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In Vitro Evaluation of Antimicrobial and Antibiofilm Potentials of Essential Oil of Tanacetum argenteum (Lam.) Willd. subsp. canum (K.Koch) Grierson var. canum

Year 2018, Volume: 5 Issue: 1, 34 - 41, 04.01.2018
https://doi.org/10.21448/ijsm.356601

Abstract

This study was performed to determine the antimicrobial and antibiofilm activities of essential oil of Tanacetum argenteum (Lam.) Willd. subsp. canum (K.Koch) Grierson var. canum. The plant was collected from Amasya. The antimicrobial activity was determined by disc diffusion and microdilution broth methods. Essential oil was active against indicator organisms. The maximal inhibition zone diameter were as follows: E. cloacae ATCC 28355 (14mm), P. fluorescens ATCC 55241 (16mm), P. aeruginosa ATCC 27853 (19mm), S. sonnei RSKK 8177 (18mm), E. coli ATCC 25922 (21mm), E. coli O157:H7 (13mm), Y. enterocolitica RSKK 1501 (11mm), C. jejuni ATCC 33291 (12mm), K. pneumoniae ATCC 27736 (13mm), S. enteritidis RSKK 171 (12mm), S. aureus ATCC 33862 (26mm) and S. aureus ATCC 25923 (29mm), M. luteus NRLL-B-4375 (20mm), E. faecalis ATCC 19433 (19mm), B. cereus NRRL-B-3711 (30mm), B. cereus RSKK (32mm), C. albicans ATCC 10231 (19mm) and C. tropicalis (17mm). The minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) were ranged from 62.5-900 µg/ml and 125-1000 µg/ml, respectively. Plant essential oil was the most active against B. cereus NRRL-B 3711 and B. subtilis RSKK 867. Also, the essential oil has shown a strong anti-candidal activity against two Candida species. For determining the antibiofilm effect, various dilutions of oil were studied on M. luteus, E. cloacae, P. fluorescens, Y. enterocolitica and S. enteritidis by using microplate biofilm assay. According to results, antibiofilm effect was found as weak. Maximum antibiofilm effect was determined on E. cloacae ATCC 28355 with 32.92% biofilm inhibition rate.

