Hexane neem leaf extract more potent than ethanol extract against Aspergillus flavus

Main Article Content

Jenny Hidayat
Ay Ly Margaret
Hanna Yolanda
Lies K. Wibisono


BACKGROUND Aspergillus flavus is one of the causes of aspergillosis, with a high virulence and resistance to standard antifungals, resulting in a high mortality rate. Medicinal plants are increasingly used as they are relatively safer with minimal side effects. Previously we found that the ethanol extract of neem (Azadirachta indica A Juss) leaves inhibits A. flavus growth in vitro. However, most chemical compounds with antifungal effect are nonpolar. The purpose of this research was to compare the antifungal effect of neem leaves extracted in a nonpolar solvent to that of leaves extracted in a polar solvent. METHODS An in vitro experimental research was conducted between October 2013 and January 2014. Neem leaves were extracted in ethanol or hexane at various concentrations. A macrodilution test with 48-hour incubation time was done in triplicate on 8 groups of samples. These comprised the neem leaf ethanol extract (NLEE) at 0.5, 1.0, and 2.0 g/dL, neem leaf hexane extract (NLHE) at 0.5, 1.0, and 2.0 g/dL, positive control, and negative control groups. Fungal growth was detected on Sabouroud dextrose agar. Statistical analysis used Chi square and Fisher’s exact test. RESULTS NLHE had a higher, but statistically non-significant, inhibitory effect on A. flavus than NLEE (p=0.996). At higher concentrations, the antifungal effect of NLHE is better than that of NLEE. CONCLUSION There is no significant difference in in-vitro inhibitory effectivity on A. flavus of neem leaves between extracts in polar and nonpolar solvents.

Article Details

How to Cite
Hidayat, J., Margaret, A. L., Yolanda, H., & Wibisono, L. K. (2014). Hexane neem leaf extract more potent than ethanol extract against Aspergillus flavus. Universa Medicina, 33(3), 171–178. https://doi.org/10.18051/UnivMed.2014.v33.171-178
Review Article


Rasmin M, Syarifuddin PK, Setiawati A,

Soemitro D, Hamid A. Sistem pernapasan. In:

Budimulyati U, Bramono K, Menaldi SL,

editors. Konsensus FKUI dan PMKI tentang tatalaksana mikosis sistemik. Jakarta: Balai

Penerbit FKUI;2001.p.5-16.

Thompson GR, Patterson TF. Pulmonary

aspergillosis. Semin Respir Crit Care Med 2008;


Bansod S, Rai M. Antifungal activity of essential

oils from Indian medicinal plants against human

pathogenic Aspergillus fumigatus and A. niger.

World J Med Sci 2008;3:81-8.

Hedayati MT, Pasqualotto AC, Warn PA, Bowyer

P, Denning DW. Aspergillus flavus: human

pathogen, allergen and mycotoxin producer.

Microbiol 2007;153:1677-92.

Radojevic ID, Stankovic MS, Stefanovic OD,

Topuzovic MD, Comic LR, Ostojic AM. Antiaspergillus

properties of different extracts from

selected plants. Afr J Microbiol Res 2011;5:3986-

Ziveai F, Afshari H, Moghadam MM,

Sobhanipur A. Study on antifungal effects of

herbal essences on Aspergillus flavus. Bull Env

Pharmacol Life Sci 2013;2:100-5.

Paul D, Kevin K. Pulmonary aspergillosis:

clinical presentation, diagnosis, and therapy. Brit

J Biomed Sci 2001;58:197-205.

Radhika SM, Michael A. In vitro antifungal

activity of leaf extracts of Azadirachta indica.

Int J Pharm Pharm Sci 2013;5:723-5.

Jabeen K. Antifungal activity of Azadirachta

indica against Alternaria solani. J Life Sci

Technol 2013;1:89-92.

Li MA, Yolanda H, Wibisono LK. Antifungal

activity of neem leaves ethanol extract on

Aspergillus flavus. Univ Med 2013;32:104-9.

Mondali NK, Mojumdar A, Chatterje SK,

Banerjee A, Datta JK, Gupta S. Antifungal

activities and chemical characterization of neem

leaf extracts on the growth of some selected

fungal species in vitro culture medium. J Appl

Sci Environ Manage 2009;13:49-53.

Akpuaka A, Ekwenchi MM, Dashak DA, Dildar

A. Biological activities of characterized isolates

of n-hexane extract of Azadirachta indica a juss

(neem) leaves. N Y Sci J 2013;6:119-24.

Moslem MA, El-Kholie EM. Effect of neem

(Azardirachta indica a.Juss) seeds and leaves

extract on some plant pathogenic fungi. Pak J

Biol Sci 2009;12:1045-8.

Clinical and Laboratory Standard Institute.

Reference method for broth dilution antifungal

susceptibility testing of yeasts; document M27-

A2. 2nd Ed. Pennsylvania: CLSI;2002.

Khan S, Singhal S, Mathur T, Upadhyay DJ,

Rattan A. Antifungal susceptibility testing

method for resource constrained laboratories.

Indian J Med Microbiol 2006;24:171-6.

Canton E, Espinel-Ingroff A, Peman J. Trends

in antifungal susceptibility testing using CLSI

reference and commercial methods. Expert Rev

Anti Infect Ther 2009;7:107-19.

Biswas K, Chattopadhyay I, Benerjee RK,

Bandyopadhyay U. Biological activity and

medical properties of neem (Azadirachta indica).

Curr Sci 2002;82:701-11.

Hazen KC. Influence of DMSO antifungal

activity during susceptibility testing in vitro.

Diagn Microbiol Infect Dis 2013;75:60-3.

Randhawa MA. The effect of dimethyl

sulphoxide (DMSO) on the growth of

dermatophytes. Japanese J Med Mycol 2006;47:


Rauf A, Khan A, Rasool S, Shah ZA, Saleem M.

In-vitro antifungal activity of three selected

pakistani medicinal plants. Middle-East J Med

Pl Res 2012;1:41-3.

Verma A, Joshi P, Arya A. Screening of eight

plant extracts for their antimicrobial properties.

Int J Curr Microbiol App Sci 2013;2:315-20.

Jeyasakthy S, Jeyadevan JP, Thavaranjit AC,

Manoranjan T, Srikaran R, Krishnapillai N.

Antifungal activity and qualitative

phytochemical analysis of extracts obtained by

sequential extraction of some medicinal plants

in Jaffna peninsula. Arch Appl Sci Res 2013;5:


Kumar PS, Mishra D, Ghosh G, Panda CS.

Biological action and medicinal properties of

various constituent of Azadirachta indica

(Meliaceae): an overview. Ann Biol Res 2010;1: