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Research Paper
Toxicological screening of Daouri, a polyherbal formulation used in children in the Central Region of Togo Mawutodji S. Edorh a, Sadikou Agbere a, Dorcas Osei-Safo b, Zakilatou Adam c, Amegnona Agbonon a,n, Damintoti S. Karou a, Rahamane A. Agbere c,d, Messanvi Gbeassor a a
Centre de Recherche et de Formation sur les Plantes Médicinales (CERFOPLAM), Université de Lomé, Togo Department of Chemistry, University of Ghana, Legon, Accra.P. O. Box LG56, Accra, Ghana c Faulté des Sciences de la Santé, Université de Lomé, Togo d Service de Pédiatrie du CHU Sylvanus Olympio, Lomé, Togo b
art ic l e i nf o
a b s t r a c t
Article history: Received 13 October 2014 Received in revised form 12 December 2014 Accepted 14 January 2015
Ethnopharmacological relevance: Daouri, a combination of several plants, is an old African Traditional Medicine based on ancestral knowledge transmitted from generation to generation and is used by the Kotokoli Community in Togo. The combination of several plants may potentiate or attenuate the toxicity of individual plants. The present investigation aims to study the composition and potential toxicity of Daouri used in children in the Kotokoli community. Materials and methods: Surveys were performed using a semi-structured questionnaire to determine the composition of Daouri. On the basis of these data collected, Standard Daouri was formulated, and its aqueous extract was orally administered at 300, 600 and 1200 mg/kg to rats for 28 days. On the 29th day, the rats were sacrificed and their serum were analysed to evaluate hepatic and renal toxicities. Results: Four categories of Daouri were collected. The plant combinations used in each Daouri formulation varied according to the pathological conditions, including the age of the children and the availability of each plant. The most plants cited in the four Daouri were Khaya senegalensis (Desv.) A. Juss, Odina acida (A. Rich.) Oliv.,Lophira lanceolata Tiegh, Paullinia pinnata L. and Pteleopsis suberosa Engl. & Diels. Although there was an increase in the alkaline phosphatase concentration, different doses of the aqueous extract of Standard Daouri were not toxic after 28 days of administration. In addition, the concentrations of alanine transaminase, creatinine and urea were not different between the Daouritreated and control groups. Conclusion: Daouri is plant combination used in children in the Kotokoli community as a part of African Traditional Medicine. Standard Daouri is not toxic in rats. & 2015 Published by Elsevier Ireland Ltd.
Keywords: Polyherbal Traditional medicine Toxicity Children health
1. Introduction Polyherbal formulations based on the combination of many organs of medicinal plants are used in many traditional communities worldwide (Bhushan and Raghunath, 2009) and represent the basis of ethnopharmacology practice in Africa. They are used in all aspects to treat many pathological conditions, such as malaria and diarrhoea, which particularly affects children. Treatment of malaria may involve the use of Cassia siamea and Flueggea virosa in Togo or Emblica officinalis, Terminalia bellerica and Terminalia chebula (Ponnusankar et al., 2011) in India. Available toxicological data on these plant combinations are often empirical. Although n Correspondence to: Laboratoire de Physiologie/Pharmacologie du CERFOPLAM, Faculté des Sciences, Université de Lomé, B.P. 1515 Lomé, Togo. Tel.: þ 228 22 36 75 82. E-mail address: [email protected] (A. Agbonon).
multiple ingredients in plant combinations may act synergistically, some ingredients may modulate the potential toxicity of other ingredients (Agbonon et al., 2010). It is also possible that some ingredients may potentiate the side effects of other ingredients in the context of polyherbal medicines. It has been well established that more than 80% of the population, including children living in developing countries, rely on alternative medicine (Bennett and Brown, 2000). Although traditional medicine is regarded as an alternative medicine, the broad use of this medicine through various periods has led the World Health Organisation (WHO) to recommend toxicological and pharmacological research studies on plants to evaluate patient safety. In African Traditional Medicine, remedies based on herbals are formulated via three different categories of knowledge, namely, popular knowledge, medicinal plants sold in the markets based on the herbalists' knowledge, and traditional healers' knowledge, which is kept secretly and is transmitted from generation to
http://dx.doi.org/10.1016/j.jep.2015.01.045 0378-8741/& 2015 Published by Elsevier Ireland Ltd.
Please cite this article as: Edorh, M.S., et al., Toxicological screening of Daouri, a polyherbal formulation used in children in the Central Region of Togo. Journal of Ethnopharmacology (2015), http://dx.doi.org/10.1016/j.jep.2015.01.045i
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generation within families. In the context of popular knowledge, any person can formulate a medicine based on a concoction of self-collected plant parts without any specific guidelines or verification. This practice is common in rural areas and in the peripheries of African cities. The second category concerns people living in towns who do not usually have the opportunity to collect these plant parts themselves and must therefore buy them from markets on the advice of herbalists. The last category refers to patients with specific diseases who look for traditional healers for specific treatments. In the Central Region of Togo, one of the popular polyherbal medicines used in children is called Daouri or Diuri. It can be administered to children from six months to ten years old for many purposes related to diarrhoea, gastrointestinal disorders, anaemia, fever, intestinal worms, and malaria, and it can also improve nutritional conditions. Regardless of this early administration of medicinal plant products to children, toxicological information on the individual plants or on their combination is frequently unavailable. However, it has been well established that plants can contain toxic substances (Philippe et al., 2004) and that folk remedies may cause renal and hepatic damage (Luyckx et al., 2005), particularly in children with vulnerable health. Due to the potential risk that herbal medicines pose to children, we hypothesised that Daouri may have a potential toxic effect in children and may affect their physiological parameters in the long-term when it is administered for longer periods. Another concern about Daouri is its variability according to both pathological conditions and the availability of plant parts, which makes it difficult to determine the composition of this medicinal formulation. The present study aims to investigate the composition and potential toxicity of Daouri used in Sokodé and Tchamba in Wistar rats. These towns are located in the Central Region of Togo and are included in the five Districts (Est-Mono, Blitta, Sotouboua, Tchaoudjo and Tchamba) involved in the National Programme known as PCIME (Prise en charge Intégrée des Maladies de l'Enfant).
