American Journal of Ethnomedicine, 2015, Vol. 2, No. 4 ISSN: 2348-9502 Available online at http://www.ajethno.com © American Journal of Ethnomedicine

Ethnobotany and Ethnopharmacognosy of from Central Burkina Faso: Leucas martinicensis (Jacquin) R. Brown, Hoslundia opposita Vahl and Orthosiphon pallidus Royle Ex Benth

Kansole Michelline Marie Regina*1, Hilou Adama1, Millogo Jeanne2 and Nacoulma Odile1

1University of Ouagadougou, Laboratory of Biochemistry and Applied Chemistry (LABIOCA), Burkina Faso 2University of Ouagadougou, Laboratory of Biology and Ecology (LEBEV), Burkina Faso

*Corresponding author e-mail: [email protected]

ABSTRACT

Objective: The main objective of the current work is to assess the ethnobotany and ethnopharmacognosy of Lamiaceae species from central Burkina Faso, and especially to focuss on the biological activities of Leucas martinicensis (Jacquin) R. Brown, Hoslundia opposita Vahl and Orthosiphon pallidus Royle Ex Benth. Methods: Different methods have been used to reach the above objective such as the Ethnobotanical survey that concerned 13 Lamiaceae species of the study area, the phytochemicals screening, the antiradical activities via FRAP, DPPH and ABTS methods. The Thin Layer Chromatography (TLC) was also performed. Results: The three species play an important role in the Burkinabe ethnomedicine as antibacterial since historical times. After some pharmacognostical surveys, the ethnobotanical investigation showed that these species are used in traditional medicine for different diseases treatment. All parts (stems, leaves, roots, flowers, fruits) of these species are used as antipyretic, anti-parasitic, anti-inflammatory, antiseptic, anti -diarrheic, eyes cleanser, and to heal rheumatic diseases, skin diseases, etc. Phytochemical investigation of these species showed the presence of compounds such as flavonoids (mainly for Orthosiphon pallidus Royle ex Benth), polyphenols, saponins and coumarins. Thin Layer Chromatography (TLC) analysis allowed to get more insight into the polyphenolic compounds and led to the isolation of cafeic acid and two undetermined flavonoids. The in vitro biological activity showed high bioactivity of the extracts as free radical scavenger and anti-gout.

Page 219 www.ajethno.com American Journal of Ethnomedicine ______ISSN: 2348-9502

Conclusion: The current study is a boost up to recognize and to valorize scientifically the use of these three Lamiaceae species in the traditional medicines, particularly in Burkina Faso via the results obtained.

Keywords- Ethnobotany, Lamiaceae, Phenolics, Flavonoids, Antioxidants, Gout, Burkina Faso.

INTRODUCTION constituents and biological activities of some extracts. The use of plants for medicinal purpose has a long history in many indigenous societies MATERIAL AND METHODS and they are continuing to play important Study zone and inhabitants roles in the treatment of various diseases, Burkina Faso, usually known as the mainly in rural areas in . The practices land of upright men is a landlocked country of traditional medicine are based on a long situated in the middle of West Africa and period of beliefs and empirical covers a surface of approximately 274 000 observations1. Lamiaceae or Labiatae are an km². It is located inside the loop of the River important family of plants which includes Niger between 10° and 15° north latitude about 6000 species and about 210 genera. and between 2° east and 5° 30' west This family is an important source of longitude. The climate is a dry season essential oils, infusion and natural (October to May) and a non regular rainy antibiotics for aromatherapy, perfumery season (June to September). The although synthetic fragrances tend to replace homogenous and seasonal dependent vegetal these species. Cosmetical firms use landscape is made of Parkia biglobosa Lamiaceae species usually for their (Néré), Vitellaria paradoxa (Karité), Cassia moisturizing and antiseptic properties. sp and Andasonia digitata ecosystems. Leucas martinicensis (Jacquin) R. Brown, Species such as Azadirachta indica (Neem) Hoslundia opposita Vahl and Orthosiphon and Eucalyptus sp., have been introduced to pallidus Royle ex Benth are three species of slow down desertification. The Mossi the Lamiaceae family growing in many plateau also called central plateau comprises regions of Burkina faso. In Ouagadougou, the central part of the country where is these plants are sold in medicinal drug located the Capital city, Ouagadougou. markets as ingredients of recipes for Mossi are the dominant ethnic group even treatment of infectious or inflammatory though other people with different ethnic diseases. backgrounds have settled in the area. The overall objective of the current Cultivation of crops like sorghum and study is to document the ethnobotanical pastoralism are the major economic knowledges on medicinal lamiaceae species activities of the local people2. Because of the in the region surrounding Ouagadougou. weakness of the modern health system and This region called Kadiogo Province services, but mainly due to the cultural comprises many ethnical groups of the beliefs the majority of the population use country, because Ouagadougou is the capital traditional medicine and particularly city of Burkina Faso. From the most used medicinal plants as primary health care. species, we aim to screen the phyto-

