PHYTOCHEMICAL VALUATION OF THE UMBELS OF RIDOLFIA SEGETUM (L.) MORIS OF MOROCCO

Jamila EL Karkouri 1,2 , Smail Amalich 1,2 , Aziz Drioiche 1,2 , Kamal Fadili 1,2 , Bruno Eto, Youssef Khabal and Touriya Zair 1,2 . 1. Research team of Chemistry Bioactive Molecules and the Environment, University Moulay Ismail, Faculty of Sciences, BP 11201, Zitoune, Meknès, Morocco. 2. Laboratory of Materials Chemistry and Biotechnology of Natural Products, University Moulay Ismail, Faculty of Sciences, B.P. 11201 Zitoune, Meknès, Morocco. 3. Laboratoire Trans Cell Lab, Faculté de Médecine Xavier Bichat 16, rue Henri Huchard, BP 416, 75870 Paris cedex 18 ...................................................................................................................... Manuscript Info Abstract ......................... ........................................................................ Manuscript History

In our study of medicinal plants, as sources of natural bioactive substances, we were specifically interested in the species Ridolfia segétum (L.) Moris harvested in the area of Mchraâ Bel Ksiri Gharb in Morocco during June 2015. Fresh umbels that have been extracted from the essential oils (HE), by applying the hydrodistillation method, gave an average yield about 2, 51±0, 05. The chemical composition of essential oils, determined by gas chromatography coupled with the spectroscopy of mass, shows that the main components of the outcome oil from the fresh umbels are: l'α-pinene (1,18%), l'α -Phellandrene (6,71%), p-cymene (4,91%), %), the β -Phellandrene (2.41%), γ-terpinene (8,25%), paracymene-8-ol (1,8%), and the myristicin (70,27%). The identification of the non-volatile chemical families was determined by performing the phytochimical screening on the various extracts of Ridolfia segétum (L.) Moris by colorimetric and/or precipitation tests. The results of the Phytochemical screeningshowed that this species is rich in sterols and triterpenes, in Antraquinones combined genus c-heteroside type, flavonic heterosides and flavonoïde catechols, it contains an average quantity of mucilage as well. Though, it countains low quantity of alkaloid and is devoid of the leucoanthocyanes, the saponosides, gallic and catechic tannins, oses and holosides and the free antraquinones, the richness of this species in secondary metabolites, particularly in HE, provides high therapeutic effects also explains their use in traditional medicine. Furthermore, a high percentage of myristicin (70, 27%) certainly makes it an important natural source of this compound.
Ridolfia Segetum (L.) Moris is a plant with Mediterranean distribution, extending to Anatolia and Macaronésie, adventitious in Western and central Europe (Thellung, 1926) when it was introduced especially with grains of wheat (Thellung, 1912), But quoted as a woolly weed in Great Britain, France and Germany (Thellung, 1926, Lousley, 1961; it was also found in 1961 on the gravels of the Meuse in the Netherlands, of possible wool origin (Vanoststoom and Reichgelt, 1962). Ridolfia segetum common to Morocco in the harvests especially in the Gharb. It is presented in all Morocco except Sahara, it is plentiful especially on clay soils (Tanji, 2005). In Moroccan folk medicine, the infusion of the plant is used as stomachic, and fruits sprayed of Ridofolia segetum are put in decoct with the madder and the egg. The egg is then eaten in the treatment of the jaundice (Belakhdar J, 1997).
The species has been reported as consumed raw in salads, cooked and medicinal in case of gastric acidity. It is used in medicine to prevent constipation, cough and respiratory tract infections (Lentini and Venza, 2007). It is a plant consumed by the cattle especially before the blooming. These cleaned and peeled stems are consumed (Tanji and al 1995). It regulates women's menstruation and increases milk for lactating women, infusion of the grains helps fight constipation and intestinal gas (Adouane and al on 2015).
The essential oil of Ridofolia segetum has been the main topic of many studies, based on the quantification of the main compounds of essential oils, chemical characterization studies, allowed the distinction of two types of HE; One is largely dominated by monoterpene hydrocarbons such as α-phelandrene, terpinolene and p-cymene (Bicchi and al., 2009, Fleisher and Fleisher, 1996, Marongiu and al., 2007, Palá-Paúl and al ., 2002; And the one which contains phenylpropanoids as main compounds or in large amounts, generally myristicin and dillapiol (Jannet and Mighri, 2007;Jabrane and al., 2010). The botanical identification of the species was carried out at the laboratory of Floristics of the Scientific Institute in Rabat. The plant species is found in the spontaneous state. Vegetable matter consisting essentially of umbels at the time of fruiting for hydrodistillation.
Then, they were dried in the shade and finally reduced to fine powder in order to carry out the phytochemical screening. The necessary extracts were obtained by extraction with the following solvents: petroleum ether, methanol, ethanol, chloroform and distilled water.
Phytochemical screening was also based on the use of several reagents. The search for alkaloids was carried out by the Dragendorff reagent. The characterization of catechic tannins was carried out by isoamyl alcohol and hydrochloric acid and gallic tannins by the Stiasny reagent, sodium acetate and ferric chloride. For the detection of sterols and triterpenes, acetic anhydride and concentrated sulfuric acid were used. Dilute hydrochloric alcohol, magnesium chips and iso-amyl alcohol were used to search for flavonoids. Chloroform, dilute ammonia and hydrochloric acid have made it possible to investigate quinone substances.

