INTRODUCTION:

India has the richest traditional healing system in the world. In India and other developing countries, the plants are important national resources of health care system.¹ The Indian Himalayan Region (IHR) has been recognized amongst 34 biodiversity hot spots in the world. It has been found to contain about 1,748 different species of medicinal plants.² Zanthoxylum Linn. family Rutaceae is a large genus of aromatic, prickly, dioecious or rarely monoecious trees or shrubs distributed in tropical areas.³ About 11 species have been recorded from India as reported by Hooker.⁴

Zanthoxylum armatum DC. (syn. Z. alatum Roxb.) is an important plant from this genus which is commonly known as tooth ache tree, Nepal pepper, timur and Indian prickly ash.⁵

Distribution:

The plant is widely distributed in India from Kashmir to Bhutan upto 2500m altitude. It also occurs throughout North East India, China, Nepal, Taiwan, Philippines, Malaysia, Pakistan and Japan with varying altitudes from 1300 to 1500 m.⁶

Botanical features:

It is an armed, erect shrub or a small tree, 6 m tall or more with dense foliage. The flowers are green or yellow, in dense terminal, seeds solitary in a fruit, globose, shining black. The branches are armed, the prickles flattened, up to 2 cm. long, bark is pale brown, deep-furrowed, leaves imparipinnate or trifoliate, 5-23 cm. long, with flattened prickles, leaflets up to 5 pairs, opposite, ovate to lanceolate, entire to glandular crenate, acute to obtusely acuminate.⁷ The leaves are dorsiventral, thick with broad, elliptical midrib. The fruits are small but they are produced in clusters which make harvesting easy. Each fruit contains a single seed and young leaves are used as condiments.^{8, 9}

Ethnomedicinal uses:

The fruit, branches and thorns are used to cure toothache and other diseases of teeth. Bark powder on mixing with honey gives relief against gum bleeding.⁷ Seeds and fruits are used as aromatic and tonic, in dyspepsia, fever and for expelling round worms.¹⁰ It is also considered as a condiment, stomachic tonic and carminative¹¹, insecticidal¹² abortifacient, antifertility agent.¹³ The volatile oil is employed as an antidiarrehal, antiseptic, deodorant and anticataerhal. The oil has a good tenacity and is appreciated for its fixative qualities.¹⁴ Timur fruits are generally used by the pharmaceutical companies for preparing tooth pastes.¹⁵ Medicinal use of this plant is in several chronic problems such as rheumatoid, skin diseases and varicose veins. It is also used for low blood pressure, fever and inflammation.¹⁶ It is extensively used in traditional practices of the Khasi tribe in North-Eastern India and in neighboring regions including South-East Asia. Herbal formulations prepared from various parts of the plant are reported to be anthelminthic and hypoglycaemic and are used in treatment of cholera, tonic for fever, remedy for skin diseases, diseases of the mouth and teeth and others.^17,18

Taxonomic position of Zanthoxylum armatum DC¹⁹

Kingdom	Plantae
Division	Tracheophyta
Class	Magnoliopsida
Order	Rutales
Family	Rutaceae
Genus	Zanthoxylum
Species	armatum- DC.

Common names^{3, 20, 21}

Hindi	Nepali dhaniya, tejphal, tumuru
Sanskrit	Tejovati
Nepali	Timur, Nepali pepper
English	Tooth ache tree, prickly ash
Urdu	Tamu

Z. armatum twig with unripe fruits Dried fruits

PHYTOCHEMICAL PROFILE:

The seeds were found to contain cis-9-hexadecenoic, eicosenoic, and palmitic acids along with tambuletin . Bark contains alkaloids like berberine, g-fagarine, b-fagarine (-skimmianine), magnoflorine, laurifoline, nitidine, chelerythrine, tambetarine and candicine, coumarins like xanthyletin, zanthoxyletin, alloxanthyletin, resins, tannins and volatile oil.²² Bark also yields a bitter crystalline principle, identical to berberine. Carpels of the fruit give an essential oil which is isomeric with turpentine and has odour and properties like eucalyptus oil. The essential oil from the seeds consists entirely over 85% of the hydrocarbons, phellandrene, small quantity of linalool and an unidentified sesquiterpene.¹⁹ An essential oil collected from the plants grown in Jammu and Kashmir contain linalool, linalyl acetate, citral, geraniol, methyl cinnamate and limonene.^23,
24

A volatile oil consisting mainly linalool, mono terpentriol-3, 7-dimethyl 1-octane3,6,7-triol, trans cinnemic acid, nevadensin umbelliferone, β-sitosterol and its glucoside, 3,5,dihydroxy-7,8,4 'trimethoxy flavone (tamblin) and tambulatin, 3-methoxy-11–hydroxy-6,8-dimethylcarboxylate biphenyl, 3, 5, 6, 7- tetrahydroxy-3',4'-dimethoxyflavone-5–β–d-xylo pyranoside, aramatamide, lignans, asarinin and fragesin, α and β-amyrins lupeol, and β -sitosterol β-D-glycoside have been reported from the plant.²⁵ A new geranylbenzofuranone been isolated from the bark of Zanthoxylum armatum.²⁶