References

  • Lopes-Lutz, D., Alviano, D.S., Alviano, C.S., Kolodziejczyk, P.P. (2008). Screening of chemical composition, antimicrobial and antioxidant activities of Artemisia essential oils. Phytochemistry, 1732-1738.
  • Kivrak, I., Duru, M.E., Öztürk, M., Mercan, N., Harmandar, M., Topçu, G. (2009). Antioxidant, anticholinesterase and antimicrobial constituents from the essential oil and ethanol extract of Salvia potentillifolia. Food Chem., 116, 470–479.
  • Goren, N. (1995). Sesquiterpene lactones from Tanacetum praeteritum. Phytochemistry, 38(5), 1261 1264.
  • Goren, N., Tahtasakal, E. (1997). Sesquiterpenoids from Tanacetum argenteum subsp. canum var. canum. Phytochemistry, 45(1), 107- 109.
  • Çaliskan, Z., Goren, N., Watson, W.H. (2004). Isolation and structures of eudesmanolides from Tanacetum cadmeum ssp. Cadmium. Journal of Chemical Crystallography, 34(5), 307-310.
  • Akpulat, H.A., Tepe, B., Sokmen, A., Daferera, D., Polissiou, M. (2005). Compositon of the essential oils of Tanacetum argyrophyllum (C.Koch) Tvzel. var. argyrophyllum and Tanacetum parthenium (L.) Schultz Bip. (Asteraceae) from Turkey. Biochemical Systematics and Ecology, 33, 511–516.
  • Gogus, F., Celik, A., Lewis, A.C., Ozel, M. Z. (2009). Analysis of Volatile Components From Tanacetum cadmium by Hydrodistillation and Direct Thermal Desorption Methods Using GC×GC-TOF/MS. Chemistry of Natural Compounds, 45(4), 557-559.
  • Bagcı, E., Kocak, A. (2010). Essential oil composition of two endemic Tanacetum (T. nitens (Boiss&Noe) Grierson and T. argenteum (Lam.) Willd. Subsp. argenteum) (Asteraceae) taxa, growing wild in Turkey. Industrial Crops and Products, 31, 542-545.
  • Polatoğlu, K., Demirci, B., Gören, N., Başer, K.H.C. (2011). Essential oil composition of endemic Tanacetum zahlbruckneri (Náb.) and Tanacetum tabrisianum (Boiss.) Sosn. and Takht. from Turkey. Natural Product Research, 25(6), 576–584.
  • Polatoglu, K., Karakoc, O.C., Yücel, Y.Y., Demirci, B., Gören, N., Başer, K.H.C. (2015). Composition, insecticidal activity and other biological activities of Tanacetum abrotanifolium Druce. essential oil. Industrial Crops and Products, 71, 7–14.
  • Mahdavi, M., Jouri, M.H., Mahmoudi, J., Rezazadeh, F., Mahzooni-Kachapi, S.S. (2013). Investigating the altitude effect on the quantity and quality of the essential oil in Tanacetum polycephalum Sch.-Bip. polycephalum in the Baladeh region of Nour, Iran. Chinese Journal of Natural Medicines, 11(5), 0553−0559.
  • Thomas, O.O. (1989). Anticoagulant and antifibrinolytic properties of Tanacetum macrophyllum. Fitoterapia, 15(4), 329–330.
  • Susurluk, H., Çalışkan, Z., Gürkan, O., Kırmızıgül, S., Gören, N. (2007). Antifeedant activity of some Tanacetum species and bioassay guided isolation of the secondary metabolites of Tanacetum cadmeum ssp. cadmeum (Compositae). Industrial Crops and Products, 26, 220-228.
  • Tabanca, N., Demirci, F., Demirci, B., Wedge, Başer K.H.C. (2007). Composition, enantiomeric distribution, and antimicrobial activity of Tanacetum argenteum subsp. flabellifolium essential oil. Journal of Pharmaceutical and Biomedical Analysis, 45, 714-719.
  • Polatoğlu, K., Demirci, F., Demirci, B., Gören, N., Başer, K.H.C. (2010). Essential oil composition and antibacterial activity of Tanacetum argenteum (Lam) Willd. ssp. argenteum and T. densum (Lab.) Schultz Bip. ssp. amani Heywood from Turkey. Journal of Oleo Science, 59(7), 361-367.
  • Polatoğlu, K., Demirci, F., Demirci, B., Gören, N., Başer, K.H.C. (2010). Antimicrobial activity and Essential Oil Composition of a New T. argyrophyllum (C. Koch) Tvzel var. argyrophyllum Chemotype. Journal of Oleo Science, 59(6), 307-313.
  • Polatoğlu, K., Karakoç, Ö.C., Gökçe, A., Gören, N. (2011). Insecticidal activity of Tanacetum chiliophyllum (Fisch.&Mey.) var. monocephalum grierson extracts and a new sesquiterpene lactone. Phytochemistry Letters, 4, 432-435.
  • Triana, J., Eiroa, J.L., Morales, M., Pérez, F.J., Brouard, I., Marrero, M.T., Estévez, S., Quintana, J., Estévez, F., Castillo, Q.A., León, F. (2013). A chemotaxonomic study of endemic species of genus Tanacetum from the Canary Islands. Phytochemistry, 92, 87–104.
