PHYTOCHEMISTRY, ANTI-CANCER AND ANTI-INFLAMMATORY ACTIVITIES OF SOLVENT LEAF EXTRACTS OF NYCTANTHES ARBOR – TRISTIS

PHYTOCHEMISTRY, ANTI-CANCER AND ANTI-INFLAMMATORY ACTIVITIES OF SOLVENT LEAF EXTRACTS OF NYCTANTHES ARBOR - TRISTIS

T. D. Sandhya Kumari ¹ and M. A. Singara Charya ^{* 2}

Department of Microbiology ¹, Quality Control, Strides Shasun Ltd., Cuddalore, Tamilnadu - 607002, India.

Department of Microbiology ², Kakatiya University, Warangal - 506009, Telangana, India.

ABSTRACT: Ayurveda is one of the oldest systems of medicine that uses plants and their extracts for treatment and management of various diseased states. Nyctanthes arbour - tristis Linn. (Family Oleaceae), which has long been used traditionally for various medicinal purposes in the Indian sub continent. Each part of the plant has some important medicinal value and is thus commercially exploitable. It is now considered as a valuable source of several unique products for the medicines against various diseases and also for the development of some industrial products. The present study is to focus on the potential phyto-chemicals and pharmacological activity of plant N. arbor - tristis. In view of this, the study was designed to investigate for chemical constituents, anti-cancer and anti-inflammatory activities of solvent leaf extracts.  Phytochemical screening revealed the presence of alkaloid, steroids etc. The extract exhibited significant anti-cancer and anti-inflammatory properties and the observed biological activities in this study provide scientific validation of ethnomedicinal use of this plant.

N. arbour - tristis, Pharmacological activity, Phytochemical screening, Anti-cancer, anti-inflammatory

INTRODUCTION: The Greek word for a plant is ‘Phyto’ and chemicals produced by plants are called phytochemicals/fight-o-chemicals. In plant flora, 25,000 phytochemicals were known to exist and of them 10,000 are estimated to be alkaloids and 4,000 flavonoids distributed all around ¹. The medicinal value of plants lies in chemical substances that possess physiological action. Higher plants have unique and rich collection of various biochemicals, so they are generally known as storehouses of chemo-therapeutants. As they possess lesser side effects, they can be used to develop natural drugs.

Secondary metabolites are chemically and taxonomically diverse group of compounds with obscure functions extensively used in research, agriculture, human therapy and so on ². They are produced from primary metabolites such as amino acids, carbohydrates and proteins with a series of metabolic reactions. The important phytochemicals in medicinal plants utilized both by humans and by animals are  alkaloids, flavonoids, glycosides, tannins, phenols, steroids, resins, saponins and so on ³. The disease preventing nature is due to the presence of antioxidants, such as flavonoids, hydrolysable tannins, phenolic acids and so on. This reacts against free radicals responsible for cancer, coronary heart diseases, mutagenic and inflammation reactions ⁴.

Nyctanthes arbor-tristis (Night-Flowering Jasmine) is an erect shrub or a small tree of the family of Oleaceae growing upto 10m Fig.1. Nyctanthes arbor-tristis Linn. is a widely spread plant from Northern Pakistan, Nepal, South and North India and Thailand.  N. arbor-tristis is normally planted by cuttings or generates from seeds. The plant sheds its leaves during the period March to May and it puts up flowers from September to November. It usually grows in red as well in black soils, where climatic conditions of arid and semi-arid are prevalent.

FIG. 1: FRESH TWIG OF NYCTANTHES ARBOR - TRISTIS

N. arbor-tristis is a beautiful and fragrant plant. Its flowers bloom at night, drop off and fall early next morning for this reason it is called as “sad tree”. It is mainly characterized by the presence of phenylethanoid derivatives and iridoid derivatives ⁵. It is used in traditional medicine as stomachic, carminative, intestinal astringent, expectorant, in biliousness, piles, and various skin diseases and as hair tonic ⁶. It has also been reported to possess hepato-protective, anti-viral, antifungal ⁷ and analgesic, anti-pyretic, ulcerogenic activities ⁸.

