SciELO - Scientific Electronic Library Online

 
vol.21 issue3A new approach for quantifying furanodiene and curzerene: a case study on the essential oils of Eugenia uniflora L., Myrtaceae (pitangueira) leavesDevelopment and validation of a HPLC method for standardization of herbal and commercial extracts of Myrcia uniflora author indexsubject indexarticles search
Home Pagealphabetic serial listing  

Services on Demand

Article

Indicators

Related links

  • Have no similar articlesSimilars in SciELO

Share


Revista Brasileira de Farmacognosia

Print version ISSN 0102-695X

Rev. bras. farmacogn. vol.21 no.3 Curitiba May/June 2011 Epub Mar 18, 2010

http://dx.doi.org/10.1590/S0102-695X2011005000037 

Chemical constituents from Piptadenia rigida Benth., Fabaceae, "angico"

 

 

Mário G. de CarvalhoI, *; Mário S. R. GomesI; Márcia C. C. de OliveiraI; Cleber J. da SilvaI; Acácio G. de CarvalhoII

IDepartamento de Química, Instituto de Ciências Exatas, Universidade Federal Rural do Rio de Janeiro, Br 465, km 07, 23890-000 Seropédica-RJ, Brazil
IIDepartamento de Produtos Florestais, Instituto de Florestas, Universidade Federal Rural do Rio de Janeiro, Br 465, km 07, 23890-000 Seropédica-RJ, Brazil

 

 


ABSTRACT

The phytochemical investigation of the roots of Piptadenia rigida Benth., Fabaceae, known as "angico", afforded sitosterol, lupeol, betuline, the chalcone isoliquiritigenin, the flavonoids, 7,4'-dihydroxyflavone, 7,3',4'-trihydroxyflavone, 7,8,3',4'-tetrahydroxyflavanone, 4-hydroxy-3,5-dimethoxybenzaldehyde and methyl-3,4-dihydroxy-benzoate. Both flavones were also isolated from the branches of this plant. Five derivatives of the aldehyde were obtained by diazomethane treatment. The structures of compounds were identified by IR, NMR and mass spectral data analysis of natural compounds and some derivatives, and by comparison with literature data.

Keywords: flavonoids, Piptadenia rigida, terpenoids, benzoyl aldehyde derivatives.


 

 

Introduction

The Piptadenia genus, Fabaceae, comprise eighty tropical species frequently occurring in South America. In Brazil these species are popularly known as "angico" and have been found in the North, Northeast, and Central West. This genus had been considered as heterogenous that was divided in Anadenanthera Speg., Pityrocarpa Benth. and Piptadenia Benth. On the other hand the Brazilian botanists have considered as Piptadenia sensu Bentham. The P. rigida has been classified as belonging to Leguminosae-Mimosoideae (Correa, 1984; Lorenzi, 2002) and more recently as Fabaceae family (Morin, 2010). It has been described the inhalation of narcotic snuff from some Piptadenia species by the Caribbean and by South American native Indians (Correa, 1984; Lorenzi, 2002, Stromberg, 1954). The previous phytochemical study of Piptadenia species described the presence of indole alkaloids from the seeds of P. peregrine, P. excelsa, P. macrocarpa, and P. leprostachya (Fish et al., 1955; Pachter et al., 1959; Legler & Tschesche, 1963, Yamasato et al., 1972). Steroids, heterosides and tannins have been identified in bark of P. columbrina (Bulhões et al., 1976) and the flavonoid anadantoside, 7,3',4'-trihydroxy-3-β-D-xylopiranosylflavane, was isolated from the bark of P. macrocarpa (Piacente et al. 1999). Besides the hallucinogenic properties of the seeds, the exudate from the wood has been used against respiratory infections (Shneider, 1937; Rangel, 1943). It is used in the popular medicine as anti leucorrhoea, anti diarrhea and as a cure for ulcer; the leaves have haemostatic function in cerebral disease and commotion (Bulhões et al., 1976; Piacente et al., 1999; Shneider, 1937; Rangel, 1943). This paper reports the first phytochemical study of P. rigida revealing for the first time the presence of a chalcone, three flavonoids, two benzoil derivatives, lupeol and betulin in Piptadenia genus.

