Asst. Lect. Islam Yaser AL-Hamdany Department of Biology College of Education / Mosul University
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1 Citrullus colocynthis L. HPLC Citrullus colocynthis L BA.0 NAA 40.6 HPLC 9.8 Quantitative and Qualitative Determination of Rutin and Quercetine in Callus Resulting from Parts of Citrullus colocynthis L. Using the High Performance Liquid Chromatography HPLC Asst. Lect. Islam Yaser AL-Hamdany Department of Biology College of Education / Mosul University Abstract: The present study arrived at initiated callus from Citrullus colocynthis L. namely from these parts (leaves, stems, cotyledons and hypocotyls)on the
2 solid medium supported with different concentrations of auxins and cytokinins. The originate callus average estimated at 00% in all plant parts under study in the medium provided with.0 mg/l BA+0. mg/l NAA. Therefore, this was selected as the best medium to product callus from the plant. The quantitative and qualitative measuring of alcohol extract of callus resulting from Citrullus colocynthisl.utilizing the HPLC technique showed that it contains the highest concentration of the Rutin compound. This was amounted to 40.6% before hydrolysis similarly the highest concentration of the quercetine compound was amounted to 9.8% in this extract after the hydrolysis. Key wordes: Citrullus colocynthis, plant tissue culture, high performance liquid chromatography, phenolic compound. Fell,97 Micro propagation Explants 99 Invitro Edwin et.al., 008 (Hartmann et.al.,00) Briskin,000 Najafi Murugesan Citrullus colocynthis L. Mulabagal & Tsay, 004 Cucurbitaceae et.al.,00 & Muthusamy,0
3 Colocynth diabetes Savitha et.al.,00 jaundice Gurudeeban et.al.,0 Kosanovic et.al.,0 Flavonal Lim Vit-P quercetin--0-rutinoside Flavonal et.al.,006 shophorin rutoside Wang et.al.,0 Zheng Citrus- bioflavonoids Meletin Sophretin et.al.,00 HO O OH OH HO O OH OH OH O OH OH O O Glu O Rh Quercetin Rutin (Wang et.al,0 ; Kosanovic et.al,0)
4 HPLC 70 Citrullus colocynthis L. 8 Magnatic Stirrer Hot Plate ph-meter 0 Autoclave Murashige & Skoog, Citrullus colocynthis L.
5 0 0 : : : : : 0 O 0,4 D NAA BA BA
6 ,4 D HPLC 96 Citrullus colocynthis L. 4 Mechanical Stirror HPLC Kosanovic et al., 0 RVE Acid Hydrolysis N HCl 0
7 0 Harborne, HPLC 97 Manual Shimadzo HPLC 0. µ M injection 0ALC H O 8 Acetonitrile C8 C. colocynthis L. HPLC C8 8 UV-Vis BDH 00 Citrullus colocynthis L. 0 0
8 Mahmoud C. colocynthis L C. colocynthis L. NAA BA 00 NAA 0. BA.0 988
9 Chand & Roy, 980 C.colocynthis L. NAA BA NAA BA BA Raven et.al., 986 B A
10 C.colocynthis L. A 0 B 0 A B A B A B A 4 B 4 BA,4-D NAA,4-D NAA NAA Hartmann BA.0,4-D C.colocynthis L. BA,4 D BA,4-D 4 4
11 D, Sultana et al., 004 Citrullus lanatus Totipotency Jimenez, 00; Acquaah, 004 4,4-D C.colocynthis L.,4 D,4-D
12 4,4-D Nuengchamnong -4 HPLC HPLC
13 4
14 C.colocynthis L Vrchotova (Larkin & Scowcroft,998) C.colocynthis L
15 6 7
16 9 8
17 Phani HPLC. ( Vrchotova et.al,00) C.colocynthis L
18 0
19
20 99 : ; 4-Acquaah.G(004).Understanding Biotechnology Pearson Education. Inc. U.S.A. -Briskin,D.B.(000).Medicinal plants and phytomedicines. Link Plant Biochemistry and Physiology to human health. Plant Physiol. 4: Chand,S; and Roy, S.C(980).Study of callus from different parts of Nigella sativa (Ranunculaceae) Experiential,6: Edwin,F.G;Michael,A.H;and Klerk, G.J.(008).Plant Propagation by Tissue Culture. rd Edition Volume().published by Siprenger 8-Fell,H.B.(97).Tissue culturte and its contribution to biology and medicine J.Exp.Biol, 7:-. 