Page 220

American Journal of Ethnomedicine ______ISSN: 2348-9502

Sites and informants selection names have been verified from the Based on information from the literature4. Direction Générale de la Pharmacie, du Médicament et des Laboratoires Laboratory material and methods (DGPML), the national administration in Stems with leaves of the three most charge of traditional medicine, and the cited species were collected: Leucas mayor office of Ouagadougou, sampling martinicensis Jacquin) R. (Collected at sites for ethnobotanical data collection were University of Ouagadougou and deposited selected from seven sites that are located in as LM 01.), Hoslundia opposita Vahl the administrative department of (collected at the Bangr - Weoogo urban park Ouagadougou (Koubri, patte-d’oie, of Ouagadougou and deposited as HO 01.) Gounghin, Pissy, Cissin, Tanghin dassouri, and Orthosiphon pallidus Royle ex Benth yagma). (harvested at the University of A total of 39 traditional medicine Ouagadougou and deposited as OP 01.). The practitioners and 67 users of the traditional material was washed and dried in the medicine products were systematically laboratory away from sunlight and dust. chosen according to the method of Martin3. They are then finely grinded and stored in clean plastic bags. In addition, some fresh Semi-structured interview plants were kept for botanical investigations The survey was based on a (histochemistry). questionnaire model. A confidence relationship was established with the Histochemical characterization traditional medicine practitioners. No money In order to characterize was given to informers. Interviews were biochemically the different tissues of the conducted in French or in moore (the plant (parenchyma, collenchyma, phloem, language spoken by Mossi people) xylem, sclerenchyma), histochemical sometimes with the help of translators who sections were performed on freshly were conversant with the local language. harvested leaves, stems and roots of Leucas The survey focused on following martinicensis (Jacquin) R. Brown, Lamiaceae species: leucas martinicensis (J.) Hoslundia opposita Vahl and Orthosiphon R. Br, Hoslundia opposita Vahl., pallidus Royle ex Benth. The sections were Orthosiphon pallidus R. ex B., Mentha treated with green Carmino after successive viridis L., Mentha piperita L., Mentha passages in bleach and acetic acid 1%. aquatica L., Ocimum americanum L., Tannins and alkaloids have been Ocimum basilicum L., Solenostemon characterized using aqueous solution of rotundifolius (Poir.) J. K. Morton, FeCl3 1% and lugol 3%, respectively. Solenostemon monostashyus (P.Beauv.) Specific reagents are then used to identify Brig, Hyptis spicigera Lam., Hyptis other secondary metabolites in the tissues5. suaveolens (L.) Poit.) and Tinnea barteri Sections have then been observed and Gürke. imaged under a light microscope at X 10. The mentioned plants were harvested according to the interviewed persons and Extraction identified by Prof. Millogo J., from The plant powder (25 g) was University of Ouagadougou biodiversity extracted using a Soxhlet system with 250 center where voucher herbarium specimens ml chloroform and then 250 ml methanol were deposited. The correspondences (until clear supernatant; 3 to 8 h cycling). between the Moore names and scientific

Page 221

American Journal of Ethnomedicine ______ISSN: 2348-9502

The extracts were then dried using a colorimetry of aluminium complexes. The Rotavapor system. results are expressed in mg quercetin equivalents per 100 mg of extract (mg Phytochemical screening EQ/100 mg). The phytochemical screening Flavonols were assayed using the consisted of a qualitative analysis of metal-chelation colometry method by secondary metabolites present in the plant Abarca et al.8. The results are expressed as samples using classical color or precipitation Quercetin Equivalent for 100 mg of dry reactions. Reactions were carried out (i) in extract. chloroform extracts, for sterols, triterpenes, Tannins were measured by flavonoid and coumarinaglycone; and (ii) in colorimetry of ferric complexes, calibrating methanol extracts, for flavonoids, catechic with a tannic acid solution7. Results are and gallic tannins, saponins, anthraquinones, expressed as mg of tannic acid equivalents alkaloids and coumarins. per 100 mg of extract (mg TA/100 mg).

