International Journal of Research ISSN NO:2236-6124

Phytochemical Characterization of the Anticancer Compounds in Ventilago denticulate

1Shakeel Ahmad Bhat, 2Dr Sanjay Telang 1Research Scholar, Professor Barkatullah University, Bhopal, M.P [email protected] 2Department of Zoology Govt. Science and Commerce College Benazir, Bhopal M.P. [email protected]

Abstract: The present research was carried with an aim to investigate the compound composition of the bark extract of V. denticulate , obtained as powder from Sanjeevni Ayurveda, Bhopal, M.P. The findings of the research led to the biochemical composition of the extract, which reported the presence of anthraquinones, naphthoquinones, naphthalenes, xanthone and naphthoquinone lactones in root bark. These compounds have antioxidative role, and have the potential of combating the oxidative stress related damage to kidney, liver, sperms and genetic makeup of patients receiving chemotherapy. Cisplatin induced hepatotoxity, nephrotoxicity, clastogenicity and spermiotoxicity was reported to be combated by the V. denticulate extract, which reveals its chemoprotective action.

Introduction: Ventilago is a of in the family . It includes about 35 found in the tropics of Australasia, with one species each in Africa and Madagascar. V. denticulata Wild commonly called the Red Creeper is an extensively branched, woody climber with hanging branches. The stem and root bark of this is a source of a red dye ‘ventilagin’, which is used for coloring cotton, wool and tasar. Stem bark when powdered and mixed with sesame oil, can be externally applied to treat skin diseases and sprains (Chopra, 1956).

Root bark is used for atonic dyspepsia, mild fever and debility. Sap is used for the treatment of deafness. The ethanolic extract of plant also shows anti-inflammatory activity

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(Akram et al ., 2009). The plant is rich in many pharmaceutical active ingredients. The stem bark contains friedelin and several anthraquinones. The root contains anthraquinones, ventinones A and B. Major Constituents of the root bark are emodin, its glucoside and corresponding analogues, ventiloquinones. The fruit, leaves and stem give lupeol, beta-sitosterol and its glucoside (Khare, 2007). Ventilago denticulate is reported for its antioxidant activity (Smitha et al. , 2012) . Ventilago denticulate is used in traditional system of medicine in treatment of various ailments (Jain et al. , 2010; Das and Mondal, 2012; Das and Mondal, 2013). From Sanjeevani ayurveda, Bhopal, it was also got to know that the bark of Ventilago denticulate is used in treatment of disease related to kidney. Patil and Patil (2005) also reported its traditional use in kidney problem. From literature survey it was observed that no scientific evidence is available for protective effect of Ventilago denticulate bark extract against any toxicant. Major toxicity associated with cisplatin (a chemotherapeutant) is nephrotoxicity, hepatotoxicity, clastogenecity, and deleterious effect on sperm. So, the present work was planned to investigate the photochemistry of Ventilago denticulate bark extract to investigate the compounds which have chemoprotective action. Materials &Methods: Collection of Plant materials Ventilago denticulate plants were collected from Sanjeevni Ayurveda, bhopal M.P. in powdered form. The authenticity of the plant was confirmed in Botanical Survey of India and Department of Botany, Barkatullah University Bhopal M.P. The V. denticulata bark were collected in bulk, cut in small pieces and dried for one month. The shade dried samples were powdered separately using an electrical grinder. The powder was stored in screw cap bottles until further analysis. Taxonomical classification Kingdom: Plantae Sub kingdom: Tracheobionta Super division: Spermatophyta Division: Magnoliophyta Sub division: Radiatopses Class: Magnoliopsida

