# Seasonal Variability and Effect of Sample Storage on Volatile Components in _Calypogeia azurea_
## 2 Results and Discussion
### Statistical Analysis of the Obtained Results  
#### 2.6.1 Observation of Changes Depending on the Season  
Analyses carried out within individual years of the samples taken showed that in 2019 there were no significant differences between seasons in terms of volatile compounds aliphatics (\(p\) = 0.420), aromatics (\(p\) = 0.062), monoterpenes (\(p\) = 0.097), monoterpends (\(p\) = 0.368), and sesquiterpenoids (\(p\) = 0.717). However, a statistically significant effect of the difference between seasons in terms of undefined relationships was confirmed, F = 7.87; df = 2; \(p\) = 0.020, and sesquiterpenes, F = 11.31; df = 2; \(p\) = 0.003.  
As it turned out, the level of volatile compounds not identified was significantly higher in condition C-1 (\(p\) = 0.040) and was biased higher in condition C-9 (\(p\) = 0.058) compared to condition C-13. Furthermore, condition C-13 for sesquiterpene compounds showed higher levels of volatile compounds compared to C-9 (\(p\) = 0.003). No statistically significant differences were found in the other conditions in 2019 (Figure 4).  
Similarly to 2019, samples collected in 2020 showed statistically significant differences only for volatile sesquiterpene compounds, F = 9.64; df = 2; \(p\) = 0.008, and non-identified compounds, F = 10.87; df = 2; \(p\) = 0.004. No significant differences were found between seasons in 2020 for the remaining compounds: aliphatics (\(p\) = 0.779), aromatics (\(p\) = 0.449), monoterpenes (\(p\) = 0.368), monoterpends (\(p\) = 0.368), and sesquiterpenoids (\(p\) = 0.717). Analysis of pairwise comparisons between seasons in terms of sesquiterpenes and differences not identified showed that the level of undefined compound in sample C-22 was significantly higher than in sample C-26 (\(p\) = 0.004). In contrast, the intensity of sesquiterpene compounds in sample C-22 was significantly lower than in sample C-26 (\(p\) = 0.006). No significant differences were found in the other conditions.  
In 2021, the comparisons showed statistically significant differences in volatile sesquiterpenes, F = 7.86; df = 2; \(p\) = 0.020, and non-identified compounds, F = 9.16; df = 2; \(p\) = 0.010. The results of the analyses confirmed that the compounds not identified had higher levels in C-35 compared to C-39 (\(p\) = 0.012), while the sesquiterpene compounds had significantly lower severity in C-35 compared to C-39 (\(p\) = 0.027). No differences were found between C-31 and the other seasons. There were also no significant differences between seasons in 2021 for the remaining compounds: aliphatics (\(p\) = 0.424), aromatics (\(p\) = 0.486), monoterpenes (\(p\) = 0.529), monoterpends (\(p\) = 0.368), and sesquiterpenoids (\(p\) = 0.717).  
Furthermore, an analysis of differences was performed using the Kruskal-Wallis test for the severity of individual volatile compounds in each section from C-9 to C-39 separately to confirm the possibility that individual compounds were more prevalent. As it turned out, statistically significant differences were found for C-9 in 2019, H = 13.45; df = 6; \(p\) = 0.036, also C-22 in 2020, H = 14.09; df = 6; \(p\) = 0.029, and C-35 in 2021, H = 13.45; df = 6; \(p\) = 0.036. Analysis of pairwise comparisons showed that the severity of sesquiterpene compounds was always significantly higher compared to aliphatic compounds, and these differences were significant at the level of \(p\) < 0.05 level. No significant differences were found for the other volatile compounds.