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Anticonvulsant activity of root and stem extracts of Calliandra portoricensis
205
Journul of Ethnopharmacobgy, 22 (1988) 205-210 Elsevier Scientific Publishers Ireland Ltd.,
ANTICONVULSANT ACTIVITY OF ROOT OF CALLIANDRA PORTORICENSIS
AND
STEM
EXTRACTS
PETER A. AKAH and JOHN I. NWAIWU Faculty of Phammeutical
Sciences, University
of Niger&o, Nsukka (Nige&
(Accepted December 21, 1987)
Summary
A comparative study of the anticonvulsant properties of root and stem extracts of Calliandra portoricensis was carried out in mice. The convulsion models were pentylenetetrazoleand electroshock-induced convulsions. The results showed that the aqueous extracts of both root and stem possess anticonvulsant activity when given intraperitoneally. An alkaloidal extract did not show any anticonvulsant action up to 316 mg/kg i.p.
Introduction
Many genera belonging to the Mimosaceae family are known to be of medicinal value (Daziel, 1948; Nwaiwu and Akah, 19861. Many members of this family are shrubs that grow abundantly along the West Coast of Africa. Among these, Calliandra portoricensis Benth has for a long time been the drug of choice among herbalists in the Southern states of Nigeria for gastrointestinal problems and convulsions. When used as a chewing stick the stem is locally employed as an analgesic for toothache. Some of the medicinal actions of this plant which have been reported include a molluscicidal action (Adewumi and Marquis, 1980; Kloos and McCullough, 19821, purgative effect (Daziel, 19481, relief of lumbar pain (Irvine, 19611 and a moderate anticonvulsant effect (Adesina and Akinwusi, 19841. C. portoricensis stands out as the most popular traditionally employed remedy for the treatment of convulsive episodes in infants. The mode of treatment and parts of the plant used vary from one locality to another. While the root decoction is used in one area, stem or leaf decoctions may be preferred in other regions, and the effectiveness of all have been acclaimed. This laboratory is investigating the effectiveness of the many herbal preparations traditionally used in the management of convulsions in infants Correspondence
to: Dr. John I. Nwaiwu.
03’78-8741/88/$02.45 0 1988 Elsevier Scientific Publishers Ireland Ltd. Published and Printed in Ireland
206
(Nwaiwu and Akah, 19861. In the present study, it was our intention to make a comparative study of the anticonvulsant activity of root and stem extracts of C. portoricensis with the aim of establishing which of these extracts (if any) is the more effective anticonvulsant.
Materials
and methods
Collection
and identification
of plant material
The plant was collected from Ozubulu town in Nnewi Local Government Area, Anambra State, Nigeria, and was authenticated by Mr. Ozioko of the Botany Department (Herbarium Section), University of Nigeria, Nsukka. Extraction
procedures
The root and the stem of the plant material were air-dried and reduced to coarse powder. In order to prepare the aqueous extracts, about 200 g of the coarse powder (root or stem) were macerated for 1 h with 1 1 of water. The process was repeated three times and the aqueous extracts were combined and concentrated to about one-half of the original volume by evaporation under reduced pressure. The concentrated aqueous extracts were then partitioned three times with 150 ml of petroleum ether (40-60°C) to remove any fatty materials and then evaporated to dryness. Extract yields of 6.6% and 4.3% (w/w) were obtained from the root and stem, respectively. For preparation of alkaloidal extracts, about 20 g of the coarse powder (root or stem) were defatted with petroleum ether and the mart air-dried. The dried mart was basified with ammonia solution and the alkaloids extracted several times with 40-ml portions of chloroform. The chloroform extracts were combined and a little water added to dissolve the impurities. The solution was passed through anhydrous Na,SO, and evaporated under reduced pressure. Yields of 1.9% and 1.2% of the alkaloidal extracts were obtained from the root and stem, respectively. Phytochemical
tests