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Sub class: Rosidae Order: Rhamnales Family: Rhamnaceae Genus: Ventilago Species: denticulate Preparation of plant material for extraction The dry V. denticulata bark powder was passed through sieve (100 microns). The coarse powdered Ventilago denticulate (250 g) was extracted in Soxhlet apparatus for 48 h with n- hexane+benzene+ethanol (50:25:25 ratio) (50-65°C, 2L). The extract obtained was concentrated under reduced pressure in rotatory evaporator below 60°C temperature to get semisolid sticky residue (20g). Physiochemical Examination (The Ayurvedic Pharmacopoeia of India, 1999; WHO, 1998) Foreign matter 150 gm of Ventilago denticulate powder was spread in a thin layer over petridish. The foreign material was separated by visual inspection using a magnifying lens (5–10X). The percentage of foreign matter was calculated. Ash value a) Total ash A tarred silica crucible was weighed & ignited. Approximately 3 gm of the sample was weighed into crucible, & then it was heated in a burner at 500°C till vapours almost ceased to evolve. It was heated at 1000°C until all carbon was burned off. After that it was cooled in desicator. The ash was weighed & percentage of total ash with reference to the air dried sample of the crude Ventilago denticulate powder was calculated. b) Acid insoluble ash The ash was dissolved in 25 ml dilute HCl. The solution was boiled for five minutes. After that the solution was filtered, using ashless filter paper, and washed twice with hot water. The crucible was burned in the flame, cooled & weighed. The filter paper and residue were together put into the crucible; heated in an oven until all carbon had been removed. Then it was cooled in desiccator. The residue was weighed and the acid insoluble ash of the crude Ventilago denticulate powder was determined. c) Water soluble ash

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The ash was dissolved in 25 ml water, and the solution was boiled for five minutes. The solutions were filtered through ashless filter paper, and washed twice with hot water. The crucible was burned in the flame, cooled & weighed. The filter paper and residue were together put into the crucible; heated in an oven until all carbon had been removed. Then it was cooled in desicator. The residue was weighed and the acid insoluble ash of the crude Ventilago denticulate powder was determined. d) Sulphated ash A tarred silica crucible was weighed and ignited. About 3 gm of the powdered Ventilago

denticulate powder was weighed & treated with 25 ml H 2SO 4 and boiled for 5-10 min. The solution was filtered & put into the crucible. It was then heated in a burner at 500°C till vapours almost ceased to evolve; then heated at 1000°C until all carbon was burned off. Then it was cooled in desicator and the ash was weighed & percentage of total ash was calculated with reference to the air dried sample of the crude Ventilago denticulate powder. Loss on drying About 2.0 gm of the Ventilago denticulate powder was weighed into a pre weighed flat & thin porcelain dish. The Ventilago denticulate powder was kept in oven at 100°C until unvarying weight was acquired. Afterwards, it was cooled in desicator & weighed. The reduction in weight was expressed as the moisture content. Extractive value Ethanol soluble extractive value About 3 gm of powdered Ventilago denticulate powder was weighed in a weighing bottle and transferred to a dry 250 ml conical flask. A 100 ml graduated flask was filled to the delivery mark with alcohol. The flask was corked & was shaken for 6 hours on vibrator. This was filtered into a 50 ml of measuring cylinder, when sufficient filtrate was collected; 25 ml of filtrate was transferred to a weighed thin porcelain dish. It was evaporated to dryness on a water bath & was completely dried in an oven at 100°C. This was cooled in desicator & weighed. The percentage w/w of extract was calculated with reference to the air dried Ventilago denticulate powder. Water soluble extractive value About 3 gm of powdered Ventilago denticulate powder was transferred to a dry 250 ml conical flask. The flask was filled to the delivery mark with water. The flask was corked & was shaken for 6 hours on vibrator. This was filtered into a 50 ml of measuring cylinder, when

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sufficient filtrate was collected; 25 ml of filtrate was transferred to a weighed thin porcelain dish. It was evaporated to dryness on a water bath & was completely dried in an oven at 100°C. This was cooled in desicator & weighed. The percentage w/w of extract was calculated with reference to the air dried Ventilago denticulate powder. Extraction In present study, extraction was performed using continuous hot percolation ‘Soxhlation’. Pulverised dried bark (total 1000 gm) of Ventilago denticulate was placed in thimble of Soxhlet apparatus. Extraction was performed at 40°C using petroleum ether as non polar solvent at first. Exhausted plant material was dried and afterward was extracted with chloroform, ethyl acetate, ethanol, and hydro-alcohol (70% ethanol). For each solvent soxhlation was continued till no color was observed in siphon tube and completion of extraction was confirmed till solvent from the siphon tube did not left any residue when evaporated. Obtained extracts were evaporated using rotary vacuum evaporator (Buchi type) at 40°C. Dried extract was weighed and percentage yield for each extract was determined using formulae: ℎ % yield = 100 ℎ Prepared extracts were observed for colour, odour, and texture, and were packed in air tight container and labeled till any further proceedings. Solubility of Extracts The solubility of Ventilago denticulate was observed in different solvents. Results: Phytochemical examination of Ventilago denticulate

Herbal medicine is a major element of almost all traditional systems like Ayurvedic, Homeopathic, Naturopathic, Traditional Chinese medicine, and Native American medicine (Chopra et al ., 1956). The commence of medicinal plants is flourishing globally day by day and besides there is also a huge demand of plant drugs all over the world, thus there might be a possibility of adulteration and substitution of the crude drugs with the genuine ones. Hence standardization of medicinal plants and natural products will provide useful information with regards to its correct identity and will help to differentiate from other closely related species as well as from other commercially available crude drugs.