2.4. Extract preparation Based on the analysis of different Daouri formulations, the plants that were frequently cited were selected to generate Standard Daouri. These plants are Khaya senegalensis (Desv.) A. Juss, Odina acida (A. Rich.) Oliv, Lophira lanceolata Tiegh, Paullinia pinnata L. and Pteleopsis suberosa Engl. & Diels. Different parts of the plants were washed and air-dried and the samples were lyophilised. The powders were mixed in equal proportions to obtain a total weight of 200 g and were subsequently extracted with 2 l of distilled water at 80 1C for 30 mins.The aqueous extract was cooled, filtered and then evaporated in a vacuum using a rotary evaporator (Buchi, R-210) to obtain a dry aqueous extract, which was stored at 4 1C until further use. The yield of the water-soluble extract was 9.78%. 2.5. Toxicological evaluation Twenty Wistar rats were used in this experiment. The animals were placed into four groups containing five animals per group, and the extract was administered daily for 28 days: group 1 (control group), where the rats were treated with vehicle and groups 2, 3 and 4, where the animals were treated with aqueous extract of Standard Daouri at 300, 600 and 1200 mg/kg, respectively. The animals were weighed daily, and on the 29th day, the rats were anaesthetized with ether; then, blood was collected from the retro-ocular artery and centrifuged at 3000 rpm for 15 mins. Serum was collected and stored in a freezer ( 40 1C) until further determination of the biochemical parameters, including alanine transaminase, alkaline phosphatase, creatinine, urea and total protein with the appropriate kit. Moreover, the rats were killed by cervical dislocation under anaesthesia and the organs, including the liver, kidney and heart, were weighed, and the relative weight for each organ was calculated. Histological observation was performed on the liver to detect abnormalities in the animals treated with Daouri compared to the control group.
2. Materials and methods
2.6. Statistical analysis
2.1. Chemicals
Values are expressed as the mean 7 standard deviation (SD) of five observations in each group. One-way analysis of variance (ANOVA) was performed to determine the significant difference between parameters. The Fischer Least Significant Difference (LSD) test was used to determine differences between groups at p o0.05 using SYSTAT 12.0.
Kits for alanine transaminase, alkaline phosphatase, creatinine, urea, and total protein were purchased from Humans (Germany). 2.2. Animals Male and female Wistar rats weighing between 100 and 130 g were obtained from the Department of Animal Physiology (University of Lome). The animals were housed in cages (5 rats per cage) at ambient temperature and humidity with a 12-hour day-night cycle and were provided with free access to food and water. Experimental protocols were performed based on the World Health Organisation Guidelines for the care and use of laboratory animals and according to the Ethics Committee of University of Lome, a branch of the National Ethics Committee. 2.3. Collection and identification of plant materials Using direct semi-structured questionnaires, plant parts used in a Daouri formulation were surveyed and collected from markets in Sokodé and Tchamba in the Central Region of Togo (Fig. 1).The plant names were identified in the local “Kotokoli” dialect and their scientific names were identified at the Laboratory of Botany and Vegetable Ecology (Faculty of Sciences–University of Lome). After identification, a voucher specimen number of each plant was recorded at the herbarium.
3. Results 3.1. Data collected Data collected from different markets at Sokodé and Tchamba indicated that four categories (two categories from each town) of Daouri were recommended by herbalists to patients according to the disease and conditions. The two categories of Daouri from Sokodé are indicated in Tables 1 and 2, while those obtained from Tchamba are presented in Tables 3 and 4. Daouri 1 (Table 1) contained 11 different plants, and the formulation was based on herbalists' guidelines. The proportion of each plant in the Daouri formulation was the same, with the exception of two species, Piper guineense and Xylopia aethiopica. However, depending on the severity of the pathological conditions affecting the children, the proportions may vary. In the Kotokoli community, Daouri 1 is preferably used to relieve malaria symptoms. As shown in Table 2, Daouri 2 contained six plants and was mainly used to treat gastro-intestinal affections, such as diarrhoea.
Please cite this article as: Edorh, M.S., et al., Toxicological screening of Daouri, a polyherbal formulation used in children in the Central Region of Togo. Journal of Ethnopharmacology (2015), http://dx.doi.org/10.1016/j.jep.2015.01.045i
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Fig. 1. Map showing the area called Kotokoli land in the Central Region of Togo.
Table 1 Daouri 1 collected from Sokodé. Scientific names (Herbarium number)
Families
Local names
Part used
Acacia abyssinica Benth (Tg 04844) Adenodolichos baumii Harms (Tg 05607) Eugenia aromatica. Berg (Tg 05274) Lophira lanceolata Tiegh (Tg 05370) Odinaacida (A. Rich.) Oliv. (Tg 01771) Paullinia pinnata L. (Tg 0879) Piper guineense Schumach. & Thonn (Tg 06863) Pteleopsis suberosa Engl. & Diels (Tg 00671) Saba florida (Benth.) Bullock (Tg 02125) Sterculia setigera Delile (Tg 08661) Xylopia aethiopica (Dunal) A. Rich. (Tg 01987)
Leguminosae Leguminosae Myrtaceae Ochnaceae Anacardiaceae Sapindaceae Piperaceae Combretaceae Apocynaceae Stem bark Annonaceae
Kikokopi Boussoussouboutchi Kanafrou Kparakpara Otchowoe Fatimagoro Djeyiyamou Sissinon Bolothi Malvaceae Souzi
Stem bark Leaves Fruit Stem bark Stem bark Leaves Fruit Stem bark Stem bark Digmatori Fruit
Table 2 Daouri 2 collected from Sokodé. Scientific names (Herbarium number)
Families
Local names
Part used
Crossopteryx febrifuga Benth (Tg 07200) Khaya senegalensis (Desv.) A. Juss (Tg 04674) Piper guineense Schumach. & Thonn (Tg 06863) Pteleopsis suberosa Engl. & Diels (Tg 00671) Paullinia pinnata L. (Tg 0879) Xylopia aethiopica(Dunal) A. Rich. (Tg 01987)
Rubiaceae Meliaceae Piperaceae Combretaceae Sapindaceae Annonaceae
Kouessa Frimou Djeyiyamou Sissinon Fatimagoro Souzi
Stem bark Stem bark Fruit Stem bark Leaves Fruit
Table 3 Daouri 3 collected from Tchamba. Scientific names (Herbarium number)
Families
Local names
Part used
Adenodolichos baumii Harms (Tg 05607) Anogeissus leiocarpa (DC.) Guill. & Perr.(Tg 00485) Cicca discoidea Baill. (Tg 03430) Khaya senegalensis (Desv.) A. Juss (Tg 04674) Lophira lanceolata Tiegh (Tg 05370) Odina acida (A. Rich.) Oliv.(Tg 01771) Odina alata Engl. (Tg 01784) Parinari alvimii Prance (Tg 00427) Pterocarpus erinaceus Poir (Tg 06459) Xeroderris chevalieri (Dunn) Roberty (Tg 06772)
Leguminosae Combretaceae Phyllanthaceae Meliaceae Ochnaceae Anacardiaceae Anacardiaceae Chrysobalanaceae Leguminosae Leguminosae
Bousoussouboutchi Bousse Kitipouaki Frimou Kparakpara Otchowoe Ditchetcheteri Koupotopoto Bouto Bougbe
Leaves Stem bark Stem bark Stem bark Stem bark Stem bark Stem bark Stem bark Stem bark Stem bark
Daouri 3 consisted of 10 medicinal plants and its composition was different from Daouri 1.Medicinal plants, such as Anogeissus leiocarpa (DC.) Guill. &Perr.,Odina alata Engl. (Anacardiaceae), Cicca discoidea Baill. (Phyllanthaceae), Parinari alvimii Prance (Chrysobalanaceae) and Pterocarpus erinaceus Poir (Leguminosae), were found in Daouri 3 and were absent in Daouri 1.
