data showed 35.79 (3.54), 31.29 (4.07) and 30.92 (3.68) for minutes 10, 20 and 30, with signicant dierences (P < 0.05) between minutes 10 and 20 and minutes 10 and 30. This may result from the fact that our measurements were performed using capillary blood.7,10 Lactate, bicarbonate and the other variables measured at steady state (except pH and PCO2) remained constant, demonstrating that lactate is by itself not the best indicator of a steady state. Our data agree with other authors. Nevertheless, it has to be assumed that we did not determine MLSS, but a steady state of lactate between the VT1 and VT2, which reduces in a signicant way the number of trials to performance, if further work conrms that it is indeed MLSS. Baron et al. used six trials to locate the MLSS, while our procedure will only need two.7 In conclusion, during a steady-state test at a load corresponding to approximately 80% of VO2max, the acid-base status analysed in capillary blood remained

constant even though lactate concentration was over 4 mmol/L. However, it should be kept in mind that, despite the maintenance of a constant acid-base status, other physiological variables did not behave in the same fashion. The utilization of ventilatory threshold as a rst approximation to MLSS should allow the reduction of the number of tests needed for its determination. ACKNOWLEDGEMENTS This study was nanced by Radiometer Espana. We are grateful for the support given by Sportsmed(UK) Ltd. REFERENCES
1. Foxdal P, Sjodin A, Sjodin B. Comparison of blood lactate concentrations obtained during incremental and constant intensity exercise. Int J Sports Med. 1996;17:360365. 2. Billat V. Marathon race is run around the maximal lactate steady state velocity (vMLSS). 10 Annual Congress of the European College of Sport Science-2005, Belgrade, 2005. 3. Kanauchi M, Ishikawa H. Acid-base balance during exercise. Nippon Rinsho. 1992;50:22542259. 4. Beneke R, von Duvillard SP. Determination of maximal lactate steady state response in selected sports events. Med Sci Sports Exerc. 1996;28:241246. 5. Harnish CR, Swensen TC, Pate RR. Methods for estimating the maximal lactate steady state in trained cyclists. Med Sci Sports Exerc. 2001;33:10521055. 6. Swensen TC, Harnish CR, Beitman L, et al. Noninvasive estimation of the maximal lactate steady state in trained cyclists. Med Sci Sports Exerc. 1999;31:742746. 7. Baron B, Dekerle J, Robin S, et al. Maximal lactate steady state does not correspond to a complete physiological steady state. Int J Sports Med. 2003;24:582587. 8. Myers J, Ashley E. Dangerous curves. A perspective on exercise, lactate, and the anaerobic threshold. Chest. 1997;111:787795. 9. Gaskill SE, Ruby BC, Walker AJ, et al. Validity and reliability of combining three methods to determine ventilatory threshold. Med Sci Sports Exerc. 2001;33:18411848. 10. Cabrera ME, Saidel GM, Calan SC. Lactate metabolism during exercise: analysis by an integrative systems model. Am J Physiol. 1999;277:R1522R1536.

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