Thin-layer chromatography profiling of DPPH radical scavenging assay polyphenols Radical scavenging properties of the Methanol extract of Orthosiphon plant extracts against stable DPPH• (2, 2’- pallidus (10mg/ml) was evaporated until diphenyl-1- picrylhydrazyl, Fluka) were dryness, dissolved in methanol then applied determined spectrophotometrically at 517 (10 µl) on silica gel plates (FLUKA, TLC- nm using the method of Vélazquez et al.9. PET foils). The plate (5cmx 10 cm) was Extract solutions were prepared by eluted up to 8.5 cm with glacial acetic acid - dissolving dry extract (10 mg) in methanol ethyl acetate - formic acid - distilled water (10 ml). The samples were homogenized [0.9: 1.1: 2: 9.5], dried, treated with a 10 g/L using an ultrasonic bath. For different solution of diphenylboric acid aminoethyl concentrations, aliquots of 0.5 ml from all ester in methanol and then with a 50 g/L samples were added to methanolic DPPH• macrogol 400 solution diluted in methanol solution (1 ml, 20 mg/ml). After 15 min in and examined in ultraviolet light at 254 and the dark at room temperature, the decrease 365 nm. Different reference compounds in absorption was measured. The blank (rhamnetin, rutin, caffeic acid, myricetin) sample was composed of the same amount have been co-eluted. of methanol and DPPH• solution. All experiments were performed in triplicate. Determination of total polyphenols, Radical scavenging activities were flavonoids, flavonols and tannins calculated by the following formula: The determinations were made on Inhibition (%) = (1 − A/A0) × 100 where A0 methanol extracts (10 mg/ml). is the absorption of blank sample and A is The concentrations of total the absorption of tested extract solution. The polyphenols were estimated via the extract concentration allowing a 50% measurement of the phosphomolybdo- scavenging activity (IC50) was determined tungstic reagent (Folin-Ciocalteu reagent) and expressed in μg/ml. reduction, then the concentrations were expressed in mg gallic acid equivalents per FRAP assay 100 mg of extract [mg GA/100 mg]6. FRAP (Ferric Reducing Antioxidant Flavonoids were determined using Power) assay was carried out as described the method of Dowd that has been adapted by Hinneburg et al.10. A volume of an by Arvouet-Grand et al.7, based on the extract solution (0.5 mL, 0.1 mg/ml) was

Page 222

American Journal of Ethnomedicine ______ISSN: 2348-9502 added and mixed together with phosphate of ascorbic acid (264.15 mg/l). The buffer (1.25 mL, 0.2 M, pH 6.6) and concentration of compounds having a aqueous potassium hexacyanoferrate (1.25 reducing effect on the radical cation ABTS•+ mL, K3Fe (CN)6, 1%). After 30 min (antiradical compounds) is expressed in incubation at 50°C, 10% trichloroacetic acid equivalent mmol of Ascorbic Acid (EAA)/g (1.25 mL) was added. The mixture was of dry extract according to the formula: centrifuged at 2000 rpm for 10 min. The c × C C(%) = × 100 upper layer (0.625 mL) was mixed with 1% M × Ci aqueous FeCl3 (0.125 mL). A blank without C: Concentration of antioxidant compounds extract was prepared under the same in EAA mmol/g of dry extract. conditions. The absorbance was recorded at c: Concentration of the sample been read. 700 nm. All determinations were carried out D: Dilution factor of the sample. in triplicate. Ascorbic acid was used as a Ci: Concentration of the extract. standard and the Iron (III) reducing capacity M: Molecular weight of ascorbic acid (176.1 was expressed as mmol ascorbic acid g/mol). equivalents (AAE)/g of extract (dry weight). Determination of Xanthine Oxidase (XO) By ABTS method Inhibition The method of Re et al.11 was used. The XO activity, using xanthine as a It is based on the decolorization of a stable substrate, was measured via radical cation ABTS•+ (2, 2 '- azynobis-[3- spectrophotometry according to the method ethylbenzothiazoline -6-sulfonic acid]) in of Ferraz Filha et al.12 with some the presence of scavenging ABTS radical modifications. The extracts were compound. The ABTS•+ (blue-green color) investigated for their XO inhibitory activity produced by direct reaction between ABTS at a final concentration of 100 μg/mL. For and potassium persulfate has a maximum the assay, the mixture contained phosphate absorption at 734 nm. buffer (150 μL, 1/15 M, pH 7.5), extract solution (50 μL) and enzyme solution (50 Preparation of the solution of ABTS•+ μL, 0.28 U/mL in phosphate buffer). The A mass of 10 mg ABTS was reaction was initiated by adding substrate dissolved in 2.6 ml of distilled water and solution (250 mL, 0.15 mM in water). The 1.7212 mg of potassium persulfate was absorbance stability at 295 nm was verified added and the mixture kept in the dark at at for 1 min. Enzymatic kinetics were recorded room temperature for 12 hours. The mixture at 295 nm for 2 min. A negative control (0% was then diluted with ethanol until an XO inhibition activity) was prepared with absorbance of 0.70 ± 0.02 at 734 nm. 1% methanol solution instead of the extract solution. All experiments were performed in Samples’ test triplicate. XO inhibitory activity was In three Eppendorf tubes containing expressed as the percentage inhibition of 15 µl of sample solution (1mg /ml) were XO, calculated as: % inhibition = (1− V/V0) added 985 µl of ABTS•+ solution (used × 100. immediately after preparation). The same Where V0 is the linear change (blank operation is performed for the ascorbic acid inclination) in absorbance per minute of used as a reference. The tubes are then negative control, and V as the reaction protected from light for 15 minutes and the kinetic’s rate. absorbance read at 734 nm in a spectrophotometer against a standard curve