Bansal et al., (2015) ⁹ reported that the bioactive compounds of Nyctanthes arbor-tristis  are very useful to provoke menstruation, treatment of scabies and other skin infections, as hair tonic, chalogogue, laxative, diaphoretic, diuretic, treatment of arthritis, malaria, bronchititis and anti-helminthic.

In view of these reports, the present study was undertaken to investigate the phytochemistry and, anti-cancer, anti-inflammatory activities of solvent leaf extracts of Nyctanthes arbor-tristis for its diverse pharmacological applications in ancient and modern system of medicine.

MATERIAL AND METHODS:

Plant Material Collection and Authentication:

Fresh plant leaves   of Nyctanthes arbour - tristis were collected from Warangal, Telangana state, India and were identified and authenticated by Botanical Survey of India, Coimbatore, Voucher Reference No. BSI/SRC/5/23/2011-12/Tech. 1443.

Organic Extraction: The fresh leaves of Nyctanthes arbor-tristis were shade-dried for 10 days at 37 ⁰C, and then finely powdered. The powder was exhaustively extracted with solvents in 1:5 ratio using soxhlet apparatus for 6-8 hours and then   centrifuged. From the centrifuged extract the supernatant was filtered over Whatman No. 1 filter paper. The filtered extract was then subjected to dryness under reduced pressure at 37 ⁰C (not exceeding 40 ^oC), until usage DMSO was added and stored at -80 ⁰C. Extracts diluted with culture media were sterilized by 0.22 µm membrane filters prior to cancer cell lines treatment. In culture medium, less than 0.2% DMSO final concentration was used. All the extracts were stored in a dessicator for further evaluation. The crude extracts were analysed for their phytochemical, thin layer chromatography and pharmacological activities.

The leaf extracts were used for phytochemical analysis as per the following standard procedures for the identification of various classes of active plant constituents ^{10, 11}.

Medium: 10% MEM possessing l-glutamine (4 mmol/L), 10% fetal bovine serum (FBS) and antibiotics of 100 μg /mL streptomycin and 100 units/mL of penicillin must be added and incubated in a humidified atmosphere containing 5% CO₂ with 95 % air at 37 °C.

Cell Lines: MDA-MB 231 (Breast Cancer Cell Lines free of pathogens) and A 549 (Lung Cancer Cell Lines) purchased from ATCC.

Anticancer activity: A colored formazan product is formed by the yellow coloured 3-(4,5-dimethythiazol-2-yl) - 2, 5 - diphenyl tetrazolium bromide (MTT) reduction, which were taken up by the cells with the detection of the readings  using spectrophotometry (λ_max = 562 nm). Based on the mitochondrial respiratory function, the MTT is reduced, by measuring the number of viable cells in the culture.

Cancer cells were treated with various concentrations (0, 10, 15, 20 and 30 μg) of the compound for 48h, washed with Dulbecco’s phosphate buffered saline (DPBS), the media is aspirated at the end of the treatment and incubated with 20μl of 5 mg/ml MTT solution in 500 μl of culture medium for 1 h at 37 ^oC. In order to solubilize the cells, 500 μl of formazan DMSO was used. By using automated TECAN multimode reader the absorbance was measured at 562 nm. The experiment is conducted in duplicates and triplicates by comparing the test results with the control in which the drug is omitted ¹².

Anti-inflammatory Activity:

Experimental Animals: Using standard laboratory conditions (12: 12 h light/dark cycle at 24 ^oC) the experiment was conducted on Male albino wistar rats of 130 to 150 g in weight. The rats were kept in polypropylene cages. Water ad libitum and commercial rat diet (NIN, Hyderabad) were provided to the rats. Approval has been given by Institutional Animal Ethical Committee of IICT to conduct experiments. The animals are quarantined first, and then acclimatized to laboratory conditions for a week. During this period of testing, the animals were observed for general health and suitability.