 

Materials and Methods

The roots and branches of Piptadenia rigida Benth., Fabaceae, were collected in the forest garden of the Instituto de Florestas (IF) of Universidade Federal Rural do Rio de Janeiro (UFRRJ), Seropédica-RJ, by Dr. Acácio G. de Carvalho, Departamento de Produtos Florestais, IF, UFRRJ. It was identified by Prof. Dr. José Aguiar Sobrinho, Departamento de Ciências Ambientais, IF, UFRRJ. Voucher specimen (No RBR-21438) is deposited at the Herbarium of Instituto de Biologia, UFRRJ.

The powdered dried root (6.3 kg) and branches (2.0 kg) were extracted exhaustively with CH2Cl2 and MeOH at room temperature. The solvents were removed under vacuum to yield the residues PRBD (P. rigida branches dichloromethane, 1.0 g) and PRBM (P. rigida branches methanol, 83.0 g) from the branches, and PRRD (P. rigida root dichloromethane, 19.2 g) and PRRM (P. rigida root methanol, 310.5 g) from the root. The methanol extract from the branches PRBM (60.0 g) was partitioned with CHCl3 and MeOH:H2O (9:1). The fraction obtained with chloroform (13.4 g) was chromatographed on silica gel column and eluted with CH2Cl2 and mixture of this solvent with EtOAc and MeOH in increasing polarity. 140 fractions were collected. Fractions 64-73 and 75-79 yielded the flavonoids 9 (90.0 mg, mp 289-290 ºC) and 8 (80.0 mg, mp 212-213 ºC), respectively. The residue PRRD was dissolved in methanol and filtered to obtain two fractions, a solid (PRRD-1, 6.9 g) and solution (PRRD-2, 12.0 g). The solid was chromatographed on silica gel column using CHCl3 increasing the polarity with methanol to methanol 100%. Twenty fractions were collected, and fractions 8-15 were reunited and crystallized from methanol yielding the solid 1 (440.0 mg, mp 132-133 ºC). The fraction PRRD-2 was fractionated on silica gel column using the same systems of solvent and 27 fractions were collected and analyzed by TLC plate. Fractions 5-12 afforded 1 (522.0 mg). Preparative TLC of the fractions 13-25 with hexane:ethyl acetate (8:2) yielded 3 (53.6 mg, mp 258-259 ºC) and 4 (26.4 mg, gum). The extract PRRM was dissolved in MeOH:H2O (8:2) and partitioned with hexane (PRRMH, 2.24 g), chloroform (PRRMC, 11.9 g) and methanol. Chromatography fractionation and TLC analysis of PRRMH were used to isolate 2 (138.0 mg, PF 194-195 ºC) from fractions 4-16, and 1 (210.0 mg) from the fractions 19-23. PRRMC was filtered on silica gel column with dichlorometane, ethyl acetate and methanol to yield PRRMCD (270.0 mg), PRRMCE (6.3 g), PRRMCM (5.3 g). The IR and 1H NMR spectra of fraction PRRMCD let to identify mixture of aliphatic acids. Fraction PRRMCE was fractionated on silica gel CC using chloroform as initial solvent and increasing the polarity with methanol and fifty fractions were collected. Chromatography fractionation and TLC analysis of fractions 8-13 (294.0 mg) afforded 5 (20.0 mg, oil) and 6 (15.0 mg, mp 158-159 ºC). Fractions 27-33 afforded the flavanone 7 (22.0 mg, oil); fractions 45-53 were fractionated by preparative TLC using chloroform:ethyl acetate (1:3) to obtain the crystalline material identified as the flavone 8 (20.0 mg, mp 213-214 ºC). The purity of the compounds was checked by silica gel TLC plate, revealed by visualization with UV (254 and 366 nm), AlCl3-ETOH (1%) or exposure to iodine vapor, and 1H spectral analysis.

