GJRMI - Volume 5, Issue 10, October 2016 by Dr. Hari Venkatesh.K.Rajaraman

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INDEX – GJRMI - Volume 5, Issue 10, October 2016 MEDICINAL PLANT RESEARCH Ethno-Botany – Short Communication EXPLORATION OF KNOWLEDGE ON NEW ETHNO-BOTANICAL VALUE OF JUSTICIA SPICIGERA SCHLTDL. Sonali Dey Sengupta, Rita Paul*

261–266

INDIGENOUS MEDICINE Dravya Guna – Short Communication - Ayurveda A PRELIMINARY MORPHOLOGICAL AND PHYTOCHEMICAL SCREENING OF BULB OF ELEUTHERINE BULBOSA (MILL.) URB. Preethy AS*, Vivek P, R Remadevi

267–273

Cover Page Photography: Dr. Hari Venkatesh K.R. Plant ID: Fruits of Vaaluluvai (Siddha) / Jyotishmati (Ayurveda) [Celastrus paniculatus Willd.]* of the family Celastraceae; Place: Shivamogga, Shimoga District, Karnataka, India *Botanical Name validated from www.theplantlist.org as on 31/10/2016

Short communication EXPLORATION OF KNOWLEDGE ON NEW ETHNO-BOTANICAL VALUE OF JUSTICIA SPICIGERA SCHLTDL. Sonali Dey Sengupta1, Rita Paul 2* 1

Assistant Professor, Department of Botany, Acharya Prafulla Chandra Roy Government College, Himachal Vihar, Matigara, Siliguri 734 010, West Bengal, India. 2 Assistant Professor, Department of Botany, Charuchandra College, 22- Lake Road, Kolkata 700 029, West Bengal, India. *Corresponding author: Email: ritapaul2000@gmail.com

Received: 29/07/2016; Revised: 10/10/2016; Accepted: 28/10/2016

ABSTRACT Since ages, herbs are being used for treating different ailments in different parts of world by different communities. The present ethno-botanical exploration was conducted within two tribal communities of four districts of West Bengal. Survey and extensive interaction resulted in new information about very effective medicinal uses of Justicia spicigera Schltdl. (family- Acanthaceae). The plant is already known in traditional medicine from ancient times and researches have been conducted about its other potential applications. But to the best of our knowledge the information gathered from tribal people is a new one which may put emphasis to explore its significant green chemistry. KEY WORDS: Justicia spicigera, Tribal people, Documentation

Cite this article: Sonali Dey Sengupta, Rita Paul* (2016), EXPLORATION OF KNOWLEDGE ON NEW ETHNO-BOTANICAL VALUE OF JUSTICIA SPICIGERA SCHLTDL. Global J Res. Med. Plants & Indigen. Med., Volume 5 (10): 261–266

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INTRODUCTION: Tribal people possess good knowledge about the potential uses of different plants in and around them in curing different ailments. Their knowledge of making medicines using natural resources is transferred from one generation to another through oral traditions. Such traditional knowledge may vary from one place to another. Some of this knowledge is not documented even in literature. The main aim of this communication is to explore one of such traditional knowledge about ethno-botanical value of Justicia spicigera Schltdl. (familyAcanthaceae). J. spicigera is an evergreen perennial shrub, native to Mexico and South America (Alonso-Castro et al., 2012; BaqueiroPeña and Guerrero-Beltrán, 2013; Vega-Avila et al., 2012); grows in wild conditions in India, but known to common people mainly as a garden plant (Tabish, 2016). It is well known in Mexican traditional medicine for healing different diseases such as dysentery, diabetes, leukaemia, anaemia; for relief of pain, wounds, fever, inflammation and others (Baqueiro-Peña and Guerrero-Beltrán, 2014; Zapata-Morales et al., 2016). Several researches have added more information about its further therapeutic values; few reports (Pavón-García et al., 2011; Casanova-González et al., 2012; BaqueiroPeña and Guerrero-Beltrán, 2014) are there in literature about its other potential applications. Through this endeavor, the authors report about the effective use of J. spicigera by tribal people of West Bengal plains in curing specific problems apart from its known uses. Further the species is taxonomically described to make it familiar to general people. MATERIAL AND METHODS: Present plant of interest Justicia spicigera Schltdl. [family – Acanthaceae; common names – Spicy Justicia, Mexican Honeysuckle, Orange Plume Flower, Muicle, Mohintli, Firecracker Flower, Hummingbird Plant (Euler and Alam, 1982; Anonymous, 2016a); Synonyms: Jacobinia spicigera (Schltdl.) L. H. Bailey, Justicia ghiesbreghtiana Lem., Sericographis mohintli Nees. (Lorenz et al., 1999; Singh, 2011;

