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Griffith's Plum Yew (Cephalotaxus Griffithii Hook. F.)- a Gymnosperm

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Griffith's Plum Yew (Cephalotaxus Griffithii Hook. F.)- a Gymnosperm International Journal of Research ISSN NO : 2236-6124 NEWTONS 2018 Griffith’s Plum Yew (Cephalotaxus griffithii Hook. f.)- A gymnosperm from Kumaun Himalaya: As a source of total phenolic and total flavonoid compound, Antioxidant potential and Antimicrobial agents Urvashi Verma Department of Botany, D. S. B. Campus, KU, Nainital (Uttarakhand) India- 263002 E-mail- [email protected] Abstract Keywords: Cephalotaxus griffithii, phytochemical, Phytochemicals in medicinal plants have received a phenol, flavonoid, antioxidant, antimicrobial. great deal of attention mainly on their role in preventing diseases caused by microbial attack and I. Inroduction oxidative stress. Many plant species found in Himalaya The present study aimed to evaluate the biochemical area well known for their medicinal value properties of Griffith s plum Yew Cephalotaxus ’ for a long time. The importance of medicinal griffithii Hook. f. needles using two different extracts (hexane and aqueous) by qualitative and quantitative plants is being highlighted as a source of phytochemical methods along with their antibacterial natural antioxidant and functional foods [1]. activity against some gram-positive and gram- The antioxidant property of the plants negative bacteria responsible for many diseases considered the best as it reduces the including human. oxidation processes in the living system [2] From the observed results it is clear that aqueous extract was found more suitable to extract out the and plays an important role in maintaining primary as well as secondary metabolites but hexane health by reducing harmful reactive oxygen was found better for secondary metabolites extraction species (ROS) [3]. In recent years, and also showed good amount of total phenolic and phytochemicals in medicinal plants have total flavonoid content ranged from 210-44 mg/ gm received attention on their role in preventing dry extract weight of the plant with low IC50 value for free radical scavenging activity. diseases caused due to oxidative stress In antimicrobial screening the hexane extracts was which releases ROS such as singlet oxygen found effective against only gram positive bacteria and various radicals as a damaging side- tested with zone of inhibition (ZOI) upto 14 mm at effect of aerobic metabolism. These radicals 1000 µg/ml concentration. These results reveal that are possibly involved in a number of the extracts of C. griffithii is a possible good source of phenolic and flavonoid content as well as good disorders including cardiovascular free radical scavenger along with having good malfunctions, tissue injury, DNA damage, antimicrobial potential and new antibiotics. tumor promotion, kidney and liver disease, 1 Volume 7, Issue VII, JULY/2018 Page No:778 International Journal of Research ISSN NO : 2236-6124 fibrosis, atherosclerosis, arthritis, Northern India, western Sichuan Provience neurodegenerative disorders and aging. in China, Myanmar, Japan and Thailand [15, Several studies suggested that antioxidants 16]. The needles are 2-3 inch long and 3 mm could prevent accumulation of these reactive wide. Bark is blackish green. The plant is oxygen species and beneficial for treatment traditionally used in Manipur, India in the of these pathologies [4,5] Many research treatment of cancer [17]. The major studies have demonstrated that medicinal component of its volatile oil are α-pinene, plants, fruits and vegetables contain various caryophyllene, β-pinene, mytcene and components with antioxidant activity, which limonene [18]. are responsible for their beneficial health II. MATERIAL AND METHODS effects. Due to their natural origin, the antioxidants obtained from plants are of Sample collection greater benefit in comparison to synthetic Leaves (needles) of selected plant was ones. collected during the month of March 2016, Further, antimicrobials of plant origin from district Nainital (Uttarakhand), India have enormous therapeutic potential [6]. and authenticated by Prof. Y. P. S. Pangtey, Human infections particularly those Department of Botany of the Kumaun involving microorganisms i.e. bacteria, University. A voucher specimen was fungi, viruses, cause serious infections in deposited in the departmental herbarium. tropical and subtropical countries of the world. In recent years, multiple drug Solvent Extraction of Plant Material resistance in human pathogenic microorganisms has been developed due to Plant material was thoroughly washed indiscriminate use of commercial off under running tap water and then shade antimicrobial drugs [7]. Over the past dried at room temperature for three-four twenty years, there has been a lot of interest weeks. The air-dried