INTRODUCTION:

The family Violaceae consists of 22 genera and about 900 species. Hybanthus is one of the genus consisting of 150 species, they have great medicinal value in Ayurveda it contains volatile oil, anthocyanin, flavonoids (rutin) and carotenoid pigments¹.

Hybanthus enneaspermus is a small suffrutescent perennial herb, 15-30cm in height, with many diffuse or ascending branches, glaborous or more or less pubescent. The leaves are linear or lanceolate with 4.5cm and 3-8mm length and width respectively, margins are either sub sessile, entire or serrate, the stipules gland is tipped subtule. Flowers are pink to dark pink, axillary, solitary with pedicles shorter than the leaves, 6-12mm long erect, slender, bracts small above the middle of the pedicel. Sepals are 2.5mm long, lanceolate, very acute, keeled. Petals appear unequal the two upper ones are oblong, slightly longer than the sepals the 2 lateral longer, falcate the lowest much larger than the other, having an orbicular or obovate limb with a long claw which is curved behind into a short spur. Capsules about 6mm diameter subglobose.

Seeds are ovoid, acute, longitudinally striate, yellowish white about 1.5mm long. The leaves are seen as tender stalks. They are demulcents and are used in decoction and as an electuary. They are employed in conjunction with some mild oil in preparing a cooling liniment for the head.

Fig:1 Hybanthus enneaspermus

The entire plant is used medicinally in Ayurveda, Siddha and other traditional systems of medicine for curing various ailments. It consists of steroids, triterpenes, sugars, alkaloids, phenols, flavones, catechins, tannins, anthraquinones and amino acids².

History:²

Hybanthus enneaspermus is an important plant in traditional system of medicine. The plant is bitter and acrid; easily digested, removes kapha and pitta, urinary calculi, stangury, pain, dysentery, vomiting, burning, wandering of the mind, urethral discharge, blood troubles, asthma, epileptic fits, cures cough, gives tone to the breast, alexetric (Ayurveda).

In south west Nigeria, Hybanthus enneaspermus locally known as Abiwere, is used by traditional birth attendants to make delivery less laborious.

In tamil it is called orilaithamarai or orithalthamarai, in Hindi it is called ratanpurus, in Sanskrit as amburuha, atichara, avyatha, charati, Padma, padmacharini, padmavati, padmavah, ramya, sthalapadmini, supushkara.

In Ayurveda it is called Sthalakamala.

Taxonomic classification:³

Botanical name : Hybanthus enneaspermus

Kingdom : Plantae

Phylum : Tracheophyta

Class : Magnoliopsida

Order : malphigiales

Family : Violaceae

Genus : Hybanthus

Species : enneaspermus

Traditional uses:¹

The fruit in combination with other drugs has been recommended as an antidote to snake and scorpion venoms. The Santals employ the root in bowel complaints of the children. The entire plant is used as nutritive and tonic, the roots are diaphoretic, diuretic and in large doses act as emetic and cathartic.

Microscopic characters:⁴

Leaf:

It is a dorsiventral, mesomorphic and amphistomatic leaf. Prominent midrib projecting equally on adaxial and abaxial sides and on the adaxial sides the midrib it is blunt conical and on abaxial side it is hemispherical. Appearance of small collateral vascular bundle in the middle of the midrib. Across the adaxial hump the palisade tissues are transcurren

Fig.2 Fig.3

Thick adaxial epidermis with rectangular or squarish cells; some of the cells contain mucilage. The abaxial cells are thin and small; some of the cells are dilated and contain mucilage. The mesophyll cells differentiated into palisade cells with two to three layers and spongy parenchymatous cells with three to five layers. Absence of chloroplast in some of the mesophyll cells and contain brownish contents. Solid multicellular elliptic secretory glands with dark contents are present at the tip of the lateral serration along the margins of the lamina.

a) Stomata:

The stomata are anisocytic. Stomata measures 30-33 mm long and 20-28 mm broad. Frequency of stomata is found to be 4-6/mm² on the adaxial side and 19-20/mm² on the abaxial side. Stomatal index on adaxial side is 12.5-15.75 and 19.25-22.5 on abaxial side. The adaxial epidermal cells are straight walled with prominent cuticular striations and the abaxial cells are slightly wavy and lack cuticular striations.

