## 2 Results and Discussion
### Qualitative and Semiquantitative Analysis of Area Nuts
A qualitative analysis of crushed areca nuts was performed under optimized conditions using HS-SPME/GC-MS. A total of 41 compounds were identified (Table 3). There are different groups of identified compounds, and most of them belong to terpenes and higher fatty acids. Arecolin, a typical compound in area nuts [25; 26], has also been identified. In total, 77 compounds were identified by GC-MS analysis in three fractions (SHDE extract, distillation residue, and hydrolate). The list of identified compounds is summarized in Table 3. Based on the evaluation of the relative content of individual substances, it can be said that fatty acids are the dominant volatile compounds in areca nuts. The biggest difference among the three fractions was observed in the content of terpenes, the SHDE extract was rich in terpenes more than the other two fractions, and HS-SPME extract. T terpenes (namely monoterpenes, sesquiterpenes, and terpenoids) made up more than half of the compounds identified in the SHDE extract. The highest relative amount of the SHDE extract had terpinen\(-\)4-ol. This compound belongs to the group of monoterpenoids, which is known to be a potent bactericidal agent [27]. Camphor had the second highest relative amount in the SHDE extract, and this terpenoid has a very strong aroma. Camphor exhibits many biological properties; one of them is the antimicrobial effect [28]. The main differences among the extracts are in the profiles of individual groups of compounds. Figure 3 shows the comparison of the chemical composition of areca nut extracts using bubbles whose sizes correspond to the representation of the individual compounds (the percentage peak area method uses the peak area of the target component \begin{table}
\begin{tabular}{c c c c c c} \hline \hline
**GROUP OF COMPOUNDS** & **Retention Index** & **CAS Number** & \begin{tabular}{c} **HS-SPME** \\ **Extraction** \\ \end{tabular} & \
\begin{tabular}{c} **SHDE Extract** \\ **Resolute** \\ \end{tabular} &
\begin{tabular}{c} **Distillation** \\ **Resolute** \\ \end{tabular} & **Hydrolate** \\ \hline \(\Sigma\) **TERFNES and TERFNESOIDS** & & - & **4.74 (22)** & - & **35.18 (8)** \\ \hline
**OTHER COMPOUNDS** & & & & & \\ \hline
2,6,6-trimethyl-2-ethernyletrahydroproyran & 969 & 7392-19-0 & - & 0.01 & - & - \\
2-2-penylfuran & 989 & 3777-69-3 & - & 0.16 & - & - \\
4-ethoxy-1,5,5-trimethylcyclopentene & 1058 & 1727-69-1 & - & 0.16 & - & - \\
4-Acetyl-1-methylcyclohexene & 1130 & 6090-09-1 & - & 0.01 & - & - \\ Arecoline & 1225 & 63-75-2 & 0.80 & - & 1.85 & - \\ Arecoline & 1287 & 4180-23-8 & 0.76 & 0.42 & - & - \\
4-Nonletrahydro-2H-prran-2-one & 1950 & 2721-22-4 & - & 0.05 & - & - \\ \hline \(\Sigma\) **OTHER COMPOUNDS** & & **1.56 (2)** & **0.83 (7)** & **1.85 (1)** & \\ \hline \hline \end{tabular}
\end{table} 
Table 3: \(Cont.\)  
Figure 2: Venn diagram: comparison of the identified compounds in the four fractions obtained using different treatment methods for the areca nut sample. as a proportion of the total area of all detected peaks). In terms of the number of compounds identified, the SHDE extract was the richest, as can be seen in Figure 3. In this extract, a broad spectrum of different compounds was identified, most of them terpenes and terpenoids (32 out of 72 identified compounds) with RI ranging from 925 to 1547. Fatty acids have the highest relative amount in all four extracts, concretely 65.51 rel.% in HS-SPME extraction, 85.22 rel.% in SHDE extract, 96.55 rel.% in distillation residue, and 48.53 rel.% in hydrolate. In the case of identified acids, fatty acids with RI were observed ranging from 982 to 2135 in the 4 extracts. Dodecanoic acid was identified in all four extracts. Compounds with RI <90 were not observed in any extract chromatograms due to solvent cuts during chromatographic analysis. The fewest compounds were identified in the distillation residue. Only eight compounds were detected, and fatty acids (dodecanoic acid and tetradecanoic acid) had the highest relative amounts.