The alcohol function in 1, 2 and 3 was confirmed by the IR and 1H and 13C NMR spectral analysis of the products with PCC oxidations. An ethereal solution of diazomethane was added to the methanol solution of 4 (13.0 mg) and 5 (15.0 mg) to confirm the phenol groups. The unexpected additional signals in the 1H and 13C NMR spectra of 4 product let us to analyze it by CG-MS and a mixture of 4a, 4b, 4c, 4d and 4e was identified, Rt [%; m/z: (%)]: 4a: 6.38 [4.4; m/z: 196 (100, M+.), 181 (57), 165 (4), 153 (10), 123 (5), 93 (26)]; 4b: 7.18 [2.87; m/z: 210 (25, M+.), 181 (100), 167 (5), 151 (5), 148 (17)]; 4c: 7.56 [19.44; m/z: 210 (60), 195 (100), 167 (11), 153 (2), 139 (15), 137 (8), 107 (5)]; 4d: 8.10 [64.44; m/z: 224 (37, M+.), 182 (12), 181 (100), 167 (5), 151 (2), 137 (4), 121 (2), 107 (3)]; 4e: 9.02 [8.81; m/z: 224 (100, M+.), 209 (52), 195 (12), 179 (10), 181 (62), 151 (10)], (Scheme 1). The 1H NMR spectral analysis confirmed 5a as expected.

 

Results and Discussion

Chromatographic fractionation of the extracts from the roots and branches of Piptadenia rigida and analysis of the fractions allowed to identify sitosterol (1), lupeol (2), betulin (3), the chalcone isoliquiritigenin (6), the flavonoids 7,8,3',4'-tetrahydroxyflavanone (7), 7,3',4'-trihydroxyflavone (8), and 7,4'-dihydroxyflavone (9), besides 4-hydroxi-3,5-dimethoxybenzaldehyde (4) and methyl 3,4-dihydroxy-benzoate (5). The structures were established by IR, NMR and mass spectral data analysis of the natural compounds and some derivatives. The steroid 1 and the triterpenes 2 and 3 were identified by mp, IR, 1H and 13C NMR spectra analysis of natural substances and of the products obtained by PCC oxidation, sitostenone, lupenone and 3-oxo betulinic acid, respectively, and by comparison with values described in the literature (Chaurasia & Wichtl, 1987; Reynolds et al. 1986; Mahato & Kundu, 1994; Siddiqui et al., 1988).

 

 

The IR spectra of 4 and 5 showed band at cm-1: 3377, 3076, 1605, 1512, 1462, 1251, 1680 (νC=O in 4) and 1692 ((νC=O in 5). The signals at δH 9.79 (s, 1H, aldehyde), 7.12 (s, 2H, Ar-H), 3.94 (s, 6H, 2x OCH3) were identified in the 1H NMR spectrum of 4. The 13C NMR spectra (BBD and DEPT) of this aldehyde showed signals at 190.7 (HC=O, C-7), 147.8, 147.3x2 (C-4 and C-3,5), 128.3 (C-1), 106.6 (C-2,6) and 56.4 (2xOCH3). The same analysis of the 1H NMR spectra of 5 let to identify the signals of 3H of the ABC system in aromatic ring by δH 7.38 (brs, H-2), 7.32 (brd, J=8 Hz, H-6) and 6.79 (d, J=8 Hz, H-5) besides the signal at δH 3.69 (s, 3H, OCH3). The 13C NMR data confirmed this proposed benzoyl methyl esther structure by the signals at δC 166.5 (C=O of ester), 150.2, 145.0, 122.4 (C-4, C-3 and C-1, respectively), 122.0, 116.1, 115.0 (CH-6, 2 and 5) and 51.2 (OCH3 of ester). The 1H NMR spectrum of the residue obtained from treatment of 4 with diazomethane did not shows signals of aldehyde and showed signals at δH 6.38 (s), 7.19 (s), 3.82 (s), 3.89(s), and multiples signals at δH 3.7-4.0, 2.8-3.12, and 2.15-2.5. The 13C NMR spectrum (BBD and DEPT) of this product showed δCH at 52.6, 60.6, 77.0, 102.1, 105.8, 105.9, 106.3, δCH2 at 51.3, 34.0, 32.0, 27.4, and of OCH3 at 56.11, 60.9. These data let us to analyze this product by the GC-MS whose chromatogram showed five peaks and corresponding mass spectrum of each one [4a-4e, Rt, % and m/z (%), see experimental] was detected. Those NMR spectral data besides the proposed ions structures (Scheme 1) for: m/z 181 (4a1) and 153 (4a2) from 4a, m/z 181 (4b1), 167 (4b2), 148 (4b3) from 4b; m/z 195 (4c1), 167 (4c2), 153 (4c3) from 4c; m/z 181 (4d1), 167 (4d2), 148 (4d3) from 4d, and m/z 181 (4e1), 153 (4e2) from 4e, were compatible with the proposed structures in the mixture of 4 derivatives (Scheme 1).