  • Ali, A., Tabanca, N., Kurkcuoglu, M., Duran, A., Blythe, E.K., Khan, I.A., Baser, K.H.C. (2014). Chemical Composition, Larvicidal, and Biting Deterrent Activity of Essential Oils of Two Subspecies of Tanacetum argenteum (Asterales: Asteraceae) and Individual Constituents Against Aedes aegypti (Diptera: Culicidae). Journal of Medical Entomology, 51(4), 824-830.
  • Davis, P.H. 1965-1985. Flora of Turkey and the East Aegean Islands. vol. 1-9. Edinburgh: Univ. Press.
  • Goren, N., Demirci, B., Baser, K.H.C. (2001). Composition of the essential oils of Tanacetum spp. from Turkey. Flavour Fragr. J., 16, 191–194.
  • Cansaran, A., Peker, S., Yıldırım, C. (2007). Floristic Characters of the Area between the Direkli (Göndes) Village, Yassıçal (Ebemi) Town and Abacı Village (A5/6 Amasya–TURKEY). International Journal of Botany, 3(3), 240-250.
  • CLSI. (2012). Performance Standarts for Antimicrobial Disk Susceptibility Test; Approved standard-Eleventh Edition. (M02-A11), Vol. 32, No. 1, Replaces M02-A10, Vol. 29, No. 1.
  • Koneman, E.W., Allen, S.D., Janda, W.M., Scherckenberger, P.C., Winn, W.C. (1997). Color Atlas and Textbook of Diagnostic Microbiology. Philadelphia: Lippincott-Raven.
  • Merritt, J.H., Kadouri, D.E., O'Toole, G.A. (2005). Growing and analyzing static biofilms, Curr Proto Microbiol. John Wiley and Sons, Inc, pp. 1B.1.1-1B.1.17.
  • Chen, Z., He, B., Zhou, J., He, D., Deng, J., Zeng, R. (2016). Chemical compositions and antibacterial activities of essential oils extracted from Alpinia guilinensis against selected foodborne. Industrial Crops and Products, 83, 607-613.
  • Bokhari, N., Perveen, K., Al Khulaifi, M., Kumar, A., Siddiqui, I. (2016). In Vitro Antibacterial Activity and Chemical Composition of Essential Oil of Mentha arvensis Linn. Leaves. Journal of Essential Oil Bearing Plants, 19(4), 907–915.
  • Pattnaik, S., Subramanyam, V.R., Bapaji, M., Kole, C.R. (1997). Antibacterial and antifungal activity of aromatic constituents of essential oils. Microbios, 89, 39–46.
  • Tzakou, O., Pitarokili, D., Chinou, I.B., Harvala, C. (2001). Composition and antimicrobial activity of the essential oil of Salvia ringens. Planta Medica, 67, 81–83.
  • Tabanca, N., Kirimer, N., Demirci, B., Demirci, F., Baser, K.H.C. (2001). Composition and antimicrobial activity of the essential oils of Micromeria cristata subsp. phyrgia and the Enantiomeric Distribution of Borneol. Journal of Agricultural and Food Chemistry, 49, 4300–4303.
  • Vardar-Unlu, G., Candan, F., Sokmen, A., Daferera, D., Polissiou, M., Sokmen, M., Donmez, E., Tepe, B. (2003). Antibacterial and antioxidant activity of the essential oil and methanol extracts of Thymus pectinatus Fisch. et Mey var. pectinatus (Lamiaceae). Journal of Agricultural and Food Chemistry, 51, 63–67.
  • Sabulal, B., Dan, M., John, J.A., Kurup, R., Pradeep, N.S., Valsamma, R.K., George, V. (2006). Caryophyllene-rich rhizome oil of Zingiber nimmonii from South India: Chemical characterization and antimicrobial activity. Phytochemistry, 67, 2469-2473.
  • Nikolic, B., Vasilijevic, B., Mitic-Culafic, D., Vukovic-Gacic, B. (2015). Comparative study of genotoxic, antigenotoxic and cytotoxic activities of monoterpenes camphor, eucalyptol and thujone in bacteria and mammalian cells. Chemico-Biological Interactions, 242, 263-271.
  • Andersson, A., Ronner, U., Granum, P.E. (1995). What problems does the food industry have with the spore-forming pathogens Bacillus cereus and Clostridium perfringens? International Journal of Food Microbiology, 28(2), 145-155.
  • Lim, J.Y., Yoon, J.W., Hovde, C.J. (2010). A Brief Overview of Escherichia coli O157:H7 and Its Plasmid O157. J. Microbiol Biotechnol., 20(1), 5–14.
  • Allos, B.M. (2001). Campylobacter jejuni Infections: Update on Emerging Issues and Trends, Food Safety, 32, 1201-1206.
  • Otto, M. (2008). Staphylococcal biofilms. Curr. Top. Microbiol. Immunol., 322, 207-228.

