Fitoterapia 76 (2005) 484 – 489

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Short report

Antimicrobial and cytotoxic constituents from the

seeds of Annona squamosa
M. Mukhlesur Rahman a,*, Shahnaj Parvin a, M. Ekramul Haque a,
M. Ekramul Islam a, Mohammad A. Mosaddik b
a
Phytochemistry Research Laboratory, Department of Pharmacy, University of Rajshahi,
Rajshahi-6205, Bangladesh
b
Centre for Phytochemistry, Southern Cross University, Lismore, NSW-2480, Australia

Received 22 July 2003; accepted 28 April 2005

Abstract

Annotemoyin-1, Annotemoyin-2, squamocin and cholesteryl glucopyranoside were isolated from

the seeds of Annona squamosa. These compounds and plant extracts showed remarkable
antimicrobial and cytotoxic activities.
D 2005 Elsevier B.V. All rights reserved.

Keywords: Annona squamosa; Antimicrobial activity; Cytotoxicity

1. Plant

Annona squamosa L. (Annonaceae) [1–4] seeds were collected from Chapai

Nawabganj district of Bangladesh in September 2000 and identified by Prof. A.T.M.
Naderuzzaman, Department of Botany, University of Rajshahi, Bangladesh where a
voucher specimen has been deposited.

* Corresponding author. Fax: +880 721 750064.

E-mail address: mukhlesurrah@yahoo.co.uk (M. Mukhlesur Rahman).

0367-326X/$ - see front matter D 2005 Elsevier B.V. All rights reserved.
doi:10.1016/j.fitote.2005.04.002
 M. Mukhlesur Rahman et al. / Fitoterapia 76 (2005) 484–489 485

2. Uses in traditional medicines

Seeds, fruits and leaves are effective as insecticide, fish poison, powerful irritant of the
conjunctiva and abortifacient. The root are effective as a drastic purgative and in acute
dysentery [1–4].

Fig. 1. Compounds 1–4.

 486
Table 1
Antibacterial activities of the A. squamosa extracts PE, CE and EE and compounds 1–4
Bacteria Diameter of the zone of inhibition (mm)

M. Mukhlesur Rahman et al. / Fitoterapia 76 (2005) 484–489

PE CE EE 1+2 3 4 Kanamycin
200 400 200 400 200 400 200 400 200 400 200 400 30
Ag/disc Ag/disc Ag/disc Ag/disc Ag/disc Ag/disc Ag/disc Ag/disc Ag/disc Ag/disc Ag/disc Ag/disc Ag/disc
Gram (+)
B. subtilis 18 28 16 18 18 20 8 10 12 13 8 10 28
B. cereus 20 21 15 17 16 19 10 11 13 15 8 10 30
B. megaterium 11 15 17 19 12 15 12 13 14 17 10 13 21
Staphylococcus aureus 12 14 12 14 8 11 13 15 15 18 10 13 40
S. b-haemolytica 30 34 20 23 17 19 15 18 16 18 12 15 40
Sarcina lutea 18 22 22 26 18 22 10 12 13 16 10 13 30
Gram ( )
E. coli 21 23 16 19 12 14 12 14 15 18 12 15 30
S. dysenteriae 12 15 10 12 12 14 14 15 12 13 15 18 15
S. shiga 14 15 10 12 10 12 15 17 11 13 14 16 20
S. flexneriae 13 15 12 13 11 11 11 13 13 16 8 11 18
S sonnei 15 17 13 14 12 12 8 10 14 17 12 14 22
Salmonella typhi 18 20 13 16 14 16 13 16 17 19 15 17 25
P. aeruginosa 24 26 20 22 14 18 14 16 18 20 17 19 29
Klebsiella spp. 23 25 18 21 14 18 18 21 21 22 14 16 25
 M. Mukhlesur Rahman et al. / Fitoterapia 76 (2005) 484–489 487

3. Previous isolated class of constituents

Acetogenins [5–18], terpenoids [19,20], alkaloids [21], cyclopeptides [22–25], steroids [26].

4. Tested materials

Petroleum ether extract (PE), CHCl3 extract (CE), EtOH extract (EE), annotemoyin-
1 (1) [27], annotemoyin-2 (2) [27], squamocin (3) [28] and cholesteryl glucopyranoside
(4) (Fig. 1) [29].

5. Studied activity

Antibacterial and antifungal activities by disc diffusion method [30–32] and

cytotoxicity by Brine shrimp bioassay [33].

6. Used microorganisms

Listed in Tables 1 and 2.

7. Results

All compounds as well as crude extracts showed antibacterial activities against all test
organisms. Petroleum ether extract and compounds 3 and 4 did not show any antifungal
activities, chloroform and ethanol extracts and compounds (1 and 2) showed reasonable
activities against all test fungi except Candida albicans. The minimum inhibitory
concentrations (MIC) of the compounds 1–4 were observed by serial dilution technique.
The MIC value of compounds 1 and 2 was found to be 64 Ag/ml against Bacillus
subtilis and Shigella shiga. The MIC values of compound 3 was 64 Ag/ml against

Table 2
Antifungal activity of the A. squamosa extracts PE, CE and EE and compounds 1–4
Fungi Diameter of zone of inhibition (mm)
PE CE EE 1+2 3 4 Nystatine
200 200 200 200 200 200 200
Ag/disc Ag/disc Ag/disc Ag/disc Ag/disc Ag/disc Ag/disc
Aspergillus flavus – 6 7 13 – – 8
A. niger – 7 5 18 – – 28
A. fumigatus – 6 6 10 – – 25
C. albicans – – – – – – –
488 M. Mukhlesur Rahman et al. / Fitoterapia 76 (2005) 484–489

Streptococcus-h-haemolyticus and Pseudomonas aeruginosa. In case of compound 4,

the MIC values were 64 Ag/ml against Escherichia coli and 128 Ag/ml against Bacillus
subtilis.
The cytotoxicity of both extracts and compounds was studied by brine shrimp lethality
bioassay. The LC50 values of petroleum ether and chloroform extracts were 0.81 and 0.83
Ag/ml, respectively. In case of compounds, the LC50 values were found to be 2.34 Ag/ml
mixture of 1 and 2, 2.39 Ag/ml of 3 and 2.81 Ag/ml of 4.

Acknowledgments

The authors are grateful to Prof. Naderuzzaman, Department of Botany, University of

Rajshahi, Bangladesh for the identification of the plant.

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