1. Compound Dose – 250mg/kg
2. STD (Indomethacin) -10 mg/kg

Carrageenan Induced Rat Paw Edema Model: By employing a slight modification in the method described by Winter et al., (1962) ¹³ the evaluation of test compounds in wistar rats was screened for anti-inflammatory activity. The overnight fasted animals were divided into three groups–standard, control and different test groups. A dosage of 10 mg/Kg of Indomethacin was administered orally to the standard group animals. Before this, a dosage of 250 mg/Kg/day of test compound for 10 days was administered by oral route. A suspension was made with gum acacia and weighed test compound in required quantity. Every day, at a specific time interval, the prepared suspension was orally administered to all the rats throughout the study. The control group of rats received the vehicle solution without test compound. On termination day, the test drugs are administered to the entire group of rats. An hour later they are challenged with 0.1ml of 1% Carrageenan in the sub plantar region of right hind paw. After this, the volumes of the rats’ paws were measured before and after an hour each for 3h using digital plesthysmometer (Ugo Basile, Italy). The mean paw volume of control group was compared with treated groups for calculating the percent inhibition of paw volume.

% Inhibition = Vc – Vt /Vc

(Vc: Control Mean paw volume and Vt: Test paw volume)

RESULTS: The phytochemical analysis of the dried leaves of N. arbor-tristis revealed that the alkaloids were present in the solvent leaf extracts of hexane, chloroform, petroleum ether and ethyl acetate (Table 1).  Flavoinods were only traced in the extract of petroleum ether, while steroids were identified only in methanol. Saponins were observed in the solvent extracts of chloroform and ethyl acetate. Glycosides, tannins, phenols, resins were completely absent in all the solvent extracts of N. arbor-tristis. 

The TLC analysis clearly indicated that the solvent extracts of dried leaves of hexane was with brown, blue, purple and yellow colours having R_f values ranging from 0.20 to 0.96, while chloroform extracts were  with green colour and R_f values were 0.50 and 0.53 (Table 2). The dried leaf extract with petroleum ether of N. arbor-tristis was with green and violet colours and R_f range was 0.18 to 0.74.  The ethyl acetate extracts were with green colour and R_f range was 0.11 to 0.53. The methnol extracts of dried leaves of N. arbor-tristis showed green and ash colours with R_f values between 0.21 to 0.75. The chemical elucidation of the methonolic leaf extract of Nyctanthes arbor-tristis was carried out and presented.  There are several peaks observed in chromatograms for the presence of various compounds between Rt 2.474 to 15.294 min Fig. 2.

A peak observed at Rt 2.781 min corresponded to be (1S,4aS,5R,6S,7S,7aS)-ethyl1,5,6-trihydroxy-7- (propionyloxy)- 1, 4a, 5, 6, 7, 7a- hexahydrocyclo -penta[c] pyran-4-carboxylate, characterized as glycoside in NAT leaves and a second peak observed at Rt 2.530 min corresponded to be Methylenenona-2,5-dien-4-yl) - 4, 4, 10, 13, 14-pentamethyl-2,3,4,7,8,9,10, 11,12,13,14,15,16,17 – tetradecahydro - 1H cyclo penta [a] phenanthren-3- ol, characterized as steroid in  leaves of methanolic extract.

The HPLC and mass spectrometry of leaf extract I revealed glycoside isolated structure had the molecular weight of 317 which corresponds to a molecular formula, C₁₄H₂₀O₈ as represented in Fig. 3. Mol. wt = 317 (C₁₄H₂₀O₈), (M+= 317, 100%)

Predicted structures: (1S, 4aS, 5R, 6S, 7S, 7aS)-ethyl1,5,6-trihydroxy-7-(propionyloxy)-1,4a, 5,6, 7,7a-hexahydrocyclopenta[c]pyran-4-carboxylate. Mol. Wt: (M+H) = 317 (C₁₄H₂₀O₈)

GLYCOSIDE

The HPLC and mass spectrometry of leaf extract II revealed steroid isolated structure had the molecular weight of 477 which corresponds to a molecular formula, C₃₄H₅₄O₁ as represented in Fig. 4. Mol. wt = 477 (C₃₄H₅₄O₁), (M-= 477, 100%)

Predicted structures: Methylenenona-2,5-dien-4-yl)-4,4,10,13,14-pentamethy 2,3,4,7,8, 9, 10, 11, 12, 13,14,15,16,17-tetradecahydro- 1H - cyclopenta [a] phenanthren-3-ol. Mol. wt: (M-H) = 477 (C₃₄H₅₄O₁)

STEROID

The HPLC and mass spectrometry revealed phenol isolated structure had the molecular weight of 331 which corresponds to a molecular formula, C₂₁H₁₄O₄. Mol. wt = 331 (C₂₁H₁₄O₄), (M+= 331, 100%).

