CHAPTER

138

Biological Activity of Seeds of Wild Banana (Ensete superbum Cheesm, Family Musaceae)
1

Monica Kachroo1, Shyam S. Agrawal2 Al-Ameen College of Pharmacy, Bangalore, India 2 HOD Department of Pharmacology, Delhi Institute of Pharmaceutical Sciences and Research, New Delhi, India 1165

CHAPTER OUTLINE
Introduction 1166 Botanical Description 1166 Historical Cultivation and Usage 1167 Present-Day Cultivation and Usage 1167 Applications to Health Promotion and Disease Prevention 1168
Traditional uses 1168 Chemistry 1168 Pharmacology 1169

Adverse Effects and Reactions (Allergies and Toxicity) 1171 Summary Points 1171 References 1172

LIST OF ABBREVIATIONS
BAP, 6-benzylaminopurine FTIR, Fourier transform infrared spectroscopy GA3, gibberellic acid HPLC, high performance liquid chromatography HPTLC, high performance thin layer chromatography NMR, nuclear magnetic resonance UV, ultra violet spectoscopy

Nuts & Seeds in Health and Disease Prevention. DOI: 10.1016/B978-0-12-375688-6.10138-0 Copyright 2011 Elsevier Inc. All rights reserved.

PART 2 Effects of Specic Nuts and Seeds

INTRODUCTION
Traditional systems of medicine continue to be widely practiced. Of the 2,500,000 higher plant species on earth, more than 80,000 are medicinal. India is one of the worlds 12 biodiversity centers, with the presence of over 45,000 different plant species. One such plant, a wild banana, Ensete superbum (Cheesm), is an ornamental plant indigenous to northern India and the higher elevations of Thailand. Since the seeds of the plant have many traditional uses, the extracts and its constituents have been studied for various pharmacological activities. The present chapter provides a review of the investigations carried out on the seeds of this traditional/folkloric plant.

BOTANICAL DESCRIPTION
Ensete superbum (Cheesman, 1947) is a member of the Musaceae family of the Order Zingiberales, and has the synonym Musa superba. Its common names include banakadli (Hindi) and wild banana (English). The plant grows to 3e4 m high, and is non-stoloniferous with a stout pseudostem 1.2e1.8 m tall, with an enormous swollen base of 2e2.4 m in circumference at the base, narrowing to 1 m below the leaves (Figure 138.1). The leaves are bright green on sides, 1.5 m long, and 45 cm broad, with a very short, deeply channeled free petiole. The leaf sheath is persistent at the base, and leaves closely set scars on the corm. The inorescence is globose at rst, 30 cm in diameter, later drooping and elongating to one-third the length of the trunk. The bracts are orbicular, dark brownish-red (dull claret-brown), reaching 30 cm in length and breadth, and subtend dense biseriate rows each of 10e15 owers. The ovary is white, cylindrical, and about 2.5 cm long. The outer perianth is whitish, situated along the ovary, and three-lobed, or formed of three loosely coherent segments. The inner perianth is shorter than the outer, and is tricuspidate with a long, linear central cusp. The rootstock and young inorescences are eaten, and the fruit is pickled when young. The fruit is subcoriaceous, 7.5 cm long and 3.5 cm in diameter, and more or less triangular in cross-section, has a thick

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FIGURE 138.1
Ensete superbum Cheesm (whole plant) with a massive baeband red ower head with broad leaves outlined in deep red and with deep mid-ribs. Source: www.centraloridafarms.com/ensete-superbum.htm

CHAPTER 138 Biological Activities of the Seeds of E. superbum

FIGURE 138.2
Ensete superbum Cheesm seeds. Source: Photographed by author.

skin, and contains numerous dark brown seeds. The seeds are brownish-black, subglobose but angled by pressure, and 8e12 mm in diameter (Figure 138.2). The plant dies down to an underground corm in the dry season, and forms new leaves at the beginning of the monsoons (Baker 1893, 1894). The plant has a massive baseband red ower head with broad leaves outlined in deep red, and deep mid-ribs are specic morphological features that add its appeal and esthetic value (http:// www.iiitmk.ac.in/wjrnair/ensetepapernal.pdf). The genus Ensete, proposed by Horananious in 1862, was further established by Cheesman (1947) as a genus separate from Musa. This separation of the two genera was based mainly on the differences in chromosome numbers, and on pseudostem morphology. The basic chromosome number in Ensete was reported to be 9, whereas that of Musa was observed to be 9, 10, and 11 (Agharakar & Bhaduri, 1933).