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Ventilago denticulate (Rhamnaceae) commonly known as Rang Dhang, is found throughout India in hotter parts as a climbing shrubs or , on the trees (Zhang et al ., 2010; Khare, 2007). Flowering from October to December and fruiting December to April of following year. Previous phytochemical studies of Ventilago denticulate have reported the presence of several anthraquinones, naphthoquinones, naphthalenes, xanthone and naphthoquinone lactones in root bark (Hanumaiah et al ., 1984; Rao et al ., 1983, 1984; Rastogi et al ., 1999). Different parts of this plant possess multifarious medicinal properties. Until now, no scientific investigation has been carried out for standardization of Ventilago denticulate bark. Hence, there was a need to provide scientific proof for standardization by the pharmacognostical and phytochemical evaluation of the bark of Ventilago denticulate .

Fluorescence analysis

Fluorescence characteristic of powdered stem bark of Ventilago denticulate were observed for resolution of doubtful specimen. Powdered material was analyzed under visible light, short ultra-violet light, long ultraviolet after treatment with organic and inorganic reagents (Table 1).

Physicochemical parameters

Physicochemical parameters of any drug give an idea of the earthy matter and/or inorganic composition and/or other impurities present along with a drug. The total ash value, acid insoluble ash value, water soluble ash value and sulphated ash value of stem bark were found to be not more than 11.61, not more than 4.2, not more than 7.41 and not more than 2.1 respectively. The moisture content of the powdered drug was evaluated using loss on drying method and value observed was 3.18% w/w (Table 2). The dried and coarse powder of stem bark of Ventilago denticulate was extracted with the solvents of increasing polarity successively by soxhlet apparatus, while the aqueous extract was obtained by cold maceration method.

The bark of Ventilago denticulate has many phytochemicals, which have proven to be beneficial health wise. Table 3 depicts the different phytoconstituents in the bark extract of Ventilago denticulate . Results showed the presence of alkaloids (Wagner’s test), carbohydrates

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(Molish test), flavinoids (Shinoda’s test), phenolics (phenol test), polysterols (Salkowski’s test), saponins (Frothing test), steroids (Libermann-Buchard’s test) and terpenes (Salkowski’s test).

Percentage yield of various extracts of dried powder of Ventilago denticulate was done, the results of which are presented in table 4. Petroleum ether was a reddish brown powder, with percentage yield of 2.77% w/w. Similarly Chloroform extract was a brownish powder, with percentage yield of 2.66% w/w. On the same hand, ethyl acetate extract was a brownish powder, with percentage yield of 3.8% w/w. Ethanol extract was a dark brown syrup, with percentage yield of 5.8% w/w. While as water extract was a blackish brown powder with percentage yield of 8.0% w/w.

Phytochemical analysis

The dried and coarse powder of stem bark was tested for the presence of phytoconstituents using reported methods mentioned in the standards and results are given in Table 4. The preliminary phytochemical screening of various extracts was carried out and it was found that petroleum ether extract showed the presence of glycosides, flavonoids, phytosterols and phenolic compounds, chloroform extract showed the presence of glycosides, phenolic compounds, ethyl acetate extract showed the presence of alkaloids, flavonoids, glycosides, phenolic compounds and carbohydrates, ethanol extract showed the presence of carbohydrates, glycosides, phenolic compounds and flavonoids and aqueous extract showed the presence of alkaloids, carbohydrates, phenolic compounds, saponins and flavonoids. Total phenolic content of dried powder of Ventilago denticulate was done, the results of which are presented in table 5. The results were expressed as the mean±standard deviation of three readings and presented as gallic acid equivalent (mg/g). Petroleum ether extract content was 2.16±0.04, chloroform extract was 4.16±1.04, ethyl acetate extract was 9.12±1.14, while as ethanol extract was 7.16±1.16, with aqueous extract of 1.16±1.02.