Anonymous, 2016b); Botanical description – evergreen, perennial shrub (Fig. 1A) of about 1–1.5 meters tall; leaf (Fig. 1B) simple, petiolate, petioles slightly winged, lamina lanceolate, velvety pubescent, bright green, 15– 20 cm long and 4–4.5 cm wide, margins undulate, apex acuminate, exstipulate. Flower (Fig. 1E) sessile, bright orange, narrow, tubular, bisexual, zygomorphic (Fig. 1F), 3–3.5 cm long, born in cluster (Fig. 1D) at the end of branch from March to November; sepals 5, gamosepalous, persistent; petals 5, gamopetalous, bilabiate with long slender corolla tube; stamens two, epipetalous, anther is dithecous, unequally lobed; carpels are two, united, ovary superior, style simple, slender, terminal, stigma shortly bilobed; Fruit is a capsule.] Methodology The present ethno-botanical exploration was conducted mainly within two tribal communities (Saontal and Lodha) of four districts (N- 24th Parganas, Nadia, Hooghly and Burdwan) of West Bengal plains during the year 2014–2015. A door to door survey was done and a total of 1462 persons were interviewed. Thorough discussion was done with several questions involving whether the plant is familiar or not, its description, distribution, uses, method of use, doses, risk factors etc. All the information gathered were noted down, studied and scrutinized later. RESULTS AND DISCUSSION: Extensive field survey and interactions especially with senior members revealed that many of them are well familiar with J. spicigera and are aware of its many potential ethnic values that have been already documented in literature. They use this plant from time immemorial to contemporary times also in a regular basis. But the most interesting information for the first time gathered in this survey is that the tribal people use the decoction of J. spicigera by boiling its leaf in water together with some other ingredients for curing cough and cold (as expectorant), catarrhal problems, sore throat, bronchitis and bronchial asthma (as bronchodilator).

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Figure 1 (A-F): Justicia spicigera plant with its different parts and leaf decoction

(A) A portion of whole plant with highlighted flower, (B) Single leaf, (C) Leaf decoction, (D) Inflorescence, (E) Single flower, (F) L.S. through flower.

Preparation of medicine – Ingredients: Leaves of J. spicigera, ginger (Zingiber officinale Roscoe), liquorice (Glycyrrhiza glabra L., locally known as yashtimadhu), tailed pepper (Piper cubeba L., locally known as kababchini), black pepper (Piper nigrum L., locally known as golmarich), and tejpatra (Cinnamomum tamala (Ham.) Nees & Eberm). Preparation: For preparation of a tea cup full of decoction, all the ingredients (10–12 full sized leaves of J. spicigera, 2–3 gm of graded ginger, 1–2 gm of liquorice, 5–6 crushed tailed pepper and 6–8 crushed black pepper) are boiled in two tea cups of water for 15–20 minutes to reduce it to almost half, then allowed to cool and strained. On boiling with the ingredients, the decoction turns into blood red colour (Fig. 1C). The decoction is taken as

oral drink by the patients in tolerable warm condition. Dose – 2–3 tea spoon full, twice or thrice daily before meal for about 2–3 days consecutively as per severity of the problems. Biosafety– According to the tribal people, the decoction is absolutely safe for application for any people of any age – so far, no reports are there to produce any undesirable side effects. Another species of Justicia namely, J. adhatoda L. (Synonym: Adhatoda vasica Nees, commonly known in English as Malabar nut, Adulsa, Adhatoda, Vasa or Vasaka) is a very well-known plant to common people in our country for curing cough and cold. But according to the tribal people, decoction of J. spicigera (they called it as ‘Lal Basaka’) is not only used as a substitute for J. adhatoda but

