Measurement of sweat lactate levels in exercise and non-exercise activities using capillary electrophoresis system with contactless conductivity detection and cyclodextrin-modified buffer

Abstract

This study presents the analysis of lactate in sweat using capillary electrophoresis with contactless conductivity detection. Two cyclodextrin (CD) compounds, carboxymethyl-β-cyclodextrin sodium salt (CM-β-CD) and heptakis (2,3,6-tri-O-benzoyl)-β-cyclodextrin (TRIME-β-CD), were evaluated as buffer modifiers for the separation of chloride, nitrate, sulfate, oxalate, maleate, acetate, lactate, and phosphate anions. The concentrations of these modifiers were tested at 0.05, 0.1, and 0.5 mM for eight anions separation. The buffer was 40.0 mM MES/L-His (pH 6.0) with 0.05 mM CTAB, resulting in anodic separation. Due to separation efficiency in resolving lactate and phosphate peaks, TRIME-β-CD at a concentration of 0.1 mM was selected as the suitable buffer additive for sweat lactate analysis. The calibration of the developed method, using maleate as an internal standard, resulted in the linear ranges of 0.1–5.0 mM, r2 of 0.9999, and an instrumental LOD of 0.042 mM. The intra-day and inter-day precisions of the relative migration time (RMT) were 0.3–0.4% and 3–4% RSD, respectively. Lactate levels in sweat samples from the same group of volunteers were measured after two distinct activities: exercise (after running) and non-exercise (after sauna). For non-exercise activities, lactate concentrations ranged from 15 ± 1 mM to 36 ± 2 mM. While, after exercise, the concentrations were between 26 ± 1 mM and 136 ± 1 mM. Further evaluation of the stability of sweat storage for monitoring lactate contents, it was found that the changes in lactate levels were insignificant when stored for up to 24 weeks at 4 °C. This was confirmed by paired t-tests (t-stat ragnes from -0.59 to 2.49; t-critical = 2.57, P = 0.05). The measured lactate concentrations from volunteers were significantly higher, approximately 2–6 times, in exercise activities compared to non-exercise activities. This substantial difference in lactate secretion indicates a clear distinction in the factors that stimulate sweat lactate production. To evaluate the intensity of exercise based on lactate levels in sweat, a proposed cut-off level of 36.0 mM, with a decision limit of 40.0 mM, can be proposed.

Keywords

• Lactate
• Sweat
• Exercise
• Cyclodextrin
• Capillary electrophoresis
• C4D