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A new flavonoid from the aerial parts of Chrysanthemum viscidehirtum
Fitoterapia 71 Ž2000. 413᎐416
A new flavonoid from the aerial parts of Chrysanthemum ¨iscidehirtum F. Khallouki a,b,c,U , M. Hmamouchi a , C. Younosb, R. Soulimani b, E.M. Essassi c a
Departement de Chimie-Biochimie, U.F.R. des Substances Naturelles, Faculte´ de Medecine et de Pharmacie, Rabat, Maroc b Laboratoire d’Ethnobotanique et de Pharmacologie, U.F.R. SciFa, 57000 Metz, France c Laboratoire de Synthese ` Organique, Faculte´ des Sciences Mohammed V, Rabat, Maroc
Received 8 November 1999; accepted in revised form 26 January 2000
Abstract From the aerial parts of Chrysanthemum ¨ iscidehirtum, a new flavonoid, 2Y-glucosyl-8-Cglucosyl-4⬘-O-methylapigenin Ž1. was isolated. 䊚 2000 Elsevier Science B.V. All rights reserved. Y
Keywords: Chrysanthemum ¨ iscidehirtum; Flavonoids; 2 -glucosyl-8-C-glucosyl-4⬘-O-methylapigenin
1. Introduction Numerous species of the genus Chrysanthemum ŽAsteraceae ., commonly known as ‘uqhuwan’ ˆ in the local language of Morocco, are used in traditional medicine: C. coronarium and C. macrocarpum as scabicides and against intestinal parasitic infections; C. trifurcatum and C. segetum in the treatment of liver and biliary disorders w1,2x. We present here the isolation and spectral characterisation of the new flavonoid 1 isolated from the aerial parts of Chrysanthemum ¨ iscidehirtum ŽSchott.. Tell Žsyn.: Heteranthemis ¨ iscidehirta Schott.. U
Corresponding author. Tel.: q33-387754114; fax: q33-387364198. E-mail address: [email protected] ŽF. Khallouki..
0367-326Xr00r$ - see front matter 䊚 2000 Elsevier Science B.V. All rights reserved. PII: S 0 3 6 7 - 3 2 6 X Ž 0 0 . 0 0 1 4 7 - 7
414
F. Khallouki et al. r Fitoterapia 71 (2000) 413᎐416
2. Experimental 2.1. Plant material C. ¨ iscidehirtum aerial parts were collected near Rabat, Morocco in May 1995 and identified by Prof. M. Bentata at Department of Botany, Scientific Institute of Rabat, Morocco. 2.2. Extraction and isolation The dry powdered plant material Ž800 g. was defatted with petrol Ž60᎐80⬚C. and extracted with 80% EtOH Žvrv.. The concentrated extract was treated with Et 2 O, EtOAc and n-BuOH. Twenty grams out of the 48 g obtained from the concentrated n-BuOH extract were chromatographed by DCCC Žascending method; CHCl 3-MeOH-H 2 O-n-BuOH 10:10:6:1. to give four fractions. The residue of fraction 2 Ž40 mg. was subjected to cellulose TLC, with AcOH-H 2 O 15:85 ŽS1 ., then with n-BuOH-AcOH-H 2 O 4:1:5 ŽS 2 ., to give apigetrin Ž12 mg., vitexin Ž7 mg. and compound 1 Ž15 mg..