On the basis of the availability of the plants throughout the year; Khaya senegalensis (Desv.) A. Juss (Meliaceae), Odina acida (A. Rich.) Oliv. (Anacardiaceae) Lophira lanceolata Tiegh (Ochnaceae), Paullinia pinnata L. (Sapindaceae) and Pteleopsis suberosa Engl. & Diels (Combretaceae) remained the most species used in all four categories of Daouri. On the basis of the frequency of plant
Please cite this article as: Edorh, M.S., et al., Toxicological screening of Daouri, a polyherbal formulation used in children in the Central Region of Togo. Journal of Ethnopharmacology (2015), http://dx.doi.org/10.1016/j.jep.2015.01.045i
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Table 4 Daouri 4 collected from Tchamba. Scientific names (Herbarium number)
Families
Local names
Part used
Dichrostachys cinerea (L.) Wight & Arn. (Tg 04918) Eugenia aromatica. Berg (Tg 05274) Khaya senegalensis (Desv.) A. Juss (Tg 04674) Lophira lanceolataTiegh(Tg 05370) Monodora myristica (Gaertn.) Dunal (Tg 04786) Paullinia pinnata L. (Tg 0879) Pteleosis suberosa Engl. & Diels (Tg 00671) Tinnea barteri Gürke (Tg 04243) Uvaria chamae P.Beauv. (Tg 01955) Xylopia aethiopica (Dunal) A. Rich. (Tg 01987)
Mimosaceae Myrtaceae Meliaceae Ochnaceae Annonaceae Sapindaceae Combretaceae Lamiaceae Annonaceae Annonaceae
Tchikili Kanafrou Frimou Kparakpara Azegbebia Fatimagoro Sissinon Kegbelewiya Molomolo Souzi
Fruit Fruit Stem bark Stem bark Fruit Leaves Stem bark Leaves Leaves Fruit
Table 5 Citation of the frequency of plants in Daouri.
Table 6 Effect of Standard Daouri on the serum concentration of alanine transaminase and alkaline phosphatase on rats treated with three doses for 28 days.
Scientific names
Number of citation in Daouri
Acacia abyssinica Benth Adenodolichos baumii Harms Anogeissus leiocarpa (DC.) Guill. & Perr. Cicca discoidea Baill. Crossopteryx febrifuga Benth Dichrostachys cinerea(L.) Wight & Arn. Eugenia aromatica. Berg Khaya senegalensis (Desv.) A. Juss Lophira lanceolata Tiegh Monodora myristica (Gaertn.) Dunal Odina acida (A. Rich.) Oliv. Odina alata Engl. Parinari alvimii Prance Paullinia pinnata L. Pteleopsis suberosa Engl. & Diels Pterocarpus erinaceus Poir Saba florida (Benth.) Bullock Sterculia setigera Delile Tinnea barteri Gürke Uvaria chamae P.Beauv. Xeroderris chevalieri (Dunn) Roberty
1 2 1 1 1 1 2 3 3 1 2 1 1 3 3 1 1 1 1 1 1
Treatments
citations (Table 5) and the availability of species, Standard Daouri (SDa) contained five medicinal plants: Khaya senegalensis, Odina acida, Lophira lanceolata, Paullinia pinnata and Pteleopsis suberosa. 3.2. Toxicological evaluation The standard Daouri (SDa) analysed in the present investigation consisted of 5 medicinal plants: Khaya senegalensis, Odina acida, Lophira lanceolata, Paullinia pinnata and Pteleopsis suberosa. Administration of the SDa extract for 28 days did not significantly (p 4 0.05) affect the body weight of the rats. However, low body variation was observed compared with the control group (data not shown). The SDa did not affect the serum concentration of alanine transaminases, creatinine and urea (Tables 6 and 7), but the serum concentration of alkaline phosphatase was increased (Table 6). Histological observation did not reveal any macroscopic abnormality in the livers of the SDa-treated groups compared to the control group.
