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Antioxidant and antimicrobial activities of Onosma argentatum and Rubia peregrina
Fitoterapia 74 (2003) 682–685
Short report
Antioxidant and antimicrobial activities of Onosma argentatum and Rubia peregrina a c ¨ U. Ozgen , P.J. Houghtonb,*, Y. Ogundipeb, M. Coskun ¸ a
¨ University, Faculty of Pharmacy, Pharmacognosy Department, Erzurum 25240, Turkey Ataturk b Pharmacognosy Research Laboratories, Department of Pharmacy, King’s College London, 150 Stamford Street, London SE1 8WA, UK c Ankara University, Faculty of Pharmacy, Pharmaceutical Botany Department, ˘ TandoganyAnkara 06100, Turkey Received 17 March 2003; accepted 2 April 2003
Abstract The n-hexane–dichloromethane (1:1) extract of the roots of Onosma argentatum and the methanol extract (partitioned between water and chloroform, ethyl acetate and n-butanol, respectively), of the underground parts (roots and rhizomes) of Rubia peregrina were tested in vitro for their antioxidant and antimicrobial activities. The highest antioxidant activity (98%) was observed at 0.1% concentration for the roots of O. argentatum. It was 96% at 0.25% concentration on the ethyl acetate fraction of R. peregrina. O. argentatum extract was effective on Staphyloccoccus aureus, Bacillus subtilis and Escherichia coli. The ethyl acetate and chloroform fractions of R. peregrina were effective on S. aureus and E. coli, respectively. These two species did not have any antifungal activity. 䊚 2003 Elsevier B.V. All rights reserved. Keywords: Onosma argentatum; Rubia peregrina; Antioxidant activity; Antimicrobial activity
Plant. Onosma argentatum Hub.-Mor. (Boraginaceae) roots collected from Dilimli village (Ilica District, Erzurum Province, Turkey) in March 2001 and identified by ¨ University, Art and Science Faculty, Dr Yusuf Kaya (Assistant Professor in Ataturk ¨ ¨ Universitesi Department of Biology). A voucher specimen was deposited at Ataturk ¨ Fen Fakultesi Herbaryumu (ATA), (ATA 9729). *Corresponding author. E-mail address: [email protected] (P.J. Houghton). 0367-326X/03/$ - see front matter 䊚 2003 Elsevier B.V. All rights reserved. doi:10.1016/S0367-326X(03)00161-8
¨ U. Ozgen et al. / Fitoterapia 74 (2003) 682–685
683
Rubia peregrina L. (Rubiaceae) underground parts (roots and rhizomes), collected ¨ from Bartın Province, Turkey in August 1992 and identified by U. Ozgen. A ¨ ¨ voucher specimen was deposited at Ankara Universitesi Eczacılık Fakultesi Herbaryumu (AEF), (AEF 21228). Uses in traditional medicine. O. argentatum is an endemic species in Turkey w1x. Some Onosma species (O. sericeum Willd., O. microcarpum Steven ex D.C.) are used for treatment of wounds in rural areas in Turkey w2x. R. peregrina grows in Northwest Europe, North Africa and Northwest Turkey w1x. The underground parts of R. peregrina have been used as a natural dye in Turkey w3 x . Previously isolated classes of constituents. Onosma species contain naphthoquinones w4x. Rubia species contain quinonic compounds comprising of anthraquinones, naphthoquinones, naphthohydroquinones and their glycosides w5–10x. Tested material. The roots of O. argentatum were powdered and extracted with boiling n-hexane–dichloromethane mixture (1:1) (yield: 2%). Preliminary phytochemical screening w11x gave positive tests for quinonic compounds and steroids andyor triterpenoids. The underground parts of R. peregrina were powdered and extracted with methanol. The concentrated extract (yield: 30%) was dissolved in 10% MeOH and partitioned with CHCl3 (yield: 2%), EtOAc (1.2%) and n-butanol (4%). Preliminary phytochemical screening w11x gave positive tests for quinonic compounds, steroids andyor triterpenoids on methanolic extract. Studied activity. Antioxidant by the TBA method w12x. Antimicrobial by the holein-plate bioassay method w13x.
