Toxicity of crude rhizome extract of Kaempferia galanga L. (Proh Hom)
Introduction
Kaempferia galanga (“Proh hom” in Thai; Zingiberaceae) is an acaulescent perennial that grows in Southern China, Indochina, Malaysia and India. The rhizomes of the plant, which contains essential oils, have been used in a decoction or powder for indigestion, cold, pectoral and abdominal pains, headache and toothache. Its alcoholic maceration has also been applied as liniment for rheumatism (Keys, 1976, Lieu, 1990). In Chinese medicine, Kaempferia galanga rhizomes have been used as an aromatic stomachic, and also as incense. The constituents of this rhizome, hitherto reported, have included cineol, borneol, 3-carene, camphene, kaempferol, kaempferide, cinnamaldehyde, p-methoxycinnaamic acid, ethyl cinnamate, and ethyl p-methoxycinnamate (Nakao and Shibu, 1924). Ethyl p-methoxycinnamate was reported to inhibit monoamine oxidase (Noro et al., 1983). The methanolic extract of Kaempferia galanga, which identified as ethyl cinnamate, ethyl p-methoxycinnamate and p-methoxycinnamic acid, showed larvicidal activity against the second stage larva of dog roundworm, Toxocara canis (Kiuchi et al., 1988). Evaluation for amebicidal activity in vitro against three species of Acanthamoeba: Acanthamoeba culbertsoni, Acanthamoeba castellanii, and Acanthamoeba polyphaga; the causative agents of granulomatous amebic encephalitis and amebic keratitis, found that the Kaempferia galanga extract possessed an effective amebicidal for all three species (Chu et al., 1998). Vimala et al. (1999) found that the rhizome extract of Kaempferia galanga exhibited Epstein-Barr virus (EBV) activiation inhibitory activity when screened for anti-tumour promoter activity using the short-term assay of inhibition of 12-O-tetradecanoyl phorbol-13-acetate (TPA)-induced EBV early antigen in Raji cells.
Recently, Pitasawat et al. (1998) demonstrated significant larvicidal activity against Culex quinquefasciatus in three of ten plant extracts, including those from Kaempferia galanga, Illicium vernum and Spilanthes acmella, which had LC50values of 50.54, 54.11 and 61.43 ppm, respectively. Subsequently, larvicidal and repellent activities of Kaempferia galanga fractions, i.e. the hexane fraction, dichloromethane fraction 1, dichloromethane fraction 2, and methanolic fraction have been affirmed by Choochote et al. (1999). They declared that the hexane fraction possessed larvicidal potency against Culex quinquefasciatus (LC50=42.33 ppm), repelled Aedes aegypti (ED50=30.73 ug/cm2), and provided biting protection for 3 h in the laboratory. It could also protect in the field against Armigeres suballbatus, Anopheles barbirostris, Anopheles aconitus, Mansonia uniformis, Culex quinquefascitus, Culex gelidus, Culex tritaeniorhynchus and Aedes aegypti. Additionally, it did not cause dermal irritation when applied on human skin. Before applying the effective formula of Kaempferia galanga for mosquito control, it is important to evaluate the toxic effects of the extract, which is essential when a safe dose has to be selected. Therefore, the present study was initiated for analysing the acute and subacute toxicites and dermal irritation of the extract in laboratory animals.
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Laboratory animals
Adult Sprague–Dawley rats of either sex, aged 6–8 weeks with a weight of 250–300 g were purchased from the National Laboratory Animal Center, Salaya Mahidol University, Nakorn Pathom, Thailand. Adult albino rabbits of either sex, weighing between 4 and 5 kg were obtained from the Animal Unit of the Faculty of Medicine, Chiang Mai University, Thailand. The animals were kept in an animal room where the temperature was maintained at 22±3 °C under a 12 h light–dark cycle. They were provided with food
Hippocratic screening test
The general behavioral changes of the rats were observed following intraperitoneal injections of the ethanolic extract of Kaempferia galanga at 25, 100, 250, 800 and 2000 mg/kg doses, which were graded through time. Doses of 25, 100 and 250 mg/kg did not cause any detectable changes, whereas, a dose of 2000 mg/kg seemed to be lethal and caused two out of four rat deaths within 24 h. Signs and symptoms, which occurred in response to Kaempferia galanga extract, decreased in motor activity and
Discussion
In toxicity studies, including the Hippocratic screening test, acute, subacute and dermal toxicities were elucidated in small laboratory animals. Ethanolic extract of Kaempferia galanga was used in most of these tests with the exception of the dermal irritation test, since this form was convenient to prepare, could be easily applied, and its larvicidal potency (LD50=50.54 ppm) was comparable to that of the hexane fraction (LD50=42.33 ppm) (Choochote et al., 1999). In the dermal irritation test,
Acknowledgements
The authors are thankful to Dean of Faculty of Medicine for providing necessary research facilities. Acknowledgment is extended to the Chulabhorn Research Institute for financial support, and the Faculty of Medicine Endowment Fund for Research Publication for helping to defray the publication cost.
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