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HISTORICAL CULTIVATION AND USAGE

As early as 1893, Baker noted that E. superbum was in common cultivation, and it was introduced into the Calcutta Botanical Gardens in the year 1800 (Baker, 1893). The plant dies down in the dry season and forms new leaves at the beginning of the monsoon season. This species can be propagated by seeds, but cannot produce vegetative side suckers naturally like other Ensete species. It has the capacity to withstand severe drought, and has therefore been used during famines in Ethiopia. It is excellent for outdoor cultivation, where weather permits, and grows well with liberal fertilization when established.

PRESENT-DAY CULTIVATION AND USAGE

Since conventional propagation of Ensete is time consuming, there was a need for optimization of tissue culture techniques for its rapid propagation. Research revealed that the encapsulated shoot tips can be handled like a seed, and could be useful in minimizing the cost of

PART 2 Effects of Specic Nuts and Seeds

production, as 1 ml of medium is sufcient for encapsulation of a single shoot tip, compared to 15e20 ml for conversion of shoot tips into plantlets. By directly sowing the encapsulated shoot tips in soil, the two-stage process (such as rooting and hardening) can be eliminated. As compared to suckers, encapsulated shoot tips are inexpensive, and easier and safer material for germplasm exchange, maintenance, and transportation (Rao et al., 1993). Recently multiple shoots were induced from in vitro cultures of male oral apices of Ensete superbum Cheesm. BAP (6-benzylaminopurine) in combination with GA3 (gibberellic acid) was found to be benecial for the establishment of cultures, and the multiplication and elongation of shoots, while auxins were shown to promote rooting (Kulkarni et al., 1997). Thus, somatic embryogenesis offers an ideal system for the production of somatic embryos on a large scale for use in the preparation of synthetic seeds, propagation, and genetic transformation (Kulkarni, 1997).

APPLICATIONS TO HEALTH PROMOTION AND DISEASE PREVENTION

Traditional uses
Under Ayurvedic practice, the esh of the fruit is given to diabetics. The ground seeds are also used as Ayurvedic medicine; the powdered seed of this plant is popularly prescribed in the prevention and treatment of smallpox and chickenpox by physicians who practice the indigenous system of therapy. The seeds are fried, the inner pulp soaked in water, boiled and again soaked in water, and the resultant solution given orally to pregnant women in the third trimester to develop immune resistance in the baby. Processed seeds are consumed for hip pain by Kadars and Malasars. Seed paste is applied to the body for fever with body pains, and for scabies, by the Malasar (Ravishankar et al., 1994). In southern India, the ripe ground seeds of Ensete superbum are eaten to cure dysentery. The seed and the stem are also reported to be given for dog bites. Seed powder is used for treating kidney stones and painful urination (Yeshodharan & Sujana, 2007). Powdered seeds are also given with milk for diabetes (Sreedharan, 2004), and approximately 5 g seeds are crushed and taken with water for stomachache (Jagtap, 2008). The seeds of Ensete superbum are used for the treatment of kidney and vesical calculi, urinary retention, burns and scalds, and menstrual disorders (www.botanical.com/site/column_ poudhia/144_kela.html).

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Chemistry
The petroleum ether extract of dried seed powder yielded fatty oil and triterpenoid esters. The subsequent aqueous methanol (50 : 50) extract, on concentration and maceration with absolute methanol, yielded a white solid, which tested positive for proanthocyanidin and glucoside. Detailed study showed that it consisted of pro-pelargonidin glucosides of varying degrees of polymerization. A 1-year-old sample of the seeds also gave a violet compound with the properties of the pelargonidin color base, but not identical with it (Mahey et al., 1971). The three fractions, VIDR-2T, VIDR-2GC, and VIDR-2GD, isolated from the seeds of E. superbum Cheesm showed presence of phosphates, and traces of alkaloids, glycoside, and sugars (Dutta et al., 1968). The extractive values of the aqueous extract, ethanolic extract, and isolated compound of the seeds of Ensete superbum Cheesm were found to be 25.0, 1.92, and 0.1%, respectively. The moisture content, total ash, water-soluble ash, acid-insoluble ash, alcohol-soluble extractive value, and water-soluble extractive values were found to be 0.8, 2.8, 1.49, 0.55, 5.8, and 22.92%, respectively (Table 138.1).