Fig.4

b) Venation pattern and distribution:

Appearance of rhomboidal and broadly rectangular distinct vein-islets. Occurrence of marked calcium oxalate crystals on either side of the veins along the network of veins. The crystals are “clinorhombic” type and are exclusively restricted to the path of veins. The crystals are 50-60μm long and 10-15μm broad. Occurence of epidermal trichomes along the margins and veins are seen. The trichomes are either unicellular or bicellular, thick walled and lopsided.

Fig.5

C) Petiole:

The petiole is plano-convex in outline in its cross sectional view. Three vascular bundles are present in proximal part of which median bundle is larger and other two bundles are smaller. The middle region of the petiole is slightly winged and the three vascular strands fuse to form two strands. The vascular bundles are fused to form a single shallow arc of xylem and phloem in the basal part of the petiole. The distal part of the petiole is spindle shaped with two lateral wings.

Fig.6

Stem:

The epidermis is uni-stratose and thin; the cells are squarish with thin cuticle striations. The stem is circular with uneven outline in its cross sectional view. Hypodermis is 1-2 layered and is cholenchymatous. Cortex 5-6 layers of thin walled, parenchymatous cells and contain prismatic crystals of calcium oxalate. The vascular cylinder consists of a single layer of discontinuous patches of perivascular sclerides, a narrow zone of phloem and closed dense cylinder of xylem. Xylem consists of narrow circular, thick walled radially arranged vessels 20-30μm diameter and thick walled fibres usually vary from 5-15μm in diameter. The size of sclereids usually vary from 30-45μm in diameter. Pith is wide and parenchymatous.

Fig.7

Root:

Root has narrow uniform outer zone of cork followed internally by cortex. Appearance of less compact parenchyma cells and secondary phloem in small radial masses in cortex. Dense and formation of compact circular cylinder secondary vessels; it consists of scattered circular vessels of 30-60μ in diameter. Calcium oxalate crystals are also present in the cortex as prismatic crystals thick walled and wide lumened fibres are reported.

Fig.8

Phytochemistry:⁵

Phytochemical analysis of Hybanthus enneaspermus have been carried out and have revealed the presence of dipeptide alkaloids, aurantiamine acetate, isoarborineol, sitosterol, sugars, flavonoids, steroids, triterpenoids, phenols, catechins, tannins, anthraquinones, amino acids andß-sitosterol.

The FTIR spectrum confirmed the presence of phenols, alcohols, alkanes, alkyl halides, carboxylicacids, aromatics, nitro compounds, and amines in ethanolic extract. The results of GC-MS analysis provide different peaks indicating the presence of phytochemical compounds with different therapeutic activities like antioxidant activity, etc. The major phyto constituents were (5E, 13E) 5, 13-docosadienoic acid (20.90%) and cedarn-diol, 8s, 14-(13.02) which possess many biological activities.

Medicinal properties of Hybanthus enneaspermus

Antioxidant⁶ and Free radical scavenging activity:⁷

Evaluation of antioxidant property of Hybanthus enneaspermus by 2,2-Diphenyl-1-PicrylHydrazyl (DPPH) assay. DPPH assay gives a strong absorption band at 517nm in visible region. The degree of reduction in absorbance measurement is indicative of the radical scavenging power of the extract. The ethanolic extract of Hybanthus enneaspermus appeared to be as potent as Butylated Hydroxy Toluene (BHT) which is used as a standard.

Better free radical scavenging activity was observed on alcoholic extract of Hybanthus enneaspermus on the concentration dependent scavenging of DPPH, nitric oxide, ABTS, superoxide and lipid peroxidation studies with the concentrations of 2μg/ml to 1024 μg/ml. This showed potent free radical scavenging activity.

Nephro-protective activity:⁷

Nephro protective activity was observed in cisplatin induced renal injury. Single administration of cisplatin at 5mg/kg bodyweight produced significant increase in blood urea, serum concentration and protein level followed by significant loss in body weight of the animals. Antioxidant defense system were impaired as indicated by the increase in TBARS level and decrease in GST, GSH, and SOD level in renal tissues.