Page 223

American Journal of Ethnomedicine ______ISSN: 2348-9502

Statistical analysis Route of administration and dosage Data are expressed as mean ± SD The recipes are used orally or by from three separate observations. For in application on skin. vitro antioxidant assays one way ANOVA test followed by Tukey’s test (P < 0.05) was Informants’ consensus used to analyze the differences among EC50 From the 106 Lamiaceae users of of various fractions for different antioxidant these eleven species, fifty nine (59) have a assays. 100% informant’s consensus, and four (04) have 75%, twenty seven (27) have 50% and RESULTS sixteen (16) have 10%.

Ethnobotanic knowledge Pharmacognostic results During the present ethnobotanical Knowledge of the chemical survey thirteen (13) plants species were composition of the plant cells or tissues is reported by the informants for their useful for the selection of the best suited medicinal usages (table 1). organ for extraction. It can help for Efforts have been made to find threatened species protection by using equivalent medical terms in English for each leaves instead of stem bark or root when of the local disease names. Analysis of data enough phytoconstituents could be revealed that Leucas martinicensis (Jacquin) recovered using quickly renewable organs R. Brown, Orthosiphon pallidus Royle ex like leaves. Histochemical cuttings made Benth and Hoslundia opposita Vahl are showed that only the roots of Leucas applied to a wide range of diseases and are martinicensis (J.) R. Br are rich in alkaloids, found everywhere in the region. Ocimum and tannins as compared to the two other basilicum L., Hyptis suaveolens (L), and species investigated. Leaves and stems of Ocimum americanum L. are mainly used the three species have unicellular hairs, against psychological or parasitisis diseases which are essential characteristics of and for body cleaning. Tinnea barteri Lamiaceae plants. Screening of the main Gürke, Hyptis spicigera L. and phytoconstituents of the tree species gave Solenostemon monostachyus are used as the results summarized in the table 2. antimicrobial and antalgic. Solenostemon The results show a better presence of rotundifolius and Mentha aquatica are used phytochemical substances in methanol as nutraceuticals. extracts rather than chloroform ones. Methanolic extract of Hoslundia opposita is Plant parts used and methods of preparation the richest of the six extracts. Phenolic For most of these species leaves compounds are the most frequents as and/or stems are the most frequent parts compared to saponins and alkaloids. Leucas used. Other parts used are respectively martinicensis seems more closed to seeds, whole plant, leaves with flowers, Hoslundia opposita as the tree species are roots and tubers. The main preparation compared for their phytoconstituents methods are respectively in decreasing composition. order: decoction, infusion (as tea), raw (dried seed) and cooking (tubers). Main phytoconstituents dosage Among the three plants investigated, only Orthosiphon pallidus Royle ex Benth showed the best results in all assays: 33.25 ± 0.65 mg EQ/100mg of extract for total

