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PHYTOTHERAPY RESEARCH
Phytother. Res. 17, 254 – 258 (2003)
Published online in Wiley InterScience (www.interscience.wiley.com). DOI: 10.1002/ptr.1122
Toxicological Effects of α-Solamargine in
Experimental Animals
Lina Al Chami1, Ramón Méndez1, Bernardo Chataing1*, James O’Callaghan1,
Alfredo Usubillaga2 and Luis LaCruz3
1Departamento de Biología, Facultad de Ciencias, Universidad de Los Andes, Mérida, Venezuela
2Instituto de Farmacia, Facultad de Farmacia, Universidad de Los Andes, Mérida, Venezuela
3Departamento de Toxicología, Facultad de Medicina, Universidad de Los Andes, Mérida, 5101, Mérida Venezuela
α-Solamargine isolated from the fresh fruits of Solanum americanum Miller was studied for its toxicity.
Lethality studies in rats showed a dose-mortality relationship with a LD of 42 mg/ kg body weight intraperi-
toneally. The chronic and subchronic toxicity investigations indicated that the size of the glycoalkaloid dose
was more important than the total glycoalkaloid intake. No appreciable toxic effects were observed at doses
below 35 mg / kg body weight as indicated by blood parameters, enzyme levels and histological sections of
kidney, liver and cardiac muscle. α-Solamargine did not affect the weight of the testes and epididymis or the
number of spermatozoa but produced a slight irritation and congestion in the epididymis and testis at doses
up to 50 mg / kg body weight. Copyright 2003 John Wiley & Sons, Ltd.
Keywords: Solamargine; Solanaceae; glycoalkaloids; toxicology; pharmacology.
INTRODUCTION
MATERIALS AND METHODS
The Solanaceae family include many species of plants Animals. The recipients, 6 to 8 week-old male Wistar
important to man as diverse agricultural crops and rats with a body weight of approximately 150 –200 g, medicinal treatments (Fewell et al., 1994; Cham and were purchased from the Animal House Center of La Meares, 1987; Cham et al., 1987; Chataing et al., 1996).
The primary components are steroidal alkaloids of thespirosolane or solanidane types, which generally occur Solamargine isolation. Fresh fruits (770 g) 1–2 weeks
as glycosides (Fig. 1). Toxicological studies indicate that old of Solanum americanum Miller were harvested the solanidanes seem to be more toxic than their from the Botanical Garden at the Pharmacy Faculty, corresponding spirosolanes – α-solamargine, solasonine The University de los Andes. They were washed with and solasodine (Friedman and McDonald, 1997).
250 mL of distilled water, minced finely and blended.
However, the toxicological profile has been centred upon After boiling the mixture for 5 min, 500 mL glacial solanine, α-chaconine and other potato components.
acetic acid (5% total volume) and 500 mL of methanol Also, most of the alkaloids from the Solanaceae inhibit were added and left at room temperature for 3 days.
acetylcholinesterase activity (Roddick, 1989). The tera- The mixture was filtered, the filtrate was heated at togenic effects of these alkaloids are inconclusive 70°C, and the alkaloids precipitated with NH OH at pH (Friedman and McDonald, 1997). In animals, adverse 12. The solids were resuspended in 5% acetic acid and physiological effects of steroidal glycoalkaloids are reprecipitated with NH OH at pH 12. The crude alka- manifested in a number of ways, such as reduced loids (1.15 g) were washed with cold water, dried at respiratory activity or blood pressure, bradycardia and 45°C, and dissolved in MeOH. The glycoalkaloids haemolysis which are thought to stem mainly from were chromatographed on an open silica gel column membrane disruption, inhibition of acetylcholinesterase which was eluted with CHCl /MeOH/H O mixtures activity or interference with sterol /steroid metabolism starting with 140:10:1, and 100 mL fractions were col- or a combination (Roddick, 1989; Kusano et al., 1987).
lected. Fractions 37– 46 eluted with CHCl /MeOH/H O In this work, experiments are reported that test some 90:10:1 afforded 128 mg of solasonine and fractions toxicological properties of solamargine and compare 69 –100 eluted with the same solvent mixture afforded them with data reported for other solanum alkaloids 152 mg of solamargine. The fractions were inspected such as solanine and glycoalkaloids present in food by TLC on silica gel plates using CHCl MeOH/1% NH OH (2:2:1, lower phase) as solvent. Solasonine and solamargine showed R 0.32 and 0.22, respectively.
Analysis of the samples by HPLC, using a µ-Bondapak * Correspondence to: Dr B. Chataing, Departamento de Biología Fac- CN column revealed the existence of two main peaks ultad de Ciencias, Universidad de Los Andes, Mérida 5101, Mérida, which correspond to solamargine and solasonine. The smaller peaks correspond to mono- and di-glycosides E-mail: chataing@ciens.ula.veContract/grant sponsor: CDCHT-ULA.
The identity of the glycoalkaloids was confirmed by 13C-NMR spectra by comparison with values reported Copyright 2003 John Wiley & Sons, Ltd.
Figure 1. Structure of Solanaceae glycoalkaloids.
Figure 2. Determination of lethal dose 50 (LD ).
by Mahato et al. (1980). The glycoalkaloid was dis- deaths in each group were calculated. The percentages solved in 0.1% acetic acid and 0.9% NaCl, filtered, were converted to probit and the LD was calculated and sterilized until its inoculation to the recipient from a statistical linear regression analysis which include the standard deviation (S.D.).
Lethal dose 50 (LD ). Ninety Wistar rats, differing less
Acute toxicity. A total of 20 Wistar rats were injected
than 25 g in weight, were randomly divided into nine i.p. with doses of 10 mg/kg BW α-solamargine over 5 groups of ten individuals each. Single doses of solamar- continuous days. A group of 10 animals served as a gine, ranging from 10 to 60 mg/kg body weight (BW), were administered intraperitoneally (i.p.) to the rats. Asa control group 10 Wistar rats were inoculated with a Subchronic toxicity. A total of 50 Wistar rats were
single i.p. dose of solvent. All groups were watched separated into groups of 10 rats each. Two groups each 3 h for 24 h, and the number and the percentage of served as the control by inoculation of anaesthesia and Copyright 2003 John Wiley & Sons, Ltd.
Phytother. Res. 17, 254 – 258 (2003)
TOXICOLOGICAL EFFECTS OF X-SOLAUCARGINE IN EXPERIMENTAL ANIMALS solvent, respectively, and the other three groups were injected i.p. with either 15, 25 or 35 mg/kg BW of α- solamargine. The animals were observed for general symptoms of toxicity, external symptoms and mortality.
All animals were killed 24 h after the last exposure.
Vital organs (liver, kidney, spleen, cardiac and skeletalmuscle, testes) were extracted and sampled for histo- logical analysis. Tissues were fixed in Bouin’s solution, dehydrated in alcohol and embedded in paraffin.
Microscopic sections were stained with haematoxylinand eosin. Blood was isolated from the rats andanalysed for haemoglobin estimation (Hb), packed cellvolume (PCV), mean corpuscular haemoglobin (MCH), mean cell volume (MVC) and red blood cell count (RBC). Levels of creatinine (CREA), glutamate- oxaloacetate transaminase (GOT), glutamate-pyruvatetransaminase (GTP) and alkaline phosphatase (AP)were also determined.
Spermatogenesis. Male Wistar rats were inoculated i.p.
with doses of α-solamargine of 15, 25 and 35 mg/kgBW. The rats were killed 24 h after the last exposure < 0.01 was considered statistically significant. Data shown p and the epididymes of rats were isolated to determinethe number of spermatozoa. Also, sections of the right and left testis were isolated to determine their weight Statistical treatment of data. Data were analysed by a
statistical non-parametric Kruskal–Allis ANOVA test
with the Statistica 6.0 program. Application of the test
was used to determine the statistical significance of data. p < 0.01 was considered statistically significant.
Toxicity test
Lethality studies showed a dose-mortality relationshipwhich was apparently sigmoidal. A plot of probit values(% mortality) versus log-dose of α-solamargine gave a straight line (Fig. 2). From the straight line graph, the LD value of solamargine was extrapolated. This value No animal died earlier than 6 h. However, the high dose rats (over 60 mg/kg BW) died within 3 h after asingle i.p. injection. Otherwise, multiple i.p. doses of 10 mg/kg animal weight of α-solamargine administered over5 consecutive days did not cause any mortality or dif- ference in the body weight of the animals in that period.
This observation confirmed the results of Chaube andSwinyard (1976) that the dose size seems to be more important than the total glycoalkaloid intake. Alsoobserved were muscular contraction and dizziness at = 10. Significant differences were determined with respect to the control (anaesthesized only). also observed by Aldous et al. (1980) in rats injected i.p.
with α-chaconine. No bleeding from the eyes or nose -solamargine on blood cells constituents, creatinine and enzyme levels
Subchronic toxicity
The blood of Wistar rats inoculated with α-solamarginewas analysed by blood components. Chemical deter- Table 1. Effect of
indicated non statistical significance, except for GTP and AP mination of GOT, GTP, creatinine and AP indicated no Copyright 2003 John Wiley & Sons, Ltd.
Phytother. Res. 17, 254 – 258 (2003)
Table 2. Testes, epydidimis and spermatozoa analysis of Wistar rats treated with different α-solamargine doses
Data shown are mean ± SD, n = 10. Significant differences were determined with respect to the control (anaesthesized only). p < 0.01was considered statistically significant. Data shown indicated non statistical significance.
appreciable differences in values, except for a slight, but similarity of LD values on rats was observed in com- not significant, increase in these variables measured at parison with values determined in mice. However, the doses of 35 mg /kg BW (Table 1). Examination of the studies performed with solanaceae alkaloids have vital organs tissues did not depict any toxic effect (data revealed differences in LD values depending on the experimental animal used, the conditions of the assaysand the purity of the alkaloids. The LD i.p. for α- solanine, α-chaconine and tomatine in mice were 27, 30 Spermatogenesis analysis
and 34 mg/kg BW, respectively and for most animals,the i.p. LD of the various glycoalkaloids were around α-solamargine at doses of 15, 25 and 35 mg/kg BW did 30 to 60 mg/kg BW (Friedman and McDonald, 1997).
not affect the number of spermatozoa or the weight of Although the cause of death in the high dose recipient the testicle and epididymis (Table 2). Examination of animals could be attributed mainly to acetylcholines- the testes at doses 45 and 50 mg /kg BW, respectively, terase (AchE) inhibition and cell membrane disruption, indicated subcapsular congestion possibly produced by these effects do not appear to be pronounced in the hyperaemia. Similarly, epididymis at the same doses case of α-solamargine. In effect, Roddick (1989) reported slight AchE inhibition by this glycoalkaloid,which confirms the importance of the aglycone moietyin the AchE activity. The absence of bleeding in theeyes and nose, together with the histology which did DISCUSSION
not show renal and hepatic congestion, indicated a loweffect on cell membrane disruption of α-solamargine Most studies of glycoalkaloids of Solanaceae have been in doses below the LD value. In contrast, even centred on rats, mice, hamster and rabbits (Friedman though the number of spermatozoa were not affected and McDonald, 1997). The toxicological properties at the doses administered congestion of the epididymis of α-solamargine, a glycoalkaloid with biological activity and testicle tissues was observed at doses of 45 and against Herpes simplex I, Herpes zoster and genital Her- pes (Chataing et al., 1996) and Trypanosoma cruzi(Chataing et al., 1998) has been tested. This glycosidehas a toxicity range in the order of other Solanaceae Acknowledgements
glycoalkaloids considering its LD value which is similar to those obtained for solanine (Nishie et al., The present work has been supported by funds from CDCHT-ULA 1975) and BEC mixtures in rats (Cham et al., 1991). A REFERENCES
Aldous CN, Sharma RP, Salunkhe DK. 1980. Effects of α-chaco- Cham BE, Meares HM. 1987. Glycoalkaloids from Solanum nine on brain biogenic amines, electroencephalogram, sodomaeum are effective in the treatment of skin cancer in cardiac rates and respiratory response in rats. J Food man. Cancer Lett 36: 111–118.
Safety 2: 20–33.
Chataing B, Buitrago de CN, Concepcion JL, Usubillaga A.
Cham BE, Daunter B, Evans RA. 1991. Topical treatment of 1996. Estudio clínico de la efectividad de los extractos malignant and premalignant skin lesions by very low alcaloideos obtenidos de los frutos de Solanum americanum concentrations of a standard mixture (BEC) of solasodine Miller sobre Herpes simple, Herpes zoster y Herpes genital.
glycosides. Cancer Lett 59: 183 –192.
Rev Facult Farm ULA 32: 15 –28.
Cham BE, Gilliver M, Wilson L. 1987. Antitumor effects of gly- Chataing B, Concepcion JL, Lobaton R, Usubillaga A. 1998.
coalkaloids isolated from Solanum sodomaeum. Planta Inhibition of Trypanosoma cruzi growth by Solanum Med 1: 34 –36.
alkaloids. A comparison with ketoconazole. Planta Med 64:
31–36.
Copyright 2003 John Wiley & Sons, Ltd.
Phytother. Res. 17, 254 – 258 (2003)
TOXICOLOGICAL EFFECTS OF X-SOLAUCARGINE IN EXPERIMENTAL ANIMALS Chaube SJ, Swinyard CA. 1976. Teratological and toxicol- num alkaloids as inhibitors of enzymatic conversion of dihy- ogical studies of alkaloidal and phenolic compounds from drolanosterol in cholesterol. Chem Pharm Bull 35: 4321–4323.
Solanum tuberosum L. Toxicol Appl Pharmacol 36: 227–
Mahato SB, Sahu NP, Ganguly AN, Kasai R, Tanak O. 1980.
Steroidal alkaloids from Solanum khasinum. Application Fewell AM, Roddick JG, Weissenberg M. 1994. Interactions of 13C-NMR spectroscopy to their structural elucidation.
between the glycoalkaloids solanine and solamargine in Phytochemistry 19: 2017–2010.
relation to inhibition of fungal growth. Phytochemistry 37:
Nishie K, Norred WP, Swain AP. 1975. Pharmacology and toxi- cology of chaconine and tomatine. Res Commun Chem Friedman M, McDonald G. 1997. Potato glycoalkaloids: chem- Pathol Pharmacol 13: 161–167.
istry, analysis, safety, and plant physiology. Crit Rev Plant Roddick JG. 1989. The acetylcholinesterase-inhibitory activity Sci 16: 55–132.
of steroidal glycoalkaloids and their aglycones. Phyto- Kusano G, Takahashi S, Nazoe Y, Sonoda K, Sato Y. 1987. Sola- chemistry 28: 2631–2634.
Copyright 2003 John Wiley & Sons, Ltd.
Phytother. Res. 17, 254 – 258 (2003)

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