Analysis of bioactive chemical compounds of Euphorbia lathyrus using gas chromatography-mass spectrometry and Fourier-transform infrared spectroscopy

The aim of this study was determination the phytochemical composition of methanolic seeds extract of Euphorbia lathyrus . Gas chromatography-mass spectrometry (GC-MS) analysis of E. lathyrus revealed the existence of the Carbonic acid, (ethyl)(1,2,4-triazol-1-ylmethyl) diester, 1H-Pyrrole,2,5-dihydro-1-nitroso, Hexanal dimethyl acetal, Isosorbide dinitrate, DL-Arabinose, Cyclopropane,1-fluoro-1-(2-bromoethenyl)-2,2,3,3-tetramethyl, α-D-Glucopyranoside, O-α-D-glucopyranosyl-(1.fwdarw.3)-ß-d-fruc, Desulphosinigrin, D-Glucose, 6-O-α-D-galactopyranosyl, Octanoic acid, Benzofuran,2,3-dihydro, 6-Acetyl-ß-d-mannose, Estragole, Ascaridole epoxide, 3-Allyl-6-methoxyphenol, 4-Amino-1,5,pentandioic acid, l-Gala-l-ido-octonic lactone, y-Sitosterol, Tetradecanoic acid, l-(+)-Ascorbic acid 2,6-dihexadecanoate, Estra -1,3,5(10)-trien-17ß-ol, Propanoic acid,2-(3-acetoxy-4,4,14-trimethylandrost-8-en-17-yl), Cis-13-Eicosenoic acid, Eicosanoic acid, 3-Pyrinecarboxylic acid, 2,7,10-tris(acetyloxy)-1,1a,2,3,4,6,7,10, Oleic acid, eicosyl ester, Butanoic acid, 4-chloro-,1,1a,1b,4,4a,5,7a,7b,8,9-decahydro-4a, Ethyl iso–allocholate, Ethyl iso –allocholate, Olean-12-ene-3,15,16,21,22,28-hexol, (3ß,15α,16α,21ß,22α)- and 2,4,6-Decatrienoic acid,1a,2,5,5a,6,9,10,10a-octahydro-5,5a-dihy. The Fourier-transform infrared spectroscopy (FTIR) analysis of E. lathyrus seeds proved the presence of alkenes, aliphatic fluoro compounds, alcohols, ethers, carboxlic acids, esters, nitro compounds, alkanes, hydrogen bonded alcohols, phenols.


INTRODUCTION
Medicinal plant parts (roots, leaves, branches/stems, barks, flowers, and fruits) are commonly rich in phenolic compounds, such as flavonoids, phenolic acids, stilbenes, tannins, coumarins, lignans and lignins (Cai et *Corresponding author. E-mail: imad_dna@yahoo.com. Tel: 009647716150716.reative Commons Attribution C Author(s) agree that this article remain permanently open access under the terms of the License 4.0 International License al., 2004;Altameme et al., 2015a;Al-Marzoqi et al., 2015).The seed of Euphorbia lathyris is a traditional Chinese medicine which has been used for the treatment of hydropsy, ascites, anuresis and constipation, amenorrhea, and scabies (Liu et al., 2011).Nowadays, traditional medicinal practices form an integral part of complementary or alternative medicine.Although their efficacy and mechanisms of action have not been tested scientifically in most cases, these simple medicinal preparations often mediate beneficial responses due to their active chemical constituents (Park and Pezzutto, 2002;Corro et al., 2014;Hameed et al., 2015a).In recent years, it was reported that the seeds of Euphorbia had a significant effect on leukemia, esophageal carcinoma, and skin cancer (Tapiero et al., 2002;Liu et al., 2011;Al-Marzoqi et al., 2016).The seed of E. lathyris is a kind of toxic traditional Chinese medicine, which is characterized by pungent, warm and poisonous in drug properties.It shows several side effects, such as irritation and inflammation intense on the skin, mouth and gastrointestinal tract irritation, carcinogenic, etc. (Buenz et al., 2004;Altameme et al., 2015b).The objective of this study was to analyse the chemical composition of seeds extract from methanol.The phytochemical compound was screened by gas chromatography-mass spectrometry (GC-MS) and Fourier-transform infrared spectroscopy (FT-IR) technique.

Plant and preparation of extracts
E. lathyrus dried seeds were purchased from local market in hilla city, middle of Iraq.After thorough cleaning and removal of foreign materials, the fruits were stored in airtight container to avoid the effect of humidity and then stored at room temperature until further use.About 30 g of the plant sample powdered were soaked in 100 ml methanol for 16 h in a rotatory shaker (Hamza et al., 2015;Hussein et al., 2016a).Whatman No.1 filter paper was used to separate the extract of plant.The filtrates were used for further phytochemical analysis.It was again filtered through sodium sulphate in order to remove the traces of moisture (Altameme et al., 2015c;Hameed et al., 2015b).

Identification of component by GC-MS analysis
The physicochemical properties of E. lathyrus are shown in Table 1.Interpretation of mass spectroscopy (GC-MS) was conducted by using data base of National Institute Standard and Technology (NIST) having more than 62000 patterns.The spectrum of the unknown component was compared with the spectrum of the known component stored in the NIST library.The identity of the components in the extracts was assigned by the comparison of their retention indices and mass spectra fragmentation patterns with those stored on the computer library and also with published literatures (Hadi et al., 2016;Hameed et al., 2015c;Hussein et al., 2016b).The GC-MS analysis of the plant extract was made in an Agilent 7890 A instrument under computer control at 70 eV.About 1 μl of the methanol extract was injected into the GC-MS using a micro syringe and the scanning was done for 45 min.The fragments obtained were actually charged ions with a certain mass (Hameed et al., 2015d;Hussein et al., 2016c).Helium gas was used as a carrier as well as an eluent.The flow rate of helium was set to 1 ml/min.The electron gun of mass detector liberated electrons having energy of about 70 eV.The column employed here for the separation of components was Elite 1 (100% dimethyl poly siloxane).

Conclusion
E. lathyrus is native plant of Iraq.It contains chemical constitutions which may be useful for various herbal formulation as anti-inflammatory, analgesic, antipyretic, cardiac tonic, and antiasthamatic.

Table 1 .
Major phytochemical compounds identified in methanolic extract of Euphorbia lathyrus.

Table 2 .
FT-IR peak values of Euphorbia lathyrus.