Page 224

American Journal of Ethnomedicine ______ISSN: 2348-9502 polyphenols, 10.83 ± 0.58 mg EAT/100mg DISCUSSION of extract for total tannins and 5.37 ± 0.39 mg EQ/100mg of extract for total Ethnobotany review from table 1 flavonoids. The total polyphenols are also shows that the whole plant of Hoslundia important in the other two plants: 14.90 ± opposita or various parts of it are used. In 0.66 mg of EQ/100mg for Leucas traditional medicine the plant is used as a purgative, diuretic, febrifuge, antiepileptic martinicensis (Jacquin) R. Brown and 15.02 13 ± 0.030 mg EQ/100mg in Hoslundia and antiseptic in west and southern Africa . opposita Vahl. Total flavonoids are quite Such usages are likewise to those obtained important in Hoslundia opposita Vahl (2.41 from our survey. ± 0.26 mg EQ/100mg extract). All the three Leucas martinicensis is known as plants had a very low amount in total “Mat-burisa” in Yabellozone of Ethiopia and is well used for its medicinal properties. flavonols. 14 It is taken mainly to prevent diarrhea . Our Free radicals scavenging capacity results results however show the use of this species The following results summarized in to treat fever, pain, hypogalactia and asthma. figure 1 have been obtained through testing From pharmacology data on Labiatea, it is with three methods (FRAP, ABTS and kown that: menthol which is used in DPPH), after graphic determinations: medicine and as a flavoring material is one Orthosiphon pallidus Royle ex Benth of the most important constituents of the species showed the best results with the oils. Thymol, extracted from thyme, is used medicinally as a very powerful antiseptic15. DPPH method with an IC50 of 4.98 ± 0.20 mg/ml. A reduction of ferric ions to ferrous Usually, extracts obtained via methanol or ethyl acetate fractions show higher ions in the range of 2.27 ± 0.06 µmol EAA/g 16 has been observed for the same species. antifungal activities . Using ABTS methods, Orthosiphon pallidus The nature of the main chemicals Royle ex Benth species still showed the best groups highlighted by the phytochemical results by reducing ascorbic acid at 88.65 ± screening foreshadows interesting 5.20 µmol EAA/g. pharmacological activities of the methanol extracts. It is basically: The Xanthine Oxidase Inhibition efficiency We got the best inhibition efficiency Coumarins (24.97 ± 3.47% for 1 µg/ml of extract) with They are beneficial in cases of skin diseases, as they are responsible for wound Orthosiphon pallidus methanol extracts. 17 Least significant results have been obtained healing . They could explain the traditional with the two other species. use of these three plants as anti-infective agent against stomach ache, dead bodies’ The Thin-Layer Chromatography (TLC) toilet, fever, convulsions, and diseases of the The elution system: Formic acid/ skin. Water/Ethyl Acetate Acid/Glacial Acetic Acid/distilled water (9.5 / 0.9 / 1.1 / 2, v / v / Saponins v/v) gave the separation of some compounds Their main property is to be antifungal, anti-inflammatory and from the methanol extract of Orthosiphon 1 pallidus Royle ex Benth and we isolated anthelmintic . Using Hoslundia opposita from the figure 2 one phenolic acid (caffeic Vahl in treating dermatoses, as a cosmetic acid) and two non-determined flavonoids for skin and as an anti- inflammatory agent compounds.

Page 225

American Journal of Ethnomedicine ______ISSN: 2348-9502 may be due to the presence of saponins in its investigated for their potential effects on the extracts. central nervous system (CNS). The neuropharmacological screening revealed Tannins and polyphenols the presence of CNS depression. abietane- They are identified in all the three type esters, 3-O-benzoylhosloppone, 3-O- species. Their presence could explain the cinnamoylhosloppone, 3-O-benzoylhinokiol species use in the treatment of skin diseases, and 3-O-benzoylhosloquinone were isolated for the release of insects’ bites’ pain, in from the root bark of Hoslundia opposita diarrhea treatment and the dead bodies’ which could justify the plant’s antimalarial toilet. properties. 3-O-Benzoylhosloppone inhibits the growth of the multidrug resistant strain Flavonoids K1 of Plasmodium falciparum in vitro with These are strong sources of an IC50-value of 0.4 μg ml−120. Essential oil phytoestrogen, insecticides and anti- extract from leaves of H. opposita inflammatory, they can be antibacterial and normalized Hyperglycemia-induced antiviral18; they could justify the use of dyslipidemia associated with diabetic Leucas martinicensis (Jacquin ) R. Brown as complications on sub-chronic treatment with insect repellents and for the treatment of the oil. This activity may be of benefits in inflammatory diseases (painful the management of type 2 diabetes and its menstruations, fever, ...). associated vascular complications. The From literature review, hepatoprotective potentials of the stem phytochemical analysis of Hoslundia extract of Hoslundia opposita Vahl have opposita19 allowed the isolation of tree also been investigated. Some treatments compounds: 5, 7-dimethoxy -6- conducted with methanol extract of methylflavone, Hoslunddiol and euscaphic Hoslundia opposita (100 mg/kg) acid (jaracandic acid ). The acaricidal active ameliorated effects of hepatoxins and principle of H. opposita was characterized reduced significantly (P>0.05) the high as ursolic acid, a triterpene mainly from the levels of the utilized biochemical markers leaves. The overall acaricidal effect of H. (enzymes). The extracts showed good opposita may have been due to synergy with toxicity profile with an LD50 value above other principles having acaricidal properties 5000 mg/kg for the methanol extract. because ursolic acid itself is not as efficient Phytochemical screening of the plant as the methanolic and ethyl acetate fractions showed the presence of some resins, where its isolation has been made19. The flavonoids, sterols/triterpenes, tannins, essential oil from the leaves of Hoslundia saponins, alkaloids, reducing sugars, cardiac opposita was extracted by hydrodistillation. glycosides and proteins in the extracts. GC/MS analysis of the volatile oil showed These results confirmed that the stem of that it contains mainly sesquiterpenes and Hoslundia opposita contains some bioactive sesquiterpene alcohols19. The essential oil properties leading to some hepatoprotective was tested against bacterial and fungal effects of the plant. Chemically, the major species. The results showed that the compounds obtained from the GC/MS essential oil from Hoslundia opposita has a analysis of ethanolic extract of Orthosiphon significant activity against Aspergillus niger, pallidus were Trihydroxy-16a, 17a- Acinetobacter calcoacetica, Brochothrix propylmethylenedioxypregna-1,4-diene-3,20 thermosphacta and Flavobacterium dione, 9, 12, 15 Octadecatrienoic acid, 1, 6, suaveolens. The chloroform extract of the 10-Dodecatrien-3-ol, 3, 7, 11-trimethyl, root of Hoslundia opposita has been Octadecanoic acid and Docosanoic acid21.

