PHYTOCHEMICAL, TOXICOLOGICAL AND HISTOPATHOLOGICAL OF SOME MEDICINAL PLANTS IN NIGERIA

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1 Internatinal Jurnal f Natural and Applied Sciences, 2(3): , 2006 ~apas Institute f Scientific Research and Develpment, 2006 PHYTOCHEMICAL, TOXICOLOGICAL AND HISTOPATHOLOGICAL OF SOME MEDICINAL PLANTS IN NIGERIA STUDIES F. C. Onwuliri', J. D. Mawak2, D. L. Wnang and E. A. Onwuiiri' 'Apptied Micrbilgy Unit, Department f Btany, University f Js, Nigeria 2Department f Micrbilgy, University f Js, Nigeria Crrespnding authr: Or F. C. Onwuliri; facnwuliri@yah.cm ABSTRACT Phytchemical, txiclgical and hist-pathlgical studies were carried ut n the leaves and stem bark extracts f Grewia m/lis, Bswe//ia da/zie//ii, Jatrpha curcas and Ptercerpus erinaceus claimed t be f medicinal values in Nigeria. The sxhlet apparatus was used fr extractin f the crude materials. The result revealed the presence f tannins, sapnins, flavnids, glycsides, balsam, phenls, terpenes, sterids and the absence f alkalids in Grewia m/lis (bark), Bswe//ia da/zie//ii (leaf) and Ptercarpus erinaceus (bark) as well as cyangenic glycsides in all the plant materials. Txiclgical result shwed that the plant is safe fr human cnsumptin but with cautin having prduce LDsat 00 mg/kg bdy weight. The extracts shwed n structural effects n the liver and heart.dn'the kidney hwever, the cnvluted tubules were filled with fluids, which suggested that the extract shuld be used with cautin n patients with renal failure. The plants therefre pssess sme imprtant bilgical activities that culd be harnessed and emplyed beneficially in the management f viral and bacterial infectins. Keywrds: Phytchemistry, txiclgy, histvpathlqv, rat, medicinal plants, Nigeria INTRODUCTION Plants cmparatively have a much greater bisynthetic ability than animals. Nt nly d plants make all the bimlecules that are universally required in living rganisms, they als synthesize a vast array r cmplex chemicals, which are unique t them. Phytchemicals are prducts f secndary metablism defined variusly as a metablic cui de sac r pheripheral metablic pathways leading ff frm the universal (r primary) metablism. Phytchemicals ranges frm cell wall substances thrugh phtsynthesis pigments, terpenes and terpnids, the alkalids, plants phenlics t plant hrmnes, plant nn-prteins, amin acids and cyangenic glycsides (Harbrne, 973). Many phytchemicals have been shwn t be biactive. that is they exhibit prnunced bilgical activity in ther living rganisms (Harbrne, 973). Many f the biactive phytchemicals have fund usefulness as chemtherapeutic agents, pesticides, fd additives, and ther bilgicals. On the ther hand, many human and animal diseases cnditin, nutritinal and therwise have been traced t expsure t certain biactive phytchemicals in fd and fdder. Many plants used fr nutritin and as therapeutic agents have been reprted t cause instant death after use. Therefre, txicity has been ne f the majr limitatins f the use f trpical plants as therapeutic agents. The claims by the traditinal medicine practitiners in Nigeria that Grewia m/lis, Bswe//ia da/zie//i/~ Jatrpha curcas and Ptercarpus erinaceus are effective in the management f varius infectius diseases needs t be evaluated. Phytchemical analysis is a rutine investigatin in bichemical studies f plant extracts and in nutritinal studies n fdstuff f plant rigin. Sme f the phytchemical substances that are usually tested fr in plant extracts include alkalids, terpernids, tannins, flavnids and sapnins amng thers (Onwuliri and Umezurumba, 2003). Jatrpha curcas (Bubble bush) is a drught - resistant perennial, grwing well in marginal, pr sil. It is easy t establish, grws relatively quickly, prducing seeds fr 0 years (Gill, 987). Jatrpha prduces seeds with il cntent f 37%. The il can be cmbusted as fuel withut being refined. The by-prducts are press cake, which serves a gd rganic fertilizer. The plant grws in many parts f Nigeria. It is rugged in nature and can survive with minimum input. Medicinally, it is used in the treatment f diseases like cancer, snakebites, drpsy and paralysis amng thers (Gerge and Pamplma-Rger, 999). The rts, stems, leaves, seeds and fruits f the plant have been widely used in traditinal flk medicine in many parts f West Africa. The seeds f J. curcas have been used as a purgative, anthelminthic and abrtifacient as well as fr treating ascites, 22