Extraction of Essential Oils:-
Extraction of the essential oils was carried out by hydro distillation in a Clevenger device. Essential oils recovered from small opaque bottles are stored at 4 ° C. The yield of essential oil is evaluated from three extractions of fresh vegetable matter.

Yield Of Essential Oil:-
The moisture content is calculated after drying three samples of five (5) grams of plant material at 100 ° C. in an oven until a constant weight is obtained.
This rate is used to calculate the yield of essential oil. The yield is expressed as a percentage by volume of essential oil (ml) with respect to the mass of the dry plant material (g).
The calculation formulas are the following ones: With: % TH = Percentage of moisture content of the plant material (moisture content). % R = Extraction yield of essential oil expressed as volume of essential oil per mass of plant material (V / m) (mean of three yields). M1 = Initial mass of vegetable matter introduced into the furnace at time (t0). M2 = Final mass of vegetable matter in g removed from the furnace at time (tx). M = mass of plant material used for hydrodistillation (in the clevenger). V = Volume of essential oil collected (in mL). ± Standard deviation calculated by the Excel software (Microsoft Office 2007) on the basis of the different yields obtained.

Analysis and Identification of Eo Components By Gc / Ms:-
Chromatographiques analyses were carried out on a Hewlett Packard-type gas chromatograph (HP 6890 series), equipped with an HP-5 capillary column (30 mx 0.25 mm × 0.25 μm film thickness) An FID detector set at 250 ° C and lined with a mixture of H2 gas and air. The mode of injection is split, the carrier gas used is nitrogen with a flow rate of 1.7 ml / min. The temperature of the column is programmed at a rate of 4 ° C./min of 50 to 200 ° C. for 5 min. The device is controlled by an "HP Chemstation" computer system managing the operation of the apparatus and allowing the evolution of the chromatographic analyzes to be monitored. The GC-MS was performed on a Hewlett Packard chromatograph (HP 6890) coupled to a mass spectrometer (HP 5973 series). The fragmentation is carried out by electronic impact at 70 eV. The column used is an HP-5SM capillary (30 mx 0.25 mm x 0.25 μm). The temperature of the column is programmed at a rate of 4 ° C. /min of 50 to 200 ° C. for 5 min. The carrier gas used is helium whose flow rate is set at 1.7 ml / min. The injection mode is split.
The identification of the constituents of the studied HE was carried out both by the method of identification by Kovāts index (1965) and Adams (2007) and from the NIST 2002 mass spectrum data base.