Page 226

American Journal of Ethnomedicine ______ISSN: 2348-9502

From the unsaponifiable molecules isolated and Hyptis. lantanaefolia Poit. have via the petroleum ether extraction of interesting inhibition potential efficiency Orthosiphon pallidus, an alkaloid, the (40%) on XO26. About antioxidant activity, orthosiphonol (C30H50O2) has been isolated. extract of Orthosiphon pallidus showed a Of the 150 species of Leucas, about seven good reduction capacity on ferric ions (2.27 species have so far been chemically ± 0.06 EAA/g) which is better than those of investigated. A number of secondary the standard compound (ascorbic having metabolites have been isolated and 5.58± 0.32 mmol EAA/g as activity)27. The characterized. From a number of Leucas reduction activity (especially radical species compounds like diterpenes, scavenging) of extract of the species is due triterpenes, flavones, alkaloids, glycosides, to the presence of flavonoids and phenolic sitosterols, chromon, sterol, oleanolic acid, acids which are known for their strong ursolic acid, leucolacton, stigmasterol, antiradical activities28 and could justify the campesterol, isopimarane, rhamnoglucoside, use for anti-inflammatory purposes. From etc. have been isolated and characterized. TLC analysis, caffeic acid (a phenolic acid) For Leucas martinicensis, essential oils from found in extract of Orthosiphon pallidus, is leaves and flowers were found to have 1- a well-known anti- inflammatory hepten -3- ol and germacrene22. The volatile compound18 that could also justify the use of oil obtained is suitable for numerous usages this species in traditional treatment of in pharmaceutics, cosmetics and diseases with an inflammatory component. alimentation products (for example in According to a recent research, high content lotions, creams, soaps, shampoos, rinses, in phenolic acids are positively correlated to gargles, candies, etc.)23. Its antibacterial human health benefit29. activities and antifungal properties have been reported24. Chloroform extracts of the CONCLUSION leaves contain triacontane (E2’), tetracosane (E2’-C) and lutein (E45) and the methanol The study objectives were to collect extracts showed the presence of cholesterol ethnobotanical knowledge on Lamiaceae (E24)14. The phytochemical composition species in the central region of Burkina closeness of Leucas martinicensis and Faso, and to characterize pharma- Hoslundia opposita could explain why both cognostically the three most cited species. are used for pain and infection treatment The obtained results showed that at least whereas Orthosiphon pallidus has more thirteen species are usually implicated in metabolic and functional indications, as traditional medicine treatment of different indicated by our results. Our biological test diseases. Screening of the main on xanthine Oxidase Inhibition efficiency phytochemical groups present in the revealed that Methanol extract of chloroform and methanol extracts showed Orthosiphon pallidus Royle ex has the best high level of phenolic and terpenoids potential. In humans, inhibition of xanthine compounds. The antioxidant and xanthine oxidase reduces the production of uric acid. oxidase inhibitory efficiencies are in relation Thus xanthine oxidase (XO) inhibitors can with the plants chemical composition. The be used in the treatment of hyperuricemia diversity of the secondary metabolites (abnormally high level of uric acid in the within the three species could explain their blood) and in the treatment of cardiovascular usages in the treatment of inflammatory disease25. Other studies showed that ethanol diseases; diarrhea; and parasitic infections of extracts of some Lamiaceae species, the skin and could be a scientific support for especially Hyptis obtusiflora Presl ex Benth. their various usages made by traditional

Page 227

American Journal of Ethnomedicine ______ISSN: 2348-9502

healers in central region of Burkina Faso. from mesquite (Prosopis julifloral, There is need to improve the chemical and Leguminoseae). . Journal of Food pharmacological investigation on Composition and Analysis 20, 119-124 Orthosiphon pallidus Royle ex Benth. (2007). 9. Velasquez E, T. H., Mordujovichole BP, Saadevra G, Scinella GR. Antioxidant ACKNOWLEDGEMENTS activity of Paraguayan plants extracts. The authors would like to thank the Fitoterapia 74, 91-97 (2003).