4. Discussion The objective of the present investigation was to determine the composition of Daouri, a polyherbal formulation used by the Kotokoli in the Central Region of Togo (Kotokoli land) and to investigate the toxicological profile of this medicine, which is frequently used in children to treat pathological conditions and maintain health. These results indicated that Daouri composition is
Control SDa 300 mg/kg SDa 600 mg/kg SDa 1200 mg/ kg
Biochemical parameters Alanine transaminase (ALAT) (U/ L)
Alkaline phosphatise (U/ L)
56.84 7 1.54 51.577 2.16 63.157 0.42 59.477 1.31
181.25 7 12.35 275.007 5.48 237.50 7 7.66 281.25 7 4.51
Table 7 Effect of Standard Daouri on the serum concentration of creatinine, urea and total protein on rats treated with three doses for 28 days. Treatments
Control SDa 300 mg/kg SDa 600 mg/kg SDa 1200 mg/kg
Biochemical parameters Serum proteins (g/L)
creatinine (mg/L)
Urea (g/L)
68.57 2.3 70.3 7 2.5 75.4 7 3.4 76.8 7 2.8
4.2 7 0.8 5.17 0.82 5.3 7 0.75 5.8 7 0.1
0.447 0.03 0.417 0.02 0.34 7 0.01 0.357 0.01
not fixed and varies according to the pathological conditions, plant availability and the age of the child. Daouri is the concept of globalising conditions of herbal medicine utilisation in children between 0–10 years old. Even if the same medicinal plants are found in the formulation used in adults, the name Daouri is no longer used in Kotokoli for adults. Daouri has been specifically used by children in this community for centuries and today, the practice has become more widespread across the country. Data analysis indicated that the common name of Daouri is not based on its composition but rather on the popular knowledge that is transmitted from generation to generation and is mainly used by females in the community. Females buy this traditional recipe without any prior advice from traditional healers or herbalists. As many traditional medicines, Daouri is used for many purposes, such as medicine used to provide anti-diarrhoeal, anti-malarial, anti-anaemic, anti-microbial and intestinal wound-healing effects and for the children's well-being. Previous investigations have indicated that Daouri has potential anti-microbial effects and has been shown to inhibit the growth of Candida albicans, Escherichia coli, Staphylococcus aureus, Salmnonella sp., Pseudomonas aeruginosa (unpublished data). These microbes have been shown to be differentially involved in fatal childhood diseases or children morbidity. However, for anti-diarrhoeal, anti-malarial and anti-anaemic purposes, no scientific studies have been performed. Thus, the use of Daouri in the treatment of malaria should be performed with caution
Please cite this article as: Edorh, M.S., et al., Toxicological screening of Daouri, a polyherbal formulation used in children in the Central Region of Togo. Journal of Ethnopharmacology (2015), http://dx.doi.org/10.1016/j.jep.2015.01.045i
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because further clinical studies are needed to confirm its effectiveness in providing an anti-malarial effect. Khaya senegalensis, Odina acida, Pteleospsis suberosa, Paulinia pinnata and Lophira lanceolata are the most important species cited in the four Daouri compositions analysed. Previous investigations have shown that Pteleopsis suberosa has anti-microbial, fungicide and antioxidant activities (Germanò et al., 1998); Khaya senengalensis has antipyretic (Lompo et al., 1998), antiinflammatory and anti-parasitic properties (Abreu et al., 1999). Odina acida and Lophira lanceolata have anti-microbial effects (Almagboul et al., 1988; Okine et al., 2004) and they are used traditionally as anti-malarial symptoms. Paulinia pinnata has been not been specifically studied for diseases affecting children, but as an antioxidant and vascular relaxant (Zamble et al., 2006; Jimoh et al., 2007). Several studies have shown that medicinal plant products are not completely safe, particularly in the liver as demonstrated by Teschke et al., 2013, 2014. However, the SDa tested at the three doses in the present study exhibited the following results: it did not kill any rats, and hepatic enzymes, such alanine transaminase, were not significantly increased. Although the results showed that the concentrations of alkaline phosphatase in the serum of animals treated with SDa increased compared to control rats, this increase did not necessarily result in liver damage because alkaline phosphatase was also present in other tissues, such as the intestinal epithelium and bone. Furthermore, this enzyme increased with an increase in the concentration of total protein as described in the present study. Despite the absence of macroscopic histological abnormalities, further investigations are needed for microscopic histological analysis. Some individual plant components of Daouri have been studied for their toxicological properties. A previous investigation has demonstrated that the aqueous extract of Khaya senegalensis leaves administered at 600–3000 mg/kg to rats for 28 days had no significant toxicological effect on renal and liver physiology (Nwosu et al., 2012). For many other species of Daouri medicines, such as Odina acida, Lophira lanceolata, Paullinia pinnata, Pteleopsis suberosa and Tinnea barteri, full toxicological investigations have not yet been undertaken to the best of our knowledge. Independent of the absence of toxicological data on individual plants, the results obtained from the present investigation may indicate that their combination in a single formulation, such as Daouri, does not induce significant toxic effects in rats. Moreover, Daouri treatment appears to increase the concentration of serum protein, which may be useful for children growth. This may be a result of the nutritional potential of Daouri as demonstrated by the Kotokoli community when using Daouri to improve immune and nutritional conditions of children. 5. Conclusion Daouri has been used in Kotokoli communities for several centuries, and the present study shows that it is not toxic to Wistar rats
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56 57 58 59 60 61 62 Acknowledgements 63 64 This work was financially supported by Plan Togo through Q2 65 PCIME in the five Districts in the Central Region of Togo. Professor 66 Agbere A. D. Rahamane, Chief of Pediatric Service of CHU Sylvanus 67 Olympio, is the Focal Point of PCIME. 68 69 References 70 71 72 Abreu, P.M., Martins, E.S., Kayser, O., Bindseil, K.U., Siems, K., Seemann, A., Frevert, J., 1999. antimicrobial, antitumor and antileishmania screening of medicinal 73 plants from Guinea-Bissau. Phytomedicine6, 187–195. 74 Agbonon, A., Eklu-Gadegbeku, K., Aklikokou, K., Gbeassor, M., Akpagana, K., Tam, 75 T.W., Arnason, J.T., Foster, C.B., 2010. In vitro inhibitory effect of West African medicinal and food plants on humancytochrome P450 3A subfamily. Journal of 76 Ethnopharmacology 128, 390–394. 77 Almagboul, A.Z., Bashir, A.K., Salih, A.K., Farouk, A., Khalid, S.A., 1988. Antimicrobial 78 activity of certain Sudanese plants used in folkloric medicine; screening for antibacterial activity (V). Fitoterapia 59, 57–62. 79 Bennett, J., Brown, C.M., 2000. Use of herbal remedies by patients in a health 80 maintenance organization. Journal of American Pharmacists Association 40, 81 353–358. Bhushan, P., Raghunath, A., 2009. Traditional medicine-inspired approachesto drug 82 discovery: can ayurveda show theway forward? Drug Discovery Today 14, 83 804–811. 