Bioactive Compounds in the Different Extracts of Flowers of Rhododendron Arboreum Sm
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Available online www.jocpr.com Journal of Chemical and Pharmaceutical Research, 2016, 8(5):439-444 ISSN : 0975-7384 Research Article CODEN(USA) : JCPRC5 Bioactive compounds in the different extracts of flowers of Rhododendron arboreum Sm. Vandana Gautam, Anket Sharma, Saroj Arora and Renu Bhardwaj * Department of Botanical and Environmental Sciences, Guru Nanak Dev University, Amritsar - 143005, Punjab, India _____________________________________________________________________________________________ ABSTRACT In the present study, chloroform, hexane, ethyl acetate and petroleum ether extracts of flowers of Rhododendron arboreum Sm. were investigated for their chemical composition using GC-MS. Different extracts showed the existence of various bioactive compounds with an extensive range of countless therapeutic values. The results showed that chloroform extract contained 34 metabolites, hexane extract contained 23 metabolites, ethyl acetate extract contained 31 metabolites and petroleum ether extract contained 20 metabolites. Presence of these compounds in flowers of R. arboreum proves the promising utilization of this food plant in the therapeutics. Keywords: GCMS, Rhododendron arboreum Sm., Chemical composition. _____________________________________________________________________________________________ INTRODUCTION Every plant makes a distinctive blend of secondary metabolites. Thousands of these metabolites have been discovered in different classes of plants. Those compounds are classified into different categories i.e. phenolic compounds, flavonoids, proanthocyanidins, glycosides, saponins, glucosinolates, terpenes, alkaloids, tannins, resins, liganins, proteins and peptides etc. Conventional remedies are imperative resource of latent valuable compounds for the intensification of chemotherapeutic representatives [1]. A broad variety of remedial plant components is used for hauling out as unprocessed drugs and they have diverse medicinal properties and low side effect profile [2]. Plant products and their man-made derivatives constitute one half of all the indexed medicines all around the globe [3]. Polyphenols possess antimicrobial, anticarcinogenic, antioxidant, anti-inflammatory and antioxidant properties [4]. Foods rich in polyphenoilc compounds reduce the possibility of cancer, heart attack, cardiovascular disease and neurological disorders [5,6]. Intake of flavonoids is beneficial in dementia [7]. Alkaloids are used in treatment of Alzheimer’s Disease [8]. Rhododendron arboreum Sm. is regarded as one of the most beautiful flowers wearing, evergreen medicinal plant of higher altitudes [9].Its flowers are used to make pickle by the native people of Himachal Pradesh. The flower juice of R. arboreum is also very popular drink amongst the hilly people. Flowers of R. arboreum contain phenols, saponins, xanthoprotein, steroids, tannin, and coumarin [10]. Presence of quercetin, rutin and coumaric acid in methanolic extract R. arboreum flowers have been reported by Swaroop and coworkers [11] using HPTLC. Some other HPTLC, NMR, IR and spectroscopic studies revealed the presence of phenolics, epicatechin, syringic acid, quercitin, terpenoids and flavonoids in methanolic, alcoholic, aqueous and hexane extracts of R. arboreum leaves [12,13]. Nisar and coworkers [14] reported alkaloids, terpenoids, tannins, reducing sugars, steroids and saponins in roots of R. arboreum . Traditionally the plant is used for the treatment of various ailments like diarrhea, headache [15]. Recent studies and experiments show that Rhododendron arboreum has many 439 Renu Bhardwaj et al J. Chem. Pharm. Res., 2016, 8(5):439-444 ______________________________________________________________________________ effective and efficient phytochemical activities in curing human diseases like diabetes [16], inflammation [17], bacterial and fungal infections [13, 18] and diarrhea [19]. Thus further studies can be conducted to investigate the unexploited potential of Rhododendron arboreum . So as a part and as a basis for further exploitation, the phytochemicals in the flowers of Rhododendron arboreum were identified by GCMS analysis. EXPERIMENTAL SECTION Sample preparation Fresh flowers were washed with double distilled water and dried in shade. The dried flowers were ground to fine powder using mixer-grinder. 