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also is far more effective in curing the problems than Adhatoda. In Mexican traditional medicine J. spicigera is well-known for the relief of pain, wounds, fever, inflammation (Zapata-Morales et al., 2016); for healing diseases such as dysentery, leukaemia, anaemia (Baqueiro-Peña and Guerrero-Beltrán, 2014); for treatment of infectious diseases such as gonorrhoea (Cáceres et al., 1995), giardiasis (Peraza-Sánchez et al., 2005); even for some emotional disorders (Cassani et al., 2014). In literature reports about the immuno stimulatory (Del Carmen et al., 2013), antidiabetic (Andrade-Cetto and Heinrich, 2005; Ortiz-Andrade et al., 2012), antitumor and immunomodulatory (AlonsoCastro et al., 2012), anticancerous (CáceresCortés et al., 2001; Sepúlveda-Jiménez et al., 2009; Vega-Avila et al., 2009; FernándezPomares et al., 2010; Jacobo-SalcedoMdel et al., 2011), antihypertensive (Esquivel-Gutierrez et al., 2013), antinociceptive (Zapata-Morales et al., 2016), antioxidant and hepatoprotective (Sepulveda-Jiménez et al., 2009; BaqueiroPeña and Guerrero-Beltrán, 2013; Awad et al., 2015), antibacterial and antifungal (Vega-Avila

et al., 2012) effects of J. spicigera are there. Investigation about the non-medicinal use of the plant utilizing its dyeing properties has also been done (Pavón-García et al., 2011; Casanova-González et al., 2012). So, in addition to these, documentation of this tribal knowledge from our country will definitely put emphasis in cultivation and conservation of the plant not only for its ornamental value but also as a very potential medicinal natural resource. CONCLUSION: Present study is an attempt to provide information regarding tribal knowledge on J. spicigera to the future researchers for potential exploration and modernization of this herbal medicine in the field of therapeutic research. Further, proper therapeutic validation, documentation and conservation of elite plants may bring the rich heritage knowledge on indigenous as well as non-indigenous plants into focus more. ACKNOWLEDGEMENTS: The authors are grateful to all the tribal and other local people in the studied area.

REFERENCES: Alonso-Castro AJ, Ortiz-Sánchez E, Domínguez F, Arana-Argáez V, JuárezVázquezMdel C, Chávez M, CarranzaÁlvarez C, Gaspar-Ramírez O, Espinosa-Reyes G, López-Toledo G, Ortiz-Andrade R, García- Carrancá A (2012). Antitumor and immunomodulatory effects of Justicia spicigera Schltdl. (Acanthaceae). J. Ethnopharmacol. 141(3):888–894.

Anonymous (2016a), Mexican honeysuckle, Orange plume flower, desert-tropicals database, accessed at www.deserttropicals.com/Plants/Acanthaceae/Justic ia_spicigera.html on 25 June 2016

Andrade-Cetto A, Heinrich M (2005). Mexican plants with hypoglycaemic effect used in the treatment of diabetes. J. Ethnopharmacol. 99:325–348.

Awad NE, Abdelkawy MA, Hamed M, Ramadan NS (2015). Antioxidant and hepatoprotective effects of Justicia spicigera ethyl acetate fraction and characterization of its anthocyanin content. Int. J. Pharm. Pharm. Sci. 7(8):91–96.

Anonymous (2016b), The plant list, A working list of all plant species, Justicia spicigera, accessed at http://www.theplantlist.org/tpl1.1/recor d/kew-2330023, on 15 October 2016