84 Germanò, P.M., Sanogo, R., Guglielmo, M., de Pasquale, R., Crisafi, G., Bisignano, G., 85 1998. Effects of Pteleopsis suberosa extracts on experimental gastric ulcers and Helicobacter pylori growth. Journal of Ethnopharmacology 59, 167–172. 86 Jimoh, F.O., Sofidiya, M.O., Afolayan, A.J., 2007. Antioxidant properties of the 87 methanol extracts from the leaves of Paullinia pinnata. Journal of Medicinal 88 Food10, 707–711. Lompo, M., Nikiema, J.B., Guissou, I.P., Moes, A.J., Fontaine, J., 1998. The topical 89 antiinflammatory effects of chloroform extract from Khaya senegalensis stem 90 barks. Phytotherapy Research 2, 448–450. 91 Luyckx, V.A., Steenkamp, V., Stewart, M.J., 2005. Acute renal failure associated with the use oftraditional folk remedies in South Africa. Renal Failure 27, 35–43. 92 Nwosu, C.U., Hassan, S.W., Abubakar, M.G., Ebbo, A.A., 2012. Anti-diarrhoeal and 93 toxicological studies of leaf extracts of Khaya senegalensis. Journal of Pharma94 cology and Toxicology 7, 1–10. Okine, L.K.N., Nyarko, A.K., Osei-Kwabena, N., Oppong, I.V., Barnes, F., Ofosuhene, M., 95 2004. The antidiabetic activity of the herbal preparation ADD-199 in mice: a 96 comparative study with two oral hypoglycaemic drugs. Journal of Ethnopharma97 cology 97, 31–38. Philippe, G., Angenot, L., Tits, M., Frédérich, M., 2004. About the toxicity of some 98 Strychnos species and their alkaloids. Toxicology 44, 405–416. 99 Ponnusankar, S., Subrata, P., Ramesh, B., Arun, B., Pulok, K.M., 2011. Cytochrome 100 P450 inhibitory potential of Triphala—a Rasayana from Ayurveda. Journal of 101 Ethnopharmacology 133, 120–125. Teschke, R., Frenzel, C., Glass, X., Schulze, J., Eickhoff, A., 2013. Herbal hepatotoxi102 city: a critical review. British Journal of Clinical Pharmacology 75, 630–636. 103 Teschke, R., Wolff, A., Frenzel, C., Schulze, J., 2014. Review article: herbal hepato104 toxicity—an update on traditional Chinese medicine preparations. Alimentary Pharmacology & Therapeutics 40, 32–50. 105 Zamble, A., Carpentier, M., Kandoussi, A., Sahpaz, S., Petrault, O., Ouk, T., Hennuyer, 106 N., Fruchart, J.C., Staels, B., Bordet, R., Duriez, P., Bailleul, F., Martin-Nizard, F., 107 2006. Paullinia pinnata extracts rich in polyphenols promote vascular relaxation via endothelium-dependent mechanisms. Journal of Cardiovascular Pharma108 cology 47, 599–608. 109 when administered for 28 days. However, administration of this traditional medicine in children should be performed under the supervision of competent women who have mastered the concept of Daouri. Further investigations are needed to evaluate the pharmacological effects of Daouri.
Please cite this article as: Edorh, M.S., et al., Toxicological screening of Daouri, a polyherbal formulation used in children in the Central Region of Togo. Journal of Ethnopharmacology (2015), http://dx.doi.org/10.1016/j.jep.2015.01.045i
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Research Paper
Toxicological screening of Daouri, a polyherbal formulation used in children in the Central Region of Togo Mawutodji S. Edorh a, Sadikou Agbere a, Dorcas Osei-Safo b, Zakilatou Adam c, Amegnona Agbonon a,n, Damintoti S. Karou a, Rahamane A. Agbere c,d, Messanvi Gbeassor a a
Centre de Recherche et de Formation sur les Plantes Médicinales (CERFOPLAM), Université de Lomé, Togo Department of Chemistry, University of Ghana, Legon, Accra.P. O. Box LG56, Accra, Ghana c Faulté des Sciences de la Santé, Université de Lomé, Togo d Service de Pédiatrie du CHU Sylvanus Olympio, Lomé, Togo b
art ic l e i nf o
a b s t r a c t
Article history: Received 13 October 2014 Received in revised form 12 December 2014 Accepted 14 January 2015
Ethnopharmacological relevance: Daouri, a combination of several plants, is an old African Traditional Medicine based on ancestral knowledge transmitted from generation to generation and is used by the Kotokoli Community in Togo. The combination of several plants may potentiate or attenuate the toxicity of individual plants. The present investigation aims to study the composition and potential toxicity of Daouri used in children in the Kotokoli community. Materials and methods: Surveys were performed using a semi-structured questionnaire to determine the composition of Daouri. On the basis of these data collected, Standard Daouri was formulated, and its aqueous extract was orally administered at 300, 600 and 1200 mg/kg to rats for 28 days. On the 29th day, the rats were sacrificed and their serum were analysed to evaluate hepatic and renal toxicities. Results: Four categories of Daouri were collected. The plant combinations used in each Daouri formulation varied according to the pathological conditions, including the age of the children and the availability of each plant. The most plants cited in the four Daouri were Khaya senegalensis (Desv.) A. Juss, Odina acida (A. Rich.) Oliv.,Lophira lanceolata Tiegh, Paullinia pinnata L. and Pteleopsis suberosa Engl. & Diels. Although there was an increase in the alkaline phosphatase concentration, different doses of the aqueous extract of Standard Daouri were not toxic after 28 days of administration. In addition, the concentrations of alanine transaminase, creatinine and urea were not different between the Daouritreated and control groups. Conclusion: Daouri is plant combination used in children in the Kotokoli community as a part of African Traditional Medicine. Standard Daouri is not toxic in rats. & 2015 Published by Elsevier Ireland Ltd.
Keywords: Polyherbal Traditional medicine Toxicity Children health
1. Introduction Polyherbal formulations based on the combination of many organs of medicinal plants are used in many traditional communities worldwide (Bhushan and Raghunath, 2009) and represent the basis of ethnopharmacology practice in Africa. They are used in all aspects to treat many pathological conditions, such as malaria and diarrhoea, which particularly affects children. Treatment of malaria may involve the use of Cassia siamea and Flueggea virosa in Togo or Emblica officinalis, Terminalia bellerica and Terminalia chebula (Ponnusankar et al., 2011) in India. Available toxicological data on these plant combinations are often empirical. Although n Correspondence to: Laboratoire de Physiologie/Pharmacologie du CERFOPLAM, Faculté des Sciences, Université de Lomé, B.P. 1515 Lomé, Togo. Tel.: þ 228 22 36 75 82. E-mail address: [email protected] (A. Agbonon).
multiple ingredients in plant combinations may act synergistically, some ingredients may modulate the potential toxicity of other ingredients (Agbonon et al., 2010). It is also possible that some ingredients may potentiate the side effects of other ingredients in the context of polyherbal medicines. It has been well established that more than 80% of the population, including children living in developing countries, rely on alternative medicine (Bennett and Brown, 2000). Although traditional medicine is regarded as an alternative medicine, the broad use of this medicine through various periods has led the World Health Organisation (WHO) to recommend toxicological and pharmacological research studies on plants to evaluate patient safety. In African Traditional Medicine, remedies based on herbals are formulated via three different categories of knowledge, namely, popular knowledge, medicinal plants sold in the markets based on the herbalists' knowledge, and traditional healers' knowledge, which is kept secretly and is transmitted from generation to
http://dx.doi.org/10.1016/j.jep.2015.01.045 0378-8741/& 2015 Published by Elsevier Ireland Ltd.