CHAPTER 138 Biological Activities of the Seeds of E. superbum

TABLE 138.1 Pharmacognostical Evaluation of Powdered Seeds of Ensete superbum

S. No. Foreign Matter Moisture Content (%) 0.8 0.8 0.8 Total Ash Water-soluble (%) Ash (%) 2.8 2.8 2.8 1.58 1.40 1.49 Acid-insoluble Alcohol-soluble Water-soluble Ash (%) Extractive Extractive Value (%) Value (%) 0.54 0.55 0.55 5.8 5.8 5.8 22.65 23.19 22.92

1 NMT* 2% 2 NMT 2% Mean NMT 2%

*NMT, not more than.

Pharmacology
Many studies have been carried out, but the exact detailed pharmacology of Ensete superbum is not yet clear. Some of the reported and established activities are discussed below. 1. Antiviral effect. The three fractions (VIDR-2T, VIDR-2GC, and VIDR-2GD) isolated from the seeds of Ensete superbum Cheesm inhibited the growth of vaccinia and variola viruses on chick chorioallantoic membrane. In variola-infected mice, they prevented the disease from appearing and signicantly cured infected mice even when the drug treatment had begun at a late stage (Dutta et al., 1968). 2. Cardiovascular activity. VIDR-2GD at the dose of 0.5 e7.5 mg did not produce any effect on the isolated perfused heart of frog. The fraction VIDR-2GD, in dose of 100e150 mg/kg, caused prolonged and sustained hypotension in cats. It caused hypotension in cases of induced hypertension produced by occlusion of the carotid artery. Topical and intravenous administration of the drug produced congestion of the mesenteric blood vessels in rats and mice. Perfusion of frogs vessels with the drug showed a dilatory effect, with an increase in outow (Roy et al., 1968). 3. Respiratory activity. VIDR-2GD did not have any effect on the bronchial musculature. It neither enhanced nor antagonized the effect of histamine on the bronchus, and it did not produce any action on the tracheal chain (Roy et al., 1968). 4. Cholinergic activity. A dose of 30 mg/ml caused immediate contraction of the ileum, but after atropine the drug effect was antagonized, showing a cholinergic-like effect. The amplitude of peristaltic movement was increased with 5 mg/ml of the drug, and adrenaline, in large doses, removed the tonicity. Tonicity of the uterus was increased, and adrenaline checked the tonicity. It has also shown a slight diuretic effect (Roy et al., 1968). 5. Hypnotic and sedative effects. The drug potentiated phenobarbitone-induced sleep, but had no sedative effect when given alone (Roy et al., 1968). 6. Coagulant and anticoagulant activity. In dogs and rabbits, the drug reduced the whole blood clotting time, recalcied plasma clotting time, and bleeding time (Roy et al., 1968). 7. Hypoglycemic and antiglycemic effects. VIDR-2GD caused hypoglycemia (15%), at the end of the fth hour, with a dose of 100 mg/kg. After 24 hours, the blood sugar remained lowered by 8.5%. It did not have any inhibitory effect on the growth of E. histolytica in vitro up to the concentration of 50 mg/ml. Toxicity studies showed that doses of up to 5 g/kg orally, 2.5 g/kg intraperitoneally, and 1 g/kg intravenously did not produce any deleterious effects during 7 days observation (Roy et al., 1968). 8. Antifertility activity. The antifertility effect of VIDR-2GD, a substance isolated from the seeds of the Musaceae banakadali, was studied in rabbits, mice, rats, hamsters, and guinea pigs. In mice and rats, pregnancy was inhibited if the drug was given during the preimplantation period; single doses of the drug within 3 days after mating inhibited pregnancy. In rabbits, pregnancy was prevented in 50e60% of the animals. The uteri of all the animals receiving the drug were congested and enlarged. No implantation sites were found. Rabbit endometrium showed hyperplasial cells. Uterine stimulation was noted in ovariectomized rats treated with VIDR-2GD. There was no antagonism to estradiol, and

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TABLE 138.2 Comparative Anti-implantation Activity of the Ethanolic Extract and Isolated Compound of Ensete superbum Seeds
Treatment Ethanolic extract Control 100 mg 125 mg 250 mg Isolated compound Control 2.5 mg 5.0 mg 27.6 65.9 72.6 25.95 87.5 96.0 28.9 53.65 65.36 82.29 32.8 86.9 94.4 98.77 Percentage Anti-implantation, Day 4 (Post-coital) Percentage Anti-implantation, Days 1e7 (Post-coital)

