Journal University of , Vol. 4(A) , No.1, Pp 81-86, 2016 ISSN: 2410-7549

PRELIMINARY PHYTOCHEMICAL SCREENING OF PERSICA L. (FLOWERS, LEAVES, AND RHIZOMES) COLLECTED IN KURDISTAN REGION-.

Hawraz Ibrahim M. Amin1, Ahmed Anwar Amin2, Faiq Hama Saeed Hussain1 and Giovanni Vidari 3 1 Department of Chemistry, College of Science, University of Salahaddin, Kurdistan Region – Iraq. 2 Department of Chemistry, College of Education, University of Salahaddin, Kurdistan Region – Iraq. 3 C.I.St.R.E., University of Pavia-Italy, Via Taramelli 10, 27100 Pavia,- Italy. (Accepted for publication: May 8, 2016)

ABSTRACT: The curative properties of medicinal are perhaps due to the presence of various secondary metabolites. This paper reports the first investigation of phytochemical constituents present in the methanolic extracts of flowers, leaves, bulbs and rhizomes of Iris persica L. (), collected in Korek Mountain (Rawanduz) in the Kurdistan Region-Iraq, which is used by local people for the treatment of wound inflammation and tumor. The phytochemical analysis was performed to detect the presence of flavonoids, polyphenols, terpenoids, protiens and reducing sugar in all extracts of Iris persica. While tannins and saponins were found in and rhizome extracts only, alkaloids, steroids, aminoacids and anthraquinons were found to be absent in all extracts.

KEYWORDS: Iris persica, Phytochemical Constituents, Qualitative Analysis.

INTRODUCTION: and De Boer, 2011). The presense of Iris persica L., have been reported in Iraq (Townsend, and edicinal plants are a rich source of Guest, 1985) and in the Kurdistan region numerous pharmacologically active M (Kaššák, 2012) especially in Halgurd mountain molecules. Scientists are currently focusing on (Choman district) and Korek mountain the phytochemicals to treat numerous ailments (), which is commonly affecting the mankind (Rajesh, et al., 2013). The employed in the Kurdish traditional medicine for genus Iris belongs to the family Iridaceae, which the treatment of wound inflammations and comprises over 300 species. The phytochemical tumor. These applications are reported by the screening and chemical investigations of various traditional herbal healers, locally called Baytars, species of Iris have resulted in the isolation of that are highly recognized as experts in herbal variety of secondary metabolites. Approximately medicinal uses. To this aim, the was more than two hundred compounds have been collected in Korek mountain and the reported from the genus Iris which includes phytochemical investigation was performed. To flavonoids, isoflavonoids and their glycosides, the best of our knowledge, this is the first report benzoquinones triterpenoids and stilbenes on the qualitative phytochemical screening of the glycosides (Guo-Yong, et al., 2013). Iris species flowers, leaves, bulbs and rhizomes of Iris have an immense medicinal importance and are persica. used in the treatment of cancer, inflammation, bacterial and viral infections. The compounds EXPERIMENTAL: isolated from these species were reported to have piscicidal, anti-neoplastic, antioxidant, 1. PLANT MATERIAL: antitumor, anti-plasmodial, molluscicidal, and Iris persica L. (flowers, leaves, bulbs and anti-tuberculosis properties, in addition to rhizomes) was collected in April 2014 from protein kinase C activation activity (Sabrin, et Korek Mountain (Rawanduz) in the Kurdistan al., 2012) and (Wirginia, et al. 2013). Iris species region (IRAQ). The plant was identified by two have been used as ornamental plants in botanists Prof. Dr. A. H. Al-khayyat and Dr. vegetative landscape of the parks and gardens in Abdullah Sh. Sardar at the Biology Department, many countries since ancient times because of College of Education, Salahaddin University- very beautiful and colorful flowers (Nezahat, et /Iraq. A voucher specimen (No. 7229) was al., 2011). Herbal remedies in deposited at Education Salahaddin University can be classified into 133 different uses; for the Herbarium (ESUH), Kurdistan. The plant raw most part they are of medicinal type, but some materials (flowers, leaves, bulbs and rhizomes) are also of cosmetic and ritual relevance (Mati,

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were shade dried at room temperature (20-25oC). (20min) then macerated for (3hrs) with After drying, the plant parts were grounded in to continuous stirring at room temperature. The fine powder using a laboratory grinding mill, to procedure repeated three times for each part provide homogeneous powder for the analysis. separately. Then the mixtures were filtered and Powdered materials were stored in bottles in a the solvent was removed under “vacuum” using dark room temperature and then used. rotary evaporator affording a crude methanol extracts (Raphael I.). The percentage yields of 2. EXTRACTION WITH METHANOL: different crude extracts are reported in The defatted flowers, leaves, bulbs and Table 1. rhizomes (each 200g) were extracted with (500 mL) of methanol using ultrasonic bath for

Table (1): Extraction yields of Iris persica flowers, leaves, bulbs and rhizomes.