Please cite this article as: Edorh, M.S., et al., Toxicological screening of Daouri, a polyherbal formulation used in children in the Central Region of Togo. Journal of Ethnopharmacology (2015), http://dx.doi.org/10.1016/j.jep.2015.01.045i
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generation within families. In the context of popular knowledge, any person can formulate a medicine based on a concoction of self-collected plant parts without any specific guidelines or verification. This practice is common in rural areas and in the peripheries of African cities. The second category concerns people living in towns who do not usually have the opportunity to collect these plant parts themselves and must therefore buy them from markets on the advice of herbalists. The last category refers to patients with specific diseases who look for traditional healers for specific treatments. In the Central Region of Togo, one of the popular polyherbal medicines used in children is called Daouri or Diuri. It can be administered to children from six months to ten years old for many purposes related to diarrhoea, gastrointestinal disorders, anaemia, fever, intestinal worms, and malaria, and it can also improve nutritional conditions. Regardless of this early administration of medicinal plant products to children, toxicological information on the individual plants or on their combination is frequently unavailable. However, it has been well established that plants can contain toxic substances (Philippe et al., 2004) and that folk remedies may cause renal and hepatic damage (Luyckx et al., 2005), particularly in children with vulnerable health. Due to the potential risk that herbal medicines pose to children, we hypothesised that Daouri may have a potential toxic effect in children and may affect their physiological parameters in the long-term when it is administered for longer periods. Another concern about Daouri is its variability according to both pathological conditions and the availability of plant parts, which makes it difficult to determine the composition of this medicinal formulation. The present study aims to investigate the composition and potential toxicity of Daouri used in Sokodé and Tchamba in Wistar rats. These towns are located in the Central Region of Togo and are included in the five Districts (Est-Mono, Blitta, Sotouboua, Tchaoudjo and Tchamba) involved in the National Programme known as PCIME (Prise en charge Intégrée des Maladies de l'Enfant).
2.4. Extract preparation Based on the analysis of different Daouri formulations, the plants that were frequently cited were selected to generate Standard Daouri. These plants are Khaya senegalensis (Desv.) A. Juss, Odina acida (A. Rich.) Oliv, Lophira lanceolata Tiegh, Paullinia pinnata L. and Pteleopsis suberosa Engl. & Diels. Different parts of the plants were washed and air-dried and the samples were lyophilised. The powders were mixed in equal proportions to obtain a total weight of 200 g and were subsequently extracted with 2 l of distilled water at 80 1C for 30 mins.The aqueous extract was cooled, filtered and then evaporated in a vacuum using a rotary evaporator (Buchi, R-210) to obtain a dry aqueous extract, which was stored at 4 1C until further use. The yield of the water-soluble extract was 9.78%. 2.5. Toxicological evaluation Twenty Wistar rats were used in this experiment. The animals were placed into four groups containing five animals per group, and the extract was administered daily for 28 days: group 1 (control group), where the rats were treated with vehicle and groups 2, 3 and 4, where the animals were treated with aqueous extract of Standard Daouri at 300, 600 and 1200 mg/kg, respectively. The animals were weighed daily, and on the 29th day, the rats were anaesthetized with ether; then, blood was collected from the retro-ocular artery and centrifuged at 3000 rpm for 15 mins. Serum was collected and stored in a freezer ( 40 1C) until further determination of the biochemical parameters, including alanine transaminase, alkaline phosphatase, creatinine, urea and total protein with the appropriate kit. Moreover, the rats were killed by cervical dislocation under anaesthesia and the organs, including the liver, kidney and heart, were weighed, and the relative weight for each organ was calculated. Histological observation was performed on the liver to detect abnormalities in the animals treated with Daouri compared to the control group.
2. Materials and methods
2.6. Statistical analysis
2.1. Chemicals
Values are expressed as the mean 7 standard deviation (SD) of five observations in each group. One-way analysis of variance (ANOVA) was performed to determine the significant difference between parameters. The Fischer Least Significant Difference (LSD) test was used to determine differences between groups at p o0.05 using SYSTAT 12.0.
Kits for alanine transaminase, alkaline phosphatase, creatinine, urea, and total protein were purchased from Humans (Germany). 2.2. Animals Male and female Wistar rats weighing between 100 and 130 g were obtained from the Department of Animal Physiology (University of Lome). The animals were housed in cages (5 rats per cage) at ambient temperature and humidity with a 12-hour day-night cycle and were provided with free access to food and water. Experimental protocols were performed based on the World Health Organisation Guidelines for the care and use of laboratory animals and according to the Ethics Committee of University of Lome, a branch of the National Ethics Committee. 2.3. Collection and identification of plant materials Using direct semi-structured questionnaires, plant parts used in a Daouri formulation were surveyed and collected from markets in Sokodé and Tchamba in the Central Region of Togo (Fig. 1).The plant names were identified in the local “Kotokoli” dialect and their scientific names were identified at the Laboratory of Botany and Vegetable Ecology (Faculty of Sciences–University of Lome). After identification, a voucher specimen number of each plant was recorded at the herbarium.
3. Results 3.1. Data collected Data collected from different markets at Sokodé and Tchamba indicated that four categories (two categories from each town) of Daouri were recommended by herbalists to patients according to the disease and conditions. The two categories of Daouri from Sokodé are indicated in Tables 1 and 2, while those obtained from Tchamba are presented in Tables 3 and 4. Daouri 1 (Table 1) contained 11 different plants, and the formulation was based on herbalists' guidelines. The proportion of each plant in the Daouri formulation was the same, with the exception of two species, Piper guineense and Xylopia aethiopica. However, depending on the severity of the pathological conditions affecting the children, the proportions may vary. In the Kotokoli community, Daouri 1 is preferably used to relieve malaria symptoms. As shown in Table 2, Daouri 2 contained six plants and was mainly used to treat gastro-intestinal affections, such as diarrhoea.
Please cite this article as: Edorh, M.S., et al., Toxicological screening of Daouri, a polyherbal formulation used in children in the Central Region of Togo. Journal of Ethnopharmacology (2015), http://dx.doi.org/10.1016/j.jep.2015.01.045i
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Fig. 1. Map showing the area called Kotokoli land in the Central Region of Togo.