Results and Discussion:-Phytochemical Screening:-
The results of each characterization reaction of the different chemical groups present in the umbels of Ridolfia segetum are summarized in Table 1.

Interpretation and Discussion:-
According to the table of results we find that Ridolfia segetum is rich in sterols and triterpenes, Antraquinones combined genus c-heteroside type, flavonic heterosides and flavonoid catechols, as it contains an average amount of mucilage. But it is poor in alkaloid and devoid of leucoanthocyanins, saponosides, gallic and catechic tannins, oses and holosides and free antraquinones. The yellowish flowers of Ridolfia segetum indicate that the umbels are rich in flavonoids. These were the first compounds studied in the genus Anethum. They are present in almost all organs of the plant and play an important role in the defense system as antioxidants. These secondary metabolites are known for their various biological properties: antioxidant, anti-inflammatory, antithrombic, antibacterial, antihepatotoxic, anti-tumor, antihypertensive, antiviral and antiallergic.
Essential oil yield:-Three test plugs of 5 g were introduced into three previously tared boxes. After 24 hours in the oven at a temperature of 103 ° C. ± 2 ° C. and then cooling in a desiccator, we reweighed again and we determined the new masses of the test samples. Table 3 summarizes the results relative to the yield of the essential oil extracted from the fresh umbels of Ridolfia segetum.  The chemical composition of the essential oil of Ridolfia segetum is described in the table 3. Table 3:-Chemical composition of the essential oil of Ridolfia segetum The coupling of gas chromatography with the mass spectrometry (CG / MS) allowed identifying in the essential oil of Ridolfia segetum 18 compounds. All these compounds (100 %) are distributed in monoterpenes (25, 34 %) and monoterpenes mono oxygenated (3, 49 %), monoterpenes trioxygenés (70, 94 %) ( Table 4). This essential oil is dominated in 91, 73 % by ten compounds compounds whose proportion is greater than 1%. These include : myristicin (70,27 %) , γ-terpinene (8,25 %), α -Phellandrene (6,71), p-cymène(4,91 %), α-pinène (1,18 %), β -Phellandrene (2,41%), and Para-cymene-8-ol (1,8 %). We notice the presence of myristicin, which is a peculiarity in the chemical composition of the essential oil of Ridolfia segetum both from a qualitative point of view (its presence) rather than a quantitative point of view (70.27%).

Main Chemical Compounds of Essential Oil:
The main chemical families of Ridolfia segetum's essential oil are described in the table 4.

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The chemical profile of the essential oil of Ridolfia segetum L) Moris varies significantly according to the regions of origin, plant material and extraction process . The chemical compounds that are found in the essential oil are also linked to the stage of development of the species as well as to organs selected for the extraction. And according to the product desired during the exploitation of the species, the selection of organs, vegetative stage and the region, turned out to be very useful to promote the obtaining of the very precise chemotypes.

Conclusion:-
In our work, we have studied the chemical composition of Ridolfia segetum (L) Morris's essential oil which allowed us to make the following observations: According to the bibliographical study, the essential oil obviously represents a considerable interest of the fact that they possess an apparent curative potential and have applications in the industrial sector.
The hydrodistillation, chosen method for the extraction of essential oil, has allowed us to show that the Ridolfia segetum L) Moris plant is in fact rich in gasoline.
The extraction of HE by hydrodistillation during three hours, allowed to obtain a yield of 2, 51%. The results of the qualitative and quantitative analysis of the chemical composition obtained using CG / MS, reveal that the gasoline studied is 70,27 % by aromatic majority compound the miristicin, the results of the phytochemistry shows that the species is rich in sterols and triterpenes, Antraquinones combined genome type c-heterosides, flavonoid heterosides and flavonoid catechols.
These highlighted bioactive molecules also contribute, to the virtues of the plant seen in particular the benefits of polyphenols and flavonoids.