Table 6 depicts the results for total flavinoid content of bark extract of Ventilago denticulate . The results were expressed as the mean±standard deviation of three readings and presented as gallic acid equivalent (mg/g). Petroleum ether extract content was 4.5±0.55, chloroform extract was 8.20±1.12, ethyl acetate extract was 10.1±0.26, while as ethanol extract

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was 6.5±1.3, with aqueous extract of 0.66±0.13. The present study dealt with the pharmacognostic and phytochemical investigation of stem bark of Ventilago denticulate . Ash values, extractive values can be used as reliable aid for detecting adulteration. The information obtained from preliminary phytochemical screening will be useful in finding out the genuineness of the drug. These simple but reliable standards will be useful to a lay person in using the drug as a home remedy. Also the manufacturers can utilize them for identification and selection of the raw material for drug production.

n-hexance constituents in Ventilago denticulate bark extract

The phytochemical constituent of bark of Ventilago denticulate are presented in Table 7. The study was not carried out during the current research period, however, the material was sought from the available literature to have an idea about the constituents in the concerned extract and their chemoprotective action.

Discussion: Ventilago denticulate

India is known for its wealth of medicinal plants which are found in its diverse climatic and physiographic conditions. Medicinal plants are widely used for the management of different disease conditions and to play a beneficial role in human health (Krishna et al ., 2017; Raaman, 2006). Plant derived drugs have a market of about 20 billion annually in the United State alone. It is also estimated that only 5-15% of potential useful plants have so for been systematically explored for useful chemical (Kalimuthu et al ., 2014). Plants are found to be sources of many chemical compounds, most of which account for their various uses by man. The most important of these compounds are alkaloids, terpenoid, steroid, phenolic compound, glycosides and tannin (Zahir and Kumaresan, 2014). Traditional folk treatment from wild plants has always guided researchers to search for novel medications to develop healthy life for humans and animals (Rahman et al ., 2017). In addition, some medicinal plants are still obscured within the plant which needs to be scientifically evaluated. The phytochemical research based on ethano- pharmacological information is considered an effective (Kalimuthu et al ., 2014).

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The chemical substances used by plants for approach in the discovery of new agents from higher plants defense system and serve as the bioactive principle for various drugs in modern chemotherapy (Cragg & Newman, 2005). Ventilago denticulata Willd is a Climbing shrub, stems sometimes twining. Leaves alternate, pinnately veined. V. denticulata Willd commonly called the Red Creeper is an extensively branched, woody climber with hanging branches. The stem and root bark of this plant is a source of a red dye ‘ventilagin’, which is used for coloring cotton, wool and tasar. Stem bark when powdered and mixed with sesame oil, can be externally applied to treat skin diseases and sprains. Root bark is used for atonic dyspepsia, mild fever and debility. Sap is used for the treatment of deafness. The ethanolic extract of plant also shows anti- inflammatory activity (Akram et al ., 2009). The plant is rich in many pharmaceutical active ingredients. The stem bark contains friedelin and several anthraquinones. The root contains anthraquinones, ventinones A and B. Major constituents of the root bark are emodin, its glucoside and corresponding analogues, ventiloquinones. The fruit, leaves and stem give lupeol, beta-sitosterol and its glucoside (Khare, 2007). The ethanolic extract from Rhang Dang leaves exhibited a strong antioxidant activity and prevented hemolysis. It showed the highest amount of phenolics (91.03 ± 12.43 mg of gallic acid equivalents/g extract) and flavonoid compound (69.76 ± 10.84 mg of catechin equivalents/g). Interestingly, this extract was more cytotoxic to HepG2 cells than PBMC (Pongjanta et al ., 2016). Considering the medicinal importance of Ventilago denticulata Willd, an attempt has been made to investigate the phytochemical composition from stem bark of V. denticulate . Natural products, either as pure compounds or as standardized plant extracts, provide unlimited opportunities for the new drug leads because of the unmatched availability of chemical diversity (Ujjwal et al ., 2008). The fact that the anticancer drugs used nowadays cause undesirable side effects, offer considerable potentials for the development of new effective chemoprotective antioxidant agents; medicinal plants are a prolific source (Teke et al ., 2011). In recent years, there has been a gradual revival of interest in the use of medicinal and aromatic plants in the developed as well as in developing countries, because plant derived drugs are reported to be safe and without any side effects especially when compared with synthetic drugs (Pandey et al ., 2003; Iniaghe et al ., 2009). During the present study, the preliminary phytochemical screening of various extracts was carried out and it was found that petroleum ether extract showed the presence of glycosides,