In Vitro Evaluation of Antimicrobial and Antibiofilm Potentials of Essential Oil of Tanacetum argenteum (Lam.) Willd. subsp. canum (K.Koch) Grierson var. canum

Year 2018, Volume: 5 Issue: 1, 34 - 41, 04.01.2018
https://doi.org/10.21448/ijsm.356601

Abstract

This
study was performed to determine the antimicrobial and antibiofilm activities
of essential oil of Tanacetum argenteum
(Lam.)
Willd. subsp. canum (K.Koch) Grierson var.
canum
. The plant was collected from Amasya. The antimicrobial activity was
determined by disc diffusion and microdilution broth methods. Essential oil was
active against indicator organisms. The maximal inhibition zone diameter were
as follows: E. cloacae ATCC 28355
(14mm), P. fluorescens ATCC 55241 (16mm), P. aeruginosa ATCC 27853 (19mm), S. sonnei RSKK 8177 (18mm), E.
coli
ATCC 25922 (21mm), E. coli
O157:H7 (13mm), Y. enterocolitica
RSKK 1501 (11mm), C. jejuni ATCC
33291 (12mm), K. pneumoniae ATCC
27736 (13mm), S. enteritidis RSKK 171
(12mm), S. aureus ATCC 33862 (26mm)
and S. aureus ATCC 25923 (29mm),  M.
luteus
NRLL-B-4375 (20mm), E.
faecalis
ATCC 19433 (19mm), B. cereus
NRRL-B-3711 (30mm), B. cereus RSKK
(32mm), C. albicans ATCC 10231 (19mm)
and C. tropicalis (17mm). The minimum
inhibitory concentration (MIC) and minimum lethal concentration (MLC) were
ranged from 62.5-900 µg/ml and 125-1000 µg/ml, respectively. Plant essential
oil was the most active against B. cereus
NRRL-B 3711 and B. subtilis RSKK 867.
Also, the essential oil has shown a strong anti-candidal activity against two Candida species. For determining the
antibiofilm effect, various dilutions of oil were studied on M. luteus, E. cloacae, P. fluorescens, Y.
enterocolitica and S. enteritidis
by using microplate biofilm assay.
According to results, antibiofilm effect was found as weak. Maximum antibiofilm
effect was determined on E. cloacae
ATCC 28355 with 32.92% biofilm inhibition rate.