The ethanolic extract of the seeds of Ensete superbum at the dose of 250 mg/kg on Day 4 and Days 1e7 post-coital possess signicant anti-implantation activity, and its isolated fraction (5 mg/kg), a chroman derivative, at the doses of 2.5 mg/kg and 5 mg/kg body weight on Day 4 and Days 1e7 post-coital also possess signicant anti-implantation activity.

only a weak antigonadotropic activity was shown (Dutta et al., 1970). The antiimplantation activity of the ethanolic extract of the seeds of Ensete superbum Cheesm (collected from the forest area of Waynard district in Kerala, India) at doses of 100, 125, and 250 mg/kg body weight when administered on days 1e7 post-coitally exhibited signicant anti-implantation activity, and also very signicant anti-estrogenic activity at the dose of 250 mg/kg body weight (Table 138.2). 1170 A compound was also isolated from the ethanolic extract of the seeds of Ensete superbum Cheesm (Family Musaceae), from the variety found in Kerala; thin layer chromotography of this reported a single peak with Rf at 0.69 (Figure 138.3). On the basis of physical and spectral parameters (i.e., HPTLC, UV, FTIR, NMR, and mass spectrometry), the isolated compound was identied possibly as a chroman derivative, with the molecular formula C16O4H22 e a nonsteroidal phytosterol (Kachroo et al., 2008). This isolated fraction, at doses of 2.5 and 5 mg/kg body weight, was found to exhibit signicant anti-implantation activity (Table 138.2), and

FIGURE 138.3
HPTLC chromatogram of the isolated chroman derivative. The HPTLC prole of the compound isolated from the ethanolic extract of E. superbum showed a single spot with Rf of 0.69 using mobile phase (toluene : ethyl acetate : formic acid, 5.0 : 4.5 : 0.5).

CHAPTER 138 Biological Activities of the Seeds of E. superbum

TABLE 138.3 Comparative Anti-estrogenic Activity of the Ethanolic Extract and Isolated Compound of Ensete superbum
Treatment Ethanolic extract Drug (oral) Oestradiol Benzoate Oestradiol Benzoate Drug (125 mg/kg) oral Isolated Compound Drug Oestradiol Benzoate Oestradiol Benzoate Drug Control 125mg/kg (0.05 mg) s.c (0.05 mg) s.c Control 5mg/kg(oral) (0.05 mg) s.c (0.05 mg) s.c 5mg/kg(oral) Mean Weight of Ovaries 27.02 0.38 25.03 0.59 36.17 1.05 30.87 0.66 26.45 0.82 25.16 0.73 41.9 15.7 35.91 0.99 Wet Weight of Uteri 78 132 65.03 0.80 154.53 194 107.5 0.70 35.94 1.05 34.93 0.98 115.74 4.76 77.64 4.24

The ethanolic extact of E.Superbum and its isolated chroman derivative when administered alone and with estradiol benzoate shows signicant anti-estrogenic activity. source of data personal research work.

also exhibited signicant anti-estrogenic activity in immature female rats at the dose of 5 mg/kg (Table 138.3). Histopathological studies revealed signicant anti-ovulatory activity in immature female rats at a dose of 5 mg/kg (Kachroo & Agrawal, 2009).

ADVERSE EFFECTS AND REACTIONS (ALLERGIES AND TOXICITY)

No health hazards or side effects are known with the proper administration of designated therapeutic dosages. The seeds of Ensete superbum have a long history of use for food and medicinal purposes, and no adverse or side effects have been reported when used within the recommended quantities. The LD50 (intraperitoneally) of VIDR-2GT was 174 mg/kg, while VIDR- 2GD and VIDR-2GC were non-toxic to mice (Dutta et al., 1968). The oral LD50 value of the ethanolic extract of Ensete superbum was found to be 3235.9 mg/kg, which appears to be safe (Kachroo & Agrawal, 2009). 1171