Hydrocarbon fraction (17.35%) of the oil was much lower and oxygenated compounds comprised fairly high portion of essential oil (39.21%). Percentages of monoterpenes and sesquiterpenes found were 47.33% and 10.83% respectively. Oxygenated monoterpenes comprised major profile of chromatogram of essential oil of Zanthoxylum armatum i.e. 37.23% where as monoterpenes hydrocarbons were 10.09%. Alcoholic percentage was much higher i.e. 26.76% and 15-hexadecanoloide (6.58%) was the only cyclic ester found in relatively high percentage.²⁷

A new amide armatamide along with two lignans, asarinin and fargesin, alpha- and beta-amyrins, lupeol, and beta-sitosterol-beta-D-glucoside has been isolated from the bark of Zanthoxylum armatum. The structure of the new compound was deduced by spectral and chemical analysis as N-(4'-methoxyphenyl ethyl)-3, 4-methylenedioxy cinnamoyl amide.^{28,
29, 30} The prominent monoterpene hydrocarbons are α-pinene (0.685%), sabinene (2.329%) and α-terpinolene (2.148%) and others are reported as myrcene, α-pinene, sabinene and limonene. Two other compounds were identified as sesequiterpene i.e. trans-caryophyllene and β-elemene.¹⁴ From the methanol extract of fruits of Zanthoxylum armatum a new flavonoid 3’, 3-dimethoxy-4’, 5, 7- trihydroxy flavone was isolated.³¹ It has been reported that sanshool amides might be responsible for biological activities of seeds, whereas furofuran lignans and cinnamoyl amides are thought to be responsible for biological activities of bark. The leaves may have the biological activities due to furofuran lignans.³² HPLC qualitative analysis carried out in methanol extracts of Z. armatum bark and leaves has shown the presence of rutin and gallic acid.³³

BIOLOGICAL ACTIVITIES:

ANTIBACTERIAL ACTIVITY:

The ethanol and n-hexane extracts of leaves, fruits and bark of Zanthoxylum armatum were evaluated for their antimicrobial potential against bacteria Micrococcus leutus, Escherichia coli, Staphylococcus aureus, Pasturella multcidia, Pseudomonas aeruginosa, Bacillus subtilis, and Streptococcus viridans. The minimum inhibitory concentration (MIC) values for most of the bacterial species were found to be 0.65 μg/ml. Highest content of flavonoids was found in ethanol extract of Z. armatum fruit followed by ethanol extract of Z. armatum bark.^6,13

The main flavonoid 3,5- diacetyltambulin reported from the plant was found to be active against both gram negative as well as gram positive bacteria. The MIC values ranged between 8-64 μg/ml.⁵

In another study, antibacterial activity of the plant was carried out by agar well diffusion method. The plates were observed for zones of inhibition. Leaf essential oil of Z. armatum was tested against various gram positive and gram negative bacteria i.e. Micrococcus leutus, Escherichia coli, Staphylococcus aureus, Pasteurella multocida, Pseudomonas aeruginosa, Bacillus subtilis, and Streptococcus viridians. Ciprofloxacin was used as standard drug. It was found effective against all tested bacterial strains. B. subtilis, and S. viridans were found more susceptible as compared to other tested strains.^{1,
20, 34}

In another study, antibacterial activity was evaluated by determination of zone of inhibition, minimum inhibitory concentration. Methanol fraction exhibited more potent antibacterial activity than ethyl acetate fraction whereas hexane fraction was found to be inactive against bacteria.³⁵ Another study suggested that the ethanol extract of Z. armatum leaf has a board spectrum of antimicrobial activity as compared to hot water extract. The extracts were tested by well diffusion method against Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia.³⁶

The essential oil obtained from the fresh leaves of Zanthoxylum armatum was analysed by gas chromatography/mass spectrometry. The essential oil showed significant antibacterial activity against Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus and Escherichia coli.³⁷

ANTIFUNGAL ACTIVITY:

The essential oil of leaves of Zanthoxylum armatum were evaluated for antimycotic potential against various fungal strains like Fusarium solani, Microsporum canis, Aspergillus flavus, Candida glabrataas, Trichophyton longifusis and Candida albicans.^{5, 6, 13} A general pattern of dose dependency was observed i.e. the effect became more prominent with increasing concentration of the tested samples.. Best antifungal effect was observed against C. albicans followed by A. flavus and F. solani. Other fungal strains were also affected. Overall results depicted that all fungal strains were inhibited by the essential oil obtained from the leaves of Zanthoxylum armatum.^{19, 20, 34}

In another project, the antifungal activity of essential oil and methanol extract from Z. armatum leaves was tested against three agriculturally important phytopathogenic fungi, A. brassicae, A. alternate and C. lunata. The methanol extract gave significant results for antifungal activity. It could be due to the presence of phenolic and flavonoid compounds. The antifungal activity of individual chemical constituents of essential oils such as 𝛽-caryophyllene, caryophyllene oxide, and linalool have been reported by many researchers.³⁸ The compounds which are present in small amount such as 𝛽-pinene, sabina ketone, and 𝛼-selinene in Z. armatum essential oil may also contribute to antimicrobial activity either directly or in synergy with some other bioactive compounds.³⁷ In another study, ethanol extract was tested for its fungicidal activity against Fusarium oxysporum. The study provided significant results for the activity. It was concluded that the antifungal activity may be due to the presence of phenolic and flavonoid compounds.³⁹