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flavonoids, phytosterols and phenolic compounds, chloroform extract showed the presence of glycosides, phenolic compounds, ethyl acetate extract showed the presence of alkaloids, flavonoids, glycosides, phenolic compounds and carbohydrates, ethanol extract showed the presence of carbohydrates, glycosides, phenolic compounds and flavonoids and aqueous extract showed the presence of alkaloids, carbohydrates, phenolic compounds, saponins and flavonoids. Various plant extracts contain many active compounds, which include alkaloids, tannins, flavonoids and phenolic compounds. Most of these secondary metabolites, in addition to possessing antimicrobial potential, can also act as potent antioxidants. Antioxidants are closely related to cancer and oxidative stress dysfunctions (Diplock, 1995; Black, 2000). While doing phytochemical screening of traditional medicinal plants in India, including V. denticulata Willd, Savithramma et al . (2011) reported secondary metabolites like saponins, triterpenoids, steroids, anthraquinons, coumarins, fatty acids, tannins, lignins, leucoanthocyanins, emodins, alkaloids, glycosides, flavonoids and phenols. The antioxidant properties of the bark of V. denticulate have been explored by Smitha et al . (2012). The ethanolic extract of V. denticulate plant shows anti-inflammation and anti-microbial activity, which was documented by Anursara et al . (2016). The authors reported that the ethanolic extract from Rhang Dang leaves exhibited a strong antioxidant activity and prevented hemolysis. It showed the highest amount of phenolics (91.03+12.43 mg of gallic acid equivalents/g extract) and flavonoid compound (69.76+10.84 mg of catechin equivalents/g). Kongara (2017) worked on the phytochemical analysis on leafs of Ventilago denticulata Willd . The authors reported that the leaves indicated the presence of steroids, resins, steroids, tannins, glycosides, reducing sugar, carbohydrates, saponins, terpenoids, acidic compounds, phenols, alkaloids, flavonoids. During the same work, GC-MS analysis of the Ventilago denticulata Willd extract result showed the presence of 14 compounds in the leaf extract by comparing their retention time and by interpretation of their mass spectra. Natthakaln et al . (2017) enumerated the antiacetylcholinesterase activity of Ventilago denticulata extracts and its chemical constituents and concluded that isolation and purification of V. denticulata root extracts afforded seven known quinone compounds: ventiloquinone 1, 2– hydroxyislandicin, chrysophanol, physcion, emodin, questin and ventilatone A, along with a large quantity of a mixture of common plant sterol, long chain fatty acids and tannin. Rajendra Kumar et al. (2018) explored the primary phytochemical study using gas chromatography-mass

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spectroscopy (GC-MS) and in vitro antimicrobial study (against gram positive bacteria and gram negative bacteria) on n-hexane(50%)+Benzene (25%)+25% ethanol stem extract of Ventilago denticulata . Preliminary phytochemical screening revealed that plant contains 17 bio active compounds with different concentrations. Qualitative analysis of the plant parts revealed the presence of various components of therapeutic importance including tannins, saponins, phenolic compounds, glycosides, flavonoids etc.

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[2] Anursara Pongjanta, Kanjana Pangjit, Somdet Srichairatanakool (2016):Antioxidant Activity and Cytotoxic Effect of Ventilago denticulata Willd Leaves Extracts. J Med Assoc Thai. 99 (Suppl. 1): S51-S57.

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[6] Das PK and Mondal AK. (2012): Biodiversity and conservation of some dye yielding plants for justification of its economic status in the local areas of lateritic zone of West Bengal, India. Advances in Bioresearch . 3 (1): 43 – 53. [7] Das PK and Mondal AK. (2013): Studies on the application of some less known ethnomedicinally dye yielding plants for dyeing in cotton cloth with focus on conservation Strategies in paschim me Dinipur district, west Bengal, India: a new approach. International Journal of Phytopharmacology . 4: 54-59.

[8] Diplock A (1995): Antioxidant nutrients – efficiency in disease prevention and safety. Biochemistry . 17: 16-18.