References

  • Lopes-Lutz, D., Alviano, D.S., Alviano, C.S., Kolodziejczyk, P.P. (2008). Screening of chemical composition, antimicrobial and antioxidant activities of Artemisia essential oils. Phytochemistry, 1732-1738.
  • Kivrak, I., Duru, M.E., Öztürk, M., Mercan, N., Harmandar, M., Topçu, G. (2009). Antioxidant, anticholinesterase and antimicrobial constituents from the essential oil and ethanol extract of Salvia potentillifolia. Food Chem., 116, 470–479.
  • Goren, N. (1995). Sesquiterpene lactones from Tanacetum praeteritum. Phytochemistry, 38(5), 1261 1264.
  • Goren, N., Tahtasakal, E. (1997). Sesquiterpenoids from Tanacetum argenteum subsp. canum var. canum. Phytochemistry, 45(1), 107- 109.
  • Çaliskan, Z., Goren, N., Watson, W.H. (2004). Isolation and structures of eudesmanolides from Tanacetum cadmeum ssp. Cadmium. Journal of Chemical Crystallography, 34(5), 307-310.
  • Akpulat, H.A., Tepe, B., Sokmen, A., Daferera, D., Polissiou, M. (2005). Compositon of the essential oils of Tanacetum argyrophyllum (C.Koch) Tvzel. var. argyrophyllum and Tanacetum parthenium (L.) Schultz Bip. (Asteraceae) from Turkey. Biochemical Systematics and Ecology, 33, 511–516.
  • Gogus, F., Celik, A., Lewis, A.C., Ozel, M. Z. (2009). Analysis of Volatile Components From Tanacetum cadmium by Hydrodistillation and Direct Thermal Desorption Methods Using GC×GC-TOF/MS. Chemistry of Natural Compounds, 45(4), 557-559.
  • Bagcı, E., Kocak, A. (2010). Essential oil composition of two endemic Tanacetum (T. nitens (Boiss&Noe) Grierson and T. argenteum (Lam.) Willd. Subsp. argenteum) (Asteraceae) taxa, growing wild in Turkey. Industrial Crops and Products, 31, 542-545.
  • Polatoğlu, K., Demirci, B., Gören, N., Başer, K.H.C. (2011). Essential oil composition of endemic Tanacetum zahlbruckneri (Náb.) and Tanacetum tabrisianum (Boiss.) Sosn. and Takht. from Turkey. Natural Product Research, 25(6), 576–584.
  • Polatoglu, K., Karakoc, O.C., Yücel, Y.Y., Demirci, B., Gören, N., Başer, K.H.C. (2015). Composition, insecticidal activity and other biological activities of Tanacetum abrotanifolium Druce. essential oil. Industrial Crops and Products, 71, 7–14.
  • Mahdavi, M., Jouri, M.H., Mahmoudi, J., Rezazadeh, F., Mahzooni-Kachapi, S.S. (2013). Investigating the altitude effect on the quantity and quality of the essential oil in Tanacetum polycephalum Sch.-Bip. polycephalum in the Baladeh region of Nour, Iran. Chinese Journal of Natural Medicines, 11(5), 0553−0559.
  • Thomas, O.O. (1989). Anticoagulant and antifibrinolytic properties of Tanacetum macrophyllum. Fitoterapia, 15(4), 329–330.
  • Susurluk, H., Çalışkan, Z., Gürkan, O., Kırmızıgül, S., Gören, N. (2007). Antifeedant activity of some Tanacetum species and bioassay guided isolation of the secondary metabolites of Tanacetum cadmeum ssp. cadmeum (Compositae). Industrial Crops and Products, 26, 220-228.
  • Tabanca, N., Demirci, F., Demirci, B., Wedge, Başer K.H.C. (2007). Composition, enantiomeric distribution, and antimicrobial activity of Tanacetum argenteum subsp. flabellifolium essential oil. Journal of Pharmaceutical and Biomedical Analysis, 45, 714-719.
  • Polatoğlu, K., Demirci, F., Demirci, B., Gören, N., Başer, K.H.C. (2010). Essential oil composition and antibacterial activity of Tanacetum argenteum (Lam) Willd. ssp. argenteum and T. densum (Lab.) Schultz Bip. ssp. amani Heywood from Turkey. Journal of Oleo Science, 59(7), 361-367.
  • Polatoğlu, K., Demirci, F., Demirci, B., Gören, N., Başer, K.H.C. (2010). Antimicrobial activity and Essential Oil Composition of a New T. argyrophyllum (C. Koch) Tvzel var. argyrophyllum Chemotype. Journal of Oleo Science, 59(6), 307-313.
  • Polatoğlu, K., Karakoç, Ö.C., Gökçe, A., Gören, N. (2011). Insecticidal activity of Tanacetum chiliophyllum (Fisch.&Mey.) var. monocephalum grierson extracts and a new sesquiterpene lactone. Phytochemistry Letters, 4, 432-435.
  • Triana, J., Eiroa, J.L., Morales, M., Pérez, F.J., Brouard, I., Marrero, M.T., Estévez, S., Quintana, J., Estévez, F., Castillo, Q.A., León, F. (2013). A chemotaxonomic study of endemic species of genus Tanacetum from the Canary Islands. Phytochemistry, 92, 87–104.
  • Ali, A., Tabanca, N., Kurkcuoglu, M., Duran, A., Blythe, E.K., Khan, I.A., Baser, K.H.C. (2014). Chemical Composition, Larvicidal, and Biting Deterrent Activity of Essential Oils of Two Subspecies of Tanacetum argenteum (Asterales: Asteraceae) and Individual Constituents Against Aedes aegypti (Diptera: Culicidae). Journal of Medical Entomology, 51(4), 824-830.