Compound 1, no melting up to 268⬚C, R F 0.66 ŽS1 ., 0.45 ŽS 2 .; UV max ŽMeOH.: 332, 275 Žlog 3.58.; MeOHrNaOAc 331, 281; MeOHrNaOAcrH 3 BO 3 331, 274; MeOHrAlCl 3 335, 383, 278, 298; MeOHrAlCl 3rHCl 340 Žsh., 275 296; MeOHrNaOH 338, 270 nm; 1 H-NMR Ž300 MHz, DMSO-d6 .: ␦ 8 Ž2H, d, J 8.74Hz, H-2⬘, H-6⬘., 6.98 Ž2H, d, J 8Hz, H-3⬘, H-5⬘., 6.88 Ž1H, s, H-3., 6.47 Ž1H, s, H-6., 3.85 Ž3H, s, OMe.; 13 C-NMR ŽDMSO-d6 .: fla¨ one ᎏ 182.2 ŽC-4., 164.2 ŽC-7., 163.4 ŽC-2., 161.4 ŽC-4⬘., 161.1 ŽC-5., 155 ŽC-9., 128.9 ŽC-2⬘ and C-6⬘., 121.5
F. Khallouki et al. r Fitoterapia 71 (2000) 413᎐416
415
ŽC-1⬘., 115.8 ŽC-3⬘ and C-5⬘., 104.7 ŽC-8., 104.2 ŽC-10., 102.5 ŽC-3., 95 ŽC-6., 56.58 Ž4⬘-OCH 3 .; C-glucosyl-60 ŽC-6Y ., 70 ŽC-4Y ., 71.3 ŽC-1Y ., 76 ŽC-3Y ., 80 ŽC-5Y ., 81.7 ŽC-2Y .; O-glucosyl-60.8 ŽC-6Y ⬘., 69.7 ŽC-4Y ⬘., 74.4 ŽC-2Y ⬘., 76 ŽC-3Y ⬘., 78 ŽC-5Y ⬘., 104.8 ŽC-1Y ⬘.; ES-MS m r z: 609 ŽM᎐Hq, 100%., 447 Ž57., 327 Ž18., 251 Ž23., 150 Ž17.. 2.3. Acid hydrolysis of compound 1 In a sealed test tube, 5 mg of 1 in 7 ml of 50% MeOH was heated for 2 h in the presence of 5 ml of 5 N HCl. Water was added Ž5 ml. and the genine was extracted with 5 ml of n-BuOH after neutralization with BaCO 3 .
3. Results and discussion Acid hydrolysis of compound 1 gave glucose and two products. The two products are suggested to be regioisomers obtained by Wessely᎐Moser transposition, isomerization specifically characteristic of C-glucosylflavones and proving that the sugar related to the flavone in position 8 is glucose w3x. The UV spectrum of compound 1 in MeOHrAlCl 3rHCl showed four absorption peaks, which is characteristic of 5-hydroxyflavones. The sodium acetate spectrum exhibited a 6-nm bathochromic shift of band II, indicating the presence of a 7-OH group. The 1 H-NMR spectrum exhibited the H-6 and H-3 protons at 6.47 and 6.88 ppm, respectively, and the pattern characteristic for a 4⬘-methoxy B-ring w4x. The presence of an OMe group at C-4⬘ was proved by the UV spectrum in MeOH᎐NaOH Ž ⌬ BI s 8 nm, ⌬ BII s 0 nm.. The assignment of the 6.47 signal to H-6⬘ was proved by the 13 C-NMR spectrum which excluded the presence of a proton at C-8. The 13 C-NMR of 1 was reminiscent of that of 8-glucosylapigenin w5x, the only differences being the downfield shift of the C-2Y resonance Ž10 ppm., an upfield shift of the C-1⬘ Ž1.5 ppm., and the presence of an extra glucose moiety. Therefore, the structure of compound 1 was deduced to be 2Y-O--D-glucosyl-8-C--D-glucosyl-4⬘-O-methylapigenin.
Acknowledgements Authors present their gratefulness to the head of the Analytical Chemistry Department of the Perm State Pharmaceutical Academy, Professor Vladislav O. Kozminykh, for his help in structure determination.
References w1x Bellakhdar J, Claisse R, Fleurentin J, Younos C. J Ethnopharmacol 1991;35:123.
416 w2x w3x w4x w5x
F. Khallouki et al. r Fitoterapia 71 (2000) 413᎐416 Bellakhdar J. These ` Doct Univ Metz 1997;1157. Wessely F, Moser GH. Monaths Chem 1930;56:97. Chatterjee A, Sarkar S, Saha SK. Phytochemistry 1981;20Ž7.:1760. Horowitz RM, Gentili B. Chem Ind 1964;498.