5 gram of flower’s powder was extracted with chloroform, hexane, ethyl acetate and petroleum ether separately and the extracts were then dried at 50 °C under reduced pressure using vacuum rotary evaporator. The residue was dissolved in 2.0 ml of respective solvents. Analysis using GC-MS Shimadzu GCMS-QP2010 Plus was used to analyze the chloroform, hexane, ethyl acetate and petroleum ether extracts of flowers of R. arboreum Sm. The carrier gas used was Helium, with an initial column oven temperature of 70°C for five minutes. The carrier gas was helium. At first, the column oven temperature was set at 70 °C and held for 5 minutes, then increased to 250°C at 10°C / min and held for 10 minutes, again increased to 300°C at 10°C / min and held for 10 minutes. Injection temperature; 280°C, injection mode; splitless, sampling time; 1 minute, flow control mode; linear, pressure; 110.8 k Pa, total flow 38.9 ml/min, column flow; 1.71 ml/min, linear velocity; 47.9 cm/sec, purge flow; 3 ml/min, sample injection volume; 4 µl, ion source temperature; 250°C, interface temperature 290°C, solvent cut time; 3.5 minute and detector gain mode; relative. Analytical column used was DB- 5ms with 30 m length and 0.025 mm id. Identification of compounds All the detected compounds were identified by comparing mass spectra with National Institute of Standard and Technology (NIST08s) and Wiley 7 library. RESULTS AND DISCUSSION Total 70 compounds were identified in four extracts of flowers of R. arboreum . Individually, the chloroform extract contained 34 compounds (Table: 1), hexane extract contained 23 compounds (Table: 2), ethyl acetate extract contained 31 compounds (Table: 3) and Petroleum ether extract contained 20 compounds (Table: 4). Linoleic acid, 9-octadecenoic acid (Z)-oleic acid, methyl commate B, flavone 4'-OH,5-OH,7-di-O-glucoside and octadecane are common in four extracts, whereas, 17-pentatriacontene and eicosanoic acid were common in chloroform, hexane and ethyl acetate extracts. Presence and absence of different compounds in the four extracts has been listed in Table:5. Amongst those compounds, palmitic acid, linoleic acid and eicosanoic acid are fatty acids; heptadecanoic acid is saturated fatty acid; methyl commate B and D are triterpine glycosides; dodecane is alkane hydrocarbon; phthalic acid is diterpene; hexadecenol is terpene alcohol; globulol is sesqueterpene [20] and octadecadienoic acid is livolic acid. Docosanoic acid, also known as behenic acid, is a carboxylic acid. Palmitic acid possesses free radical scavenging activity [21]. Vitamin E possess antioxidant, anti-inflammatory, antiageing, analgesic, antidiabatic, antidermatitic, antileukemic, antitumor, anticancer, hepatoprotective, hypocholesterolemic, antiulcerogenic, vasodilator, antispasmodic and antibronchitic, antiplasmodic, antimicrobial and anti-inflammatory properties [22,23]. 9-octadecenoic acid exhibits antimicrobial and antibacterial potential [24]. Phthalic acid is used in neurodegenerative diseases. 1,2-Benzenedicarboxylic acid is a plasticizer compound possessing antimicrobial properties [23]. 9,12-Octadecadien-1-ol possess anti-inflammatory hepatoprotective, antiarthritic properties [23]. Linoleic acid, heptadecanoic acid and oleic acid possess antimicrobial activity [25]. Octadecene was found to exhibit anticancer, antioxidant and antimicrobial activity [26,27]. Farnesol is used by the commensal, opportunistically pathogenic fungus Candida albicans as a quorum sensing molecule that inhibits filamentation [28]. 9,12 octadecadienoic acid possess anti-inflammatory, antiarthritic and antimicrobial properties [29]. (3b)-stigmast-5-en- 3-ol is of great therapeutic value as it possess cholesterol lowering effects [30] and antiproliferative activities [31]. (3b)-Stigmast-5-en-3-ol also acts anti-diabetic agent and regulates the glucose transport. It restores the glucose uptake activity without the stimulation of insulin, which proves its insulin-like property [32]. 440 Renu Bhardwaj et al J. Chem. Pharm. Res., 2016, 8(5):439-444 ______________________________________________________________________________ Table:1 Phytochemicals present in chloroform extract of Rhododendron arboreum flowers Sr. No. Name of Compound Retention Time Peak Area (%) 1 1-Dodecene adacene 8.074 0.49 2 1-Tetradecene n-tetradec-1-ene 12.040 1.02 3 9-Octadecene, (E) 14.825 1.46 4 9-Octadecenoic acid (Z)-oleic acid 16.730 0.27 5 9-Eicosene, (E) 17.153 1.58 6 Phthalic acid, 17.539 0.20 7 Heptadecanoic acid 18.916 9.09 8 3-Eicosene, (E) 19.221 1.45 9 9,12-Octadecadienoic acid (Z,Z)-linoleic acid 20.466 13.77 10 Docosanoic acid 20.753 1.91 11 1-Docosanol 21.096 1.07 12 Linoleic acid 21.226 0.21 13 Eicosanoic acid 22.463 0.46 14 1-Octacosanol 22.815 0.72 15 Octadecanal stearaldehyde 22.968 0.26 16 Tetratetracontane n-tetratetracontane 23.690 1.43 17 1,2-Benzenedicarboxylic acid 23.750 0.28 18 Thiosulfuric acid (H 2S2O3) 24.079 0.21 19 Heptafluorobutyric acid 24.422 0.48 20 Docosane n-docosane 24.488 0.55 21 Nonacosane n-nonacosane 25.426 10.26 22 17-Pentatriacontene 26.374 0.28 23 Octadecane 26.462 0.46 24 Acetic acid, chloro-, octadecyl ester 27.529 0.82 25 Pentadecane, 8-hexyl-8-n-hexylpentadecane 27.780 3.45 26 Dodecanoic