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Baqueiro-Peña I, Guerrero-Beltrán JA (2013). Antioxidant and Coloring Characteristics of Muicle (Justicia spicigera). In Proceedings of a Conference: International Food Technology, At Chicago, Illinois, USA, Volume: 13. Baqueiro-Peña I, Guerrero-Beltrán JA (2014).Uses of Justicia spicigera in medicine and as a source of pigments. Funct. Foods. Health Dis. 4(9):401– 414. Cáceres A, Menéndez H, Méndez E, Cohobón E, Samayoa BE, Peralta E, Carrillo G (1995). Antigonorrhoeal activity of plants used in Guatemala for the treatment of sexually transmitted diseases. J. Ethnopharmacol. 48:85–88. Cáceres-Cortés JR, Cantú-Garza FA, MendozaMata MT, Chávez-González MA, Ramos-Mandujano G, ZambranoRamírez IR (2001). Cytotoxic Activity of Justicia spicigera is inhibited by bcl2-Proto-oncogene and Induces Apoptosis in a Cell Cycle Dependent Fashion. Phytother. Res. 15:691–697. Casanova-González E, García-Bucio A, Ruvalcaba-Sil JL, Santos-Vásquez V, Esquivel B, Falcón T, Arroyo E, Zetina S, Roldán ML, Domingo C (2012). Surface enhanced Raman spectroscopy spectra of Mexican Dyestuffs. J. Raman Spectrosc. 43(11):1551–1559. Cassani J, Dorantes-Barrón AM, Novales LM, Real GA, Estrada-Reyes R (2014). Anti-depressant-like effect of kaempferitrin isolated from Justicia spicigera Schltdl. (Acanthaceae) in two behavior models in mice: evidence for the involvement of the serotonergic system. Molecules 19(12):21442–21461.

vitro. J. Ethnopharmacol. 148(1):337– 340. Esquivel-Gutiérrez ER, Noriega-Cisneros R, Arellano-Plaza M, Ibarra-Barajas M, Salgado-Garciglia R, Saavedra-Molina A (2013). Antihypertensive effect of Justicia spicigera in L-NAME-induced hypertensive rats. Pharmacologyonline 2:120–127. Euler KL, Alam M (1982). Isolation of Kaempferitrin from Justicia spicigera. J. Nat. Prod. 45 (2):220–221. Fernández-Pomares C, Bravo-Ávila M, MuñozMuñiz O, Juárez-Aguilar E, Domínguez–Ortíz MA, HernándezAguilar ME (2010). Antiproliferative activity of the polar extract of Justicia spicigera on LNCaP cells. Cancer Prev. Res. 3(12):B50. Jacobo-SalcedoMdel R, Alonso-Castro AJ, Salazar-Olivo LA, CarranzaAlvarez C, González-Espíndola LA, Domínguez F, Maciel-Torres SP, García-Lujan C, GonzálezMartínezMdel R, Gómez-Sánchez M, Estrada-Castillón E, Zapata-Bustos R, Medellin-Milán P, García-Carrancá A (2011). Antimicrobial and cytotoxic effects of Mexican medicinal plants. Nat. Prod. Commun. 6(12):1925–1928. Lorenz P, Stermitz FR, Ismail LD (1999). An amide of L-threo-γ-hydroxyglutamic acid from Justicia ghiesbreghtiana. Phytochemistry 52:63–66. Ortiz-Andrade R, Cabañas-Wuan A, AranaArgáez VE, Alonso-Castro AJ, ZapataBustos R, Salazar-Olivo LA, Domínguez F, Chávez M, CarranzaÁlvarez C, García- Carrancá A (2012). Antidiabetic effects of Justicia spicigera Schltdl. (Acanthaceae). J. Ethnopharmacol. 143(2):455–462.

Del Carmen JVM, Alonso-Castro JA, GarcíaCarrancá A (2013). Kaempferitrin induces immunostimulatory effects in

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Pavón-García LM, Pérez-Alonso C, OrozcoVillafuerte J, Pimentel-González DJ, Rodríguez- Hueso ME, Vernon-Carter EJ (2011). Storage stability of the natural colorant from Justicia spicigera microencapsulated in protective colloids blends by spray-drying. Int. J. Food Sci. Technol. 46:1428–1437. Peraza-Sánchez SR, Poot-Kantún S, TorresTapia LE, May-Pat F, Simá-Polanco P, Cedillo-Rivera R (2005). Screening of Native Plants from anducatan for antiGiardia lamblia Activity. Pharm. Biol. 43(7):594–598. Sepúlveda-Jiménez G, Reyna-Aquino C, Chaires-Martinez L, Bermúdez-Torres K, Rodriguez-Monroy M (2009). Antioxidant Activity and Content of Phenolic Compounds and Flavonoids from Justicia spicigera. J. Biol. Sci. 9(6):629–632. Singh G. (2011). Acanthaceae Week: Justicia spicigera from Delhi blog in efloraofindia; original citation: Linnaea. 7: 395–396 395 1832.