Plant parts Solvent Extraction residue (g) Percentage yield (w/w) Hexane 1.87 0.93 Flowers Methanol 31.33 15.66 Hexane 1.17 0.58 Leaves Methanol 26.47 13.23 Hexane 2.16 1.08 Bulbs Methanol 37.28 18.64

Hexane 1.89 0.94 Rhizomes Methanol 51.16 25.58

3. IDENTIFICATION TESTS: presence of flavonoids. (c) A few drops of 1% aluminium solution was added to a portion of the Qualitative phytochemical analysis of the filtrate. A yellow colouration indicated the methanolic extract of Iris persica were carried presence of flavonoids (Saxena, and Sahu, out using standard procedures to assess the 2012). (d) A portion of the extract was heated different types of phytochemical constituents with 10 mL of ethyl acetate over a steam bath for present in the flowers, leaves, bulbs and 3 min. The mixture was filtered, and 4 mL of the rhizomes of Iris persica using different chemical filtrate was shaken with 1 mL of dilute ammonia tests. Screenings were carried out for flavonoids, solution. A yellow colouration indicated the polyphenols, tannins, alkaloids, terpenoids, presence of flavonoids. (e) A small piece of saponins, protiens, amino acids, steroids, magnesium ribbon was added to the alcohol anthraquinnones and reducing sugar (Sawant, solution of the extract followed by dropwise and Godghate, 2013, Mohammad, and Arun, addition of concentrated hydrochloric acid. The 2009, Saxena, and Sahu, 2012, Minakshi, and colour changing from red-crimson indicates Sushma, 2006, Amin Mir, 2013). flavonols, crimson to magenta indicates

flavonones and green blue indicates the test is 3.1. TEST FOR FLAVONOIDS: positive (Sawant, and Godghate, 2013). Five methods were used to test for flavonoids. (a) Alkaline reagent test: Extract was 3.2. TEST FOR PHENOLIC COMPOUNDS: treated with (5 mL) 10 % NaOH solution, Two methods were used to test for Phenolic formation of intense yellow colour indicates Compounds: (a) lead acetate test: The extract (50 presence of Flavonoid. (b) Diluted ammonia (5 mg) were dissolved in distilled water and to this; mL) was added to a portion of an aqueous 3 mL of 10% lead acetate solution was added. filtrate of the extract. Concentrated sulphuric Formation of a bulky white precipitate indicated acid (1 mL) was then added. A yellow the presence of phenolic compounds. (b) FeCl discolouration that on standing indicated the 3 test: A small quantity of extract was diluted with

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water and tested with Dilute FeCl3 solution (5%), Rochelle salt) made in a strong alkali, commonly intense blue, green colour indicated the presence with sodium hydroxide. of phenolic compounds (Mohammad, and Arun, 2009). 3.8. TEST FOR ANTHRAQUINONES:

The extract (0.5g) was boiled with 10 mL of 3.3. TEST FOR TANNINS: sulphuric acid (H2SO4) and filtered while it was Gelatin test: 50 mg of extract dissolved in 5 hot. The filtrate was shaken in 5 mL of mL of distilled water and to this; 2 mL of a 1% chloroform. The chloroform layer was pipetted solution of gelatin containing 10% sodium into another test tube, and 1 mL of diluted chloride was added. The formation of white ammonia was added. The resulting solution was precipitates indicated the presence of phenolic observed for colour changes (Mohammad, and compounds (Sawant, and Godghate, 2013). Arun, 2009).

