Isolation and identification of the chemical composition of Salvia Species growing in Iraq by GC-MS

ABSTRACT


Introduction
The Salvia is colloquially known as sage and is considered the largest genus in the family Lamiaceae according to [1], the Lamiaceae is well-known throughout the world for its economic significance in the food and pharmaceutical industries, there are more than 700 species of Salvia plants that are found worldwide [2,3], in the flora of Iraq, Salvia is present 33 species [4], also recorded a new species, so the number of species in Iraq is 34 [5]. The name Salvia means to save and is derived from the Latin word salvere. This probably refers to its healing properties, as some of its species have historically been used in traditional medicine.
Salvia is an important and useful plant, some are famous for their medicinal, aromatic, and antioxidant where they have been used as volatile compounds and aqueous extracts of some types of Salvia for the treatment of characteristics [6], some Salvia species such as S. officinalis, S. viridis, and S. multicaulis possess antioxidant properties sore mouth and throat, excessive sweating, indigestion, and stomach disorders [1,7].
According to [8] Salvia species exhibit great chemical diversity, [9] pointed out Salvia is chemically rich in various kinds of natural compounds, especially flavonoids, monoterpenoids, triterpenoids, polysaccharides, and phenolic acids. This may be due to the nature of Salvia growth, as some species of Salvia can be grown in gardens as ornamental plants, and some of them grow in the wild [10].
One of the most crucial techniques for figuring out the taxonomic placements of taxa is chemotaxonomic research. Today, even individual genotypes and low and high taxonomic levels can have their chemical profiles studied [11].
The purpose of the current study was to evaluate the chemical diversity of the Salvia genus and its usefulness in the chemotaxonomic of Salvia.

Material and methods: 2.1 Plant material collection:
Samples were gathered in several locations around the Iraqi Kurdistan area: the wild grown of Salvia was S. indica, S. multicaulis, S. spinosa, S. reuterana, S. russellii, and S. officinalis was cultivated in Ramadi, during the months of May and June ( Figure 1).
After cleaning and drying the samples at room temperature, they were identified using specimens kept in the College of Science's herbarium at the University of Baghdad for subsequent use.

Preparation of plant extract:
The samples (aerial part) were extracted at the Desert Studies Center University of Anbar, according to the method mentioned by [12]. Where the dry samples were crushed by the electric grinder, 2g of the dry powder samples were taken with 20 ml of absolute ethyl alcohol, and the samples remained for 48 hours at room temperature, then added 10 ml distilled water to each sample, and put in an Ultrasonic bath for 20min to increase the extraction efficiency, the samples were filtered with filter papers and evaporated using a hot plate at 50‫ﹾ‬C.

Analysis of extracts by GC-MS:
The examination of Chemical compounds present in the sediment was carried out using the GC-MS device in the laboratories of Ibn Al-Baytar Center of the Ministry of Industry and Minerals, Gas Chromatograph: Agilent (7820A) USA GC Mass Spectrometer Analytical Column: Agilent HP-5ms Ultra unit (30 m length x 250 µm inner diameter x 0.25 µm film thickness), Injection volume 1 µl, Pressure 11.933 psi, GC Inlet Line Temperature: 250 ˚C, Aux heaters Temperature 300 ˚C, Carrier Gas: He 99.99%. (Figure 2).
The degree of similarity between the studied species was determined by analyzing the hierarchical cluster using the Past program ( Figure 3).

Results and Discussion:
Table (1) shows some of the important chemical compounds that were obtained from the analysis of plant sample extracts with the (GC-MS) device.
In S. spinosa and S. russellii, the number of the chemical compounds have been identified as (18,22) in S. spinosa and S. russellii respectively.
Depending on the presence of chemical compounds between the studied species, a diagram was drawn showing the degree of convergence and similarity between the studied species in Figure (3), where it was divided into three clusters: the first cluster (S. multicaulis, S. indica, and S. russellii), the second cluster (S. spinosa and S. reuterana) and finally the third cluster (S. officinalis).
Chemical characteristics have been adopted in the diagnosis of plant families, genera, and species, such as the presence or absence of various chemical compounds that were important taxonomic characteristics in the diagnosis of genus and species, and from these studies [13,14,15]. This study showed clear and important variations in the content of chemical compounds and their concentration, in all samples, it was found that n-Hexadecanoic acid was the dominant component, but in a varying percentage between the studied species. some chemical compounds appeared in one species only and did not appear in the other species, so they may be the distinguishing signs of this type, and we believe that it is possible to rely on them in distinguishing morphologically similar species.
In addition, the species of the genus Salvia, especially the most famous S. officinalis, have been proven to possess a wide range of biological and pharmaceutical substances and have antimicrobial activity for many bacteria and fungi [16,17,18,19].

Conclusion
we have significant differences in terms of the presence or absence of different chemical compounds between species, which indicates that it is possible to rely on these results to support the classification between species, especially as there is a morphological similarity between the S. reuterana and S. spinosa.