Aqueous extracts of both root and stem were chemically tested for the presence of glycosides, alkaloids, saponins, tannins, flavonoids, sterols and triterpenoids using standard procedures (Trease and Evans, 1983; Agrawal and Nwaiwu, 19841. Animals
Inbred adult albino mice of both sexes weighing 20-30 g, obtained from stock maintained at the Department of Veterinary Pathology, University of
Nigeria, Nsukka were used. The animals were fed on Pfizer mice cubes with free access to drinking water. Food was withdrawn 12 h before experimentation (Davenport and Davenport, 1948). Pentylene te trazole-induced convulsions Thirty-six albino mice of either sex were randomly divided into six groups of 6 mice per group. One group (control) received 25 ml/kg of normal saline intraperitoneally (Lp.). Groups 2-6 received different doses (mg/kgI of the root or stem extracts i-p. Thirty minutes later, ‘70 mg/kg of pentylenetetrazole (PTZ, Sigma) was administered i.p. to each group. The animals were observed for 60 min for seizures. An episode of clonic spasm that persisted for a minimum of 30 s was taken as a threshold convulsion. Animals devoid of a threshold convulsion during the 60 min of observation were considered protected. Electroshock-induced convulsions This test was carried out as described by Dikshit et al. (19721,Thirty-six albino mice of either sex were divided randomly into six groups of 6 mice per group. Five groups were given i.p. injection of different doses of extracts of root or stem (rng~g). The control group received normal saline 25 ml/kg i.p. Thirty minutes after drug administration, each mouse was given an electroshock Wgo Basile eleetr-convulsive treatment unit) using ear-clip electrodes previously dipped in normal saline to make better contact. The stimulation parameters used (45 mA, 0.2 s, 100 Hz) produced maximum shock without being lethal. ED, de termination The median effective doses (ED,) of both the root and stem extracts were determined in mice along with their respective 95W confidence limits (Miller and Tainter, 19441. Results
Phytochemical analysis Both the root and stem aqueous extracts gave positive reactions to glycosides, alkaloids, saponins, tannins, flavonoids, sterols and triterpenoids. The root extracts, however, showed the presence of greater alkaloidal content than the stem extracts. The thalleioquin test (Trease and Evans, 1983) revealed the presence of quinine-like alkaloids.
208 TABLE
1
EFFECT OF PTZ (70 mg/kg i.p.) ON MICE PRETREATED AQUEOUS ROOT EXTRACT OF C. PORTORICENSIS
30 min EARLIER
Dose of extract (mg.kg)
Onset of convulsion (min)
Quanta1 protection
Protection (Oh)
Mean duration
Control 62.5 187.5 312.5 437.5 562.5
8.9 12.0 16.9 18.7 22.0 27.1
O/6 O/6 116 416 5/6 516
0 0 16.7 66.7 83.3 83.3
9.4 38.2 27.3 12.6 9.1 6.0
TABLE
r?: 2.0 * 1.2 + 1.0 2 0.8 zt 1.6 2 2.0
WITH
THE
of convulsion S.E.M. (min) f f + +
f
3.1 2.1 1.6 3.1
2
EFFECT OF PTZ (70 mg/kg i.p.1 ON MICE PRETREATED AQUEOUS STEM EXTRACT OF C. PORTORICENSIS
30 min EARLIER
WITH
THE
(mg.kg)
Onset of convulsion (min)
Quanta1 protection
Protection (O/b)
Mean duration of convulsion i: S.E.M. (min)
Control 57.5 172.5 287.5 402.5 517.5
8.7 11.2 16.0 18.3 19.7 24.0
016 016 l/6 216 416 5/6
0 0 16.7 33.3 66.7 83.3
8.8 40.1 29.0 17.4 10.1 6.3
Dose of extract
+ + f + + f
1.8 1.6 2.0 1.4 1.0 2.2
+ + f f =
2.4 3.1 2.6 1.2 2.0
TABLE 3 EFFECT AQUEOUS
OF ELECTROSHOCK ROOT EXTRACT
ON MICE
PRETREATED
30 min EARLIER
WITH
THE
OF C. PORTORICENSIS
Onset of convulsion (min)
Quanta1 protection
Protection (Oh-l)
Mean duration
(m&kg)
Control 62.5 187.5 312.5 437.5 562.5
0.6 0.90 1.4 1.7 2.1 2.8
O/6 l/6 216 316 416 516
0 16.2 33.3 50.0 66.7 83.3
34.7 35.8 24.8 9.7 9.0 5.0
Dose of extract
f f 2 f f f
0.2 0.1 0.3 0.4 0.2 0.7
of convulsion S.E.M. (s) f f f -t 2
3.2 2.8 1.2 1.1 1.4
+
209 TABLE 4 EFFECT OF ELECTROSHOCK ON MICE PRETREATED AQUEOUS ROOT EXTRACT OF C. PQRTORICENSZ3
30 min EARLIER WLTH THE
Dose of extract (m&kg)
Onset of convulsion (min)
Quanta1 protection
Protection (O/o)
Mean duration of convulsion f S.E.M. (~1
Control 57.5 172.5 287.5 402.5 517.5
0.6 0.7 1.0 1.5 1.7 2.3
016 010 l/8 216 316 518
0 0 16.7 33.3 50.0 83.3
35.2 37.3 29.4 14.8 11.3 7.0