  • Davis, P.H. 1965-1985. Flora of Turkey and the East Aegean Islands. vol. 1-9. Edinburgh: Univ. Press.
  • Goren, N., Demirci, B., Baser, K.H.C. (2001). Composition of the essential oils of Tanacetum spp. from Turkey. Flavour Fragr. J., 16, 191–194.
  • Cansaran, A., Peker, S., Yıldırım, C. (2007). Floristic Characters of the Area between the Direkli (Göndes) Village, Yassıçal (Ebemi) Town and Abacı Village (A5/6 Amasya–TURKEY). International Journal of Botany, 3(3), 240-250.
  • CLSI. (2012). Performance Standarts for Antimicrobial Disk Susceptibility Test; Approved standard-Eleventh Edition. (M02-A11), Vol. 32, No. 1, Replaces M02-A10, Vol. 29, No. 1.
  • Koneman, E.W., Allen, S.D., Janda, W.M., Scherckenberger, P.C., Winn, W.C. (1997). Color Atlas and Textbook of Diagnostic Microbiology. Philadelphia: Lippincott-Raven.
  • Merritt, J.H., Kadouri, D.E., O'Toole, G.A. (2005). Growing and analyzing static biofilms, Curr Proto Microbiol. John Wiley and Sons, Inc, pp. 1B.1.1-1B.1.17.
  • Chen, Z., He, B., Zhou, J., He, D., Deng, J., Zeng, R. (2016). Chemical compositions and antibacterial activities of essential oils extracted from Alpinia guilinensis against selected foodborne. Industrial Crops and Products, 83, 607-613.
  • Bokhari, N., Perveen, K., Al Khulaifi, M., Kumar, A., Siddiqui, I. (2016). In Vitro Antibacterial Activity and Chemical Composition of Essential Oil of Mentha arvensis Linn. Leaves. Journal of Essential Oil Bearing Plants, 19(4), 907–915.
  • Pattnaik, S., Subramanyam, V.R., Bapaji, M., Kole, C.R. (1997). Antibacterial and antifungal activity of aromatic constituents of essential oils. Microbios, 89, 39–46.
  • Tzakou, O., Pitarokili, D., Chinou, I.B., Harvala, C. (2001). Composition and antimicrobial activity of the essential oil of Salvia ringens. Planta Medica, 67, 81–83.
  • Tabanca, N., Kirimer, N., Demirci, B., Demirci, F., Baser, K.H.C. (2001). Composition and antimicrobial activity of the essential oils of Micromeria cristata subsp. phyrgia and the Enantiomeric Distribution of Borneol. Journal of Agricultural and Food Chemistry, 49, 4300–4303.
  • Vardar-Unlu, G., Candan, F., Sokmen, A., Daferera, D., Polissiou, M., Sokmen, M., Donmez, E., Tepe, B. (2003). Antibacterial and antioxidant activity of the essential oil and methanol extracts of Thymus pectinatus Fisch. et Mey var. pectinatus (Lamiaceae). Journal of Agricultural and Food Chemistry, 51, 63–67.
  • Sabulal, B., Dan, M., John, J.A., Kurup, R., Pradeep, N.S., Valsamma, R.K., George, V. (2006). Caryophyllene-rich rhizome oil of Zingiber nimmonii from South India: Chemical characterization and antimicrobial activity. Phytochemistry, 67, 2469-2473.
  • Nikolic, B., Vasilijevic, B., Mitic-Culafic, D., Vukovic-Gacic, B. (2015). Comparative study of genotoxic, antigenotoxic and cytotoxic activities of monoterpenes camphor, eucalyptol and thujone in bacteria and mammalian cells. Chemico-Biological Interactions, 242, 263-271.
  • Andersson, A., Ronner, U., Granum, P.E. (1995). What problems does the food industry have with the spore-forming pathogens Bacillus cereus and Clostridium perfringens? International Journal of Food Microbiology, 28(2), 145-155.
  • Lim, J.Y., Yoon, J.W., Hovde, C.J. (2010). A Brief Overview of Escherichia coli O157:H7 and Its Plasmid O157. J. Microbiol Biotechnol., 20(1), 5–14.
  • Allos, B.M. (2001). Campylobacter jejuni Infections: Update on Emerging Issues and Trends, Food Safety, 32, 1201-1206.
  • Otto, M. (2008). Staphylococcal biofilms. Curr. Top. Microbiol. Immunol., 322, 207-228.
There are 37 citations in total.

Details

Primary Language English
Subjects Structural Biology
Journal Section Articles
Authors

Nazime Mercan Doğan

Arzu Cansaran

Miraç Yılmaz

Naime Nur Bozbeyoğlu This is me

Publication Date January 4, 2018
Submission Date July 17, 2017
Published in Issue Year 2018 Volume: 5 Issue: 1

Cite

APA Mercan Doğan, N., Cansaran, A., Yılmaz, M., Bozbeyoğlu, N. N. (2018). In Vitro Evaluation of Antimicrobial and Antibiofilm Potentials of Essential Oil of Tanacetum argenteum (Lam.) Willd. subsp. canum (K.Koch) Grierson var. canum. International Journal of Secondary Metabolite, 5(1), 34-41. https://doi.org/10.21448/ijsm.356601
International Journal of Secondary Metabolite

e-ISSN: 2148-6905