Table 1 Daouri 1 collected from Sokodé. Scientific names (Herbarium number)
Families
Local names
Part used
Acacia abyssinica Benth (Tg 04844) Adenodolichos baumii Harms (Tg 05607) Eugenia aromatica. Berg (Tg 05274) Lophira lanceolata Tiegh (Tg 05370) Odinaacida (A. Rich.) Oliv. (Tg 01771) Paullinia pinnata L. (Tg 0879) Piper guineense Schumach. & Thonn (Tg 06863) Pteleopsis suberosa Engl. & Diels (Tg 00671) Saba florida (Benth.) Bullock (Tg 02125) Sterculia setigera Delile (Tg 08661) Xylopia aethiopica (Dunal) A. Rich. (Tg 01987)
Leguminosae Leguminosae Myrtaceae Ochnaceae Anacardiaceae Sapindaceae Piperaceae Combretaceae Apocynaceae Stem bark Annonaceae
Kikokopi Boussoussouboutchi Kanafrou Kparakpara Otchowoe Fatimagoro Djeyiyamou Sissinon Bolothi Malvaceae Souzi
Stem bark Leaves Fruit Stem bark Stem bark Leaves Fruit Stem bark Stem bark Digmatori Fruit
Table 2 Daouri 2 collected from Sokodé. Scientific names (Herbarium number)
Families
Local names
Part used
Crossopteryx febrifuga Benth (Tg 07200) Khaya senegalensis (Desv.) A. Juss (Tg 04674) Piper guineense Schumach. & Thonn (Tg 06863) Pteleopsis suberosa Engl. & Diels (Tg 00671) Paullinia pinnata L. (Tg 0879) Xylopia aethiopica(Dunal) A. Rich. (Tg 01987)
Rubiaceae Meliaceae Piperaceae Combretaceae Sapindaceae Annonaceae
Kouessa Frimou Djeyiyamou Sissinon Fatimagoro Souzi
Stem bark Stem bark Fruit Stem bark Leaves Fruit
Table 3 Daouri 3 collected from Tchamba. Scientific names (Herbarium number)
Families
Local names
Part used
Adenodolichos baumii Harms (Tg 05607) Anogeissus leiocarpa (DC.) Guill. & Perr.(Tg 00485) Cicca discoidea Baill. (Tg 03430) Khaya senegalensis (Desv.) A. Juss (Tg 04674) Lophira lanceolata Tiegh (Tg 05370) Odina acida (A. Rich.) Oliv.(Tg 01771) Odina alata Engl. (Tg 01784) Parinari alvimii Prance (Tg 00427) Pterocarpus erinaceus Poir (Tg 06459) Xeroderris chevalieri (Dunn) Roberty (Tg 06772)
Leguminosae Combretaceae Phyllanthaceae Meliaceae Ochnaceae Anacardiaceae Anacardiaceae Chrysobalanaceae Leguminosae Leguminosae
Bousoussouboutchi Bousse Kitipouaki Frimou Kparakpara Otchowoe Ditchetcheteri Koupotopoto Bouto Bougbe
Leaves Stem bark Stem bark Stem bark Stem bark Stem bark Stem bark Stem bark Stem bark Stem bark
Daouri 3 consisted of 10 medicinal plants and its composition was different from Daouri 1.Medicinal plants, such as Anogeissus leiocarpa (DC.) Guill. &Perr.,Odina alata Engl. (Anacardiaceae), Cicca discoidea Baill. (Phyllanthaceae), Parinari alvimii Prance (Chrysobalanaceae) and Pterocarpus erinaceus Poir (Leguminosae), were found in Daouri 3 and were absent in Daouri 1.
On the basis of the availability of the plants throughout the year; Khaya senegalensis (Desv.) A. Juss (Meliaceae), Odina acida (A. Rich.) Oliv. (Anacardiaceae) Lophira lanceolata Tiegh (Ochnaceae), Paullinia pinnata L. (Sapindaceae) and Pteleopsis suberosa Engl. & Diels (Combretaceae) remained the most species used in all four categories of Daouri. On the basis of the frequency of plant
Please cite this article as: Edorh, M.S., et al., Toxicological screening of Daouri, a polyherbal formulation used in children in the Central Region of Togo. Journal of Ethnopharmacology (2015), http://dx.doi.org/10.1016/j.jep.2015.01.045i
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Table 4 Daouri 4 collected from Tchamba. Scientific names (Herbarium number)
Families
Local names
Part used
Dichrostachys cinerea (L.) Wight & Arn. (Tg 04918) Eugenia aromatica. Berg (Tg 05274) Khaya senegalensis (Desv.) A. Juss (Tg 04674) Lophira lanceolataTiegh(Tg 05370) Monodora myristica (Gaertn.) Dunal (Tg 04786) Paullinia pinnata L. (Tg 0879) Pteleosis suberosa Engl. & Diels (Tg 00671) Tinnea barteri Gürke (Tg 04243) Uvaria chamae P.Beauv. (Tg 01955) Xylopia aethiopica (Dunal) A. Rich. (Tg 01987)
Mimosaceae Myrtaceae Meliaceae Ochnaceae Annonaceae Sapindaceae Combretaceae Lamiaceae Annonaceae Annonaceae
Tchikili Kanafrou Frimou Kparakpara Azegbebia Fatimagoro Sissinon Kegbelewiya Molomolo Souzi
Fruit Fruit Stem bark Stem bark Fruit Leaves Stem bark Leaves Leaves Fruit
Table 5 Citation of the frequency of plants in Daouri.
Table 6 Effect of Standard Daouri on the serum concentration of alanine transaminase and alkaline phosphatase on rats treated with three doses for 28 days.
Scientific names
Number of citation in Daouri
Acacia abyssinica Benth Adenodolichos baumii Harms Anogeissus leiocarpa (DC.) Guill. & Perr. Cicca discoidea Baill. Crossopteryx febrifuga Benth Dichrostachys cinerea(L.) Wight & Arn. Eugenia aromatica. Berg Khaya senegalensis (Desv.) A. Juss Lophira lanceolata Tiegh Monodora myristica (Gaertn.) Dunal Odina acida (A. Rich.) Oliv. Odina alata Engl. Parinari alvimii Prance Paullinia pinnata L. Pteleopsis suberosa Engl. & Diels Pterocarpus erinaceus Poir Saba florida (Benth.) Bullock Sterculia setigera Delile Tinnea barteri Gürke Uvaria chamae P.Beauv. Xeroderris chevalieri (Dunn) Roberty
1 2 1 1 1 1 2 3 3 1 2 1 1 3 3 1 1 1 1 1 1
Treatments
citations (Table 5) and the availability of species, Standard Daouri (SDa) contained five medicinal plants: Khaya senegalensis, Odina acida, Lophira lanceolata, Paullinia pinnata and Pteleopsis suberosa. 3.2. Toxicological evaluation The standard Daouri (SDa) analysed in the present investigation consisted of 5 medicinal plants: Khaya senegalensis, Odina acida, Lophira lanceolata, Paullinia pinnata and Pteleopsis suberosa. Administration of the SDa extract for 28 days did not significantly (p 4 0.05) affect the body weight of the rats. However, low body variation was observed compared with the control group (data not shown). The SDa did not affect the serum concentration of alanine transaminases, creatinine and urea (Tables 6 and 7), but the serum concentration of alkaline phosphatase was increased (Table 6). Histological observation did not reveal any macroscopic abnormality in the livers of the SDa-treated groups compared to the control group.
