# Metabolomics Approach for Sweat Mapping the Performance of Soccer Players in Pretoria, South Africa
## 3 Results
### Metabolic Pathway Analysis  
The Pathway Analysis Module employed the KEGG _Homo sapiens_ (human) for analysis, the global test for enrichment analysis, and relative-betweenness centrality for topological analysis (Figure 5). To determine the distinct sweat metabolic pathways engaged in soccer players' pre- and post-exercise, the conduct test used \(p<0.05\) and Impact \(>0.1\) as screening parameters. Eighteen pathways were found when significantly different metabolites were employed in KEGG. Alanine, aspartate and glutamate metabolism, Valline, leucine, and isoleucine degradation as well as nicotine and nicotinamide metabolism are the primary pathways implicated in the key significantly altered metabolites (\(p<0.05\)) of sweat in soccer players following high-intensity interval training, as is also clear from Table 3. The pathway employed by KEGG _Homo sapiens_ for Alanine, aspartate, and glutamate metabolism is presented in Figure 6.  
Figure 5: Global test for enrichment analysis and a relative-betweenness centrality topology analysis for significant metabolite measurement in particular pathways of sweat samples. The y-axis signifies pathway impact, and the x-axis signifies pathway enrichment. Darker colors and larger sizes denote greater pathway enrichment and higher pathway impact values, respectively.  
Figure 6: Primary metabolite pathway of Alanine, aspartate, and glutamate metabolism for sweat samples induced from pre- and post-exercise. Red lines: Carbohydrate metabolism and amino acid metabolism. Green part: Represents the interconversion of alanine, aspartate, and glutamate, facilitated by transamination reactions.  
\begin{table}
\begin{tabular}{l c c c c}  & **Hits \({}^{a}\)** & \(p\)**-Value** & **Holm \(p\)\({}^{b}\)** & **Impact Value** \\ \hline Alanine, aspartate and glutamate metabolism & 3 & 0.001 & 0.080 & 0.224 \\ Valine, leucine and isoleucine biosynthesis & 2 & 0.001 & 0.115 & 0.000 \\ Valine, leucine and isoleucine degradation & 3 & 0.003 & 0.222 & 0.023 \\ Nicotinamide and nicotinamide metabolism & 2 & 0.005 & 0.407 & 0.194 \\ Pantotheate and CoA biosynthesis & 2 & 0.009 & 0.712 & 0.000 \\ Galactose metabolism & 2 & 0.017 & 1.000 & 0.356 \\ Glyoxylate and dicarboxylate metabolism & 2 & 0.023 & 1.000 & 0.074 \\ Argin biosynthesis & 1 & 0.102 & 1.000 & 0.000 \\ D-Amino acid metabolism & 1 & 0.109 & 1.000 & 0.000 \\ Histidine metabolism & 1 & 0.116 & 1.000 & 0.000 \\ Glycerolipid metabolism & 1 & 0.116 & 1.000 & 0.237 \\ Selenocon compound metabolism & 1 & 0.143 & 1.000 & 0.000 \\ Citrate cycle (TCA cycle) & 1 & 0.143 & 1.000 & 0.090 \\ beta-Alanine metabolism & 1 & 0.149 & 1.000 & 0.000 \\ Pentose phosphate pathway & 1 & 0.162 & 1.000 & 0.000 \\ Glutathione metabolism & 1 & 0.194 & 1.000 & 0.007 \\ Sphingolipid metabolism & 1 & 0.219 & 1.000 & 0.000 \\ Cysteine and methionine metabolism & 1 & 0.225 & 1.000 & 0.022 \\ Glycine, serine and threonine metabolism & 1 & 0.225 & 1.000 & 0.215 \\ \hline \end{tabular}  
* Hits \({}^{a}\): represents the number of metabolites in one pathway. Holm \(p\)\({}^{b}\): indicates the statistical \(p\)-values that were further adjusted using the Holm–Bonferroni method for multiple tests.  
\end{table}
Table 3: Outcomes from the sweat metabolomic pathway analyses of soccer players pre- and post-exercise.