ANTHELMINTIC ACTIVITY:

Aqueous, petroleum ether and methanol extracts of the seeds of Zanthoxylum armatum were tested against Pheretima posthuma (earthworm). Piperazine citrate was taken as the standard. The extracts caused paralysis as well as death of the worms even at low concentration of 10 mg/ml. Methanol extract was found to be most potent.⁷

LOUSICIDAL ACTIVITY:

The lousicidal potential of the plant was tested against tropical hen louse, Lipeurus lawrensis tropicalis. The seed extracts were diluted and the activity varied in proportion to the dilution and exposure time. The undiluted extract showed 100% mortality after12 hours.⁵

PISCICIDAL ACTIVITY:

The ethyl alcohol extract of the fruits was tested for piscicidal activity in different tissues of a carnivorous catfish Heteropneustes fossilis. The extract significantly inhibited enzyme activity in brain, liver and muscle tissues. The inhibition was reversible and both dose and time dependent. The activity of Mg²⁺-ATPase was inhibited more than that of Na⁺, K⁺-ATPase. As piscicide, it is a non-competitive inhibitor.^{1, 5, 6, 20, 40}

INSECTICIDAL ACTIVITY:

The insecticidal activity of ethanol extract was evaluated against mustard aphid (Lipaphis erysimi).The food poisoning method was used for the determination of insecticidal activity of the extract. The insects were put in a petriplate sized 6 inch with mustard leaf and then plant extract was sprayed with chromatographic sprayer. After 6, 12 and 24 hours of spraying, the observations were recorded with number of insect mortality. The results showed that Zanthoxylum armatum could be used as a source of insecticidal compounds which may find applications in food and pesticide industries.³⁹

Z. armatum seed oil, vanillin and fruit oil of Z. piperitum, in combination were able to enhance repellent activity against female Aedes aegypti. The effect was compared with N,N diethyl-3-methylbenzamide (DEET) repellent.¹

The plant has been found to show insecticidal activity against Aedes albopictus, Aedes aegypti Culex pipiens, Culex quinquefasciatus, Anopheles stephensi and Pieris brassicae.^6,19

LARVICIDAL ACTIVITY:

The essential seed oil singly showed larvicidal activity against three medically important species of mosquito vectors Culex quinquefasciatus, Aedes aegypti, Anopheles stephensi and was also effective against Aedes albopictus and Culex pipiens.¹ Study of essential oil yielded at least 28 compounds, consisting mainly of oxygenated monoterpenes and monoterpenes. The larvae of Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus were very much susceptible to the essential oil composition. This presents a potential for the development of alternative plant based larvicides.^{12, 19}

MOSQUITO REPELLANT ACTIVITY:

The mosquito repellent property of oil of Zanthoxylum armatum was performed against mosquitoes in mustard and coconut oil base. The results were compared with synthetic repellent dimethyl phthalate (DMP) which was used as standard. The oil has shown better protection in both the bases at all the concentrations. Repellents in mustard oil gave longer protection time than those in coconut oil. At 0.57 mg/cm² concentration the oil gave significant higher protection both in coconut oil (404 min) as well as in mustard (445min) than DMP.^{1, 12, 19}

LEECH REPELLANT:

Volatile oil of Z. armatum has leech repellent action. Investigations on perseverance of anti-agents properties of N,N diethyl phenyl acetamide (DEPA) N,N diethyl phenyl m-toluamide (DEET), 3-acety 2 (2,6-dimethyl-5-heptenyl) oxazolidine (citronyl), Dimethyl phthalate (DMP) and N-benzoyl piperidine (NBP) on material were tried against land leeches in evergreen rain and deciduous backwoods of Assam. Results obtained were contrasted with unpredictable oil of it to assess its adequacy as parasite repellant.^{5, 18, 20}

SNAKEBITE REMEDY:

The plant has been used against snakebites.^41,
42

ANTINOCICEPTIVE ACTIVITY:

The ethyl acetate fraction of ethanol extract of the plant has significantly decreased acetic acid induced writhing numbers and suppressed formalin induced licking times. These observations have shown that ethanol extract of Z. armatum has powerful antinociceptive activity. Major components of this fraction were found to be eight lignans, These lignans were namely eudesmin, horsfieldin, fargesin, kobusin, sesamin, asarinin, planispine A, and pinoresinol-di-3,3-dimethylallyl.⁴³

PHYTOTOXIC ACTIVITY:

The volatile oil of the leaves of Zanthoxylum armatum was screened for phytotoxic activity using Lamina minor as test species. The standard drug used was Atrazine that served as negative and positive control. Plants were observed on a daily basis and the number of fronds were counted on each seventh day. All the parts showed significant dose dependant phytoinhibition. The result of the study has suggested that Zanthoxylum armatum has phytotoxic potential and can be a good herbicide or weedicide.³⁴