Anticonvulsant
~fr0.8 f 0.1 t 0.3 f 0.2 It 0.1 + 0.7
+ + 4 rt s
2.4 2.1 1.0 1.2 1.0
activity
Tables l-4 show that both the root and stem aqueous extracts of C. ~orto~ce~~~ offered protection against PTZ- and electroshock-induced convulsions in mice. The results did not show any overwhelming advantage that the root or stem extract might have over each other. There was no significant difference Cp < 0.05) in the dosage of the two extracts which offered maximum protection in the two convulsion models. However, the ED, values (calculated within 95% confidence limits) for the root extract (279.6 f: 33 mg/kg and 312 + 2.1 mgikg) were lower than those for the stem extract (338 f 4.2 mgkg and 402 rt 2.6 mg/kg) for chemo- and electroshock convulsions, respectively. The average duration of convulsions was consistently shorter in the root extract-treated animals. The alkaloidal extracts of both root and stem up to 316 mg/kg offered no protection against both chemo- and electroshock convulsions. At the dosage of 312 mgikg, the percentage protection offered by the aqueous root extracts, were 66.1 and 50.0% against PTZ and electroshock convulsions, respectively. On the other hand, the percentage protection offered by 281.5 mgjkg of the aqueous stem extracts against PTZ and electroshock convulsions were 33.3 and 33.?%, respectively. Discussion This study provides evidence for the anticonvulsant activity of the aqueous extracts of the root and stem of C. portoricensis using two animal models. The results did not show any marked difference in the anticonvulsant effect of one plant part relative to the other since both the aqueous root and stem extracts appeared equipotent in the two experimental
210
in addition to the models. The apparent lower ED,, of the root extracts, shorter duration of convulsions in root extract-treated animals, may indicate some advantage for the root extract over stem extract. In our present state of knowledge of the chemical constituents of the extracts, it is not possible to identify with certainty the main anticonvulsant principle in the extracts. This is supported by the fact that the alkaloidal fraction did not show any anticonvulsant activity. Efforts to isolate and identify the anticonvulsant but until then we support the principle(s) of the extracts will continue, medicinal use of the aqueous extract. Acknowledgement The authors wish to thank Mr. Ozioko for the identification and Mr. D.U. Egwu for his laboratory assistance.
of the plant
References Adesina, S.K. and Akinwusi D.D. (1984) Biological effects of Calliundra portoricensis and Lagenaria breviflora extracts. Fitoterapia 56 (6). 75-76. Adewumi, S.O. and Marquis, V.O. (1980) Laboratory and field trials of the molluscicidal properties of Calliandra portoricensis Benth. In: Proceedings of the 10th International Congress of Tropical Medicine and Malaria, Ile-Ife, Nigeria, p. 354. Agrawal, S.P. and Nwaiwu, J.I. (1984) Dimethylsulphoxide as a spray reagent for the detection of triterpenoids and some steroids on thin-layer plates. Journal of Chromatography 295, 537 - 543. Davenport, V.A. and Davenport, H.W. (1948) The relationship between starvation, metabolic acidosis and convulsive seizures. Journal of Nutrition 36, 139- 151. Daziel, J.M. (1948) The Useful Plants of West Tropical Africa. The Crown Overseas Agents of the Colonies, London, pp. 223- 224. Dikshit, S.K., Tewan, P.V. and Dixit. S.P. (1972) Anticonvulsant activity of Causoru de&sate. Indian Journal of Physiology and Pharmacology 16, 81- 82. Irvine F.R. (1961) Woody Plants of Ghana Oxford University Press, London, pp. 336-337. Kloos, H. and McCullough, F.S. (1982) Plant molluscicidals. Pkznta Medica 46, 195-209. Miller, L.C. and Tainter, M.L. (1944) Estimation of ED, and it,s error by means of log probit graph paper. Proceedings of the Society for Experimental Biology and Medicine 57, 261264. Nwaiwu, J.I. and Akah, P.A. (1986) Anticonvulsant activity of the volatile oil from the fruit of Tetrapleura tetraptera Journal of Ethnopharmacology 18. 103- 107. Trease, C.E. and Evans, W.C. (1983) Pharmacognosy, 12th edn. Bailliere Tindall, London, pp. 383 - 385.