‘Africa and beauty Forum’ 2nd edition and 10. Iris Hinneburg, H. J. D. D., Raimo Hiltunen. Antioxidant activities of extracts from the West African Economic and Monetary selected culinary herbs and spices. Food Union (WAEMU) for their financial Chemistry 97, 122-129 (2006). supports. 11. Re R, P. N., Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity REFERENCES applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med. 1. Nacoulma-Ouedraogo, O. G. Plantes 26, 9-10 (1999). médicinales et pratiques médicales 12. Ferraz-Filha Z.S., I. F. V., L.G. Fietto, J.A. traditionnelles au Burkina Faso, cas du Lombardi, D.A. Saúde-Guimarães. Xanthine plateau central, Tome I et II Thèse de oxidase inibitory activity of Lychnophora Doctorat d’Etat. thesis, Université de species from Brazil (“Arnica”). Journal of Ouagadougou., (1996). Ethnopharmacology 107, 79–82 (2006). 2. Institut National de la Statistique et de la 13. Tonzibo Z.F., J. C. C. Y. T. N. G. Chemical Démographie Ministère de l’Économie et du Composition of Essential Oils of Ocimum Développement Ouagadougou, B. F. canum Sims from Côte d’Ivoire. Journal of Enquête Démographique et de Santé 2003. Essential Oil Bearing Plants 11, 530-535 (ORC Macro Calverton, Maryland, USA (2013). 2004). 14. Estifanos, E. Chemical Studies of Leucas 3. Martin, G. J. Ethnobotany: A Methods martinicensis. M.Sc. Thesis submitted to the Manual. Vol. Volume 1 de People and Addis Ababa University,, (2003). plants conservation serie (1995). 15. Hooker W.J.. Niger Flora. 489 (Wheldon & 4. Maydell, H.-J. v. Arbres et arbustes du Wesley Ltd., 1996). Sahel : leurs caractéristiques et leurs 16. Perera, D. F. T. N., Fernando, K.M.E.P., utilisations. 531 (1990). Wijendra, W.A.S. Efficacy of 5. Ciulei, I. Methodology of analysis of phytochemicals present in leaves of Punica vegetable drug. Practical manual on granatum against Malassezia species. industrial utilization of medicinal and American Journal of pharmacology and aromatic plants. the Ministry of chemical pharmacotherapeutics 2 (2015). industry. BUCAREST, 16-27 (1982). 17. Kontogiorgis, C. A., Xu, Y., Hadjipavlou- 6. Singleton, L. V., Orthofer, R., Lamuela- Litina, D., Luo, Y. Coumarin derivatives raventos, R.R. Analysis of total phenol and protection against ROS production in other oxidation substances and antioxidants cellular models of Abeta toxicities. Free by mean of Folin-Ciocalteu Reagent. radical research. 41, 1168-1180 (2007). Method in Enzymology., 547-551 (1999). 18. Bruneton, J. Pharmacognosy: 7. Arvouet-Grand, A. B., Vennat, A., Legret, Phytochemistry, Medicinal Plant., (1999). P. Standardization of propolis extract and 19. Gundidza G. M., S. G. D., K. P. Svoboda identification of key components. Journal de and S. Mavi Antimicrobial Activity of Pharmacie de Belgique 49, 462-429 (1994). Essential Oil from Hoslundia opposite. 8. Abarca, N. A., Campos, M.G., Reyes, J- Central African Journal of Medicine 38, A.A., Jimenez, N.N., Corral, J.H., Valdez, 290-300 (1992). S.G. Antioxidant activity of phenolic extract of monofloral honeybee-collected pollen

Page 228

American Journal of Ethnomedicine ______ISSN: 2348-9502

20. Achenbach, W. R. N. M. H. H. W. H. in the prevention of cardiovascular disease? Antimalarial compounds from Hoslundia Br. J. Clin. Pharmacol 62, 633-644 (2006). opposita. Phytochemistry 31, 3781-3784 26. Antonio, G., González, I.L., Bazzocchi, (1992). L.M., Angel, G.R., Mireya, D.C., Mahabir, 21. Ashokan, K. M., Meenakshi Sundaram. P.G. Xanthine oxidase inhibitory activity of nticancer studies on Orthosiphon pallidus some Panamanian plants from Celastraceae Royle. And Peristrophe bicalyculata Nees. and Lamiaceae. Journal of Journal of Pharmacy Research 4, 2654 Ethnopharmacology. 46, 25-29 (1995). (2011). 27. Dastmalchi, K., Dorman, H.J.D., Kosar, M., 22. Ancilla Muhayimanaa, J.-C. C. R.-P. G. Hiltunen, R. Chemical composition and in Chemical Composition of Essential Oils of vitro antioxidant evaluation of a water Some Medicinal Plants from Rwanda. soluble Moldavian balm (Dracocephalum Journal of Essential Oil Research 10, 251- moldavica L.) extract.. Lebensmittel- 259 (1998). Wissenschaft & Technology. 40, 239-248 23. Muhammad S., A. F., M. M. Yahaya. The (2007). Phytochemical Components of Leucas 28. Nijveldt, R. J., van Nood, E., van Hoorn, Martinicensis that Cause Repellence of D.E., Boelens, P.G., van Norren, K., van Adult Mosquito. International Journal of Leeuwen, P.A. Flavonoids: a review of Modern Botany 2, 1-5 (2012). probable mechanisms of action and potential 24. Uche A. Eze, S. O. B., Emmanuel U. Etuk, applications. . American Journal of Clinical George, Ameh, Oguejiofor M. Ugwah, Nutrition. 74, 418-425 (2001). Chinenye J. Ugwah-Oguejiofor. 29. Amitabh Singh, S. M., Mandavi Singh, Udai Phytochemical and preliminary Pratap Singh Studies on the phenolic acid toxicological studies of the aqueous leave contents in different parts of raw and ripe extract of Leucas martinicensis in wistar jackfruit and their importance in human rats. International Journal of Medicinal health. International Annals of Avanced Plants Research 2, 166-169 (2013). Scientific Research 2 (2015). 25. Dawson, J., and Walters, M.. Uric acid and xanthine oxidase: future therapeutic targets

