Phytochemical Analysis of Methanolic Extract of Jordanian Melissa officinalis L

The methanolic extract obtained from leaves, steam and seeds of M. officinalis growing in Karak city-Jordan, was investigated, discussed for its phytoconstituents for the first time and analyzed by GC-MS instrument with BSTFA and Heptan solvents. Plant parts were collected during the spring semester 2018. Results showed that leaves extract have twenty one major and minor natural compounds available in all parts. Six of them which were above 1 % have been found in the extract of leaves more than other parts of the plant. Leaves extract are a good source of various phytoconstituents. These natural compounds gave leaves an important role to use it as anticancer and sedative due to the presence of Palmitic acid and polar compounds. Leaves are much interesting part due to the availability of these bioactive compounds as major and minor compounds more than other parts. Thus, the isolation of leaves phytochemical compounds will give fruitful results for further detailed study.


Introduction
In recent years, fruits and plants are classified as an important source of main active biologically compounds. There are 85-90% of the world's populations use plants as traditional herbal medicines according to the World Health Organization (WHO) reports (Selvamangai & Bhaskar, 2012).
Many people in Jordan use traditional medicinal plants as additional, alternative or complementary medicine (Aldal'in, 2018). However, most of them haven't detailed phytochemical and main biologically active compounds. One of these plants is called Jordanian Melissa (Al-Khalil, 1995). Melissa officinalis L. (M. officinalis) is a member of the Lamiaceae family, known as Lemon balm. It is distributed and commonly in Mediterranean region and western Asia. This plant is cultivated in Jordan and largely used as an important folk medicinal plant species (Schultze et al., 1993). Melissa has been traditionally used for the treatment of different medical purpose as tonic, colic, depression, nervousness and stress induced headaches (Beloued, 2009), antiseptic, calmative (Baytop, 1984), antimicrobial effect against Gram positive and Gram negative bacteria (Vitullo et al., 2011, Larrando et al., 1995, antifungi (Abdellatif et al., 2014). Furthermore, it can be used as antitumor activity (Galasinski, 1996), anti-HIV activity (Yamasaki et al., 1998), antiviral (Adorjan andBuchbauer, 2010), antidiabetic activity (Chung et al., 2010). M. officinalis has been reported that contain natural products that can inhibits protein biosynthesis in the cancer cells (Adorjan and Buchbauer, 2010). It has been also found that M. officinalis has stronger than vitamins B and C as antioxidant effects (Ghayoor et al., 2010). The bioactivity of M. officinalis refers to contain phenolic acids such as rosmarinic acid and caffeic acids (Constantine, 2007;Tagashira & Ohtake, 1998) and flavonoids (Ziakova et al., 2003). Triterpene (Mencherini et al., 2007) and phenylpropanoid heteroside was reported (Mulkens & Kapetanidis, 1988). Moreover, it was also found to treat externally such as gout, herpes, and sores, insect repellent (Bown, 1995).
The methanolic extract and volatile oils of aerial parts of Jordanian M. officinalis from Wadi Rujb-Ajlun were showed anti-proliferation effects against two types of colorectal (SW480), (HCT116), and prostate (PC3) cancer cell lines (Barakat et al., 2016).
The comparison of chemical composition of methanolic extract of leaves stems and seeds of Jordanian M. officinalis by GC/Ms haven't been investigated. Moreover, the contents of natural products of M. officinalis may vary based on the location. Therefore, the aim of this paper is to analyze and compare the chemical composition of

Discussions
Heptan and N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) derivatization solution were used as a nonpolar and polar solvents in GC/MS analysis, respectively. The GC-MS outputs can be analyzed to get a good information about nonpolar (fatty acids, hydrocarbons) and polar functionalities such aromatic and aliphatic alcohols, carbonyl compounds and carbohydrates. Furthermore, BSTFA derivatization can be used to improve peak shape of natural compounds by reducing tailing and this will lead to characterize unknown ketones, alcohols, and fatty acids by GC-MS (Stalling et al., 19968).
The GC-MS chromatograms of methanolic extract of M. officinalis showed twenty one major peaks available in all parts. The analysis of these peaks described that leaves have the highest contents in BSTFA of different natural products. All peaks were identified by comparing their mass with NIST08s library.
A later step of the present work was to find the phytoconstituents by using GC-MS analyses, where the composition of M. officinalis contributes significantly to the biological effects (Bakkali et al., 2008). From table 1, it was noticed that six components of which were above 1 % have been found in the extract of leaves. These compounds Cyanuric Acid 1.52%, 3-OH-Tetradecenedioic Acid 8.21%, 3-OH-Dodecenedioic Acid 9.87%, Glucuronic-1-Pent 2.19%, phenobarbitol 1.54% and α-D-Glucopyranose 9.36% were found as major compounds with the highest percentage than others. Otherwise, the extract of leaves was found also rich in the minor compounds compared with other parts of the plant.
The leaves content have several uses such as Palmitic acid which was reported as antimicrobial effects, cytotoxicity against MCF-7, WRL-68, CaCo2, and Colo-320 cancer cell lines, it can possess significantly hepatoprotection against galactosamine and cholesterolaemic (French, 2002;Barbara, 2002). Furthermore, most of these contents have traditionally been gave M. officinalis a memory enhancing properties and it is widely used as a sedative (Kennedy et al., 2004).

Conclusion
GC-MS analysis of methanolic extract of leaves, stems, and seeds of M. officinalis showed the presence of several of the important components like Fatty acids, polar compounds and their biological activities. The extract of leaves is much interesting due to having highest contents of major and minor natural compounds than other parts of the plant. These results are the first step to understand the nature of active ingredient in this medicinal plant. Thus, the presence study will be helpful for further detailed work.