IMMUNOMODULATOR ACTIVITY:

The plant has been found to have a stimulating effect upon circulatory system, lymphatic system and mucous membranes. The crude extract was useful in the treatment of respiratory, cardiovascular and gastrointestinal disorders.¹

ANTIDIABETIC ACTIVITY:

In another study, hydromethanolic extract of the bark of Zanthoxylum armatum was tested for its antidiabetic activity in streptozotocin‑induced diabetic rats. Streptozotocin (60 mg/kg i.p.) was used to induce diabetes in the rats. Blood samples were collected from overnight‑fasted rats on 7, 14 and 21 days of treatment and analyzed for blood glucose level and lipid profile. The rats were sacrificed on day 21 and liver and kidney tissues were excised. It was found that oral administration of the extract for 21 days at 200 and 400 mg/kg resulted in significant reduction in blood glucose, total cholesterol, triglycerides, low density lipoprotein, very low density lipoproteins. There was a significant increase in high density lipoproteins and body weight of streptozotocin diabetic rats. It was concluded that the plant may have therapeutic value in diabetes and related complications.^{34, 44}

ANTISPASMODIC ACTIVITY:

Z. armatum has been used as an antidiarrhoeal drug as per ethnopharmacological data. A study was undertaken to provide a scientific proof for this activity. So the samples of essential oil of leaves of Z. armatum were tested against spontaneous and potassium chloride induced contracted smooth muscle of the isolated rabbit jejunum. The tissue was pre-treated with high concentration of KCl (80 Mm in final bath solution). KCl causes depolarization and keep the tissue in a position of sustained contraction. The samples significantly relaxed the contracted smooth muscles. The spasmolytic effect of the oils started from 0.03 mg/ml and showed 100% effect at 10 mg/ml dose. The extracts relaxed the contracted muscle, suggesting that the possible mode of action of this plant is either blocking the calcium channel or blocking the release of stored calcium from the sarcoplasmic reticulum.^{6, 20, 34}

EFFECT ON GASTROINTESTINAL TRACT AND RESPIRATION:

The crude extract of Z. armatum exhibits spasmolytic effect. The effect is supposed to be mediated through Ca⁺⁺antagonist mechanism which provides pharmacological base for its medicinal use in the gastrointestinal and respiratory disorders.¹⁸

CARDIOVASCULAR EFFECTS:

The crude extract of the plant exhibited concentration- dependent relaxation of spontaneous and high K⁺(80 mM) induced contractions against isolated rabbit jejunum (in vitro). It was more effective against K⁺and suggestive of Ca²⁺antagonist effect. The extract also caused inhibition of both atria force and rate of spontaneous reactions similar to Verapamil in guinea pig atria.^{5, 18, 19}

CYTOTOXIC ACTIVITY:

The cytotoxic activity of essential oil from the leaves, bark and fruits of Zanthoxylum armatum was tested using brine shrimp assay method. Each extract was dissolved in respective solvent to obtain a concentration equivalent to 10, 100 and 1000 μg/ml, respectively. The solvents were allowed to evaporate overnight and then 5 ml of seawater solution was added to each vial. After 36 h of hatching and maturation of larvae as nauplii, 10 larvae were transferred to each vial. The vials were then placed at room temperature (25-27^oC) under illumination. Other vials were supplemented with brine solution and served as positive controls. The oil has showed outstanding mortality rate (100%) at a dose of 1000 μg/ml with 15.90 LC50 values. There is need of identification and quantification of pharmacologically bioactive specific constituents from Z. armatum.^20,
34

In another study, it has been shown that Z. armatum has potential as anticancer drug because the crude extract of leaves and fruits show cytotoxicity. It contains a monoterpene, lupeol which act as therapeutic and chemopreventive agent for the treatment of cancer.¹ Another study was undertaken to evaluate the genotoxic and cytotoxic potential of aqueous extracts of fruits of Z. armatum under in vivo conditions using the Allium test. Physiological and cellular data obtained from the study indicated that the extracts induce clumped chromosomes at metaphase stage of cell division coupled with mitotic arrest. Electron microscopy data revealed membrane damage of cellular organelles, chromatin marginalisation and chromatin condensation in cells of roots incubated in the extracts. The extracts were also found to induce concentration dependent genomic DNA degradation and protein precipitation.¹⁷

The ethyl acetate extract of Zanthoxylum armatum stem bark was evaluated for its cytotoxic activity on different cancerous cell lines such as pancreatic, lung, breast and colon cancer. The relative percentage of metabolically active cells compared with untreated controls and then determine on the basis of mitochondrial conversion of 3-(4, 5-dimethylthiazol-2-yl) 2, 5 diphenyltetrazolium bromide (MTT) to formazan crystals. The extract has shown significant cytotoxic potential on lung and pancreatic cancer cell lines. Flavonoids; apigenin and kaempferol-7-O-glucoside have been isolated from column and have shown significant cytotoxic potential. Flavonoids isolated from ethyl acetate extract were responsible for cytotoxic activity of the extract. Therefore it was concluded that Zanthoxylum armatum can be further explored for the development of anticancer drug.⁴⁵