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[9] Hanumaiah T, Marshall DS, Rao BK, Rao JUM, Rao KVJ, Thomson RH (1985): Napthoquinonelactones and extended quinones from Ventilago calyculata . Phytochemistry . 24(11): 2669-72.

[10] Iniaghe OM, Malomo SO, Adebayo, JO (2009): Proximate composition and Phytochemical constituents of leaves of some Acalypha species. Pakistan Journal of Nutrition. 8: 256-258.

[11] Jain SP, Singh SC, Srivastava S, Singh J, Mishra NP, Prakash A. (2010): Hitherto unreported ethnomedicinal uses of plants of Betul district of Madhya Pradesh. Indian Journal Of Traditional Knowledge . 9(3): 522-525.

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[14] Khare GP. (2007): Indian Medicinal Plants: An Illustrated Dictionary.Springer Private Ltd. New Delhi India; 2007. p.697.

[15] Kongara Shobha Rani (2017): Phytochemical screening and GC-MS analysis of n-hexane/ethyl acetate leaf extract of Ventilago denticulata wild . Journal of Advanced Studies in Agricultural, Biological and Environmental Sciences (JABE). 4(2): 14-23.

[16] Krishna CS, Chandra R and Khaleel KM (2017): Phytochemical screening and GC-MS studies of Syzygium dhaneshiana leaf and bark extracts. Int J Pharm Sci Res . 8(5): 2277-81.

[17] Natthakaln Lomchoey, Jannarin Nontakham, Parichat Suebsakwong and Sunit Suksamrarn (2017): Antiacetylcholinesterase activity of Ventilago denticulata extracts and its chemical constituents. KKU Sci. J . 45(4): 701-713.

[18] Pandey, AK and Rai, MK (2003): Plant derived antimycotics: potential of Asteraceous plants. In: MK Rai, Mares D (Eds) Plant derived antimicotics , The Haworth Press, Inc. Binghamton, NY, USA, pp 343-344.

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[19] Patil MV and Patil DA (2005): Ethnomedicinal practices of Nasik District, Maharashtra. Indian Journal of Traditional Knowledge. 4(3): 287-290.

[20] Pongjanta A, Pangjit K, Srichairatanakool S. (2016): Antioxidant activity and cytotoxic effect of Ventilago denticulata Willd leaves extracts. J Med Assoc Thai. 99 (1): S51-7.

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[22] Rahman Gul, Syed Umer Jan, Syed Faridullah, Samiullah Sherani, and Nusrat Jahan (2017): Preliminary phytochemical screening, quantitative analysis of alkaloids, and antioxidant activity of crude plant extracts from Ephedra intermedia indigenous to Balochistan. The Scientific World Journal . 5873648.7 pages.

[23] Rao BK, Hanumaiah T, Rao CO, Rao GSR, Rao KVJ, Thomson RH. (1983): Anthraquinones in Ventilago species. Phytochemistry. 22 (11): 2583-5.

[24] Rao BK, Hanumaiah T, Rao JUM, Rao KVJ, Thomson RH. (1984): Two anthraquinones from Ventilago calyculata . Phytochemistry . 23(9): 2104-5.

[25] Rastogi RP, Mehrotra BN. (1999): Compendium of Indian Medicinal Plants: Drug research perspectives: A CDRI series. Volume 3, National institute of science communication. CSR, New Delhi. 667-8.

[26] Savithramma, N., M. Linga Rao and S. Ankanna (2011): Phytochemical screening of traditional medicinal plants. Journal of Pharmacy Research. 4(10): 3414-3416. [27] Smitha P. Venkata, Mohan Ch. Murali (India), Jaime A. Teixeira da Silva (Japan), B. Akondi Raju, R. Sravani (India) (2012): Screening the antimicrobial and antioxidant potential of Ventilago denticulata , Scolopia crenata and Rivea hypocrateriformis from Maredumilli Forest, India. 6: 58-62. [28] Smitha P. Venkata, Mohan Ch. Murali (India), Jaime A. Teixeira da Silva (Japan), B. Akondi Raju, R. Sravani (India) (2012): Screening the antimicrobial and antioxidant

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potential of Ventilago denticulata , Scolopia crenata and Rivea hypocrateriformis from Maredumilli Forest, India. 6: 58-62.