A new flavonoid from the aerial parts of Chrysanthemum ¨iscidehirtum F. Khallouki a,b,c,U , M. Hmamouchi a , C. Younosb, R. Soulimani b, E.M. Essassi c a
Departement de Chimie-Biochimie, U.F.R. des Substances Naturelles, Faculte´ de Medecine et de Pharmacie, Rabat, Maroc b Laboratoire d’Ethnobotanique et de Pharmacologie, U.F.R. SciFa, 57000 Metz, France c Laboratoire de Synthese ` Organique, Faculte´ des Sciences Mohammed V, Rabat, Maroc
Received 8 November 1999; accepted in revised form 26 January 2000
Abstract From the aerial parts of Chrysanthemum ¨ iscidehirtum, a new flavonoid, 2Y-glucosyl-8-Cglucosyl-4⬘-O-methylapigenin Ž1. was isolated. 䊚 2000 Elsevier Science B.V. All rights reserved. Y
Keywords: Chrysanthemum ¨ iscidehirtum; Flavonoids; 2 -glucosyl-8-C-glucosyl-4⬘-O-methylapigenin
1. Introduction Numerous species of the genus Chrysanthemum ŽAsteraceae ., commonly known as ‘uqhuwan’ ˆ in the local language of Morocco, are used in traditional medicine: C. coronarium and C. macrocarpum as scabicides and against intestinal parasitic infections; C. trifurcatum and C. segetum in the treatment of liver and biliary disorders w1,2x. We present here the isolation and spectral characterisation of the new flavonoid 1 isolated from the aerial parts of Chrysanthemum ¨ iscidehirtum ŽSchott.. Tell Žsyn.: Heteranthemis ¨ iscidehirta Schott.. U
Corresponding author. Tel.: q33-387754114; fax: q33-387364198. E-mail address: [email protected] ŽF. Khallouki..
0367-326Xr00r$ - see front matter 䊚 2000 Elsevier Science B.V. All rights reserved. PII: S 0 3 6 7 - 3 2 6 X Ž 0 0 . 0 0 1 4 7 - 7
414
F. Khallouki et al. r Fitoterapia 71 (2000) 413᎐416
2. Experimental 2.1. Plant material C. ¨ iscidehirtum aerial parts were collected near Rabat, Morocco in May 1995 and identified by Prof. M. Bentata at Department of Botany, Scientific Institute of Rabat, Morocco. 2.2. Extraction and isolation The dry powdered plant material Ž800 g. was defatted with petrol Ž60᎐80⬚C. and extracted with 80% EtOH Žvrv.. The concentrated extract was treated with Et 2 O, EtOAc and n-BuOH. Twenty grams out of the 48 g obtained from the concentrated n-BuOH extract were chromatographed by DCCC Žascending method; CHCl 3-MeOH-H 2 O-n-BuOH 10:10:6:1. to give four fractions. The residue of fraction 2 Ž40 mg. was subjected to cellulose TLC, with AcOH-H 2 O 15:85 ŽS1 ., then with n-BuOH-AcOH-H 2 O 4:1:5 ŽS 2 ., to give apigetrin Ž12 mg., vitexin Ž7 mg. and compound 1 Ž15 mg..