The potential anticancer effects of the methanol extract and the crude saponins from fruit, bark and leaves of the plant on breast (MDA-MB-468 and MCF-7) and colorectal (Caco-2) cancer cell lines using MTT, neutral red uptake (NRU) and DAPI stain assays were evaluated. It has the potential to exert its cytotoxic effect on cancer cell lines isolated from human by a mechanism involving apoptosis. The overall finding demonstrate that this plant specially fruits, could be potential source of new anticancer compounds for possible drug development against cancer.⁴⁶

Clinical use of chemotherapeutic drug, cisplatin is limited by its toxicity and drug resistance. Therefore, efforts continue for the discovery of novel combination therapies with cisplatin, to increase efficacy and reduce its toxicity. ZALE induced cytotoxicity, nuclear blebbing and DNA fragmentation in HeLA cells suggesting apoptosis induction in human cervical cell line. It induced DNA damage.⁴⁷

INHIBITION OF SKIN SENSITIVITY:

A lipophilic extract of fruits of Zanthoxylum armatum was attributed diminishing mouth aggravation because of food. The extract was diluted with oleyl alcohol. This gave elements of restorative which is anything but difficult to figure and is blessed with a relieving impact in view of hindrance of tactile aggravation from sun bathing, shaving, depilation, insect bites, chemical treatments and different causes.^{5, 20}

INHIBITION OF KERATINOCYTE GROWTH:

The methanol extract of Zanthoxylum armatum was found to have antiproliferative activity against the growth of human keratinocytes.¹⁹

ANTIINFLAMMATORY ACTIVITY:

In-vivo antiinflammatory activity of ethanol extract of stem bark of the plant was evaluated in wistar species of rats by using carrageenan induced paw edema model. The plant extract at a dose 250 mg/kg caused inhibition of paw edema by 19.12%, 4 hours after carageenan administration. The edema in rats is said to be biphasic where the first phase is due to the release of serotonin or histamine and the second phase is caused by the release of bradykinin, protease, prostaglandin and lysosome. The effect could be due to the inhibition of the enzyme cyclooxygenase, leading to the inhibition of prostaglandin synthesis.^{1, 19, 48}

The ethyl acetate fraction (EAF) from the ethanol extract also inhibited xylene-induced ear swelling in mice. This animal model is an acute inflammatory model and used commonly for evaluating vascular permeability. Xylene-induced neurogenous swelling is partially associated with substance P. EAF at all tested doses, as well as acetylsalicylic acid (200 mg/kg), significantly inhibited xylene-induced ear swelling in a dose-dependent manner in mice and suggested that it might reduce the release of substance P or antagonize its action. Lignan was supposed to be responsible for its anti-inflammatory activity.⁴³

Another study was designed to investigate antiinflammatory active fraction(s) from the aqueous extract of Zanthoxylum armatum stem bark in acute and chronic models of inflammation in rats. Acute inflammation was induced by subplantar administration of carrageenan (1%) in rat hind paw whereas chronic inflammation was induced by interscapular implantation of a sterile cotton pellet (50 mg). Alkaloid fraction (100 mg/kg) was found to be more effective in comparison to flavonoid (100 mg/kg) in carrageenan induced inflammation model. Therefore alkaloid fraction was selected for further study. Pretreatment with alkaloid fraction (100, 150 mg/kg) have shown significant (p < 0.05) check on carrageenan induced paw edema and significantly (p < 0.05) decreased granuloma tissue formation, as compared to control. The alkaloid constituents are more potent in prevention of acute and chronic inflammation. The study has concluded that the alkaloid fraction is responsible for the antiinflammatory claim of the drug.⁴⁹

A coumarin extracted from the plant, bergepten, significantly inhibited the production of pro-inflammatory cytokines specifically tumour necrotic factor-α and interleukin-6, in a concentration dependent manner.^{5, 6, 20}

HEPATOPROTECTIVE ACTIVITY:

A study was carried out to evaluate the hepatoprotective activity of ethanol extract of bark of Zanthoxylum armatum DC in CCl₄ induced hepatotoxicity in rats. CCl₄ produces a damage that histologically resembles viral hepatitis. Toxicity begins with the change in endoplasmic reticulum, that results in the loss of metabolic enzymes located in the intracellular structures. The activity was assessed using various biochemical parameters like aspartate aminotransferase, alanine aminotransferase, serum bilirubin, alkaline phosphatase, total protein and serum antioxidant enzymes along with histopathological studies of liver tissue. The extract was found to have hepatoprotective effect on liver injuries induced by CCl₄ and the results suggest that this effect may be due to presence of berberine in the bark of Zanthoxylum armatum. Berberine is an isoquinoline alkaloid, which has preventive and curative effects on chemically induced hepatotoxicity in rodents.^{1, 6, 19, 50}

With ethanol extract of Z. armatum, a decrease has been recorded in the incidence of hepatic damage mainly due to the viral infection, hepatotoxic chemicals (alcohol), toxin in food (aflotoxins), peroxides (particularly peroxidised edible oil), pharmaceuticals (antibiotics, chemotherapeutics, and CNS active agents) and environment pollutants.²⁰