Page 229

American Journal of Ethnomedicine ______ISSN: 2348-9502

Table 1. Ethnobotanical survey results

Mooré Parts of the Informants scientific name applications name plant used consensus Foreign body in the eye Dried seeds 100% (insects, dust) Yuling- Leaves or whole Nightmares; psychological Ocimum basilicum L. 100% gnuuga plant trauma Leaves to drink Deworming (mouth, stomach); 50% as a tea antidiarrheal Complicated angina 10% Kinkiris- Leaves; stems; Tinnea barteri Gürke Hemorrhoids 10% kaandé seeds Toothache 50% Old wounds 100% Leaves Constipation 50% Whole plant Convulsion in children 10% Leaves + flowers Skin diseases 100 % Leaves +leaves of Ocimum Antidote against snake bite. 10% Hoslundia opposita Waamb- basilicum L Vahl kiparé Fever, stomach and lips Roots decoction 50% aches Muscles aches; rib aches; lung Leaves; stems; aches; stomachaches; 100 % flowers difficulties to pee; skin cosmetic Teeth pain; malaria; fever; Rung- Leaves; stems; Hyptis spicigera L. headaches, to conserve cereal 100% rungui flowers; seeds grains in the attics Leaves Dysentery 100 % Solenostemon Leaves+stems Ocular disorders 50 % Pèesa rotundifolius (Poir.) J. Stems Animal feed 100 % K. Morton Tubers (cooked) Eaten (food) 100 % Whole plant (tea) Toilet to dead bodies 50% Dental pain; colic; convulsions; Solenostemon fever; headache; infant cough; 50% monostachyus (P. Kiinga leaves; stems; visual disturbances Beauv.) Brig. flowers; seeds Purifies and disinfects people attending patients until their 100% death. Ruung- Hyptis suaveolens Itchy skin; insect repellent rung- Leaves; stems 100 % (L)Poit. (especially mosquitoes) gnaaga

Page 230

American Journal of Ethnomedicine ______ISSN: 2348-9502

Foreign objects fallen into the Dried seeds 100 % eye (insect, dust) Toilet to dead bodies; breast Orthosiphon pallidus Nin- pain and poor quality of milk leaves; stems 75 % Royle ex Benth. kaansega of women who newly delivered; gout Foreign objects fallen into the Dried seeds 100 % Ocimum americanum Yulin-gnu- eye (insect, dust) L. raaga Nightmares; deworming; Whole plant 100 % insect repellent. Painful menstruation; wounds; fever; stomach pain; agalactia; Leucas martinicensis Bin-wubdo Whole plant hypogalactia; insect repellent 100 % (Jacq.) R. Br. (especially mosquitoes); asthma; insect bites Mentha aquatica L. Used primarily for the aroma Mentha piperita L. Menthe Leaves; stems in drinks 100 % Mentha viridis L. Facilitates digestion (tea)

Table 2. Pharmacognostic screening of the three species

Leucas martinicensis Orthosiphon pallidus Hoslundia opposita (Jacquin) R. Brown Royle ex Benth Vahl ME CE ME CE ME CE Flavonoids + - - - + - Tannins/polyphenols + - + - + - Saponins - - - - + - Coumarins + - + - + - Alkaloids ------

ME: Methanol Extacts / CE: Chloroform Extacts.

Figure 1. Free radicals scavenging capacities of the three species

Page 231

American Journal of Ethnomedicine ______ISSN: 2348-9502

Figure 2. Thin-Layer Chromatography of Orthosiphon pallidus Royle ex Benth

Page 232