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Table 1: Fluorescence characteristics of dried powder of Ventilago denticulate

Powder + Reagent Color observed in ordinary Colour observed under UV Colour observed under UV light short (254 nm) Long (365 nm) Powder as such Brown Green Green Powder + 1N NaOH in water Dark green Brownish green with black Yellow with white spots spots Powder + 1N HCl Yellow with pinkish spots Yellow Yellow with black spots Powder + 50% H 2SO 4 Brown Yellow Yellow with pink spots Powder+Formaldehyde White Brown Yellow Powder + 5% Ferric Chloride Brown Brown Black Powder+Iodine Black Yellow Black

Table 2. Phytochemical evaluation of Ventilago denticulate

S.N. Properties Stem bark (% w/w) 1. Foreign Matter 0.62% 2. Ash Value a) Total ash Not more than 11.61 b) Acid insoluble ash Not more than 4.2 c) Water soluble ash Not more than 7.41 d) Sulphated Ash Not more than 2.1 3. Loss on drying 3.18 4. Extractive Value a) Ethanol soluble extractive value mg GAE/g extract 91.03±12.43 b) Water soluble extractive value mg GAE/g extract 21.88±1.9

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Table 3: Display the presence/absence of different phytochemicals in the bark extract of Ventilago denticulate

Phytoconstituents Test n-Hexane/Ethyl Acetate Alkaloids Wagner’s test ++ Amino Acids Ninhydrin test -- Carbohydrates Molish test ++ Cardiac glycosides Keller-Killani test -- Flavonoids Shinoda’s test ++ Phenolics Phenol test ++ Polysterols Salkowski’s test ++ Proteins Biuret test -- Saponins Frothing test ++ Steroids Libermann-Buchard’s test ++ Tannins Ferric chloride test -- Terpenes Salkwaski’s test ++

Table 4: Nature and percentage yield of various extracts of dried powder of Ventilago denticulate

Extracts Colour and consistency of Percentage yield (% w/w) extract Petroleum ether Reddish brown powder 2.77 Chloroform Brownish powder 2.66 Ethyl acetate Brownish powder 3.8 Ethanol Dark brown with syrupy 5.8 consistency Water Blackish brown powder 8.0

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Table 5: Total phenolic content of bark extracts of Ventilago denticulate

S.N. Plant extract Gallic acid equivalent (mg/g) 1. Petrolem ether 2.16±0.04* 2. Chloroform 4.16 ±1.04* 3. Ethyl acetate 9.12±1.14* 4. Ethanol 7.16±1.16* 5. Aqueous 1.16±1.02 * values are means of triplicate determination±standand deviation

Table 6: Total flavonoid content of bark extracts of Ventilago denticulate

S.N. Plant extract Gallic acid equivalent (mg/g) 1. Petrolem ether 4.5±0.55* 2. Chloroform 8.20±1.12* 3. Ethyl acetate 10.1±0.26* 4. Ethanol 6.5±1.3* 5. Aqueous 0.66±0.13* * values are means of triplicate determination±standand deviation

Volume VIII, Issue II, February/2019 Page No:270 International Journal of Research ISSN NO:2236-6124

Table 7: Components detected in n-hexane extract of Ventilago denticulate bark extract

S.N. RT % of Compound name MF MW Nature of compound Medicinal importance area Derived from literature

1. 4.4 16.27 L-(+) ascorbic acid; C6H8O6 176.03 Saturated carboxylic acid Vitamin C, Antioxidant, Vitamin C Immunomodulator

2. 8.8 8.54 9,12-Octadecadienoic C18 H32 O2 280.44 Ethyl Ester/Fatty Acid Anti-inflammatory, acne reductive, acid (Z,Z)- antioxidant

3. 11.1 14.7 Methyl hexadecanoate C17 H34 O2 270.45 A saturated fatty acid that Antioxidant or Palmitic acid is the major fat in meat methyl ester and dairy

4. 11.5 1.57 Dronabinol C21 H28 O4 344.44 Steroid Analgesic, anti-inflammatory 5. 20 10.07 Phytol Acyclic diterpene alcohol Antimicrobial, antioxidant, cancer preventive 6. 25.4 4.57 Squalene C30H50 410.7 Defydrotriterpenic Antibacterial, antioxidant, hydrocarbon antitumour, cancer preventive, chemoprotective, immunostimulants and lipoxygenase inhibitor #Source: Dr. Duke's phytochemical and ethnobotanical databases [Online database].

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