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Natural Product Research: Formerly Natural Product Letters Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/gnpl20
Cytotoxic, antibacterial and antioxidant activities of extracts of the bark of Melia azedarach (China Berry) a
a
a
Muhammad Zahoor , Manzoor Ahmed , Sumaira Naz & Musarrat a
Ayaz a
Department of Chemistry, University of Malakand Chakdara Dir (lower), P.O Box: 18000, Khyber Pakhtunkhwa, Pakistan Published online: 26 Nov 2014.
Click for updates To cite this article: Muhammad Zahoor, Manzoor Ahmed, Sumaira Naz & Musarrat Ayaz (2014): Cytotoxic, antibacterial and antioxidant activities of extracts of the bark of Melia azedarach (China Berry), Natural Product Research: Formerly Natural Product Letters, DOI: 10.1080/14786419.2014.982649 To link to this article: http://dx.doi.org/10.1080/14786419.2014.982649
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Natural Product Research, 2014 http://dx.doi.org/10.1080/14786419.2014.982649
SHORT COMMUNICATION Cytotoxic, antibacterial and antioxidant activities of extracts of the bark of Melia azedarach (China Berry) Muhammad Zahoor*, Manzoor Ahmed, Sumaira Naz and Musarrat Ayaz Department of Chemistry, University of Malakand Chakdara Dir (lower), P.O Box: 18000, Khyber Pakhtunkhwa, Pakistan
Downloaded by [York University Libraries] at 15:14 04 March 2015
(Received 19 September 2014; final version received 25 October 2014)
Nature provides a variety of drugs and medicinal agents derived from plants. This study was conducted to determine antimicrobial, antioxidant and cytotoxic activities of extracts of Melia azedarach bark with methanol/water (9:1 v/v), chloroform, butanol, hexane, water and ethyl acetate. For the determination of the antimicrobial activities, the agar well diffusion method was employed. Cytotoxicity was studied by brine shrimp lethality assay; antioxidant activities were measured using 1,1-diphenyl-2picrylhydrazyl. The chloroform extract was active against Enterobacter aerogenes and Proteus mirabilis, the ethyl acetate extract had highest antibacterial spectrum against Pseudomonas aeruginosa, the n-hexane extract had highest inhibition against E. aerogenes, the aqueous extract showed highest activities against P. mirabilis, the butanol fraction showed highest activities against E. aerogenes and the methanolic extract was highly active against P. mirabilis. Keywords: antimicrobial activities; antioxidant activities; cytotoxic activities; Melia azedarach; bark extract
1. Introduction Nature provides a variety of drugs and medicinal agents derived from the natural sources such as plants which are used as medicine. Herbal medicines are used as an integral part of modern pharmacopoeia (Shaw et al. 1997). According to the World Health Organization, for about 80% of the world’s population in developing countries, medicinal plants play an essential role (Fransworth et al. 1985). About 50% of modern drugs are isolated or derived from plants, while in Unani medicine, medicinal plant extracts dominate. The value of drugs derived from
*Corresponding author. Email: [email protected] q 2014 Taylor & Francis
2
M. Zahoor et al.
tropical forests is projected to represent about 20 million dollars a year (Sharif & Banik 2006). In general, parts of plants such as roots, stems, flowers, fruits and secretions of twigs are used for this purpose. Usually local communities and folk healers collect these raw plant materials in smaller amount, and on a larger scale these raw materials are traded in the market and used as bulk raw materials by the herbal industries for preparing herbal medicines (Uniyal et al. 2006). Medicinal plants provide antimicrobial agents that are used worldwide (Srivastava et al. 1996). The aim of this study was performed to determine whether extracts of Melia azedarach bark with aqueous methanol, ethyl acetate, chloroform, n-hexane, butanol or water exhibit antimicrobial, cytotoxic or antioxidant activities.