In another study, ethanolic extract of Zanthoxylum armatum was screened for hepatoprotective activity. The extract exhibited significant hepatoprotective activity. Preliminary phytochemical screening has shown the presence of flavonoids and phenolic substances in the extract. Flavonoids have been reported as active substances for the treatment of hepatitis induced by virus and chemicals. The study proposed that the antioxidant potential of flavonoids can scavenge free radicals and protect the cell membrane from destruction.⁷

ANTIOXIDANT ACTIVITY:

The antioxidant activity of methanol extract of the leaves was evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, reducing power and phosphomolybdate assay. Ascorbic acid was used as standard. The antioxidant potential evaluated by three assays increased in a concentration dependent manner. The results of the study suggested that Z. armatum exhibited remarkable scavenging effects on DPPH and prominent reducing power and total antioxidant activity. Various authors have reported that phenols, carotenoids, flavonoids, and ascorbic acid present in the plants are majorly responsible for strong antioxidant activity. Phytochemical screening of Z. armatum indicated the presence of various flavonoids, flavonol glycosides, alkaloids, lignans, phenolics, terpenoids, amino acids, fatty acids.^{1, 6, 36, 51}

The major chemical constituents found in the oil were 1, 8 – cineole, linalool, α– terpeniol and ß- cubebene.¹⁹

In another study, petroleum ether, ethyl acetate, methanol and chloroform extracts of the plant were evaluated for their free radical scavenging property using different in vitro models. Comparative antioxidant potential was evaluated using DPPH, nitric oxide scavenging assay and ferric reducing power assay. Petroleum ether and ethyl acetate extracts of the plant have shown potent antioxidant potential.⁵² Two flavonoids, apigenin and kaempferol-7-O-glucoside were isolated from the ethyl acetate extract of dried stem bark of Zanthoxylum armatum. The flavonoids act as good antioxidants.^{18, 45}

The free radical scavenging activity and antioxidant potential of volatile oil and methanol extract/fractions of Z. armatum were also investigated using in vitro assays including scavenging ability against DPPH, reducing power and chelating ability on Fe²⁺ ions. The results of antioxidant activity tests show that methanol extract and derived fractions of Z. armatum should be treated as efficient radical scavengers and reducing agents as compared to volatile oil.^{37, 53}

ALLELOPATHIC INFLUENCE:

Allelopathic influence of the plant was tested on some important crops like Triticum aestivum, Hordeum vulgare, Lens culinaris and Brassica compastris of Garhwal Himalaya region. Different concentrations of leaf, bark, and fruit pulp aqueous extracts were used for the purpose. Significant effects were noticed on germination and growth of all the tested crops. But the increased concentrations (5 and 10%) of leaf, bark, and fruit pulp aqueous extracts considerably reduced the plumule and radicle growth of all the tested crops as compared with control. It was concluded that L. culminaris and H. vulgare may be cultivated in proximity of Z. armatum.^{17, 54}

CONCLUSION:

On the basis of above literature, it can be stated that the plant Zanthoxylum armatum is a versatile plant that has been used for its therapeutic effects in traditional systems of medicines. The plant has been proven to have a number of biological activities. Further studies are required on the secondary metabolites of the plant in search for useful therapeutic agents. The plant can be a successful option for therapeutic and agro-industries application.

REFERENCES:

1 Brijwal L, Pandey A, Tamta S. An overview on phytomedicinal approaches of Zanthoxylum armatum DC: An important magical medicinal plant. J Med Plant Res 2012; 7(8): 366-370.

2 Samant SS, Dhar U, Palni LMS. Medicinal plants of Himalaya: diversity distribution and potential values. Himavikas Gyan Prakash, Nainital, 1998; 13:163-172.

3 Anonymous, Ayurvedic Pharmacopoeia of India, Government of India, Ministry of Health and Family Welfare and Department of Indian System of Medicine and Homoeopathy,1^st ed, volume-II, 168-169.

4 Gupta DD, Mandi SS. Species specific AFLP markers for authentication of Zanthoxylum acanthopodium and Zanthoxylum oxyphyllum. J Med Plants Stud 2013; 1(6):1-9.

5 Singh TP, Singh OM. Phytochemical and pharmacological profile of Zanthoxylum armatum DC- An overview. Indian J Nat Prod Resour 2011; 2(3):275-285.

6 Firake PD, Firake DM, Behere GT. Zanthoxylum armatum: Versatile plant species in nature. Popular Kheti 2014; 2(3):181-184.

7 Mehta DK, Das R, Bhandari A. In-vitro anthelmintic activity of seeds of Zanthoxylum armatum DC against Pheretima posthuma. Int J Green Pharmy 2012; 6:26-28.

8 Verma N, Khosa RL. Hepatoprotective effect of Zanthoxylum armatum DC. InTech 2012, Available from: http://www.intechopen.com/books/bioactive-compounds-in phytomedicine/hepatoprotective-effect-of Zanthoxylum armatum DC.

9 Gupta AK, Tandon N, Sharma M. Quality Standards of Indian Medicinal Plants. Indian Council of Medical Research, New Delhi, 2008, vol 4, p. 271-278.

