**3. Results**

The SPME analysis of scent of *H. hircinum* showed the presence of elemicin (spicy, floral scent) as the main component (61.71%) in the presence of a relevant amount of eugenol (4.50%) (Table 2). Minor products observed in the scent were 3,4,5-trimethoxybenzaldehyde, benzyl benzoate, 3-(4,8,12-trimethyltridecyl)furan, and 9-tricosene. All the values are based on per cent of the TIC area.


**Table 2.** SPME-GC-MS analysis of *Himantoglossum* species.

#### *Compounds* **2021**, *1*


#### *Compounds* **2021**, *1*

**Table 2.** *Cont.*

The scent of the sample of *H. adriaticum* collected in Abruzzo (Sample 1) showed the presence 4-amino-5-(4-morpholinylmethyl)-2-oxazolidinone, β-ocimene (green tropical woody floral vegetable scent), decyl decanoate, and 9-tricosene as main components (17.37, 8.23, 5.36, and 6.41%, respectively) (Table 2). Furthermore, several compounds are presents in relevant amounts: 1-methoxy-4-methylbenzene (1.63%), geraniol (1.49%), geranyl acetate (1.43%), methyleugenol (2.34%), pentadecane (2.98%), elemicin (2.95%), hexadecane (2.70%), heptadecane (2.11%), benzyl benzoate (2.17%), 6,10,14-trimethyl-2-pentadecanone (1.83%), and 11-hexadecen-1-ol acetate (2.42%). The sample of *H. adriaticum* collected at Marsico Nuovo (Sample 2) has an aroma where the main component was pentadecyl hexanoate (22.73%), 9-tricosene (5.06%), methyleugenol (5.93%), tetradecane (3.93%), pentadecane (3.41%), and elemicin (3.80%) (Table 2). The samples of *H. adriaticum* collected at Viggianello (samples 3 and 4) showed some similarities in the scent: the main components were 9-tricosene (31.21% in sample 3 and 40.22% in sample 4) and methyleugenol (18.06% in sample 3 and 4.73% in sample 4) (Table 2). Furthermore, β-ocimene was found in relevant amount (10.28%) only in sample 3.

## **4. Discussion**

It is interesting to note the large differences between our reported results and those reported in the introduction section for *H. hircinum*. We did not find a correlation with the results reported by Schiestl and Cozzolino [27]. While they determined the presence of high molecular weight alkanes, we did not find these compounds in the scent. We think that the observed different results depend on the different procedures used in the determination of the scent. Schiestl and Cozzolino used an extraction of labella. Probably they determined the presence of waxy compounds present in the surface of labella but not involved in the composition of the scent. Furthermore, the components determined in that article [27] are not volatile compounds. Furthermore, in a previous work, (*E*)-ocimene, elemicin, (*E*)-3-methyl-4-decenoic acid, (*Z*)-4-decenoic acid, and lauric acid were determined as the main components of the aroma [24]. While elemicin was the main component of the scent in the sample we analyzed, we did not find the other compounds determined in the work of Kaiser [24]. Finally, we did not find (*E*,*Z*)- and (*E*,*<sup>E</sup>*)-2,6-dimethyl-3,5,7-octatrien-2-ol whose presence has been claimed in [25,26].

The scent of *H. adriaticum* has not been studied until now. It is interesting to note that all the samples we analyzed have common components, although in different amounts. Thus, methyleugenol and 9-tricosene were found in all the samples. Furthermore, some linear hydrocarbons (tetradecane, pentadecane, and hexadecane) were present. Nevertheless, sample 1 of *H. adriaticum* showed the presence of an oxazolidinone as an important component, while in sample 2 we found pentadecyl hexanoate, and in sample 3 β-ocimene was one the main components.

Based on this work, we can assume that the scent of *H. hircinum* found in Basilicata has a different composition from those described elsewhere. We can assume also that *H. hircinum* has a scent showing a different composition in comparison with the compounds found in *H. adriaticum*. Finally, we can assume that the samples of *H. adriaticum* we analyzed have some common characters, with some differences depending on the place where the samples have been collected.

## **5. Conclusions**

This work shows the analysis of samples from Basilicata and Abruzzo of *H. hircinum* and *H. adriaticum*. The analyses have been performed by using the same procedure and the same fiber in SPME-GC-MS, allowing to have a homogenous data set. The analysis of *H. hircinum* showed a peculiar composition that differs from those observed in *H. adriaticum*. In fact, the analysis of the scent of *H. hircinum* showed the presence of elemicin in the presence of a relevant amount of eugenol. The scent of the sample of *H. adriaticum* showed the presence 4-amino-5-(4-morpholinylmethyl)-2-oxazolidinone, β-ocimene, decyl decanoate, 9-tricosene, pentadecyl hexanoate, methyleugenol, tetradecane, pentadecane, and elemicin.

The observed differences, when other head-space techniques are used, can depend both on different absorption rates of the analytes on the fiber and on variation of the scent due to natural adaptation of the plant to different environmental conditions. Furthermore, the observed differences can be due to different pollination insects. The analysis of the scent of *H. adriaticum* showed the presence of some common components (i.e., 9-tricosene and methyleugenol) that are not depending on the origin of the flowers.

**Author Contributions:** Conceptualization, M.D. and V.A.R.; methodology, R.R.; investigation, R.R., L.V. and M.M.; data curation, R.L.; writing—original draft preparation, review and editing, M.D., V.A.R. and R.L.; All authors have read and agreed to the published version of the manuscript.

**Funding:** This research received no external funding.

**Institutional Review Board Statement:** Not applicable.

**Informed Consent Statement:** Not applicable.

**Data Availability Statement:** Not applicable.

**Conflicts of Interest:** The authors declare no conflict of interest.