Downloaded by [York University Libraries] at 15:14 04 March 2015
2. Results and discussion 2.1. Antibacterial activities of M. azedarach extracts The mean diameter of the zone of inhibition of chloroform, hexane, aqueous, butanol, ethyl acetate and crude extract fractions of M. azedarach bark against Staphylococcus aureus was 8, 7, 4, 6, 4 and 4 mm, respectively (Table 1). The chloroform fraction showed highest antibacterial spectrum against S. aureus. For ethyl acetate, hexane, aqueous, butanol, chloroform and crude extract fractions of M. azedarach bark, the mean values of the diameter of the zone of inhibition against Escherichia coli were 6, 4, 5, 5, 3 and 4 mm, respectively (Table 1). The ethyl acetate extract was highly effective against E. coli. For the hexane, chloroform, aqueous, butanol, ethyl acetate and crude extract fractions of M. azedarach bark, the mean values of the diameter of the zone of inhibition against Enterobacter aerogenes were 17, 10, 6, 9, 8 and 7 mm, respectively (Table 1). The data show that the hexane extract fraction has highest zone of inhibition against E. aerogenes. For the hexane, chloroform, aqueous, butanol, ethyl acetate and crude extract fractions of M. azedarach, the mean values of the diameter of the zone of inhibition against Salmonella typhi were 6, 4, 4, 5, 4 and 5 mm, respectively (Table 1). The hexane fraction was highly effective against S. typhi. For the chloroform, hexane, aqueous, butanol, ethyl acetate and crude extract fractions of M. azedarach bark, the mean values of the diameter of the zone of inhibition against Proteus mirabilis were 10, 8, 8, 6, 5 and 8 mm, respectively (Table 1). The chloroform extract fraction was highly effective against P. mirabilis. For the ethyl acetate, hexane, aqueous, butanol, chloroform and crude extract fractions of M. azedarach bark, the mean values of the diameter of the zone of inhibition against Pseudomonas aeruginosa were 9, 6, 7, 7, 7 and 9 mm, respectively (Table 1). The ethyl acetate and crude extract fractions were highly effective against P. aeruginosa. For the hexane, ethyl acetate, aqueous, butanol, chloroform and crude extract fractions of M. azedarach bark, the mean values of the diameter of the zone of inhibition against Klebsiella pneumonia were 8, 3, 4, 7, 6 and 4 mm, respectively (Table 1). The hexane fraction showed high zone of inhibition against K. pneumonia. Table 1. Diameter of zone of inhibition (mm) of M. azedarach (China berry) bark extracts. Diameter of zone of inhibition (mm) Extract Hexane Chloroform Water Butanol Ethyl acetate Crude
S. aureus P. mirabilis K. pneumonia S. typhi E. coli P. aeruginosa E. aerogenes 7 8 4 6 4 4
8 10 8 6 5 8
8 6 4 7 3 4
6 4 4 5 4 5
4 3 5 5 6 4
6 7 7 7 9 9
17 10 6 9 8 7
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Table 2. Free RSA (%) of the tested extract fractions of M. azedarach relative to DPPH solution (control). Concentration
Downloaded by [York University Libraries] at 15:14 04 March 2015
%RSA %RSA %RSA %RSA %RSA %RSA
(chloroform extract) (butanol extract) (hexane extract) (ethyl acetate extract) (aqueous extract) (crude extract)
20 mg L21
40 mg L21
60 mg L21
80 mg L21
100 mg L21
93.00 94.00 40.56 96.49 56.45 98.15
94.00 84.00 62.00 96.49 73.48 95.89
94.00 97.00 83.85 94.73 81.37 95.44
94.00 94.00 95.92 91.22 96.57 97.50
96.92 94.81 98.91 89.47 97.37 98.15
2.2. Antioxidant activities of the extracts The free radical scavenging activities (RSAs) of the chloroform, crude, butanol, hexane, water and ethyl acetate extract fractions are shown in Table 2. It is evident from the table that with the increase in concentration, the antioxidant activities were pronounced for all the tested extract fractions of the M. azedarach bark. 2.3. Cytotoxic activities of the extracts The cytotoxic activities were determined for hexane, chloroform and ethyl acetate fractions of the extract. The results are graphically shown in Figures S1 – S3. Brine shrimp eggs were used to determine the cytotoxic activities. All the three tested fractions showed significant cytotoxic effects on brine shrimp. It was found that for these fractions, with the increase in concentration from 30 to 100 mg/mL, the cytotoxic activities were enhanced many folds. 3. Conclusions From this study, it was concluded that the chloroform, crude, butanol, hexane, aqueous and ethyl acetate fractions of M. azedarach bark extracts showed significant antibacterial activities. The selected extracts also showed the antioxidant and cytotoxic activities. Thus as a substitute source, these extracts should be used as a source of useful antimicrobial agents in various infectious diseases and also as antioxidant agents. Supplementary material Experimental materials relating to this article are available online, alongside Figures S1 –S3. References Fransworth NR, Akerele O, Bingel AS, Soejarto DD, Guo ZG. 1985. Medicinal plants in therapy. Bull World Health Org. 63:965–981. Sharif MDM, Banik GR. 2006. Status and u¨tilization of medicinal plants in Rangamati of Bangladesh. Res J Agri Biol Sci. 2:268– 373. Shaw D, Soleu K, Murray V. 1997. Traditional remedies and food supplements. Drug Saf. 17:342–356. Srivastava J, Lambert J, Ietmeyer NV. 1996. Medicinal plants: an expanding role in development. World Bank technical Paper. No. 320. Uniyal SK, Singh KN, Jamwal P, Lal B. 2006. Traditional use of medicinal plants among the tribal communities of Chhota Bhangal, Western Himalayan. J Ethnobiol Ethnomed. 2:1–14.