Source of Support: NIL

Tabish (2016), Mexican honeysuckle, Flowers of India database, accessed at http://www.flowersofindia.net/catalog/s lides/Mexican%20Honeysuckle.html, on 25 June 2016 Vega-Avila E, Tapia-Aguilar R, Reyes-Chilpa R, Velasco-lezama R (2012). Antibacterial and antifungal activity of Justicia spicigera. Revista. Latinoamericana. de. Quimica. 40(2):75–82. Vega-Avila E, Espejo-Serna A, AlarcónAguilar F, Velasco-Lezama R (2009).Cytotoxic activity of four Mexican medicinal plants. Proc. West. Pharmacol. Soc. 52:78–82. Zapata-Morales JR, Alonso-Castro AJ, Domínguez F, Carranza-Álvarez C, Castellanos LM, Martínez-Medina RM, Pérez-Urizar J (2016). Antinociceptive Activity of an Ethanol Extract of Justicia spicigera. Drug. Dev. Res. 77(4):180–186.

Conflict of Interest: None Declared

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Short communication A PRELIMINARY MORPHOLOGICAL AND PHYTOCHEMICAL SCREENING OF BULB OF ELEUTHERINE BULBOSA (MILL.) URB. Preethy AS1*, Vivek P2, R Remadevi3 1

P G Scholar, Department of Dravyaguna vijnana,V P S V Ayurveda College, Kottakkal, Kerala, India Assistant Professor, Department of Dravyaguna vijnana, V P S V Ayurveda College, Kottakkal, Kerala, India 3 Retd.Professor and Head, Department of Dravyaguna vijnana, V P S V Ayurveda College, Kottakkal, Kerala, India *Corresponding author: E-mail: dr.preethy.cinnish@gmail.com; Mob: +91 9497347649 2

Received: 23/08/2016; Revised: 05/10/2016; Accepted: 20/10/2016

ABSTRACT The major threat in front Ayurvedic field nowadays is nothing but drug crisis. And one of the probable solutions may be introduction of botanically known drugs with similar properties. In this view, Department of Dravyaguna vijnana,Vaidyaratnam P.S.Varier Ayurveda College, Kottakkal, Kerala, India, has taken some drugs to be explored in terms of Ayurveda. As a first step towards that, preliminary morphological and phytochemical evaluation of drugs has been done. One such drug is Eleutherine bulbosa (Mill.) Urb. E. bulbosa is known as ‘Vishanarayani’ in Malayalam language is a herbaceous, ornamental, perennial flowering plant species in the family Iridaceae. It’s a native of South America, being introduced to Indian subcontinent. Now it is naturalised in Kerala, India and is also cultivated for its medicinal properties. It has known astringent, antiepileptic, anti-diabetic, antihaemorrhagic properties and is prescribed in sprains, urinary diseases and haemorrhoids among the tribes. An attempt was made to establish the morphological features of the plant Eleutherine bulbosa and to set the preliminary phytochemical standards of bulb of the same, which can be further used, for the detailed study in future.The organoleptic studies of fresh bulbs revealed that the fresh bulb was purplish red in colour with a pungent odour and bitter taste. Powder microscopy revealed plenty of oil granules, epidermal tissues, fibres and xylem parenchyma Phytochemistry of the bulb yielded more extracts in cold methanol. Qualitative analysis indicated presence of alkaloids, tannins, flavanoids and steroids in it. TLC of the extracts showed different bands in different solvent systems. Key words: Eleutherine bulbosa, Phytochemical screening.