The 1H NMR spectrum of chalcone 6 showed signals of ABC system [ δH 6.33 (d, J=2.0 Hz, H-3), 6.45 (dd, J1=8.7, J2=2.0 Hz, H-5), 8.01 (d, J=8.7 Hz, H-6)], two doublets of J=15 Hz (δH 7.65 and 7.84) and two signals of AA'BB' system at δH 6.90 (d, J=8.7 Hz, H-3',5') and 7.67 (J=8.7 Hz, H-2'.6'). These data and the carbons chemical shift of 6 were identical to those of literature for isoliquiritigenin (Almtorp et al., 1991).

The IR spectra of 7 and 8 showed band at cm-1: 3238, 3030, 1630 (νC=O), 1600, 1510 and 1272. The analysis of 1H NMR spectra of 7 and comparison with literature data of 7,8,3',4'-tetrahydroxy-flavanone, isolated from Acacia melanoxylon (Foo,1987), were used to confirm the proposed structure of 7. The unambiguous 1H and 13C NMR chemical shift assignment, including the 1D and 2D spectra of 7, are reported in the literature by Nascimento et al. (2003). The same spectral analysis of 8 allowed to identify two ABC system of hydrogen in an aromatic ring by the signals at δH 6.96 (brd, J=8.6 Hz, 1H), 6.98 (brs, 2H), 7,48 (brs, 1H), 7.45 (d, J=8.0 Hz, 1H), 8.01 (d, J=8.6 Hz, 1H). These six hydrogens and the singlet at δH 6.8 were compatible with a trihydroxyflavone. The doublet at δH 8.1 agrees with H-5 chemical shift of the ABC system in the flavone. The 13C NMR spectrum of 8 showed signal of quaternary carbons at δC 180.4 (C=O), 165.7 (C-2), 165.3 (C-7), 157.8 (C-9), 151.0 (C-4'), 147.2 (C-3'), 124.1 (C-1'), 116.6 (C-10), and of δC-H at 127.8 (C-5), 114.2 (C-6), 116.5/116.9 (C-2'/5"), 120.2 (C-6'), 103.8/105.2 (C-3/8). The peak at m/z (%): 272 [M+., 1%, C15H10O5), 161(43), 178(100), and 133(14), confirmed the proposed structure as 7,3',4'-trihydroxyflavone for 8, which carbon-13 data are registered in the literature (Agrawal, 1989).

The 1H NMR spectra analysis of 9 allowed to identify an ABC system in the ring A of flavone [ δH 6.92 (brd, J=8.0 Hz, H-6), 7.97 (d, J=8.0 Hz, H-5), 6.97 (brs, H-8)] and two doublets of para substituted aromatic ring [ δH 6.94(d, J=8.7 Hz, H-3'.5') and 7.88(d, J=8.7 Hz, H-2',6') of a flavone with δH-3 6.71 (s). The carbon chemical shifts at ? δC 103.5 (CH-8), 105.1 (CH-3), 116.2 (CH-6) 116.4 (CH-3',5'), 117.0 (C-10), 122.9 (C-1'), 127.8 (C-5), 129.4 (CH-2',6'), 159.0 (C-9), 162.6 (C-7), 165.2 (C-2), 166.0 (C-4'), 180.0 (C-4) were compatible with 7,4'-dihydroxyflavone proposed for 9. This compound was isolated from Medicago sativa and from Vicia faba (Fabaceae) (Wollehweber, 1994).

 

Acknowledgment

The authors thank to Conselho Nacional de Desenvolvimento Científico e Tecnológico, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Fundação Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro for scholarship and financial support and to Universidade Estadual do Norte Fluminense, Campos-RJ, for the 400 MHz NMR spectra.