Used microorganisms. Listed in Table 1.
Results. Antioxidant activity. O. argentatum extract had 98% antioxidant activity at 0.1% concentration and the IC50 was 0.0076% wyv. The IC50 value for the total methanolic extract of R. peregrina was 0.149%. The MeOH, CHCl3 and EtOAc fractions had high antioxidant activities at 0.25% wyv concentration, i.e. 94.2, 95.4 and 96% inhibition, respectively. The lowest inhibition (38% at the same concentration) was observed on the n-butanol extract. Antioxidant activity results are means of four replicates. Antimicrobial activity. Reported in Table 1.
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684
Table 1 Antimicrobial activity of O. argentatum and R. peregrina extracts Tested material* a
O. argentatum R. peregrina MeOHb R. peregrina EtOAcc R. peregrina CHCl3c R. peregrina BuOHc AMP
B.s.
E.c.
P.a.
S.a.
T.t.
T.i.
M.g.
C.a.
28 18 21 – – 32
13 – – 12 – 12
– – – – – –
32 18 28 – – 28
– – – – – NT
– – – – – NT
– – – – – NT
– – – – – NT
B.s., Bacillus subtilis; E.c., Escherichia coli; P.a., Pseudomonas aeruginosa; S.a., Staphylococcus aureus; T.t., Trichophyton tonsurans; T.i., Trichophyton interdigitale; M.g., Microphyton gypseum; C.a., Candida albicans. Ns4. AMPsampicillin 50 mgyml. NTsnot tested. –sNo inhibition. * 100 mg extractydisk; inhibition zone mm. a n-Hexane–dichloromethan (1:1) extract. b Crude extract. c Extract.
Conclusions. O. argentatum extract was effective on Bacillus subtilis, Escherichia coli and Staphyloccoccus aureus, EtOAc extract of R. peregrina on S. aureus and CHCl3 extract of R. peregrina on E. coli. Antifungal activity was not observed for these two species. These findings support the use of O. argentatum in Turkish traditional medicine. The wound healing activity of O. argentatum may be due to its antimicrobial and antioxidant activities since oxygen free radicals impair the wound healing activity and antioxidants and antimicrobials partly improve the healing process w14x. Purification of the active principles of O. argentatum and R. peregrina are in progress. References w1x Davis PH. Flora of Turkey and the East Aegean Islands. Edinburgh: Edinburgh University Press, 1988. w2x Sezik E, Yesilada E, Tabata M, Honda G, Takaishi Y, Fujita T, et al. Econ Bot 1997;51:195. w3x Baytop T. Therapy with medicinal plants in Turkey. Istanbul: Istanbul University Publications, No 3255, 1984. w4x Khajuria RK, Jain SM. Indian J Chem Sect B 1993;32:390. w5x Burnett AR, Thomson RH. J Chem Soc (C) 1968:854. w6x Itokawa H, Ibraheim ZZ, Oiao Y, Takeya K. Chem Pharm Bull 1993;4:1869. w7x Itokawa H, Mihara K, Takeya K. Chem Pharm Bull 1983;31:2353. w8x Itokawa H, Qiao Y, Takeya K. Phytochemistry 1989;28:3465. w9x Itokawa H, Qiao Y, Takeya K. Phytochemistry 1991;30:637. w10x Kawasaki Y, Goda Y, Yoshimira K. Chem Pharm Bull 1992;40:1504.