Aqueous extract of Hybanthus enneaspermus at the dose of 500mg/kg body weight and alcoholic extract at the doses of 250mg/kg and 500mg/kg was found to normalize the raised blood urea, blood protein and serum creatinine. Further, the extracts were able to raise the cisplatin induced decreased GST, GSH, SOD and protect the kidney from lipid peroxidation damage. It was observed that 10^th day administration of 500mg/kg of alcoholic extract prior to cisplatin administration (5mg/kg, single dose) in prophylactic regimen, effectively prevented the cisplatin induced renal injury.

Anti-hyperlipidemic activity:⁸

The evaluation of anti-hyperlipidaemic activity was carried out in high fat diet induced hyperlipidaemia in wistar albino rats, significant effects were obtained from ethanolic extract of Hybanthus enneaspermus as evident from restoration of biochemical parameters altered from cholesterol towards the normal levels. Studies and observations on histopathology of the liver were normal in treated groups. The study was carried out using three doses and among these the dose of 400mg/kg showed the best activity comparable with that of the standard drug atorvastatin in a dose of 1.2mg/kg.

The anti-infertility activity:⁹

Infertility is an inability to conceive or inability to carry a pregnancy to give birth. It has been a major medical and social preoccupations, various parameters had been evaluated such as MDA, GSH, ɑ-tocopherol (vitamin.E), ascorbic acid, antioxidant enzyme of ALT (SGOT), AST (SGPT), urea, total cholesterol, haemoglobin, total protein, sperm count and sperm motility on ethanolic extract of Hybanthus enneaspermus. The plant is used to recover the infertility in endosulfan induced toxicity in male albino rats.

Analgesic activity:¹⁰

The analgesic activity was screened using hot plate method, tail immersion method, tail flick method, Petroleum extract and alcoholic extracts were compared alcoholic extract had significant analgesic activity.

Anti-allergic activity:¹⁰

The anti-allergic activity was screened by milk induced eosinophilia and leukocytes method. Comparing petroleum and alcoholic extracts latter showed significant anti-allergic activity. The extract possessing this activity may be due to the presence of phytochemical constituents mainly flavonoids and polar constituents¹⁰.

Anti-microbial activity:¹¹

The leaves of Hybanthus enneaspermus was evaluated for antimicrobial activity. The in-vitro antimicrobial activity was performed by agar disc diffusion method using Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Candida albicans, Pseudomonas aeroginosa and Klebsiella pneumonia. The significant and broad spectrum of inhibition was exhibited by ethanolic extract when compared with chloroform and petroleum ether extract comparing with the standards. Thus, the plant was observed to have antimicrobial activity and phytochemical compounds which can be used for medicinal purposes.

The anti-arthritic activity:¹²

The alcoholic and aqueous extracts of the entire plant of Hybanthus enneaspermus on Freund’s adjuvant induced arthritis was evaluated. The percentage of yield was found to be 12.8% and 10.6% for alcoholic and aqueous extracts respectively. Both the extracts significantly (p<0.001) decrease the paw thickness at the end of 30 days treatment, though both the extracts showed the same level of activity in the acute phase of inflammation, the alcoholic extract was found to be more potent than the aqueous extract in the chronic phase of inflammation. There was 57.4%-59.4% inhibition seen with the alcoholic extract. This results support the folklore claim of the plant against the inflammatory conditions like arthritis.

CONCLUSION:

From the review it is known that the plant is rich in several vital phytoconstituents such as dipeptide alkaloids, aurantiamine acetate, isoarborineol, sitosterol, sugars, flavonoids, steroids, triterpenoids, phenols, catechins, tannins, anthraquinones, amino acids and ß-sitosterol. Acute toxicity studies proved that this herb is safe upto 5000mg/kg in animal studies. Pharmacological evaluation have shown that the plant has anti-oxidant, anti-microbial anti-allergic, analgesic, anti-arthritis, anti-infertility, antihyperlipidaemic and nephroprotective activities. There are still several activities for the plant has been used in traditional medicine. Scientific validation of these claims will help us to obtain a complete profile on this plant.

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Received on 28.09.2015 Modified on 08.10.2015

Res. J. Pharmacognosy & Phytochem. 7(4): Oct-Dec. 2015; Page 245-249

DOI: 10.5958/0975-4385.2015.00038.2