Cite this article: Preethy AS*, Vivek P, R Remadevi (2016), A PRELIMINARY MORPHOLOGICAL AND PHYTOCHEMICAL SCREENING OF BULB OF ELEUTHERINE BULBOSA (MILL.)URB., Global J Res. Med. Plants & Indigen. Med., Volume 5 (10): 267–273

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INTRODUCTION The major threat in front Ayurvedic field nowadays is nothing but drug crisis. And one of the probable solutions may be introduction of drugs which are being used as ethnomedicine. With known Rasapanchaka (the five features of drug – taste, property, potency, metabolite and action) and site of action they can replace extinct or endangered drugs. In this view, Department of Dravyaguna vijnana, Vaidyaratnam P.S. Varier Ayurveda College, Kottakkal, Kerala, India, has taken some drugs to be explored in terms of Ayurveda. As a first step towards that, preliminary morphological and phytochemical evaluation of drugs has been done. One such drug is Eleutherine bulbosa (Mill.) Urb. (syn: Sisyrinchium bulbosum). This herb known as ‘Vishanarayani’ (J.M. Garg, 2013.), also ‘ponnulli’ and ‘neeroottokkizhangu’ in Malayalam is a herbaceous ornamental perennial flowering plant species in the family Iridaceae. It’s a native of South America, being introduced to Indian subcontinent, commonly known as ‘Dragon blood’. Now it is naturalised in Kerala, India especially in Kannore and Malappuram districts (N. Sasidharan, 2004.); Figure 1 – Bulb of Eleutherine bulbosa (Mill.) Urb.

Drying and Processing The collected bulbs were thoroughly washed and shade dried after which it was again put in hot air oven and dried. The dried bulbs were cut into small pieces (Figure 2) and they were homogenized using a mechanical

also being cultivated for its medicinal properties. The decoction of the bulb is used as a hypoglycemic agent in folklore practice in these districts. It has known astringent, antiepileptic, anti diabetic, antihaemorrhagic properties and is prescribed in sprains, urinary diseases and haemorrhoids among the tribes (Ken Fern, 2014) especially among the tribal population of Bankure District of West Bengal (Sinhababu. A., 2013). Researches on this plant revealed its antifungal (Mohanta. Y.K., 2014) and antimicrobial (B.G. Brasileiro, 2006) properties. MATERIAL AND METHODS Collection of Identification

plant

materials

and

Bulb of E. Bulbosa (Figure 1) was collected from the Botanical garden of Vaidyaratnam P.S.Varier Ayurveda College, Kottakkal, Kerala, India and authenticated in the Department of Dravyaguna vijnana, Vaidyaratnam P.S. Varier Ayurveda College, Kottakkal, Kerala, India by external morphological characters (Henry Shaw School of Botany, 1991). Figure 2: Cut pieces of Eleutherine bulbosa (Mill.) Urb.

grinder to obtain coarse powder and stored in air-tight containers for further analysis. The methods used for morphological and Phytochemical analysis were standard procedures as per Ayurvedic Pharmacopeia of India (Anonymous, 2010).

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Pharmacognosy and Phytochemical Lab, Department of Dravyaguna vijnana, V.P.S.V. Ayurveda College, Kottakkal, Kerala, India.

The commonly employed technique for separation of active substance for crude drug is called extraction, which involves the use of different solvents like water, cold and hot alcohol (Anonymous, 2010).

Pharmacognostic studies

Phytochemical screening

Organoleptic evaluation can be done by means of the organs of sense, and includes the macroscopic appearance of the plant material, its odour and taste.

The extracts obtained were subjected to qualitative tests for identification of various plant constituents (C.K. Kokate, 2013).

Setting

Powder microscopy Microscopic slides were prepared by soaking a pinch of fine powder in distilled water for 1hr and were spread on a glass slide. Physico – chemical screening Total ash, water soluble ash and acid insoluble ash, volatile oil content, moisture content, fibre content were determined by using standard methods (Anonymous, 2010). Water soluble, cold alcohol soluble, hot alcohol soluble extracts were obtained and measured (Anonymous, 2010). Ash values Ash values are determined according to the standard procedures (Anonymous, 2010). Moisture content The moisture content is determined by using Dean and Stark’s apparatus (Anonymous, 2010). The percentage of moisture content in the drug was calculated by dividing the reading of water content by the weight of the original sample taken and multiplying it by 100. Volatile oil content The volatile oil content was determined by distilling the drug with distilled water using Clevenger apparatus. The volatile oil was noted and percentage was calculated.