2 Onwuliri et al.: phytchemical, txiclgical and hist-pathlgical studies f medicinal plants paralysis and skin diseases (Gills, 987). Oil frm the seed has been used as an ingredient in the treatment f rheumatic cnditins, itches, fever, jaundice, gnrrhea, as a diuretic agent and as a muthwash. The leaf has been used as a haemstatic agent and als in the basting f irnrnunitv against sme infectius diseases. The bark is used as a fish pisn (Gerge and Pamplma-Rger, 999). Grewia mllis (False buttn weed) belngs t the family Tiliaceae and is cmpsed f 0 genera and 40 species distributed in the trpical regins. The plant is mstly trees r shrubs and rarely herbs. The stem is tall, erect, simple r branded with mucilage. The leaves are alternate, simple, entire r tthed and stipulate (Gills, 987). Grewia mllis is f great imprtance as it cntains chemical cnstituents, which have medical prperties. The plant is als knwn t yield timber. The leaves, stems and fruits are used t cure gnrrhea (Gills, 987). Bswellia dalzeieli/~ Hutch (Family: Burseraceae) is a tree f the savanna frest recgnizable by its papery bark peeling ff in a ragged manner. The bark yields a whitish gum resin, which dries readily and friable. The plant prducts (such as the gum resin) and different parts f the plant are widely emplyed in traditinal medicine. The gum resin is used alng with ther materials as stmach disturbance remedy and fr the treatment f venereal diseases. The bark is biled in large quantities and used fr fever, rheumatism and gastrintestinal trubles treatment. The rt decctin biled alng with HIbiscus sabdanffa is used fr the treatment f syphilis. The rt decctin with Damiellia liveri is used in wunds. In Nigeria (specifically in Adamawa State), the fresh bark is ften used as an anti-emetic and relief f giddiness and palpitatins. This study was therefre designed t determine the essential chemical cmpsitin f these plant materials that giv~ them their medicinal prperties as well as the txicity f the plant extracts in experimental rats: MATERIALS AND METHODS Cllectin, preparatin and extractin f plant materials: The leaves and stem barks f Grewia mths, Bswellia dalzielli/~ Jatrpha curcas and Ptercarpus erinaceus claimed t be f medicinal values in Nigeria were cllected frm Js Nrth L.G.A. f Plateau State, Nigeria and used fr the study. The plants were identified at the Department f Btany, University f Js, Nigeria. The leaves and barks were dried in pen air under shade t prevent the direct sunlight frm inactivating the chemical cnstituents. The samples were pulverized int pwders and stred in plyethylene bags until required fr analysis. Altgether, six samples made up f leave and stem backs f Bswellia dalzielli~ and Ptercarpus ennsceus, leaf f Jatrpha curcas and back f Grewia mllis. Fifteen grams ( g) each f the six-pwdered plant sample was extracted using a sxhlet extractr accrding t standard prcedure. The resultant cncentrates f extractin were then evaprated in a water bath at 60 C t btain the extract in slid. The extract was then stred in airtight cntainers and kept in the refrigeratr at 4 C until needed. Percentage yield was then calculated as % Yield = Weight f Extract X Weight f Crude Drug 00 Phytchemical screening f the plants extracts: The phytchemical screenings f the plant extracts were carried ut using standard qualitative prcedures as described by Trease and Evans (984), Sfwra (982) and Onwuliri (2004). Altgether, the parameters screened were alkalids (dragendrf reagent), flavnid (lead acetate), tannins (ferric chlride), sapnins (general test), cardiac glycside (salkwski), cyangenic glycside and terpenes/sterids (phenl, resin and balsam) Hist-pathlgical Test: A ttal f 6 adult winster rats were used determine the txicity f the plant extracts. Each f the rats was administered 00 mg/kg f n f the plant extract intraperitneally fr