Journul of Ethnopharmacobgy, 22 (1988) 205-210 Elsevier Scientific Publishers Ireland Ltd.,
ANTICONVULSANT ACTIVITY OF ROOT OF CALLIANDRA PORTORICENSIS
AND
STEM
EXTRACTS
PETER A. AKAH and JOHN I. NWAIWU Faculty of Phammeutical
Sciences, University
of Niger&o, Nsukka (Nige&
(Accepted December 21, 1987)
Summary
A comparative study of the anticonvulsant properties of root and stem extracts of Calliandra portoricensis was carried out in mice. The convulsion models were pentylenetetrazoleand electroshock-induced convulsions. The results showed that the aqueous extracts of both root and stem possess anticonvulsant activity when given intraperitoneally. An alkaloidal extract did not show any anticonvulsant action up to 316 mg/kg i.p.
Introduction
Many genera belonging to the Mimosaceae family are known to be of medicinal value (Daziel, 1948; Nwaiwu and Akah, 19861. Many members of this family are shrubs that grow abundantly along the West Coast of Africa. Among these, Calliandra portoricensis Benth has for a long time been the drug of choice among herbalists in the Southern states of Nigeria for gastrointestinal problems and convulsions. When used as a chewing stick the stem is locally employed as an analgesic for toothache. Some of the medicinal actions of this plant which have been reported include a molluscicidal action (Adewumi and Marquis, 1980; Kloos and McCullough, 19821, purgative effect (Daziel, 19481, relief of lumbar pain (Irvine, 19611 and a moderate anticonvulsant effect (Adesina and Akinwusi, 19841. C. portoricensis stands out as the most popular traditionally employed remedy for the treatment of convulsive episodes in infants. The mode of treatment and parts of the plant used vary from one locality to another. While the root decoction is used in one area, stem or leaf decoctions may be preferred in other regions, and the effectiveness of all have been acclaimed. This laboratory is investigating the effectiveness of the many herbal preparations traditionally used in the management of convulsions in infants Correspondence
to: Dr. John I. Nwaiwu.
03’78-8741/88/$02.45 0 1988 Elsevier Scientific Publishers Ireland Ltd. Published and Printed in Ireland
206
(Nwaiwu and Akah, 19861. In the present study, it was our intention to make a comparative study of the anticonvulsant activity of root and stem extracts of C. portoricensis with the aim of establishing which of these extracts (if any) is the more effective anticonvulsant.
Materials
and methods
Collection
and identification
of plant material
The plant was collected from Ozubulu town in Nnewi Local Government Area, Anambra State, Nigeria, and was authenticated by Mr. Ozioko of the Botany Department (Herbarium Section), University of Nigeria, Nsukka. Extraction
procedures
The root and the stem of the plant material were air-dried and reduced to coarse powder. In order to prepare the aqueous extracts, about 200 g of the coarse powder (root or stem) were macerated for 1 h with 1 1 of water. The process was repeated three times and the aqueous extracts were combined and concentrated to about one-half of the original volume by evaporation under reduced pressure. The concentrated aqueous extracts were then partitioned three times with 150 ml of petroleum ether (40-60°C) to remove any fatty materials and then evaporated to dryness. Extract yields of 6.6% and 4.3% (w/w) were obtained from the root and stem, respectively. For preparation of alkaloidal extracts, about 20 g of the coarse powder (root or stem) were defatted with petroleum ether and the mart air-dried. The dried mart was basified with ammonia solution and the alkaloids extracted several times with 40-ml portions of chloroform. The chloroform extracts were combined and a little water added to dissolve the impurities. The solution was passed through anhydrous Na,SO, and evaporated under reduced pressure. Yields of 1.9% and 1.2% of the alkaloidal extracts were obtained from the root and stem, respectively. Phytochemical
tests