Extractive values

Detection of steroids One ml of the extracts was taken in a clean test tube and 3 ml of chloroform was added. Then a few drops of concentrated sulphuric acid were added through the sides of the test tube. Formation of a brown ring indicated the presence of steroid (C.K. Kokate, 2013). Detection of alkaloids With Mayer’s reagent – Two ml of the extract was taken in a test tube and dried it by placing on a heating mantle. Then a few drops of dilute Hydrochloride acid was added and filtered into another test tube using a filter paper and a funnel. Then a few drops of Mayer’s reagentwas added to it. Turbidity indicated the presence of alkaloids (C.K. Kokate, 2013). With Dragendroff’s reagent – Two ml of the extract was taken in a watch glass and one drop of Dragendroff’s reagent was added and rubbed gently with a glass rod. Formation of an orange brown precipitate indicated the presence of alkaloids (C.K. Kokate, 2013). Detection of phenols Two ml of the extract was taken in a test tube and a few drops of neutral ferric chloride were added to it. A deep blue or violet colour indicated the presence of phenols (C.K. Kokate, 2013). Detection of tannins A few ml of the extract taken in a test tube was added with a few drops of lead acetate solution. A yellow or white precipitate

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indicated the presence of tannins (C.K. Kokate, 2013). Detection of flavonoids Two ml of the extract was taken in a test tube. A few drops of concentrated hydrochloric acid and a piece of magnesium ribbon were added to it. A reddish brown, magenta or pink colour indicated the presence of flavonoids (N. Raaman, 2006). TLC profile

Visualization Plate is observed in UV light at 366 nm and Rf values are noted (Anonymous, 2010). Observation The organoleptic studies of fresh bulbs revealed that the fresh bulb was purplish red in colour with a pungent odour and bitter taste. The dried powder was reddish brown in colour with a characteristic odour. RESULTS

Test solution: 1 gm of accurately weighed coarsely powdered sample was taken in a round bottom flask. 100 ml of 99% of Ethyl alcohol was added to it. It was then condensed with water condenser and boiled for one hour in electric mantle, allowed to stand for some time and then filtered. Solvent system: Chloroform: methanol - 9:1 Plates: Plates were prepared of silica gel of uniform thickness of 0.2 mm. Procedure Extract is applied over the plate with the capillary and plate is developed in the solvent system.

Powder microscopy revealed plenty of oil granules, epidermal tissues, fibres and xylem parenchyma (Figure 3 and Figure 4). Physico chemical parameters are tabulated in Table no:1. The results of qualitative analysis are tabulated in Table no: 2. Qualitative analysis suggested that steroids were found in all the solvents, alkaloids in cold methanol (Dragendroffâ&#x20AC;&#x2122;s method), flavonoids in cold methanol, Tannins were found in alcohol and water extracts, phenols were absent in all the extracts. TLC of the alcoholic extract showed 7 bands in Chloroform: methanol (9:1) solvent system. Rf values are tabulated in Table no 3.

Table no. 1 Physico-chemical analysis of E. bulbosa. Experiments Moisture content Ash content Water soluble ash Acid insoluble ash Volatile oil Water soluble extract Hot alcohol extract Cold alcohol extract Reducing sugar Total sugar

Percentage 3.97% 2.97% 1.62% 0.21% 1.97% 36.67% 10.75% 3.04% 0.6 0.9

Table no. 2 Qualitative analysis Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 10 | October 2016 | 267–273

Sl. No

Chemical constituents

Solvent

Alkaloids

2 3 4 5 6 7

Flavanoids Saponins Carbohydrates Phenols Steroids Tannins

Water ˗˗ ˗˗ ˗˗ ˗˗ + ˗˗ + +

Hot alcohol ˗˗ ˗˗ ˗˗ ˗˗ + ˗˗ + +

Cold alcohol + ˗˗ + ˗˗ + ˗˗ + +

Table no 3. Rf values of TLC Sl. no

Rf value

0.13

Colour of the band in UV light at 366 nm Brown

0.26

Light brown

0.41

Light green

0.52

Light blue

0.71

Light green

0.87

Brown

0.93

Light brown

Figure 3 – Powder microscopy

Fibers

Oil globules

Figure 4– Powder microscopy

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Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 10 | October 2016 | 267â&#x20AC;&#x201C;273