4. Discussion The objective of the present investigation was to determine the composition of Daouri, a polyherbal formulation used by the Kotokoli in the Central Region of Togo (Kotokoli land) and to investigate the toxicological profile of this medicine, which is frequently used in children to treat pathological conditions and maintain health. These results indicated that Daouri composition is
Control SDa 300 mg/kg SDa 600 mg/kg SDa 1200 mg/ kg
Biochemical parameters Alanine transaminase (ALAT) (U/ L)
Alkaline phosphatise (U/ L)
56.84 7 1.54 51.577 2.16 63.157 0.42 59.477 1.31
181.25 7 12.35 275.007 5.48 237.50 7 7.66 281.25 7 4.51
Table 7 Effect of Standard Daouri on the serum concentration of creatinine, urea and total protein on rats treated with three doses for 28 days. Treatments
Control SDa 300 mg/kg SDa 600 mg/kg SDa 1200 mg/kg
Biochemical parameters Serum proteins (g/L)
creatinine (mg/L)
Urea (g/L)
68.57 2.3 70.3 7 2.5 75.4 7 3.4 76.8 7 2.8
4.2 7 0.8 5.17 0.82 5.3 7 0.75 5.8 7 0.1
0.447 0.03 0.417 0.02 0.34 7 0.01 0.357 0.01
not fixed and varies according to the pathological conditions, plant availability and the age of the child. Daouri is the concept of globalising conditions of herbal medicine utilisation in children between 0–10 years old. Even if the same medicinal plants are found in the formulation used in adults, the name Daouri is no longer used in Kotokoli for adults. Daouri has been specifically used by children in this community for centuries and today, the practice has become more widespread across the country. Data analysis indicated that the common name of Daouri is not based on its composition but rather on the popular knowledge that is transmitted from generation to generation and is mainly used by females in the community. Females buy this traditional recipe without any prior advice from traditional healers or herbalists. As many traditional medicines, Daouri is used for many purposes, such as medicine used to provide anti-diarrhoeal, anti-malarial, anti-anaemic, anti-microbial and intestinal wound-healing effects and for the children's well-being. Previous investigations have indicated that Daouri has potential anti-microbial effects and has been shown to inhibit the growth of Candida albicans, Escherichia coli, Staphylococcus aureus, Salmnonella sp., Pseudomonas aeruginosa (unpublished data). These microbes have been shown to be differentially involved in fatal childhood diseases or children morbidity. However, for anti-diarrhoeal, anti-malarial and anti-anaemic purposes, no scientific studies have been performed. Thus, the use of Daouri in the treatment of malaria should be performed with caution
Please cite this article as: Edorh, M.S., et al., Toxicological screening of Daouri, a polyherbal formulation used in children in the Central Region of Togo. Journal of Ethnopharmacology (2015), http://dx.doi.org/10.1016/j.jep.2015.01.045i
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because further clinical studies are needed to confirm its effectiveness in providing an anti-malarial effect. Khaya senegalensis, Odina acida, Pteleospsis suberosa, Paulinia pinnata and Lophira lanceolata are the most important species cited in the four Daouri compositions analysed. Previous investigations have shown that Pteleopsis suberosa has anti-microbial, fungicide and antioxidant activities (Germanò et al., 1998); Khaya senengalensis has antipyretic (Lompo et al., 1998), antiinflammatory and anti-parasitic properties (Abreu et al., 1999). Odina acida and Lophira lanceolata have anti-microbial effects (Almagboul et al., 1988; Okine et al., 2004) and they are used traditionally as anti-malarial symptoms. Paulinia pinnata has been not been specifically studied for diseases affecting children, but as an antioxidant and vascular relaxant (Zamble et al., 2006; Jimoh et al., 2007). Several studies have shown that medicinal plant products are not completely safe, particularly in the liver as demonstrated by Teschke et al., 2013, 2014. However, the SDa tested at the three doses in the present study exhibited the following results: it did not kill any rats, and hepatic enzymes, such alanine transaminase, were not significantly increased. Although the results showed that the concentrations of alkaline phosphatase in the serum of animals treated with SDa increased compared to control rats, this increase did not necessarily result in liver damage because alkaline phosphatase was also present in other tissues, such as the intestinal epithelium and bone. Furthermore, this enzyme increased with an increase in the concentration of total protein as described in the present study. Despite the absence of macroscopic histological abnormalities, further investigations are needed for microscopic histological analysis. Some individual plant components of Daouri have been studied for their toxicological properties. A previous investigation has demonstrated that the aqueous extract of Khaya senegalensis leaves administered at 600–3000 mg/kg to rats for 28 days had no significant toxicological effect on renal and liver physiology (Nwosu et al., 2012). For many other species of Daouri medicines, such as Odina acida, Lophira lanceolata, Paullinia pinnata, Pteleopsis suberosa and Tinnea barteri, full toxicological investigations have not yet been undertaken to the best of our knowledge. Independent of the absence of toxicological data on individual plants, the results obtained from the present investigation may indicate that their combination in a single formulation, such as Daouri, does not induce significant toxic effects in rats. Moreover, Daouri treatment appears to increase the concentration of serum protein, which may be useful for children growth. This may be a result of the nutritional potential of Daouri as demonstrated by the Kotokoli community when using Daouri to improve immune and nutritional conditions of children. 5. Conclusion Daouri has been used in Kotokoli communities for several centuries, and the present study shows that it is not toxic to Wistar rats
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Please cite this article as: Edorh, M.S., et al., Toxicological screening of Daouri, a polyherbal formulation used in children in the Central Region of Togo. Journal of Ethnopharmacology (2015), http://dx.doi.org/10.1016/j.jep.2015.01.045i