Aqueous extracts of both root and stem were chemically tested for the presence of glycosides, alkaloids, saponins, tannins, flavonoids, sterols and triterpenoids using standard procedures (Trease and Evans, 1983; Agrawal and Nwaiwu, 19841. Animals
Inbred adult albino mice of both sexes weighing 20-30 g, obtained from stock maintained at the Department of Veterinary Pathology, University of
Nigeria, Nsukka were used. The animals were fed on Pfizer mice cubes with free access to drinking water. Food was withdrawn 12 h before experimentation (Davenport and Davenport, 1948). Pentylene te trazole-induced convulsions Thirty-six albino mice of either sex were randomly divided into six groups of 6 mice per group. One group (control) received 25 ml/kg of normal saline intraperitoneally (Lp.). Groups 2-6 received different doses (mg/kgI of the root or stem extracts i-p. Thirty minutes later, ‘70 mg/kg of pentylenetetrazole (PTZ, Sigma) was administered i.p. to each group. The animals were observed for 60 min for seizures. An episode of clonic spasm that persisted for a minimum of 30 s was taken as a threshold convulsion. Animals devoid of a threshold convulsion during the 60 min of observation were considered protected. Electroshock-induced convulsions This test was carried out as described by Dikshit et al. (19721,Thirty-six albino mice of either sex were divided randomly into six groups of 6 mice per group. Five groups were given i.p. injection of different doses of extracts of root or stem (rng~g). The control group received normal saline 25 ml/kg i.p. Thirty minutes after drug administration, each mouse was given an electroshock Wgo Basile eleetr-convulsive treatment unit) using ear-clip electrodes previously dipped in normal saline to make better contact. The stimulation parameters used (45 mA, 0.2 s, 100 Hz) produced maximum shock without being lethal. ED, de termination The median effective doses (ED,) of both the root and stem extracts were determined in mice along with their respective 95W confidence limits (Miller and Tainter, 19441. Results
Phytochemical analysis Both the root and stem aqueous extracts gave positive reactions to glycosides, alkaloids, saponins, tannins, flavonoids, sterols and triterpenoids. The root extracts, however, showed the presence of greater alkaloidal content than the stem extracts. The thalleioquin test (Trease and Evans, 1983) revealed the presence of quinine-like alkaloids.
208 TABLE
1
EFFECT OF PTZ (70 mg/kg i.p.) ON MICE PRETREATED AQUEOUS ROOT EXTRACT OF C. PORTORICENSIS
30 min EARLIER
Dose of extract (mg.kg)
Onset of convulsion (min)
Quanta1 protection
Protection (Oh)
Mean duration
Control 62.5 187.5 312.5 437.5 562.5
8.9 12.0 16.9 18.7 22.0 27.1
O/6 O/6 116 416 5/6 516
0 0 16.7 66.7 83.3 83.3
9.4 38.2 27.3 12.6 9.1 6.0
TABLE
r?: 2.0 * 1.2 + 1.0 2 0.8 zt 1.6 2 2.0
WITH
THE
of convulsion S.E.M. (min) f f + +
f
3.1 2.1 1.6 3.1
2
EFFECT OF PTZ (70 mg/kg i.p.1 ON MICE PRETREATED AQUEOUS STEM EXTRACT OF C. PORTORICENSIS
30 min EARLIER
WITH
THE
(mg.kg)
Onset of convulsion (min)
Quanta1 protection
Protection (O/b)
Mean duration of convulsion i: S.E.M. (min)
Control 57.5 172.5 287.5 402.5 517.5
8.7 11.2 16.0 18.3 19.7 24.0
016 016 l/6 216 416 5/6
0 0 16.7 33.3 66.7 83.3
8.8 40.1 29.0 17.4 10.1 6.3
Dose of extract
+ + f + + f
1.8 1.6 2.0 1.4 1.0 2.2
+ + f f =
2.4 3.1 2.6 1.2 2.0
TABLE 3 EFFECT AQUEOUS
OF ELECTROSHOCK ROOT EXTRACT
ON MICE
PRETREATED
30 min EARLIER
WITH
THE
OF C. PORTORICENSIS
Onset of convulsion (min)
Quanta1 protection
Protection (Oh-l)
Mean duration
(m&kg)
Control 62.5 187.5 312.5 437.5 562.5
0.6 0.90 1.4 1.7 2.1 2.8
O/6 l/6 216 316 416 516
0 16.2 33.3 50.0 66.7 83.3
34.7 35.8 24.8 9.7 9.0 5.0
Dose of extract
f f 2 f f f
0.2 0.1 0.3 0.4 0.2 0.7
of convulsion S.E.M. (s) f f f -t 2
3.2 2.8 1.2 1.1 1.4
+
209 TABLE 4 EFFECT OF ELECTROSHOCK ON MICE PRETREATED AQUEOUS ROOT EXTRACT OF C. PQRTORICENSZ3
30 min EARLIER WLTH THE
Dose of extract (m&kg)
Onset of convulsion (min)
Quanta1 protection
Protection (O/o)
Mean duration of convulsion f S.E.M. (~1
Control 57.5 172.5 287.5 402.5 517.5
0.6 0.7 1.0 1.5 1.7 2.3
016 010 l/8 216 316 518
0 0 16.7 33.3 50.0 83.3
35.2 37.3 29.4 14.8 11.3 7.0
Anticonvulsant
~fr0.8 f 0.1 t 0.3 f 0.2 It 0.1 + 0.7