10 Ramchandran R, Ali M. Chemical composition of fruit oil of Zanthoxylum armatum. Hamdard Med 1996; 42:51-54.

11 Verma N, Khosa RL. Hepatoprotective activity of leaves of Zanthoxylum armatum DC in CCl₄ induced hepatotoxicity in rats. Indian J Biochem Biophys 2010; 47(2): 124-127.

12 Tiwary M, Naik SN, Tewary DK, Mittal PK, Yadav S. Chemical composition and larvicidal activities of the essential oil of Zanthoxylum armatum DC (Rutaceae) against three mosquito vectors. J Vector Borne Dis 2007; 44 (3):198-204.

13 Ibrar BM, Muhammad M, Tahir L. Antimicrobial evaluation, determination of total phenolic and flavonoid contents in Zanthoxylum armatum DC,” J Med Plant Res 2012; 6(11): 2105-2110.

14 Waheed A, Mahmud S, Akhtar M, Nazir T. Studies on the components of essential oil of Zanthoxylum armatum by GC-MS. Am J Analyt Chem 2011; 2: 258-261.

15 Kala CP, Farooquee AN, Dhar U. Traditional uses and conversation of Timur (Zanthoxylum armatum DC) through social institutions in Uttaranchal Himalaya, India. Conservat Soc 2005; 3(1): 224-230.

16 Kayat HP, Gautam SD, Jha RN. GC-MS analysis of hexane extract of Zanthoxylum armatum DC. fruits. J Pharmacogn Phytochem 2016; 5(2): 58-62.

17 Kharshiing EV. Aqueous extracts of dried fruits of Zanthoxylum armatum DC, (Rutaceae) induce cellular and nuclear damage coupled with inhibition of mitotic activity in- vivo. Am J Plant Sci 2012; 3:1646-1653.

18 Medhi K, Deka M, Bhau BS. The Genus Zanthoxylum - A stockpile of biological and ethnomedicinal properties. Open Access Scientific Reports 2013; 2(3):1-8.

19 Bachwani M, Shrivastava B, Sharma V, Khandelwal R, Tomar L. An update review on Zanthoxylum armatum DC. Am J Pharm Tech Res 2012; 2(1):274-286.

20 Bharti S, Bhushan B. Phytochemical and pharmacological activities of Zanthoxylum armatum DC: An overview. Res J Pharm Biol Chem Sci 2015; 6(5):1403-1409.

21 Khare CP. Indian Medicinal Plants: An Illustrated Dictionary. Springer Verlog, New York, Edition1, 2007, p.730.

22 Nazir BN, Handa KL. Chemical investigation of Zanthoxylum alatum. Proc Natl Acad Sci 1961; 3:160-161.

23 Anonymous. The Wealth of India- A Dictionary of Indian Raw Materials and Industrial Products, Raw Material Series, Publications and Information Directorate, Council of Scientific and Industrial Research, New Delhi, 1976, 11, p. 18-21.

24 Jain N, Srivastava SK, Aggarwal KK, Ramesh S, Kumar S. Essential oil composition of Zanthoxylum alatum seeds from northern India. Flavour Fragr J 2001; 16:408-410.

25 Sati SC, Sati MD, Raturi R, Badoni PP. A new flavonoidal glycoside from stem bark of Zanthoxylum armatum. IJPI’s J Pharmacognosy Herbal Formulations. 2011; 1(2):29-31.

26 Bhatt V, Kumar V, Singh B, Kumar N. A new geranylbenzofuranone from Zanthoxylum armatum. Nat Prod Commun 2015;10(2):313-314.

27 Waheed A, Mahmud S, Akhtar M, Nazir T. Studies on the components of essential oil of Zanthoxylum armatum by GC-MS. Am J Analytical Chem 2011; 2 (2): 258-261.

28 Kalia NK, Singh B, Sood RP. A new amide from Zanthoxylum armatum. J Nat Prod 1999; 62(2): 311-312.

29 Prajapati N, Ojha P, Karki TB. Antifungal property of essential oil extracted from Zanthoxylum armatum (Timur). J Nutr Health Food Eng 2015; 3(1):96-100.

30 Vashist H, Sharma RB, Sharma D, Upmanyu N. Pharmacological activities of Zanthoxylum armatum- A review. World J Pharm Pharm Sci 2016; 5(12): 408-423.

31 Mehta DK, Bhandari A, Satti NK, Das R. Isolation and characterization of a flavonoid 3,3-dimethoxy-4,5,7-trihydroxy flavone from Zanthoxylum armatum DC. Int J Pharm Sci Health Care 2012; 5(2):29-32.

32 Kumar V, Kumar S, Singh B, Kumar N. Quantitative and structural analysis of amides and lignans in Zanthoxylum armatum by UPLC-DAD-ESI-QTOF–MS/MS. J Pharm Biomed Anal 2014; 94:23-29.

33 Alam F, Us Saqib QN. Pharmacognostic study and development of quality control parameters for fruit, bark and leaf of Zanthoxylum armatum (Rutaceae). Anc Sci Life 2015; 34(3):147-155.

34 Ibrar BM, Muhammad N, Rehman IU, Rehman M U, Khan A. Chemical composition and biological screening of essential oils of Zanthoxylum armatum DC leaves. J Clin Toxicol 2013; 3(5):1-6.