¨ U. Ozgen et al. / Fitoterapia 74 (2003) 682–685
685
w11x Chhabra SC, Uiso FC, Mshiu EN. J Ethnopharmacol 1984;11:157. w12x Halliwell B, Gutteridge JMC. Free radicals in biology and medicine. New York: Oxford University Press, 1999. w13x Hugo WB, Russel AD. Pharmaceutical microbiology. 3rd ed. Blackwell Scientific Publications, 1983. ¨ w14x Senel ¸ ¸ ¸ ˘ F, Bulan R. Ann Plast Surg 1997;39:516. O, Cetinkale O, Ozbay G, Ahcioglu
Short report
Antioxidant and antimicrobial activities of Onosma argentatum and Rubia peregrina a c ¨ U. Ozgen , P.J. Houghtonb,*, Y. Ogundipeb, M. Coskun ¸ a
¨ University, Faculty of Pharmacy, Pharmacognosy Department, Erzurum 25240, Turkey Ataturk b Pharmacognosy Research Laboratories, Department of Pharmacy, King’s College London, 150 Stamford Street, London SE1 8WA, UK c Ankara University, Faculty of Pharmacy, Pharmaceutical Botany Department, ˘ TandoganyAnkara 06100, Turkey Received 17 March 2003; accepted 2 April 2003
Abstract The n-hexane–dichloromethane (1:1) extract of the roots of Onosma argentatum and the methanol extract (partitioned between water and chloroform, ethyl acetate and n-butanol, respectively), of the underground parts (roots and rhizomes) of Rubia peregrina were tested in vitro for their antioxidant and antimicrobial activities. The highest antioxidant activity (98%) was observed at 0.1% concentration for the roots of O. argentatum. It was 96% at 0.25% concentration on the ethyl acetate fraction of R. peregrina. O. argentatum extract was effective on Staphyloccoccus aureus, Bacillus subtilis and Escherichia coli. The ethyl acetate and chloroform fractions of R. peregrina were effective on S. aureus and E. coli, respectively. These two species did not have any antifungal activity. 䊚 2003 Elsevier B.V. All rights reserved. Keywords: Onosma argentatum; Rubia peregrina; Antioxidant activity; Antimicrobial activity
Plant. Onosma argentatum Hub.-Mor. (Boraginaceae) roots collected from Dilimli village (Ilica District, Erzurum Province, Turkey) in March 2001 and identified by ¨ University, Art and Science Faculty, Dr Yusuf Kaya (Assistant Professor in Ataturk ¨ ¨ Universitesi Department of Biology). A voucher specimen was deposited at Ataturk ¨ Fen Fakultesi Herbaryumu (ATA), (ATA 9729). *Corresponding author. E-mail address: [email protected] (P.J. Houghton). 0367-326X/03/$ - see front matter 䊚 2003 Elsevier B.V. All rights reserved. doi:10.1016/S0367-326X(03)00161-8
¨ U. Ozgen et al. / Fitoterapia 74 (2003) 682–685
683
Rubia peregrina L. (Rubiaceae) underground parts (roots and rhizomes), collected ¨ from Bartın Province, Turkey in August 1992 and identified by U. Ozgen. A ¨ ¨ voucher specimen was deposited at Ankara Universitesi Eczacılık Fakultesi Herbaryumu (AEF), (AEF 21228). Uses in traditional medicine. O. argentatum is an endemic species in Turkey w1x. Some Onosma species (O. sericeum Willd., O. microcarpum Steven ex D.C.) are used for treatment of wounds in rural areas in Turkey w2x. R. peregrina grows in Northwest Europe, North Africa and Northwest Turkey w1x. The underground parts of R. peregrina have been used as a natural dye in Turkey w3 x . Previously isolated classes of constituents. Onosma species contain naphthoquinones w4x. Rubia species contain quinonic compounds comprising of anthraquinones, naphthoquinones, naphthohydroquinones and their glycosides w5–10x. Tested material. The roots of O. argentatum were powdered and extracted with boiling n-hexane–dichloromethane mixture (1:1) (yield: 2%). Preliminary phytochemical screening w11x gave positive tests for quinonic compounds and steroids andyor triterpenoids. The underground parts of R. peregrina were powdered and extracted with methanol. The concentrated extract (yield: 30%) was dissolved in 10% MeOH and partitioned with CHCl3 (yield: 2%), EtOAc (1.2%) and n-butanol (4%). Preliminary phytochemical screening w11x gave positive tests for quinonic compounds, steroids andyor triterpenoids on methanolic extract. Studied activity. Antioxidant by the TBA method w12x. Antimicrobial by the holein-plate bioassay method w13x.
Used microorganisms. Listed in Table 1.