+ + 4 rt s
2.4 2.1 1.0 1.2 1.0
activity
Tables l-4 show that both the root and stem aqueous extracts of C. ~orto~ce~~~ offered protection against PTZ- and electroshock-induced convulsions in mice. The results did not show any overwhelming advantage that the root or stem extract might have over each other. There was no significant difference Cp < 0.05) in the dosage of the two extracts which offered maximum protection in the two convulsion models. However, the ED, values (calculated within 95% confidence limits) for the root extract (279.6 f: 33 mg/kg and 312 + 2.1 mgikg) were lower than those for the stem extract (338 f 4.2 mgkg and 402 rt 2.6 mg/kg) for chemo- and electroshock convulsions, respectively. The average duration of convulsions was consistently shorter in the root extract-treated animals. The alkaloidal extracts of both root and stem up to 316 mg/kg offered no protection against both chemo- and electroshock convulsions. At the dosage of 312 mgikg, the percentage protection offered by the aqueous root extracts, were 66.1 and 50.0% against PTZ and electroshock convulsions, respectively. On the other hand, the percentage protection offered by 281.5 mgjkg of the aqueous stem extracts against PTZ and electroshock convulsions were 33.3 and 33.?%, respectively. Discussion This study provides evidence for the anticonvulsant activity of the aqueous extracts of the root and stem of C. portoricensis using two animal models. The results did not show any marked difference in the anticonvulsant effect of one plant part relative to the other since both the aqueous root and stem extracts appeared equipotent in the two experimental
210
in addition to the models. The apparent lower ED,, of the root extracts, shorter duration of convulsions in root extract-treated animals, may indicate some advantage for the root extract over stem extract. In our present state of knowledge of the chemical constituents of the extracts, it is not possible to identify with certainty the main anticonvulsant principle in the extracts. This is supported by the fact that the alkaloidal fraction did not show any anticonvulsant activity. Efforts to isolate and identify the anticonvulsant but until then we support the principle(s) of the extracts will continue, medicinal use of the aqueous extract. Acknowledgement The authors wish to thank Mr. Ozioko for the identification and Mr. D.U. Egwu for his laboratory assistance.
of the plant
References Adesina, S.K. and Akinwusi D.D. (1984) Biological effects of Calliundra portoricensis and Lagenaria breviflora extracts. Fitoterapia 56 (6). 75-76. Adewumi, S.O. and Marquis, V.O. (1980) Laboratory and field trials of the molluscicidal properties of Calliandra portoricensis Benth. In: Proceedings of the 10th International Congress of Tropical Medicine and Malaria, Ile-Ife, Nigeria, p. 354. Agrawal, S.P. and Nwaiwu, J.I. (1984) Dimethylsulphoxide as a spray reagent for the detection of triterpenoids and some steroids on thin-layer plates. Journal of Chromatography 295, 537 - 543. Davenport, V.A. and Davenport, H.W. (1948) The relationship between starvation, metabolic acidosis and convulsive seizures. Journal of Nutrition 36, 139- 151. Daziel, J.M. (1948) The Useful Plants of West Tropical Africa. The Crown Overseas Agents of the Colonies, London, pp. 223- 224. Dikshit, S.K., Tewan, P.V. and Dixit. S.P. (1972) Anticonvulsant activity of Causoru de&sate. Indian Journal of Physiology and Pharmacology 16, 81- 82. Irvine F.R. (1961) Woody Plants of Ghana Oxford University Press, London, pp. 336-337. Kloos, H. and McCullough, F.S. (1982) Plant molluscicidals. Pkznta Medica 46, 195-209. Miller, L.C. and Tainter, M.L. (1944) Estimation of ED, and it,s error by means of log probit graph paper. Proceedings of the Society for Experimental Biology and Medicine 57, 261264. Nwaiwu, J.I. and Akah, P.A. (1986) Anticonvulsant activity of the volatile oil from the fruit of Tetrapleura tetraptera Journal of Ethnopharmacology 18. 103- 107. Trease, C.E. and Evans, W.C. (1983) Pharmacognosy, 12th edn. Bailliere Tindall, London, pp. 383 - 385.