Predicted structures: 8-(5,7-dihydroxy-3-vinyl naphthalen-1-yl) – 2 H - chromen-2-one Mol. wt: (M+H) = 331 (C₂₁H₁₄O₄).

PHENOL

The HPLC and mass spectrometry revealed glycoside isolated structures had the molecular weights of 552.52 (2 compounds), 566.55, 568.52 and 536.53 (2 compounds) which corresponds to the molecular formulas, C₂₆H₃₂O₁₃, C₂₇H₃₄O₁₃, C₂₆H₃₂O₁₄, C₂₆H₃₂O₁₂, as represented in Fig. 5.

The TLC analysis clearly indicated that the solvent extracts of dried leaves of hexane was with brown, blue, purple and yellow colours having R_f values ranging from 0.20 to 0.96, while chloroform extracts were  with green colour and R_f values were 0.50 and 0.53. The dried leaf extract with petroleum ether of N. arbor-tristis was with green and violet colours and R_f range was 0.18 to 0.74.  The ethyl acetate extracts were with green colour and R_f range was 0.11 to 0.53.  The methnol extracts of dried leaves of N. arbor-tristis showed green and ash colours with R_f values between 0.21 to 0.75.

The methonolic leaf extract of Nyctanthes arbor-tristis was further investigated for its anti-cancer, anti-inflammatory activities.

The effect of N. arbor-tristis leaves extracted in methanol were evaluated using MTT assay on human cancer cell lines. The growth inhibitory potential of N. arbor-tristis extracts was screened against two cancer cell lines which are of epithelial origins.

The cell lines such as lung (A549) and breast (MDA MB-231) represents the major organ sites. Hence, they were selected for testing the effect of N. arbor-tristis extracts on their proliferation.

A solvent control (methanol) in different concentrations (10ml, 20ml and 30ml) were tested against MDA-MB 231 human breast cancer cell lines and checked for cell viability percentage. The percentage of cell viability decreased with increased concentration of methanol solvent (Table 3). The 10ug of methanol solvent concentration showed 83.1% and 20 ug showed 79.0% of cell viability while 30 ug caused for 71.8% cell viability. Similarly the decrease in cell viability was also examined with cell line A549 (Table 4) and noticed that the 10 ug was responsible for 91.8%, 20 ug was for 74.1% while 30 ug was showed 68% of decrese in cell viability. DMSO was used as another control showed no activity against cell lines. The cancer cell lines sensitivity was determined by using a positive control, etoposide. The potency of N. arbor-tristis extracts was compared with the conventional anti-cancer drugs by using this anti-proliferative assay. The control used to screen the cancer cell lines susceptibitility is etoposide and observed that all the cell lines were susceptible to standard drug treatment. A comparison made between anti-proliferative activity of N. arbor-tristis extracts and standard anti-cancer drug, showed that the growth arresting activity of N. arbor-tristis was higher than that of etoposide, and thus displayed its efficacy as a cancer preventing agent.

The anti-inflammatory activity of methonolic extract of Nyctanthes arbor-tristis of the carrageenan induced wistar rats in terms of their body weight index with control and NAT methanol leaf extract were compared on the 1^st and the 11^th day and found no significant change. On the 1^st day the control is 154.80±2.63 and on the termination day159.95±2.42 whereas the extract is 156.68±2.94 on the 1^st day and 158.82±0.03 on the termination day (Table 5).

TABLE 1:  PHYTOCHEMICAL ANALYSIS OF LEAF EXTRACTS OF NYCTANTHES ARBOR - TRISTIS  

TABLE 2: THIN LAYER CHROMATOGRAPHY OF SOLVENT EXTRACTS OF LEAF OF N. ARBOR-TRISTIS

TABLE 3: EFFECT OF DIFFERENT CONCENTRATIONS OF METHANOL EXTRACTS OF N. ARBOR-TRISTIS AGAINST MDA MB-231 HUMAN BREAST CANCER CELL LINES

^a All data were average (± SD) of three replicates.