Results. Antioxidant activity. O. argentatum extract had 98% antioxidant activity at 0.1% concentration and the IC50 was 0.0076% wyv. The IC50 value for the total methanolic extract of R. peregrina was 0.149%. The MeOH, CHCl3 and EtOAc fractions had high antioxidant activities at 0.25% wyv concentration, i.e. 94.2, 95.4 and 96% inhibition, respectively. The lowest inhibition (38% at the same concentration) was observed on the n-butanol extract. Antioxidant activity results are means of four replicates. Antimicrobial activity. Reported in Table 1.
¨ U. Ozgen et al. / Fitoterapia 74 (2003) 682–685
684
Table 1 Antimicrobial activity of O. argentatum and R. peregrina extracts Tested material* a
O. argentatum R. peregrina MeOHb R. peregrina EtOAcc R. peregrina CHCl3c R. peregrina BuOHc AMP
B.s.
E.c.
P.a.
S.a.
T.t.
T.i.
M.g.
C.a.
28 18 21 – – 32
13 – – 12 – 12
– – – – – –
32 18 28 – – 28
– – – – – NT
– – – – – NT
– – – – – NT
– – – – – NT
B.s., Bacillus subtilis; E.c., Escherichia coli; P.a., Pseudomonas aeruginosa; S.a., Staphylococcus aureus; T.t., Trichophyton tonsurans; T.i., Trichophyton interdigitale; M.g., Microphyton gypseum; C.a., Candida albicans. Ns4. AMPsampicillin 50 mgyml. NTsnot tested. –sNo inhibition. * 100 mg extractydisk; inhibition zone mm. a n-Hexane–dichloromethan (1:1) extract. b Crude extract. c Extract.
Conclusions. O. argentatum extract was effective on Bacillus subtilis, Escherichia coli and Staphyloccoccus aureus, EtOAc extract of R. peregrina on S. aureus and CHCl3 extract of R. peregrina on E. coli. Antifungal activity was not observed for these two species. These findings support the use of O. argentatum in Turkish traditional medicine. The wound healing activity of O. argentatum may be due to its antimicrobial and antioxidant activities since oxygen free radicals impair the wound healing activity and antioxidants and antimicrobials partly improve the healing process w14x. Purification of the active principles of O. argentatum and R. peregrina are in progress. References w1x Davis PH. Flora of Turkey and the East Aegean Islands. Edinburgh: Edinburgh University Press, 1988. w2x Sezik E, Yesilada E, Tabata M, Honda G, Takaishi Y, Fujita T, et al. Econ Bot 1997;51:195. w3x Baytop T. Therapy with medicinal plants in Turkey. Istanbul: Istanbul University Publications, No 3255, 1984. w4x Khajuria RK, Jain SM. Indian J Chem Sect B 1993;32:390. w5x Burnett AR, Thomson RH. J Chem Soc (C) 1968:854. w6x Itokawa H, Ibraheim ZZ, Oiao Y, Takeya K. Chem Pharm Bull 1993;4:1869. w7x Itokawa H, Mihara K, Takeya K. Chem Pharm Bull 1983;31:2353. w8x Itokawa H, Qiao Y, Takeya K. Phytochemistry 1989;28:3465. w9x Itokawa H, Qiao Y, Takeya K. Phytochemistry 1991;30:637. w10x Kawasaki Y, Goda Y, Yoshimira K. Chem Pharm Bull 1992;40:1504.
¨ U. Ozgen et al. / Fitoterapia 74 (2003) 682–685
685
w11x Chhabra SC, Uiso FC, Mshiu EN. J Ethnopharmacol 1984;11:157. w12x Halliwell B, Gutteridge JMC. Free radicals in biology and medicine. New York: Oxford University Press, 1999. w13x Hugo WB, Russel AD. Pharmaceutical microbiology. 3rd ed. Blackwell Scientific Publications, 1983. ¨ w14x Senel ¸ ¸ ¸ ˘ F, Bulan R. Ann Plast Surg 1997;39:516. O, Cetinkale O, Ozbay G, Ahcioglu