Natural Product Research: Formerly Natural Product Letters Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/gnpl20
Cytotoxic, antibacterial and antioxidant activities of extracts of the bark of Melia azedarach (China Berry) a
a
a
Muhammad Zahoor , Manzoor Ahmed , Sumaira Naz & Musarrat a
Ayaz a
Department of Chemistry, University of Malakand Chakdara Dir (lower), P.O Box: 18000, Khyber Pakhtunkhwa, Pakistan Published online: 26 Nov 2014.
Click for updates To cite this article: Muhammad Zahoor, Manzoor Ahmed, Sumaira Naz & Musarrat Ayaz (2014): Cytotoxic, antibacterial and antioxidant activities of extracts of the bark of Melia azedarach (China Berry), Natural Product Research: Formerly Natural Product Letters, DOI: 10.1080/14786419.2014.982649 To link to this article: http://dx.doi.org/10.1080/14786419.2014.982649
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms &
Downloaded by [York University Libraries] at 15:14 04 March 2015
Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Natural Product Research, 2014 http://dx.doi.org/10.1080/14786419.2014.982649
SHORT COMMUNICATION Cytotoxic, antibacterial and antioxidant activities of extracts of the bark of Melia azedarach (China Berry) Muhammad Zahoor*, Manzoor Ahmed, Sumaira Naz and Musarrat Ayaz Department of Chemistry, University of Malakand Chakdara Dir (lower), P.O Box: 18000, Khyber Pakhtunkhwa, Pakistan
Downloaded by [York University Libraries] at 15:14 04 March 2015
(Received 19 September 2014; final version received 25 October 2014)
Nature provides a variety of drugs and medicinal agents derived from plants. This study was conducted to determine antimicrobial, antioxidant and cytotoxic activities of extracts of Melia azedarach bark with methanol/water (9:1 v/v), chloroform, butanol, hexane, water and ethyl acetate. For the determination of the antimicrobial activities, the agar well diffusion method was employed. Cytotoxicity was studied by brine shrimp lethality assay; antioxidant activities were measured using 1,1-diphenyl-2picrylhydrazyl. The chloroform extract was active against Enterobacter aerogenes and Proteus mirabilis, the ethyl acetate extract had highest antibacterial spectrum against Pseudomonas aeruginosa, the n-hexane extract had highest inhibition against E. aerogenes, the aqueous extract showed highest activities against P. mirabilis, the butanol fraction showed highest activities against E. aerogenes and the methanolic extract was highly active against P. mirabilis. Keywords: antimicrobial activities; antioxidant activities; cytotoxic activities; Melia azedarach; bark extract
1. Introduction Nature provides a variety of drugs and medicinal agents derived from the natural sources such as plants which are used as medicine. Herbal medicines are used as an integral part of modern pharmacopoeia (Shaw et al. 1997). According to the World Health Organization, for about 80% of the world’s population in developing countries, medicinal plants play an essential role (Fransworth et al. 1985). About 50% of modern drugs are isolated or derived from plants, while in Unani medicine, medicinal plant extracts dominate. The value of drugs derived from
*Corresponding author. Email: [email protected] q 2014 Taylor & Francis
2
M. Zahoor et al.
tropical forests is projected to represent about 20 million dollars a year (Sharif & Banik 2006). In general, parts of plants such as roots, stems, flowers, fruits and secretions of twigs are used for this purpose. Usually local communities and folk healers collect these raw plant materials in smaller amount, and on a larger scale these raw materials are traded in the market and used as bulk raw materials by the herbal industries for preparing herbal medicines (Uniyal et al. 2006). Medicinal plants provide antimicrobial agents that are used worldwide (Srivastava et al. 1996). The aim of this study was performed to determine whether extracts of Melia azedarach bark with aqueous methanol, ethyl acetate, chloroform, n-hexane, butanol or water exhibit antimicrobial, cytotoxic or antioxidant activities.