plant material was in the investigation of natural materials as finely grinded and packed in self seal air sources of new antimicrobial agents [8]. The tight polythene bags for further use. use of plants extracts by extracting their Exposure to sunlight was avoided to prevent phytochemicals with known antimicrobial the loss of active components. properties can be of great significance in therapeutic treatments. In the last few years, Fine powdered plant material was a number of studies have been conducted in soaked in two different solvents (1:10 w/v) different countries to prove such efficiency. separately i.e. Hexane (highly non-polar) [9-13]. and double distilled water (i.e. aqueous) Cephalotaxus griffithii Hook.f. is a (highly polar) for 48 hours in closed shrub or small tree and grows at altitude of electrical shaker (120 rpm at 25ºC) and then 2000 m. This plant is distributed in Asia filtered with Whatman’s filter paper no.1. [14] and endemic to North east India, Only supernatant was taken and solvent was 0 especially in Mishmi Hills, Assam to evaporated using vacuum evaporator at 40 C and stored at 4ºC for further studies. 2 Volume 7, Issue VII, JULY/2018 Page No:779 International Journal of Research ISSN NO : 2236-6124 Chemicals and reagents colorimetric method given by Singleton and Rossi [20] with certain modifications. 2,2-diphenyl-1-picryl-hydrazyl (DPPH), Absorbance of the sample was measured quercetin, sodium nitrite (NaNO2), ascorbic spectrophotometrically (UV-VIS) at 765nm. acid, Ferric chloride (FeCl3), gallic acid, Quantification of total phenolic content was sodium carbonate (Na2CO3), aluminium based on standard curve of Gallic acid. The chloride (AlCl3), sodium hydroxide (NaOH), results were expressed in mg gallic acid potassium per-sulfate (K2S2O8), Ethylene equivalent (GAE)/gm dry extract weight of diamine tetra acetic acid (EDTA), were the sample. obtained from Himedia Laboratories Pvt. Ltd, Mumbai, India. Folin-Ciocalteu s ’ Determination of Total Flavonoid reagent, Molisch s reagent, conc. H SO , ’ 2 4 Content (TFC) Fehling’s reagent, glacial acetic acid, conc. Content of flavonoids of the sample HCl, NH4OH, Meyer’s reagent (potassiomercuric iodide solution), 2,2 extract were determined by AlCl3 [azinobis (3-ethyl benothiazoline- 6- colorimetric method given by Chang et al, sulphonic acid) diammonium salt] (ABTS), [21] with certain modifications. The hexane, chloroform, ethanol and. methanol absorbance was recorded at 415 nm using were obtained from Merck, Mumbai, India. UV-VIS spectrophotometer. Quantification All chemicals used were of analytical grade. of total flavonoid content was done on the basis of standard curve of Quercetin. Results Media Used : Nutrient agar (NA), were expressed in mg quercetin equivalent Nutrient Broth (NB), Muller Hinton Agar (QE)/gm dry extract weight of the sample. (MHA), Brain Heart Infusion Agar (BHIA), Brain Heart Infusion Broth (BHIB) (Hi DETERMINATION OF ANTIOXIDANT Media Laboratories Ltd., Bombay). ACTIVITY The antioxidant potential of C. griffithii QUALITATIVE PHYTOCHEMICAL was evaluated by two different methods, SCREENING DPPH (1,1-diphenyl 2-picrylhydrazyl), and To assess the chemical composition of ABTS (2,2̍ [azinobis (3 ethyl the plant qualitatively, phytochemical benothiazoline- 6 sulphonic acid). analysis was conducted using hexane and aqueous extracts following Harborne, 1998 DPPH Antioxidant Activity Assay [19]. The DPPH assay was done according to the method of Brand-Williams et al. (1995) QUANTITATIVE PHYTOCHEMICAL [22] with certain modifications. The ASSAYS reduction in absorbance was recorded at 517 Determination of Total Phenolic Content nm in UV-VIS spectrophotometer. Ascorbic (TPC) acid (AA), Butylated hydroxyl anisole (BHA) and Butylated hydroxyl toluene The total phenolic content of the sample (BHT) was used as standard and for control extract was determined by Folin-Ciocalteu’s 3 Volume 7, Issue VII, JULY/2018 Page No:780 International Journal of Research ISSN NO : 2236-6124 absorbance of DPPH cations was taken 24 hour at 37ºC in shaking condition for without adding sample extract. even suspension of bacterial colonies and maintained equivalent with Mac-Farland ABTS Antioxidant Activity Assay standard (0.5%) as recommended by WHO. The ABTS (2,2̍ -Azinobis-3- Determination of zone of inhibition (ZOI) ethylbenzotiazoline-6-sulphonic acid) assay and minimum inhibitory concentration was conducted according to [23, 24] Miller (MIC) et al. (1993) and Re et al. (1999) with minor modifications. The antioxidant activity of Antibacterial tests of selected tested sample was calculated by determining microorganisms were carried out by using . the decrease in the absorbance at different disc diffusion method [25] The freshly concentrations. prepared inoculums was swabbed on MHA plates using sterile cotton swab and left the % scavenging
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