TABLE 4: ANTICANCER ACTIVITY OF N. ARBOR-TRISTIS AGAINST A-549 HUMAN LUNG CANCER CELL LINES

TABLE 5: ANTI-INFLAMMATORY ACTIVITY OF METHANOLIC EXTRACT OF NYCTANTHES ARBOR-TRISTIS (BODY WEIGHT INDEX)

Values are mentioned as Mean ± S.E.M (n=6).

FIG. 2: HPLC CHROMATOGRAM OF METHANOL FRACTION OF N. ARBOR - TRISTIS LEAF EXTRACT

FIG. 3: MASS SPECTRUM OF METHANOLIC LEAF EXTRACT I OF NYCTANTHES ARBOR - TRISTIS

FIG. 4: MASS SPECTRUM OF METHANOLIC LEAF EXTRACT II OF NYCTANTHES ARBOR - TRISTIS

FIG. 5: MASS SPECTRUMS OF METHONOLIC LEAF EXTRACT OF NYCTANTHES ARBOR-TRISTIS

DISCUSSION: Number of reports are available on the phytochemistry and other pharmacological assessment of leaf extract of Nyctanthes arbor-tristis ^14-22. Saxena et al., (1984) ¹⁴ screened the water soluble portion of the alcoholic extract of the leaves of Nyctanthes arbor-tristis for its anti-inflammatory activity.  The extract inhibited the acute inflammatory oedema produced by different phlogistic agent viz. carrageenan, formalin, histamine, 5-hydroxytryptamine and hyaluronidase in the hind paw of rates.  Acute and chronic phases of formaldehyde induced arthritis were significantly inhibited.

Puri et al., (1994) ²³ reported the hepato-protective, anti-leishmania, anti-viral, anti-fungal activities of ethanolic extracts of flowers and leaf of Nyctanthes arbor-tristis. A strong stimulation of antigen specific and non-specific immunity and increase in humoral and delayed type of hyper sensitivity was demonstrated in mice fed with 50% ethanolic extracts. Singh et al., (1995) ²⁴ examined the leaves of Nyctanthes arbor-tristis led to isolation and identification of a new minor iridoid glucoside, arborside - D as its acetyl derivative.  They concluded that the structure of arborside D as 10-benzoylnyctanthoside.  Similar to the present observations, Khatune et al., (2001) ²⁵ recorded cytotoxic activity with chloroform and ethyl acetate flower and leaf extacts of Nyctanthes arbor-tristis.  The fractionation of an ethanol extract from the leaf and flowers of Nyctanthes arbor-tristis to the isolation of an antiplasmodial cyclohexylethanoid; a new iridoid glucoside, 6-0-trans cinnamoyl, 7-0-acetyl 6-b-hydroxyloganin and three known iridoid glucosides were elucidated and reported ¹⁵.

Singh et al.,  (2010) ²⁶ studied the immune-pharmocological properties of ethanolic extracts of Nyctanthes arbor-tristis and after administration of ethonolic leaf extracts in doses of 0.25 and 0.5 g/kg body weight, a significant increase in phagocytic index, leucocyte count and spleenic antibody cells were noticed. They further, identified through HPLC the presence of methoxylated flavonoid quercetin - 3, 3’ dimethoxy - 7 – O -rhamnogluco pyranose. The phytochemical investigations of solvent extracts of leaf of Nyctanthes arbor-tristis revealed the presence of alkaloids, carbohydrates, glycosides, phytosterols, fixed oil, tannins, flavonoids, proteins and amino acids, gums and mucilages in varied quantities (Suresh and Arunachalam, 2012) ²⁷.  Rani et al., (2012) ²⁸ recorded Nyctanthes arbor-tristis as one of the most useful traditional medicinal plant in India and reported its distribution widely in sub-himalayan regionsand southwards to Godavari. Each part of the plant has some medicinal value and is thus commercially exploitable. Further, they reported on the comprehensive information on chemical constituents, biological activities of important compounds, pharmacological actions, medical applications and micro propagations.

