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Chromatographic Study of the Ethanol Extract of Cordia Myxa L. Fruit Tarik Z

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Chromatographic Study of the Ethanol Extract of Cordia Myxa L. Fruit Tarik Z 142 Journal of Pharmaceutical, Chemical and Biological Sciences ISSN: 2348-7658 Impact Factor (GIF): 0.615 Impact Factor (SJIF): 2.092 June-August 2016; 4(2):142-152 Original Research Article Chromatographic Study of the Ethanol Extract of Cordia myxa L. Fruit Tarik Z. T. Shwaish1*, Faris J. M. Al-Imarah 2 1Gifted Students’ School of Basra, Basra, Iraq 2Dept. Chem. & Marine Environ. Poll., Marine Science Center, University of Basrah, Basra, Iraq *Corresponding Author: Tarik Z. T. Shwaish, 61001, Gifted students’ school, Arabian Gulf district, Basra, Iraq Received: 15 May 2016 Revised: 25 May 2016 Accepted: 30 May 2016 ABSTRACT The aim of this study was to perform possible phytochemical screening for the Cordia myxa L. fruit extract by some analytical techniques. Ethanol extract was obtained by treating dried fruit sample with ethanol, the extract was used for GC and the peaks were studied to determine the composition and volatility of the extract. The extract was also analyzed by using HPLC and further analysis was carried out to determine the presence, concentration, and carcinogenic effects of 16 polycyclic aromatic hydrocarbons. The GC analysis revealed the presence of 10 alkaloids with one major component comprising about 64% of the total extract. In addition, 16 compounds were also reported with HPLC analysis. The presence of the following 6 PAHs in the extract was confirmed: Flourene, Pyrene (B2 carcinogen), Chrysene (B2), Phenanthrene (A1), Flouranthene (A1), and Benzo[a]anthracene with total concentration of about 412 ng/g. From the results, we can conclude the presence of many alkaloids and we can predict that most of the medical effects of the fruits are in the major compound which needs more standardization for further identification. The second finding of the study is the probable carcinogenic and toxic effects of the fruit due to the presence of several PAHs. Keyword: GC; HPLC; chromatography; ethanol extract; plant extract; Cordia myxa; PAHs; phytochemistry; toxicity INTRODUCTION Plants are used medicinally in different The subject of phytochemistry or plant countries, and they are the source of many chemistry, has developed to be a distinct potent and powerful drugs. Plants have been an discipline somewhere in between natural important source of medicine with qualities for product chemistry and plant biochemistry. thousands of years. [1] Phytochemistry is concerned with the enormous J Pharm Chem Biol Sci, June-August 2016; 4(2):142-152 Shwaish & Al-Imarah 143 variety of organic structures that are elaborated Gastroprotective [6], antioxidant and analgesic and accumulated by plants and deals with the effects [23]. In traditional medicine, Cordia myxa chemical structure of these substances, their L. fruit is used as diuretic, demulcent and in the biosynthesis, turnover, and metabolism, their treatment of stomach aches, coughs and chest natural distribution and their biological function complaints [7]. The fruit is also used externally as [2]. an emollient plaster to maturate abscesses, to Cordia myxa is a species of flowering plant in the calm rheumatic pain and as an anti-parasitic on borage family, Boraginaceae. It is a medium- ringworm. In addition, the sticky, mucilaginous sized broad-leaved deciduous tree. Common pulp of the fruit is the source of a well-known names include Lasura, Assyrian Plum, Pidar, medicine in the near and middle east called Panugeri, Naruvilli, Geduri, Spistan, Burgund “sapistan” which is useful in the treatment of dulu wanan, Ntege, and Bumber [3] [4]. The coughs, sore throats and chest-complaints on Cordia myxa plant parts are widely used in the account of its demulcent property [8]. traditional medicine especially in the Middle The fruit of Cordia myxa L. has been determined East, India, and China. The seeds, roots, leaves, to have the probability of the presence of some and other parts of the plant have various uses in Pyrrolizidine alkaloids [9]. Moreover, prior traditional medicine [5]. studies have confirmed the presence of the non- The fruit begins appearing in July-August. It is toxic pyrrolizidine alkaloid macrophylline but no light pale to brown in color and it to tends to actual evidence of the concentration of these darken when ripening proceeds. Being full of alkaloids has been reported [24]. viscid glue like mucilage, the pulp is somewhat Concentrated ethanol was chosen for this study translucent. When fully ripe the pulp becomes because it has been widely used as a solvent in quite sweet in taste. [3] The origin of this species pharmacology and phytochemical science and is the region stretching from the eastern has been proved to be the best solvent to