3.4. TEST FOR ALKALOIDS: 3.9. TEST FOR PROTEINS Dragendroff’s test: 2 drops of Dragendroff’s To 3 mL of extracts add 3% NaOH and few reagent were added to 1 mL of the extract. The drops of 1% CuSO4. The solution turns from development of a creamy precipitate was blue to violet (purple) or to pink, indicates the indicative of the presence of alkaloids (Saxena, presence of protein (Amin Mir, 2013). and Sahu, 2012). 3.10. TEST FOR AMINO ACIDS 3.5. TEST FOR TERPENOIDS: To 5 mL of extract add few drops of 40% The chloroform (2 mL) was added to 0.5 g of NaOH and 10% lead acetate boiled the solution; the extract. Concentrated H2SO4 (3 mL) was formation of black precipitate indicate the carefully added to form a layer, and the solution presence of amino acid (Minakshi, and Sushma, was observed for a reddish brown discolouration 2006). at the interface, which indicated the presence of terpenoids (Amin Mir, 2013). 3.11. TEST FOR STEROIDS

The extract (1 mL) was dissolved in 10 mL of 3.6. TEST FOR SAPONINS: chloroform and equal volume of concentrated 5 mL extract was mixed with 20 mL of H2SO4 acid was added from the side of test tube distilled water then agitated in graduated .The upper layer turns red and H2SO4 layer cylinder for 15 min formation of foam indicates showed yellow with green fluorescence .This Saponin (Sawant, and Godghate, 2013). indicates the presence of steroid (Mohammad, and Arun, 2009). 3.7. TEST FOR REDUCING SUGARS: RESULT AND DISCUSSION: Fehling’s test: The methanol extract (0.5 g in 5 mL of water) was added to boiling Fehling’s The present study carried out on the Iris solution (A and B) in a test tube. The solution persica revealed the presence of medicinal active was observed for a colour reaction (a purple ring constituents. The phytochemical active at the junction of two liquids) (Minakshi, and compounds of Iris persica were qualitatively Sushma, 2006). analyzed for flowers, leaves bulbs and rhizomes Note: Fehling's A is aqueous solution of separately and the results are presented in Table copper (II) sulphate, which is deep blue. 2. The preliminary phytochemical screening of Fehling's B is a colorless solution of aqueous methanol extracts indicated the presence of potassium sodium tartrate (also known as flavonoids, polyphenols, tannins, trpenoids, saponins, protiens and reducing sugar.

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Table (2): Qualitative phytochemical analysis of the methanol extract of Iris persica (flowers, leaves, bulbs and rhizomes). Extracts Phytochemical constituents Flowers Leaves Bulbs Rhizomes Flavonoid: Alkaline reagent test + + + + Ammonia test + + - +

AlCl3 test + + + + Ethylacetate test + + + - Shinoda test + + + +

Polyphenol: Lead Acetate test + + + +

FeCl3 test + + + + Tannin: Gelatin test - - + + Alkaloid: Dragen droff’s test - - - - Terpenoid + + + + Saponin: Foam test - - + + Protein + + + + Amino acid - - - - Steroid - - - - Anthraquinone: Borntrager’s test - - - - Reducing sugar: Fehling’s test + + + + Key: + = Present, - = Absent

From phytochemical screening, we observed reason these plants are used for the treatment of that the methanolic extracts of (flower, leaf, bulb same diseases. and rhizome) parts gave a positive result with the Alkaline reagent test, ALCl3 test and the Shinoda CONCLUSIONS: test, which indicated the presence of flavonoids in all extracts. The Dragendorff’s reagent failed This is the first report on the phytochemical to show the presence of alkaloids in all extracts. screening of the flowers, leaves, bulbs and The Gelatin test would confirm the presence of rhizomes of Iris persica L. growing from tannin in the methanolic extracts of bulbs and Kurdistan region-Iraq. Phytochemical analysis rhizomes. Based on the general test for terpenes, revealed the presence of flavonoids, indicates the presence of terpenes in all extracts polyphenols, tannins, terpenoids, saponins, (Table 2). The borntragers test for anthraquinons protiens and reducing sugar in Iris persica. gave negative results in all extracts. Test for Further chemical analysis on the composition of saponins gave positive results with the I. persica methanol extract is necessary to isolate methanolic extract of bulbs and rhizomes only. and identify bioactive compounds.

Lead acetate test and FeCl3 test gave positive results, which indicates the presence of ACKNOWLEDGMENT: polyphenols in all extracts. The test of protein, Thanks to University of Salahaddin-Erbil for gave positive results in all extracts. The test for the finantial supporting as a part of split-side amino acids and steroids gave negative results in Ph.D. program, and many thanks to Prof. Abdul all extracts. Hussain Al Khayyat and Dr. Abdullah Sh. All these facts support the usefulness of I. Sardar for the botanical identification. persica in folklore remedies and may be the

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