Downloaded by [York University Libraries] at 15:14 04 March 2015
2. Results and discussion 2.1. Antibacterial activities of M. azedarach extracts The mean diameter of the zone of inhibition of chloroform, hexane, aqueous, butanol, ethyl acetate and crude extract fractions of M. azedarach bark against Staphylococcus aureus was 8, 7, 4, 6, 4 and 4 mm, respectively (Table 1). The chloroform fraction showed highest antibacterial spectrum against S. aureus. For ethyl acetate, hexane, aqueous, butanol, chloroform and crude extract fractions of M. azedarach bark, the mean values of the diameter of the zone of inhibition against Escherichia coli were 6, 4, 5, 5, 3 and 4 mm, respectively (Table 1). The ethyl acetate extract was highly effective against E. coli. For the hexane, chloroform, aqueous, butanol, ethyl acetate and crude extract fractions of M. azedarach bark, the mean values of the diameter of the zone of inhibition against Enterobacter aerogenes were 17, 10, 6, 9, 8 and 7 mm, respectively (Table 1). The data show that the hexane extract fraction has highest zone of inhibition against E. aerogenes. For the hexane, chloroform, aqueous, butanol, ethyl acetate and crude extract fractions of M. azedarach, the mean values of the diameter of the zone of inhibition against Salmonella typhi were 6, 4, 4, 5, 4 and 5 mm, respectively (Table 1). The hexane fraction was highly effective against S. typhi. For the chloroform, hexane, aqueous, butanol, ethyl acetate and crude extract fractions of M. azedarach bark, the mean values of the diameter of the zone of inhibition against Proteus mirabilis were 10, 8, 8, 6, 5 and 8 mm, respectively (Table 1). The chloroform extract fraction was highly effective against P. mirabilis. For the ethyl acetate, hexane, aqueous, butanol, chloroform and crude extract fractions of M. azedarach bark, the mean values of the diameter of the zone of inhibition against Pseudomonas aeruginosa were 9, 6, 7, 7, 7 and 9 mm, respectively (Table 1). The ethyl acetate and crude extract fractions were highly effective against P. aeruginosa. For the hexane, ethyl acetate, aqueous, butanol, chloroform and crude extract fractions of M. azedarach bark, the mean values of the diameter of the zone of inhibition against Klebsiella pneumonia were 8, 3, 4, 7, 6 and 4 mm, respectively (Table 1). The hexane fraction showed high zone of inhibition against K. pneumonia. Table 1. Diameter of zone of inhibition (mm) of M. azedarach (China berry) bark extracts. Diameter of zone of inhibition (mm) Extract Hexane Chloroform Water Butanol Ethyl acetate Crude
S. aureus P. mirabilis K. pneumonia S. typhi E. coli P. aeruginosa E. aerogenes 7 8 4 6 4 4
8 10 8 6 5 8
8 6 4 7 3 4
6 4 4 5 4 5
4 3 5 5 6 4
6 7 7 7 9 9
17 10 6 9 8 7
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Table 2. Free RSA (%) of the tested extract fractions of M. azedarach relative to DPPH solution (control). Concentration
Downloaded by [York University Libraries] at 15:14 04 March 2015
%RSA %RSA %RSA %RSA %RSA %RSA
(chloroform extract) (butanol extract) (hexane extract) (ethyl acetate extract) (aqueous extract) (crude extract)
20 mg L21
40 mg L21
60 mg L21
80 mg L21
100 mg L21
93.00 94.00 40.56 96.49 56.45 98.15
94.00 84.00 62.00 96.49 73.48 95.89
94.00 97.00 83.85 94.73 81.37 95.44
94.00 94.00 95.92 91.22 96.57 97.50
96.92 94.81 98.91 89.47 97.37 98.15
2.2. Antioxidant activities of the extracts The free radical scavenging activities (RSAs) of the chloroform, crude, butanol, hexane, water and ethyl acetate extract fractions are shown in Table 2. It is evident from the table that with the increase in concentration, the antioxidant activities were pronounced for all the tested extract fractions of the M. azedarach bark. 2.3. Cytotoxic activities of the extracts The cytotoxic activities were determined for hexane, chloroform and ethyl acetate fractions of the extract. The results are graphically shown in Figures S1 – S3. Brine shrimp eggs were used to determine the cytotoxic activities. All the three tested fractions showed significant cytotoxic effects on brine shrimp. It was found that for these fractions, with the increase in concentration from 30 to 100 mg/mL, the cytotoxic activities were enhanced many folds. 3. Conclusions From this study, it was concluded that the chloroform, crude, butanol, hexane, aqueous and ethyl acetate fractions of M. azedarach bark extracts showed significant antibacterial activities. The selected extracts also showed the antioxidant and cytotoxic activities. Thus as a substitute source, these extracts should be used as a source of useful antimicrobial agents in various infectious diseases and also as antioxidant agents. Supplementary material Experimental materials relating to this article are available online, alongside Figures S1 –S3. References Fransworth NR, Akerele O, Bingel AS, Soejarto DD, Guo ZG. 1985. Medicinal plants in therapy. Bull World Health Org. 63:965–981. Sharif MDM, Banik GR. 2006. Status and u¨tilization of medicinal plants in Rangamati of Bangladesh. Res J Agri Biol Sci. 2:268– 373. Shaw D, Soleu K, Murray V. 1997. Traditional remedies and food supplements. Drug Saf. 17:342–356. Srivastava J, Lambert J, Ietmeyer NV. 1996. Medicinal plants: an expanding role in development. World Bank technical Paper. No. 320. Uniyal SK, Singh KN, Jamwal P, Lal B. 2006. Traditional use of medicinal plants among the tribal communities of Chhota Bhangal, Western Himalayan. J Ethnobiol Ethnomed. 2:1–14.