35 Joshi S, Gyawali A. Phytochemical and biological studies on Zanthoxylum armatum of Nepal. J Nep Chem Soc 2012; 30:71-77.

36 Akbar S, Majid A, Hassan S, Rehman AU, Khan T, Jadoon MA, Rehman MU. Comparative in- vitro activity of ethanol and hot water extracts of Zanthoxylum armatum to some selective human pathogenic bacterial strains. Int J Biosci 2014; 4(1):285-291.

37 Guleria S, Tiku AK, Koul A, Gupta S, Singh G, Razdan VK. Antioxidant and antimicrobial properties of the essential oil and extracts of Zanthoxylum alatum grown in North-Western Himalaya. Sci. World J 2013; Article ID 790580, 1-9.

38 Cha JD, Jung EK, Kil BS, Lee KY. Chemical composition and antibacterial activity of essential oil from Artemisia feddei. J Microbiol Biotechnol 2007;17(12):2061–2065.

39 Raturi R, Badoni PP, Ballabha R. Insecticidal and fungicidal activities of stem bark of Zanthoxylum armatum (Rutaceae). World J Pharm Pharm Sci 2014: 3(3):1838-1843.

40 Ramanujam SN, Ratha BK. Effect of alcohol extract of a natural piscicide — Fruits of Zanthoxylum armatum DC on Mg²⁺ and Na⁺, K⁺ ATPase activity in various tissues of a freshwater air-breathing fish, Heteropneustes fossilis. Aquaculture 2008; 283:77-82.

41 Houghton PJ, Osibogun IM. Flowering plants used against snakebite. J Ethnopharmacol 1993; 39(1):1-29.

42 Weisberg ME, Graham JR. A Barefoot Doctor's Manual: A Guide to Traditional Chinese and Modern Medicine, Cloudburst Press, London, p 244.

43 Guo T, Deng Y, Xie H, Yao C, Cai C, Pan S, Wang Y. Antinociceptive and antiinflammatory activities of ethyl acetate fraction from Zanthoxylum armatum in mice. Fitoterapia 2011: 82: 347-351.

44 Karki H, Upadhayay K, Pal H, Singh R. Antidiabetic potential of Zanthoxylum armatum bark extract on streptozotocin‑induced diabetic rats. Int J Green Pharm 2014; 8:77-83.

45 Mukhija M, Singh MP, Dhar KL, Kalia AN. Cytotoxic and antioxidant activity of Zanthoxylum alatum stem bark and its flavonoid constituents. J Pharmacogn Phytochem 2015; 4(4):86-92.

46 Alam F, Najum Us Saqib Q, Waheed A. Cytotoxic activity of extracts and crude saponins from Zanthoxylum armatum DC against human breast (MCF-7, MDA-MB-468) and colorectal (Caco-2) cancer cell lines. BMC Compliment Altern Med 2017; 17(1):368- 375.

47 Singh TD, Meitei HT, Sharma AL, Robinson A, Singh LS, Singh TR. Anticancer properties and enhancement of therapeutic potential of cisplatin by leaf extract of Zanthoxylum armatum DC. Biol Res 2015; 48(1):46-49.

48 Sati SC, Sati MD, Raturi R, Badoni P, Singh H. Antiinflammatory and antioxidant activities of Zanthoxylum armatum stem bark. Global J Res Engineering 2011; 11(5):19-22.

49 Mukhija M, Goyal R, Kalia AN. Alkaloids from Zanthoxylum alatum stem bark with antiinflammatory potential in rats against acute and chronic inflammation in rats. Indian Journal of Novel Drug Delivery 2012; 4(2):139-144.

50 Ranawat L, Bhatt J, Patel J. Hepatoprotective activity of ethanolic extracts of bark of Zanthoxylum armatum DC in CCl₄ induced hepatic damage in rats. J Ethnopharmacol 2010;127: 777-780.

51 Kanwal R, Arshad M, Bibi Y, Asif S, Chaudhari SK. Evaluation of ethnopharmacological and antioxidant potential of Zanthoxylum armatum DC. J Chem 2015; Article ID 925654, 1-8.

52 Mukhija M, Kalia AN. Antioxidant potential and total phenolic content of Zanthoxylum alatum stem bark. J App Pharm 2014; 6(4):388-397.

53 Joshi P, Raghuvanshi RS, Sankhala A, Sharma V. Proximate composition and antioxidant potential of fruits of Zanthoxylum alatum Roxb. (Tooth Ache Tree) and its acceptability assessment in food products. Curr Res Nutr Food Sci 2016; 4(1):74-79.

54 Singh B, Uniyal AK, Todaria NP. Studies on allelopathic influence of Zanthoxylum armatum DC on important field crops seeking its sustainable domestication in existing agroforestry systems of Garhwal Himalaya. Indian J Sustain Agr 2007; 30(3): 51-56.

Received on 24.11.2017 Modified on 29.12.2017

Research J. Pharm. and Tech 2018; 11(5):2131-2138.

DOI: 10.5958/0974-360X.2018.00395.5