Compound 1, no melting up to 268⬚C, R F 0.66 ŽS1 ., 0.45 ŽS 2 .; UV max ŽMeOH.: 332, 275 Žlog 3.58.; MeOHrNaOAc 331, 281; MeOHrNaOAcrH 3 BO 3 331, 274; MeOHrAlCl 3 335, 383, 278, 298; MeOHrAlCl 3rHCl 340 Žsh., 275 296; MeOHrNaOH 338, 270 nm; 1 H-NMR Ž300 MHz, DMSO-d6 .: ␦ 8 Ž2H, d, J 8.74Hz, H-2⬘, H-6⬘., 6.98 Ž2H, d, J 8Hz, H-3⬘, H-5⬘., 6.88 Ž1H, s, H-3., 6.47 Ž1H, s, H-6., 3.85 Ž3H, s, OMe.; 13 C-NMR ŽDMSO-d6 .: fla¨ one ᎏ 182.2 ŽC-4., 164.2 ŽC-7., 163.4 ŽC-2., 161.4 ŽC-4⬘., 161.1 ŽC-5., 155 ŽC-9., 128.9 ŽC-2⬘ and C-6⬘., 121.5
F. Khallouki et al. r Fitoterapia 71 (2000) 413᎐416
415
ŽC-1⬘., 115.8 ŽC-3⬘ and C-5⬘., 104.7 ŽC-8., 104.2 ŽC-10., 102.5 ŽC-3., 95 ŽC-6., 56.58 Ž4⬘-OCH 3 .; C-glucosyl-60 ŽC-6Y ., 70 ŽC-4Y ., 71.3 ŽC-1Y ., 76 ŽC-3Y ., 80 ŽC-5Y ., 81.7 ŽC-2Y .; O-glucosyl-60.8 ŽC-6Y ⬘., 69.7 ŽC-4Y ⬘., 74.4 ŽC-2Y ⬘., 76 ŽC-3Y ⬘., 78 ŽC-5Y ⬘., 104.8 ŽC-1Y ⬘.; ES-MS m r z: 609 ŽM᎐Hq, 100%., 447 Ž57., 327 Ž18., 251 Ž23., 150 Ž17.. 2.3. Acid hydrolysis of compound 1 In a sealed test tube, 5 mg of 1 in 7 ml of 50% MeOH was heated for 2 h in the presence of 5 ml of 5 N HCl. Water was added Ž5 ml. and the genine was extracted with 5 ml of n-BuOH after neutralization with BaCO 3 .
3. Results and discussion Acid hydrolysis of compound 1 gave glucose and two products. The two products are suggested to be regioisomers obtained by Wessely᎐Moser transposition, isomerization specifically characteristic of C-glucosylflavones and proving that the sugar related to the flavone in position 8 is glucose w3x. The UV spectrum of compound 1 in MeOHrAlCl 3rHCl showed four absorption peaks, which is characteristic of 5-hydroxyflavones. The sodium acetate spectrum exhibited a 6-nm bathochromic shift of band II, indicating the presence of a 7-OH group. The 1 H-NMR spectrum exhibited the H-6 and H-3 protons at 6.47 and 6.88 ppm, respectively, and the pattern characteristic for a 4⬘-methoxy B-ring w4x. The presence of an OMe group at C-4⬘ was proved by the UV spectrum in MeOH᎐NaOH Ž ⌬ BI s 8 nm, ⌬ BII s 0 nm.. The assignment of the 6.47 signal to H-6⬘ was proved by the 13 C-NMR spectrum which excluded the presence of a proton at C-8. The 13 C-NMR of 1 was reminiscent of that of 8-glucosylapigenin w5x, the only differences being the downfield shift of the C-2Y resonance Ž10 ppm., an upfield shift of the C-1⬘ Ž1.5 ppm., and the presence of an extra glucose moiety. Therefore, the structure of compound 1 was deduced to be 2Y-O--D-glucosyl-8-C--D-glucosyl-4⬘-O-methylapigenin.
Acknowledgements Authors present their gratefulness to the head of the Analytical Chemistry Department of the Perm State Pharmaceutical Academy, Professor Vladislav O. Kozminykh, for his help in structure determination.
References w1x Bellakhdar J, Claisse R, Fleurentin J, Younos C. J Ethnopharmacol 1991;35:123.
416 w2x w3x w4x w5x
F. Khallouki et al. r Fitoterapia 71 (2000) 413᎐416 Bellakhdar J. These ` Doct Univ Metz 1997;1157. Wessely F, Moser GH. Monaths Chem 1930;56:97. Chatterjee A, Sarkar S, Saha SK. Phytochemistry 1981;20Ž7.:1760. Horowitz RM, Gentili B. Chem Ind 1964;498.