SUMMARY POINTS
l

Ensete superbum Cheesm, is essentially a plant of Western Ghats of India. It is known as banakadli in Hindi, and wild banana in English. In Kerala, the plant is known as kal vazhai, or rock banana. The ground seeds are used in Ayurvedic medicine, and certain fractions isolated from the seeds have shown to possess anti-variola and anti-vaccinia properties. A compound with the molecular formula C16O4H22, appearing to be a chroman derivative, has been isolated from seeds of Ensete superbum. The ethanolic extract, at doses of 100, 125 and 250 mg/kg body weight, and the isolated chroman, at doses of 2.5 and 5 mg/kg body weight, exhibit signicant anti-implantation activity. The oral LD50 value of the ethanolic extract of Ensete superbum was found to be 3235.9 mg/kg, which appears to be safe. Although a few promising leads have been identied by studies on seeds of Ensete superbum, none of these have been seriously pursued due to the very limited yield of the components isolated. The compound isolated and evaluated on the basis of physical and spectral data (i.e., a chroman derivative) can be further be validated by HPLC/HPTLC as a marker compound for elaborate anti-fertility studies.

PART 2 Effects of Specic Nuts and Seeds

References
Agharakar, S. P., & Bhaduri, P. N. (1933). Variation of chromosome numbers in Musaceae Agriculture Research Institute Coimbatore. Indian Journal of Agricultural Sciences, 3(IV), 1098. Baker, J. G. (1893). A synopsis of the genera and species of Musaceae. Annals Botany, 7, 189e229. Baker, J. G. (1894). Botany of the hadramaut expedition. Kew Bulletin, 1, 241. Cheesman, E. E. (1947). Classication of the bananas. The genus Ensete Horan. Kew Bulletin, 2, 97e106. Dutta, N. K., Dave, K. H., Desai, S. M., & Mhasalkar, M. Y. (1968). Anti-variola and anti-vaccinia principals from the seeds of banakadali (Ensete superbum, Cheesm, Musaceae). Indian Journal of Medical Research, 56, 735e741. Dutta, N. K., Mhasalkar, M. Y., & Fernando, G. R. (1970). Studies on the anti-fertility action of VIDR-2GD: a constituent isolated from the seeds of Ensete superbum Cheesm, Musaceae (banakadali). Fertility Sterility, 21, 247. Jagtap, S. D. (2008). Ethnomedicobotanical uses of endemic and RET plants utilised by the Korku tribe of Amaravati District, Maharastra. Indian Journal of Traditional Knowledge, 7(2), 284e287. Kachroo, M., Agrawal, S. S., & Sanjay, P. N. (2008). Characterization of a chroman derivative isolated from the seeds of Ensete superbum Cheesm. Pharmacognosy Magazine, 4, 114e117. Kachroo, M., & Agrawal, S. S. (2009). Isolation, characterization and anti-fertility activity of the active moiety from the seeds of Ensete superbum Cheesm (banakadali). Journal of Natural Remedies, 9, 12e20. Kulkarni, V. M., Ganapati, T. R., Suprasanna, P., Bapat, V. A., & Rao, P. S. (1997). In vitro propagation in Ensete superbum (Roxb.) Cheesman e a species closely related to Musa. Indian Journal of Experimental Biology, 35, 96e98. Mahey, S., Mukerjee, S. K., Saroja, T., & Seshadri, T. R. (1971). Proanthocyanidin glycosides in Musa accuminata. Indian Journal of Chemistry, 4, 381. Ravishankar, T., Vedavalli, L., Nambi, A. A., & Selvam, V. (1994). Role of tribal people in the conservation and utilization of plant genetic resource. Madras, India: MSSRF. Rao, P. S., Ganapathi, T. R., Suprasanna, P., & Bapat, V. A. (1993). Encapsulated shoot tips of banana: a new propagation and delivery system. InfoMusa, 2(2), 4e5. Roy, R. N., Bhagwager, S., & Dutta, N. K. (1968). Pharmacological observations on the fractions isolated from seed of banakadali. Indian Journal of Pharmaceutical Sciences, 30, 285.

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Sreedharan, T. P. (2004). Biological diversity of Kerala: A survey of Kalliasseri panchayat, Kannur district, Kerala. Research Programme on Local Level Development Centre for Development Studies Thiruvananthapuram. Discussion Paper No. 62. Yeshodharan, K., & Sujana, K. A. (2007). Ethno-medicinal knowledge among the Malamalsar tribe of Parambikulam wildlife sanctuary, Kerala. Indian Journal of Traditional Knowledge, 6(3), 481e485.