Epidermal tissues

Xylem parenchyma

Figure 5: TLC Plate as visualised in UV light at 366 nm

DISCUSSION Physico â&#x20AC;&#x201C; chemical and Phytochemical parameters Ash values are determined to evaluate quality and purity of the crude drug that contains inorganic radicals like phosphates, carbohydrate, potassium, magnesium and calcium. The residue after incineration is the ash content of the drug which represents the naturally occurring inorganic salts. It helps to detect and check adulteration with exhausted drugs and to ensure the absence of an abnormal proportion of external mineral matter like soil or sand. The total ash value was 3.12%. The low value indicated the purity of the powder. Moisture content of a particular drug is the amount of moisture that particularly absorbed by the drug on exposure to atmosphere after drying. Moisture content was 3.97%. Moisture

content of plant powders which are to be used as drugs should not exceed 14% (Chand S., 2014). Otherwise it may enhance microbial growth. The highest percentage of the extracts was found in water, may be due to its universal solvent property followed by hot alcohol and cold alcohol. The solvent used for extraction should be in a position to dissolve appreciable quantity of chemical constituents. Water soluble extracts of the drug mainly represents the percentage of organic constituents such as tannins, sugars, plant acids, mucilage and glycosides. Alcohol soluble extracts mainly represents the percentage of organic constituents such as alkaloids, phenols, flavonoids, steroids, and sugars present in the drug.

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Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 10 | October 2016 | 267–273

The extracts obtained by exhausting crude drugs are indicative of approximate measure of their chemical constituents. This study goes in hand with previous phytochemical researches on this plant (Rani. V.S., 2015) in a few parameters only. This may be because of the difference in the selection of solvent system, procedure and geographic or ecological variation (Wei Liu, 2015). CONCLUSION

The preliminary morphological and phytochemical analysis of the plant Eleutherine bulbosa conducted here will be useful for further investigations on the same. The study of pharmacognostic features of medicinal plants in general is an imperative process to know their quality, purity and to check for the adulterants and substitutes. Along with this, here the study was conducted as a preliminary step for adopting Eleutherine bulbosa to practice of medicine of Ayurveda.

REFERENCES Anonymous (2010). The Ayurvedic Pharmacopoeia of India, The controller of publication, Dept. of AYUSH, New Delhi, Part II, Volume III, pages 140– 147.

Ken Fern (2014). Useful Tropical Plants Database. Accessed at http://tropical.theferns.info/viewtropical .php?id=Eleutherine+bulbosa on 30 June 2016.

B.G. Brasileiro (2006). Brazilian Journal of Pharmaceutical Sciences vol.42,n.2,abr./jun.

N. Raaman (2006). Phytochemical Techniques, New India Publishing Agency, New Delhi, p.22.

C.K. Kokate (2013). Pharmacognosy, Nirali Prakashan, p. 7.18–7.19

N.Sasidharan (2004). Biodiversity documentation for Kerala, KFRI, Part 6: Flowering plants.

Henry Shaw School of Botany, Missouri Botanical Garden (1991). Annals of the Missouri Botanical Garden , v.78 Page 946–948. Accessed at http://www.biodiversitylibrary.org/item/ 89022#page/959/mode/1up. J.M.

Garg (2013). "Efloraofindia (2007 onwards). Database of Indian Plants developed by the members of Efloraofindia Google group. Accessed at https://sites.google.com/site/efloraofindi a/ on 30 June 2016."

Source of Support: NIL

Chand

S (2014). Importance of Pharmacognostic study of medicinal plants: An over view. Journal of Pharmacognosy and Phytochemistry; 2(5):69–73.

Sinhababu. A (2013). Ethnobotanical study of medicinal plants used by Tribals of Bankure districts, West Bengal. Wei Liu (2015). Influence of Ecological Factors on the Production of Active Substances in the Anti-Cancer Plant Sinopodophyllum hexandrum (Royle) T.S. Ying. PLoS One. 10(4). Conflict of Interest: None Declared

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