extract Mediterranean to eastern India. It has been the maximum components from a plant sample introduced long ago to tropical Africa, tropical compared with other solvents that might be Asia, and Australia, and more recently to the suitable for the extraction process [10] [11] [12] Americas [3] [4]. (Table 1). Cordia myxa plant has many medical uses, it has been long studied for its anti-inflammatory [5], Table 1: Phytochemical screening in various plant extracts [6] Phytochemicals Petroleum ether Chloroform Acetate Ethanol Flavonoids - - + + Alkaloids - - - + Tannins - - + + Glycosides + + + + Steroids + + + + Phenols - - + + Cardiac glycosides + + + + Saponins + + + + Carbohydrates + + + + Proteins - - - + ‘+’ Presence, ‘-’ Absence J Pharm Chem Biol Sci, June-August 2016; 4(2):142-152 Shwaish & Al-Imarah 144 min. isothermal) to 280oC (30 min isothermal). MATERIALS AND METHODS The total running time was 33.3 min. This study consists of three main stages: Ethanol The HPLC test was run by using Shimadzu UFLT extraction of some bioactive components in the equipped with a gradient pump and manual fruit, Gas chromatography for the determination injector. The separation column had internal of the volatility of the extract as well as some low diameter of 4.6 mm and total length of 25 molecular weight alkaloids, and HPLC for the meters, the bonded phase was Carbon-18 with determination of some compounds found in the pore size of 5-100 μm. The separation mode was ethanol extract and measuring their relative normal phase. The detector type was UV-Visible concentrations [13] [14]. spectrometer. The ambient temperature was o Cordia myxa totally dried black whole fruit 25 C. Shimadzu LC solution computer software sample was cultivated and shade-dried in Basra was used for control and analysis. southern Iraq during summer. The fruit sample was identified by morphological comparison RESULTS with traits described by AP database provided by The results of gas chromatography test Conservatory and Botanical Garden/ Geneva and described earlier were as shown in the gas South African National Biodiversity Institute chromatogram shown in fig. 1 and 65 peaks of [15]. The totally dried fruit sample was grinded different compounds were identified and into fine powder alongside with the seeds by described in terms of peak number, retention using Philips HL7720 mechanical grinder. time, area, height and concentration in table 2. The required amount of fruit was weighed and The chromatogram was very crowded in the was fully immersed with ethanol absolute of retention time in the range of 5-10. The 99.8% concentration at room temperature. The chromatogram was optimized and the modified extract was shaken each hour for the first 12 major peaks were as shown in fig. 2. The solvent hours and was then left for another 12 hours, the peak was removed and the relative composition process was repeated for the next 72 hours. The of the sample was obtained alongside the extract was dried over anhydrous sodium sulfate description of the modified peaks as shown in powder and was then filtered and Nitrogen gas table 3. was passed over the extract for degasification The results of the HPLC test described earlier for purpose. the ethanol extract were as shown in the The extract was diluted and directly employed chromatogram in fig. 3 and the peaks were for gas chromatography. The GC test was run characterized as in table 4 in terms of retention using Shimadzu 2014 gas chromatography time, height, area, and relative composition system coupled with FID detector equipped with shown as percentage of height and area (area is silica wax polar standard capillary tube with considered to be more accurate). length of 59 meters and internal diameter of The presence and concentration of 16 poisonous 0.25 μm. Nitrogen gas (99.999%) was used as the and carcinogenic polycyclic aromatic carrier gas with flow rate of 4 ml/min and an hydrocarbons (PAHs) was obtained from the injection volume of 2 μl was employed manually results of HPLC analysis in nanogram per gram of in the system; injector temp. was 290oC. The sample with total concentration of 411.5 ng/g as oven temp. was programmed from 40oC (0.5 shown in table 5. J Pharm Chem Biol Sci, June-August 2016; 4(2):142-152 Shwaish & Al-Imarah 145 Table 2: Gas chromatography peaks description Peak # Ret. Time Area Height 1 5.280 3498 650 2 5.494 1524 266 3 5.655 32899 5600 4 5.955 4938 2287 5 6.098 11115522 1665234 6 6.206 5012583 1639231 7 6.254 5043586 1644037 8 6.301 4659237 1676138 9 6.452 16893119 1764980 10 6.498 5382162 1756027 11 6.548 4381933 1744094 12 6.604 585212 1753413 13 6.646 5796895 1738379 14 7.311 195091024 6585856 15 7.488 75745170 6925636 16 7.536 23350365 6982197 17 7.601 25457179 7028199 18 7.645 23492366 7041280 19 7.702 21502692 7023398 20 7.750 117692344 7020179 21 8.039 28875391 6493918 22 8.122 21406896
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