9-Gurudeeban.S;Ramanathan.T and Satyavani.K(0).Characterization of volatile compounds from Bitter Apple Citrullus colocynthis using GC-.International Journal of Chemical and Analytical Science. Issn: Harborne,T.B(97).Phytochemical Methods:A Guide to Modern Technique of Plant Analysis. st ed.,cox and Wyman,London.-7. -Hartmann,H.T;Kester,D.E;Davies,F.T;andGeneve,R.L. (00).Plant Propagation Principles and Practices.7 th.ed.prentice Hall, Inc, U.S.A. -Jimenez,V.M(00).Regulation of in vitro somatic embryogenesis with emphasis on the role of endogenous hormones. Rev. Bars. Fisiol. Veg. ():96-. -Kosanovic,M.M;Samardzic.M; Malatesti, N and Bosnar, M.S(0). Electroanalytical Characterization of a Copper (II)-Rutin Complex.Int.J. Electrochem. Sci., Larkin,P.J; and Scowcroft, W.R(998).Somaclonal variations a novel source of variability from cell culture for plant improvement. theor. Appl. Genet.60: Lim,T.H;Park.J;Min.D;Choi.K andkown. T(006).Antioxidant activity of four endemic stachys Taxa.Biol.Pharma.Bull.9(4): Mahmoud, I.N; Ibrahim, A.I; Haga, M.H and Tarek, K.(008). Rapid micropropagation of diploid and tetraploid watermelon (Citrullus
21 lanatus) cultivars. Genetic Engineering and Biotechnology Research Institute (GEBRT), sadat city, cairo university. 7-Mulabagal,V.and Tsay,H.S.(004).Plant cell culture an alternative and efficient source for the production of biologically important secondary metabolites. Int.J. Appl.Sci.Eng(): Murashige,T and Skoog, F(96).A revised medium for rapid growth and bioassay with tobacco tissue culture, Physiol Plant.: Murugesan,S ; and Muthusamy, M.(0).Screening and Quantitative Estimation of Phytochemical Constituents in Citrullus colocynthis (L.)Schrad.J.Res.Phytochem.Pharmacol.,(): Najafi, S;Sanadgol,N; Nejad,B.S; Beiragi,M.A and Sanadgol, E. (00). Phytochemical screening and antibacterial activity of Citrullus colocynthis (L.) Schrad aginst Staphylococcus aureus.4(): -. -Nuengchamnong, N; Lokkerbol, A.H;and Ingkaninan,K(004).Separation and detection of the antioxidant flavonoids, Rutin and Quercetin, using HPLC coupled on-line with colorimetric detection of antioxidantactivity. Naresuan University Journal.():-7. -Phani, Ch. R.S; Vinaykumar, Ch; Umamaheswararao, K; and Sindhuja,G (00). Quantitative Analysis of Quercetin in Natural Sources by RP- HPLC- International Journal of Research in Pharmacentical and Biomedical Sciences,():9-. -Raven,P.H;Evwrt,R.F;and Eichhor, S.E(986).Biology of Plants. 4 th. ed., Worth publishers,inc. 4-Savitha,R;Shasthree,T;and Mallaiah,S.B.(00).High freguency of plantlet regeneration and multiple shoot induction from leaf and stem explants of Citrullus colocynthis(l.) Schrad, an endangered medicinal cucurbit. International Journal of Pharma and Bio Sciences.V(). - Sultana,R.S;Bari,M.A;Rahman,M.H;Rahman,M.M; Siddique,N.A; and Khatun,N(004). In vitro rapid regeneration of plantlets from leaf explants of water melon Citrullus lanatus Thumb.Biotechnology. ():-. 6-Vrchotova, N; Sera, B; and Dadakova, E (00). HPLC and CE analysis of stilbens and quercetin in flowers and stems of Polygonum cuspidatum, P. sachalinense and P. xbchemicum, J.Indian Chem. Soc., Vol. 87: Wang,J; Zhao, L.L; Sun,G.X;Liang,Y;Wu, F.A; Chen, Z. L; and Cui, S.M (0). Acomparison of acidic and enzymatic hydrolysis of rutin.african Journal of Biotechnology0(8): Zheng, Y; Haworth,I.S; Zuo,Z; Chow, M.S.S; and Chow,A.H.L(00). Physiochemical and structural characterization of quercetin-bcyclodextrin complexes.journal of Pharmaceutical Sciences.94().
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