 

References

Agrawal PK 1989. Methods in signal assignment of flavonoides In: Agrawal PK, Baural MC. Carbon-13 NMR of flavonoids, studies in organic chemistry, Amsterdam: Ed. Elsevier, 201 p.         [ Links ]

Almtorp GT, Bachmann TL, Torssel KBG 1991. Synthesis of flavonoids via the isoxazoline route. Acta Chem Scand 45: 212-215.         [ Links ]

Bulhões GCC, Silva AMN, Sá MAM 1976. Abordagem Fitoquímica de Plantas Nativas do Nordeste Brasileiro. An Depto Farm Centro de Cien Saude UFPE 15: 33-38.         [ Links ]

Chaurasia N, Wichtl MJ 1987. Sterols and steryl glycosides from Urtica dioica. J Nat Prod 50: 881-885.         [ Links ]

Correa MP 1984. Dicionário de Plantas úteis do Brasil; e das exóticas cultivadas. Ed. Ministério da Agricultura, Instituto Brasileiro de Desenvolvimento florestal, RJ, 4329 p.         [ Links ]

Fish MS, Johnson NM, Horning EC 1955. Alkaloids indole bases of P. peregrina (L.) Benth. and related species. J Am Chem Soc 27: 2892-2895.         [ Links ]

Foo LY 1987. Configuration and conformation of dihydroflavonols from Acacia melanoxylon. Phytochemistry 26: 813-817.         [ Links ]

Legler G, Tschesche R 1963. Die Isolierung von N-methyltriptamin, 5-methoxy-N-methyltryptamin und 5-O-methoxy-NN-dimethyltryptamin aus der rinde von Piptadenia peregrine Benth. Die Naturwissenschaften 50: 94-95.         [ Links ]

Lorenzi H 2002. Árvores Brasileiras: Manual de Identificação e cultivo de plantas arbóreas naturais do Brasil, Vol 1. 4ª ed. Nova Odessa, SP, Instituto Plantarum, 357 p.         [ Links ]

Mahato SB, Kundu AP 1994. 13C-NMR spectra of pentacyclic triterpenoids-a compilation and some saliente features. Phytochemistry 37: 1517-1575.         [ Links ]

Morim, MP 2010. Piptadenia in Lista de Espécies da Flora do Brasil. Jardim Botânico do Rio de Janeiro, available on http://floradobrasil.jbrj.gov.br/2010/FB103167.         [ Links ]

Nascimento IA do, Gomes MS da R, Carvalho MG de, Carvalho AG de 2003. Deslocamentos químicos de 1H e 13C de 5-H-flavanona e 5-H-flavonol isolados de Leguminosae. Rev Univ Rural Ser Cien Ex e da Terra 22: 81-87.         [ Links ]

Pachter JI, Zacharias DE, Ribeiro O 1959. Indole alkalods of Acer saccharinum (teh Silver Maple), Dictyoloma incanescens, Piptadenia columbrina, and Mimosa hostlis. J Org Chem 16: 1285-1287.         [ Links ]

Piacente S, Balderrama L, de Tommasi H, Morales L, Vargas L, Pizza C 1999. Anadanthoside: a flavanol-3-O-β-D-xilopyranoside from Anadenanthera macrocarpa. Phytochemistry 51: 709-711.         [ Links ]

Reynolds WF, Mclean S, Poplawski J, Enriquez RG, Escobar LI, Leóm I 1986. Total assignment of 13C and 1H-spectra of three isomeric triterpenol derivatives by 2D-NMR- an investigation of the potential utility of 1H chemical shifts in structural investigations of complex natural products. Tetrahedron 42: 3419-3428.         [ Links ]

Shneider HS 1937. Angico gum. Rev de Quim Ind Rio de Janeiro 6: 286-290.         [ Links ]

Rangel JL 1943. Angico gum. Rev de Quím Ind Rio de Janeiro 12: 16-18.         [ Links ]

Siddiqui S, Hafeez F, Begum S, Siddiqui DS 1988. Oleanderol, a new pentacyclic triterpene from the leaves of Nerium oleander. J Nat Prod 51: 229-233.         [ Links ]

Stromberg V 1954. The isolation of bufotenin from Piptadenia peregrine. J Am Chem Soc 20: 1707.         [ Links ]

Wollehweber E 1994. Flavones and Flavonoids in the Flavonoids-Advances in Research since 1986. Ed. J.B. Harborne, New York: Chapman & Hal.         [ Links ]

Yamasato S, Kawanishi K, Kato A, Hashimoto Y 1972. Organic bases from Brazilian Piptadenia species. Phytochemistry 11: 737-739.         [ Links ]

 

 

Received 8 Jul 2010
Accepted 27 Sep 2010

 

 

* E-mail: mgeraldo@ufrrj.br;Tel.+55 21 2682 2807; Fax: +55 21 2682 2807.