Detection of Some Fatty Acids and Phenolic Compounds in the Seeds of Plant in Coriander (Coriandrum Sativum L.) Seeds Cultivated in Iraq

Coriander sativum (Linn) is an important medicinal plant belonging to the family of Apiaceae. It is a well-recognized plant in the traditional medicine and used by people in treatment of gastrointestinal problems and rheumatism. In the present investigation, after phytochemical screening of the seeds of coriander results revealed the presence of four fatty acid compounds (Lauric, Palmatic, Stearic and Linoleic acids) in the Pet-ether extract, while two fatty acids (Palmatic and Stearic acids) were found in the Chloroform extract. Phenolic compounds and their fractions Quercetin and Benzoic extract were also identified in the Ethyl acetate . Also, Quercetin was identified in the extracts of (F2) and (F3), Moreover, IMS extract was contained of high concentration of Gallic acid while Quercetin were identified the fractions (F4, F5 and F6) of IMS extract. Other phenolic compounds were presented as low concentrations in other extracts.


Introduction
Coriander (Coriandrum sativum L.) is an annual Apiaceae herb [1]. Recent studies have also demonstrated a hypoglycaemic action and effects on carbohydrate metabolism [2]. It has also been reported the antimicrobial effect of coriander leaves and seeds against several microorganism [3][4].
The yield and chemical composition of coriander seed essential oil varies both qualitatively and quantitatively in relation to the method of extraction type of the cultivar and the area of harvest [5]. Coriander seed essential oil is mainly composed of linalool, together with some other oxygenated monoterpenes and monoterpene hydrocarbons. The phenolic compounds, apigenin, catechin, coumaric acid, aliphatic alkenals and alkanal were reported in Coriandrum sativum of aerial parts [6,7]. While linalool, geranyl acetate, and petroselinic acid were found in the fruits [8]. The fruits contain vegetable oil with a high concentration of monounsaturated fatty acids especially of petroselinic acid [9]. Moreover, multiple extracts of different polarity from leaves and seeds of coriander (Coriandrum sativum) and coriander oil were investigated for their antioxidant activity, total phenolic content was also quantified as well [2]. Also, coriander seed oil methyl esters as biodiesel fuel was investigated and contained an usual fatly acids hitherto unreported as the principle component in biodiesel fuels petroselinic acid and the remaining fatty acid profile consisted of common 18 carbon constituents such as Linoleic, Oleic and also Stearic acid [9].
vacuum rotary evaporator at 50°C. The crude of each extract was kept in Deep freez till us it for further studies.

2.3Alkaline hydrolysis (Saponification) of pet-ether (60-80)°C and chloroform extracts [11]:
Pet-ether and Chloroform extracts as resulted from preparation of plant extracts, 10 ml of each extracts were added to 100 ml of 7.5 M of solution of KOH in Methanol: water (3:2) and refluxed in a round bottom flask for 90 min at 100°C. The mixture was allowed to cool at room temperature and 50 ml of distilled water added.
The crude was extruded with diethyl ether (2×25 ml) to remove unhydrolylized lipid.
The aqueous layer was acidified with 10% H2SO4, up to PH=2. The fatty acids were extracted with diethyl ether (2×25 ml). Drying of two combined extracts with anhydrous sodium sulfate, filtered and concentrated in vacuo to give 1.92 gm of saponified pet-ether and 2.70 gm of saponified chloroform extracts respectively.

Esterification of fatty acid compounds:
To prepare of methyl esters, 0.5 ml of acetyl chloride was added to 125 ml of methanol with stirring. A sample of 2 ml of dry fatty acids was added to above mixture then boiled under reflux in water bath for 30 min, cooled then and for GLC .

GLC Analysis:
The esters were analyzed by using GLC, a type of (Shimadzo-14A) equipped with a denl flame ionization detectors held at 250°C A (3m × 1.8 inch) international diameter column packed with 3% SE-30 on Teflon (100-120 ml) was held at 100°C initially then raised at 5°C/min to 250°C. Samples of (1ml) was used for injection. The identification of fatty acids were determined by reference to a standard of a known composition [14]. After defatted of coriander seeds, using pet-ether and chloroform,seed were-extracted with IMS to get the IMS extracts and their fractions which were obtained by using column chromatography (CC) of each extracts, ethyl acetate & IMS, using different solvents to get three fractions from Ethyl acetate (F 1 , F 2 , F 3 ) and also three fractions from IMS (F 4 , F 5 , F 6 ) using silica gel (60-120 mesh) as packing malaria. Also acid hydrolyses was carried out on ethyl acetate, IMS and also on there obtained fractions using 1N HCl and reflux for 30 min.

Kirkuk University Journal /Scientific Studies (KUJSS)
After cooling the mixture, liberated free phenolic compounds were dissolving in ethyl acetate (2×10 ml), which also concentrated to 1 ml, using rotary vacuum evaporator [13].

HPLCanalysis:
Free phenolic compounds were investigated by HPLC instrument, type (Shimadzo, LC 2010 AHT), after purification with filters (0.1 μm) with 320 nm as wave length, with (Acetonitrile:H 2 O, 85-15%) as mobile phase using a column C18 (4×240 mm) at 30°C and the runs carried out at Education college in Mosul university, identification of these phenolic compounds was carried out by using standard compounds as 0.1 gm was dissolved in 10 ml ethanol [14]. Samples were injected on to the HPLC bed manually with injection volume as 20 ml.
To identify the peaks, the spectral patterns and retention time of the samples were compared with standard. Phenolic compounds were quantified by comparing the peak areas and retention times obtained for the extracted sample with the peak areas and retention times of appropriate standards of phenolic compounds.
The content of phenolic compounds in the samples was calculated, using the following equation [15],

Results and Discussion:
1-Chromatographic identification of fatty acid compounds presented in pet-ether & chloroform extracts using GLCanalysis. Petroselinic acid C 18 :1n-12 was the fatty acid marker in edible plant glycolipid (GL) substances obtained from coriander seed oil, followed by linoleic acid C 18 :2n-6 [16].
From the above result, we noticed the absence of Quercetin from the IMS extracts and their fractions while Hydroquinone was the main compound that presented and also identified of these mentioned extracts.