tw weeks. The rats were maintained n cmmercial cncentrate ratin and water ad-libitum. Thereafter, the animals were scarified and their kidney, heart and liver harvested fr hist-pathlgical studies. The tissues were prepared hist-pathlgical examinatin accrding t the methd described by Brenda and Rack (989). The sectins were cut between -0 J,Jm.And thereafter munted as permanent slides ready fr micrscpic examinatin. LDsdeterminatin: A pilt test was first carried ut using ten rats, which were gruped int five grups, tw in each grup. Different dses f the extracts were administered t the rats intraperitneally (IP) accrding t their bdy weight. The rats were then bserved fr 24 hurs, after which the dse regimen fr the LDs determinatin was drawn up t be 00 mg/kg, 000 mg/kg, and 2000 mg/kg bdy weight. 226

3 , Onwuliri et al: phytchemical, txiclgical and hist-pathlgical studies f medicinal plants Anther grup f 20 rats were divided int fur grups f ratsand each grup was administered ne f the dses f the drug extract intra-peritneally based n the bdy weight f each grup. The anlma.ls were kept fr 24 hurs and bservatins recrded. The LDs was determined by the graphical, prbit value n the herb r arithmetic methds (Trease and Evans, 983). RESULTS The results f phytchemical screening f the crude extract f plant materials are represented in Tabl~. The extracts cntained alkalids, flavnids, tannins, sapnins, terpenes, sterid, phenl, crlucgjyc~)ie) and ljajiqm wlli/e cyangenic glycside was absent The result f thf: percentage mrtality f the mice is represented in Table 2. The result revealed that grup D rats receiving 2000 mg/kg bdy weight f the test extract had the highest mrtatitv'rate. The result f the mean death (Md), dse differences (Dd), and their prducts are shwn in Table 3. The dse difference was similar (0.0 mg/kg) in all grups, while mean death (Md) was shwn t be 3,20. The lethal median dse (LDs) ccurred in grup C rats, where 0% f the ppulatin was killed by the dse (,00 mg/kg). The grup D rats with the highest lg dse had the highest percentage cumulative death (Table 4). Frm the grss and histlgical studies, the kidneys f the unmarked pilt rat had tabular necrsis (Lcalized death), which was mre prnunced, at the crtical regin. Its kidneys and liver were dark brwn and edematus while the marked rats kidneys and liver were bright red in clr. Structural differences'were als nted in the kidneys f treated rats against the untreated where the cnvluted tubules were seen t be filled with fluids. Table : phytchemical characteristics f the extracts f the leaves and barks f the plant used Chemical Cmpnents Crude Extracts ~~~~~~ A B C D E F Alkalids (dragendrf reagent) Flavnid (lead acetate) Tannins (ferric chlride Sapnin (general test) cardiac Glycside (salkwski) Cyangenic glycside Terpenes/Sterids Phenl Resin Balsam Key: A = Grewia mllis (The bark) B = Bswellia dalzielii (The leaf) C = Bswellia dalzielii (The bark) D = Jatrpha cureas (The leaf) E = Ptercarpus erinaceus (The leaf) F = Ptercarpus erinaceus (The bark) + = Present ++ = Highly Present = Nt Present DISCUSSION The phytchemical cnstituents f the test plants investigated in this study are alkalids, balsam, flavnids, tannins, sapnins, terpenes, sterids, phenls and cardiac glycsides at variable cncentratins. Sfwra and Olaniyi (979), Sfwra (982) and Onwuliri, (2004) have als bserved the presence f such cnstituents as alkalids, sapnins, favnids, tannins, glycsides amng thers in trpical plants grwing in Nigeria. Sme f which have been shwn t exhibit varying bilgical activities. These include the inhibitin f dental plaque frmatin, analgesic, anti sickling and antiviral activity. The mst significant prperty f these plants was reprted t be their anti-micrbial and anti-viral activities against cmmn pathgens. 227