Agarwal and Pal (2013) ²⁹ noticed that the crude extracts and isolated compounds from Nyctanthes arbor-tristis were shown to be pharmacologically active against inflammation, malaria, viral infection, leishmaniasis and immuo-stimulant.  They isolated arbortristoside A, B and C and other molecules such as, calceolarioside A, 4-hydroxy-hexahydrobenzofuran-7one and beta-sitosterol and demonstrated their active role as anti-leishmanial, anti-cancer and anti-inflammatory activities.   

Recently,  Santhosh and Patil (2016) ²² presented a review on Nyctanthes arbour - tristis and recorded phytochemicals like flavanol glucoside, oleanic acid, essential oils, tannic acid, carotene, friedeline, lupeol, glucose, benzoic acid for their significant hepato-protective, anti-leshmaniasis, anti-viral, anti-fungal, anti-pyritic, anti-histaminic, anti-malarial, anti-bacterial, anti-inflammatory, anti-oxidant activities.

Thus, the present study has shown the anti-cancer and anti-inflammatory activities of leaf of Nyctanthes arbor-tristis and suggested its therapeutic usefulness. The plant is rich resource of biologically active compounds which would attract the attention of drug discovery groups to discover novel bioactive molecules for safer and effective treatment of various diseases.

CONCLUSION: Nyctanthes arbour-tristis widely used in traditional system of medicine for varied ailments are supported by various studies involving its pharmacological evalutions. The above article documented and revealed phytochemicals study and pharmacological activities of Nyctanthes arbor -tristis and proved as a unique source of metabolites such as alkaloids, phytosterols, phenolics, tannins, flavonoids, glycosides and saponins which is obtained from crude extract of plant and showed effective anti-cancer and anti-inflammatory.

CONFLICT OF INTEREST: The authors declare that there is no conflict of interest regarding the publication of this paper.

REFERENCES:

1. Singh A, editor. Herbalism, phytochemistry and ethnopharmacology. CRC Press; 2011.
2. Goud JV, Suryam A, Charya MS. Biomolecular and phytochemical analyses of three aquatic angiosperms. African journal of microbiology research. 2009; 3: 418-21.
3. Okwu DE, Josiah C. Evaluation of the chemical composition of two Nigerian medicinal plants. African Journal of Biotechnology 2006; 5: 357.
4. Aruoma OI, Halliwell B. Free radicals and food additives. Taylor & Francis Ltd.; 1991.
5. Jensen SR, Franzyk H, Wallander E. Chemotaxonomy of the Oleaceae: iridoids as taxonomic markers. Phytochemistry. 2002; 60: 213-31.
6. Khatune NA, Mosaddik MA, Haque ME. Antibacterial activity and cytotoxicity of Nyctanthes arbor-tristis Fitoterapia. 2001; 72: 412-4.
7. Puri A, Saxena R, Saxena RP, Saxena KC, Srivastava V, Tandon JS. Immunostimulant activity of Nyctanthes arbor-tristis Journal of ethnopharmacology 1994; 42: 31-7.
8. Rani C, Chawla S, Mangal M, Mangal AK, Kajla S, Dhawan AK. Nyctanthes arbor-tristis (Night Jasmine): A sacred ornamental plant with immense medicinal potentials. Indian J Tradit Knowl 2012; 11: 427-35.
9. Bansal G, Suri KA, Gorver, A comprehensive review on Nyctanthes arbor-tristis, J. Drug Dev. & Res. 2015; 7:  1-8.
10. Gibbs RD. Chemotaxonomy of flowering plants. Vols I-IV. McGill-Queen's univ. Press: Montreal & London. 1974 1: 1-680.
11. Trease, G. E. and Evans, W. C. 1989. A Textbook of Pharmacognosy. Bacilliere tinal Ltd., London, 13th edition
12. Kotamraju S, Willams CL, Kalyanaraman B. Statin-induced breast cancer cell death: role of inducible nitric oxide and arginase-dependent pathways. Cancer research 2007; 67: 7386-94.
13. Winter CA, Risley EA, Nuss GW. Carrageenin-induced edema in hind paw of the rat as an assay for antiinflammatory drugs. Experimental Biology and Medicine 1962; 111: 544-7.
14. Saxena RS, Gupta B, Saxena KK, Singh RC, Prasad DM. Study of anti-inflammatory activity in the leaves of Nyctanthes arbor- tristis—an Indian medicinal plant. Journal of Ethnopharmacology. 1984; 11: 319-30.
15. Tuntiwachwuttikul P, Rayanil K, Taylor WC. Chemical constituents from the flowers of Nyctanthes arbor-tristis. Science Asia 2003; 29: 21-30.
16. Mahida Y, Mohan JS. Screening of plants for their potential antibacterial activity against Staphylococcus and Salmonella spp. Natural Product Radiance 2007; 6: 301-5.
17. Vankar PS, Shanker R, Mahanta D, Tiwari SC. Ecofriendly sonicator dyeing of cotton with Rubia cordifolia using biomordant. Dyes and pigments 2008; 76: 207-12.
18. Akki KS, Krishnamurthy G, Bhojanaik HS. Phytochemical investigations and in vitro evaluation of Nyctanthes arbor-tristis leaf extracts for antioxidant property. Journal of Pharmacy Research. 2009; 4:65-78.
19. Verma NS, Dwivedi S, Panigrahi D, Gupta SK. Anti-bacterial activity of root bark of Nyctanthes arbor-tristis Int J Drug Discov Herb 2011; 2: 61-62.
20. Michael JS, Kalirajan A, Padmalatha C, Singh AR. In vitro antioxidant evaluation and total phenolics of methanolic leaf extracts of Nyctanthes arbor-tristis Chinese journal of natural medicines 2013;11:484-7.
21. Khanapur M, Avadhanula RK, Setty OH. In vitro antioxidant, antiproliferative, and phytochemical study in different extracts of nyctanthes arbor-tristis BioMed research international. vol. 2014, Article ID 291271, 10 pages, 2014. doi:10.1155/2014/291271.
22. Santosh J, and Patil M. A review on: Nyctanthes arbortristis Rejuvinating herbs. International Journal of Research in Pharmacy and Pharmaceutical Sciences 2016; 1: 54-62.
23. Puri A, Saxena R, Saxena RP, Saxena KC, Srivastava V, Tandon JS. Immunostimulant activity of Nyctanthes arbor-tristis Journal of ethnopharmacology 1994; 42:31-7.
24. Singh KL, Roy R, Srivastava V, Tandon JS, Mishra A. Arborside D, a minor iridoid glucoside from Nyctanthes arbor-tristis. Journal of Natural Products 1995; 58:1562-4.
25. Khatune NA, Mosaddik MA, Haque ME. Antibacterial activity and cytotoxicity of Nyctanthes arbor-tristis Fitoterapia 2001; 72:412-4.
26. Singh D, Singh SK, Maurya VB, Prajapati K, Kumar H, Niranjain PS. Evaluation of anticonvulsant activity of the leaves ethanolic and aqueous extracts of Nyctanthes arbortristis against seizures induced by PTZ and electroconvulsive shock in mice. Int J Pharm Sci Res 2010; 1:63-71.
27. Suresh V, Arunachalam G. Pharmacognostical and preliminary phytochemical studies of bark of nyctanthes arbor–tristis Int J Pharm Sci 2012; 4(Suppl 1):356-63.
28. Rani C, Chawla S, Mangal M, Mangal AK, Kajla S, Dhawan AK. Nyctanthes arbor-tristis (Night Jasmine): A sacred ornamental plant with immense medicinal potentials. Indian J Tradit Knowl 2012; 11:427-35.
29. Agrawal J, Pal A. Nyctanthes arbor-tristis Linn—A critical ethnopharmacological review. Journal of ethnopharmacology 2013; 146:645-58.
    

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    How to cite this article:

    Kumari TDS  and  Charya MAS: Phytochemistry, anti-cancer and anti-inflammatory activities of solvent leaf extracts of Nyctanthes arbor-tristis. Int J Pharm Sci Res 2017; 8(4): 1654-63.doi: 10.13040/IJPSR.0975-8232.8(4).1654-63.

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