4 Onwuliri et al.: phytchemical, txiclgical and hist-pathlgical studies f medicinal plants Table 2: Number f death and percentage mrtality fr mice with plant extracts Grup N. f Mice Average weight f Dse (mg/kg) Number f (%) mice Death Mrtality A B C Table 3: Determinatin f mean death (md), dse difference (dd) and prduct fr mice tested with plant extracts ~ Grup f Mice Dse fr mice Dse diff Number f Mean Prduct f Md and (mg/kg) (Dd) mg/kg Death Death Dd (MdDD A 00 0 B C Ttal Table 4: Determinatin f cumulative (death and survived), percentage cumulative death and lg dse fr mice tested with plant extracts. Grup f Dse' Death Survived Cumulative Death % Lg Dse Mice (mg/kg) Survived Cumulative Death (mg/kg) A B C The presence f glycsides in the plant materials is expected, as glycsides are knwn t ccur universally in the higher plants (Trease and Evans, 983). It is f interesting that nne f these plant materials shwed the presence f cyangenic glycsides. This makes the plants safer fr cnsumptin since cyangenic glycsides are knwn t release n hydrlysis, the very pisnus cyanic acid (Harbne, 973). The presence f sterids in these plant materials suggests that they culd be f sme uses ther than the nes already knwn. It is pssible that these sterids ccur as part f the aglycne mieties f ther cnstituents f the plants like sapnins and alkalids (Harbne, 973). The LDsis a measure f hw lethal/txic chemtherapeutic agents can be half f the ppulatin under study. (Onwuliri and Umezurumba, 2003). The results shwed that fr all dses used, nly the dse f 00mg/kg bdy weight and abve prduced LOsand dses with values less than 000mg/kg bdy weights are f n practical interest (Lrke, 983). Frm the grss histlgical studies, the kidneys f the unmarked pilt rat had tabular necrsis (lcalized death), which was mre prnunced, at the crtical regin. Its kidneys and liver were dark brwn and edematus while the marked rats kidneys and liver were bright red in clr. This agrees with the findings f Aguiyi et al. (996) where hepatic cngestin and necrsis f the kidney f experimental rats was precipitated by an vert decmpsitin f the renal tubules. The edematus nature f the pst mrtem rgans culd be due t tissue bidegradatin leading t carbn dixide prductin and accumulatin in the rgans. Structural differences were als nted in the kidneys f treated rats against the untreated where the cnvluted tubules were seen t be filled with fluids, which suggested that the drugs shuld be used with cautin, mst especially fr patients with renal failure. CONCLUSION The traditinal claims f these plants fr the treatment f sme infectius diseases seem justified. This is because the present f chemical cmpsitins and txicity studies tend t supprt the use f these herbs. The claims by the traditinal medicine practitiners f herbs that are effective against varius diseases may prvide new research interests fr pharmaceutical industries. 228

5 Onwuliri et al: phytchemical, txiclgical and hist-pathlgical studies f medicinal plants ACKNOWLEDGEMENT The authr wishes t thank the Management f Victry Labratry fr using their labratry facilities in the curse f this prject. REFERENCES Aguiyi, J. c., Jhnsn, P. B., Obi, C.., Onwukeme, K. and Dafur, S. J. (996). Studies n the rdenticidal activities f indmethacin. West African Jurnal f Pharmaclgy and Drug Research, : ~ Gerge. P. and Parnplrna - Rger, M. D. (999). Encyclpedia f Medicinal Plants. Editrial Safeliz S. l. Aranvac. 0: 7-9; 20: Gills, l. S (987). Taxnmy f Flwering Plants. Africana Fep Publishers., Onitsha pp. Harbrne, J. B. (973). Phytchemical screening methds: A guide t mdern techniques in plants analysis. Chapman and Hal/. Lndn Pp: Lrke, D. (983). A New apprach t practical Acute txicity testing. Arch. Txical., 4: Onwuliri F. C. and Umezurumba. C. (2003). Studies n the investigatin f the claimed antimicrbial ptency f selected plant parts n Salmnella typhi. Jurnal f Medicine in the Trpics, (): Onwuliri, F. c., (2004). Antimicrbial studies f the extracts f Acalypha wllkesiana L. n.rlicrrganisms assciated with wund and skin infectins. West African Jurnal f,,' Bilgical Science, : -9. Trease, G. E. and Evans, W. C. (983). Pharmacgnsy. 2th Editin. Pub!. Bailere Tindall- Macmillan, Lndn. Sfwra, E. A., and Olaniyi, A. A. (979). Phytchemcial examinatin f the stem, bark f Dracaena manni. Prceeding f cnterpnce. Ie-Ife: University Press, 70p. Sfwra, E. A., (982). Medicinal Plants and Treduins! Practice in Africa. Pub!. Jhn Wiley and Sns, New Yrk. 229