Determination of Stomatal Index

Determination of Stomatal Index The Plant material of Viscum capitellatum Smith. parasitism on Dendrophthoe falcata which is itself parasitic on M. indica was collected from Amba Ghat, Kolhapur, Western Ghat region of Maharashtra from India in November 2009. The collection are lies [Latitude 16o 58 0.59N and Longitude 73 ° 48 36.61E at altitude 1100m]. The plant specimen (Voucher no. 550) was authenticated by Dr. Vinay Raole, Reader, Department of Botany, M.S. University, Baroda, India. Pharmacognostical Study Macroscopical Study[68] It includes the shape, size, colour, texture, surface and odour of the drug in crude or powered form and often sufficient to enable to identify the whole drugs. Microscopical Study Histochemistry It gives the idea about the colour reaction of specific chemical reagent towards plant tissues [68]. Microscopical images are given in Figure no. 2. Quantitative Microscopy [66-69] Transverse sections of scale and stems were obtained by means of a microtome and stained with different staining reagents as per standard procedures [66, 70-71]. All observations were performed using Motic Digital Photomicroscope. Histological study of leaves and stem were performed by reported method [69]. Leaves were boiled in a 5% aqueous solution of NaOH for 5 min while stems were boiled with 10% aqueous solution of NaOH for 10 min. After cooling and washing with water, pieces were treated with a 25% aqueous solution of chromic acid for 30 min at room temperature. Washed pieces of both leaf and stem were pressed in between two slides and slides coves. Determination of Stomatal Number The average number of stomata per square millimeter of epidermis is termed the stomatal number. Determination of Stomatal Index The percentage proportional to the ultimate divisions of the epidermis of a leaf, which has been converted into stomata, is termed the stomatal index. SI = S ÃÆ'- 100 E + S Where SI = Stomatal index, S = number of stomata per unit area and E = number of ordinary epidermal cells in the same unit area. Procedure [68] Pieces of leaf between margin or midrib was cleared and mounted, and the lower surface examined by means of a microscope with a 4mm objective and an eyepiece containing a 5mm square micrometer disc. Counts were made of the numbers of the epidermal cells and of stomata within a square grid, a cell being counted if at least half of its area lies within the grid. The stomata index was determined for both leaf surfaces. Results pertaining to quantitative microscopical study are given in table no. 8. Analytical Study Ash Value 1.1 Total ash Total ash gives the idea about the residue obtained after ignition. It consist of physiological ash obtain by ignition of plant tissues and non physiological ash obtain by ignition of extraneous matter adhering to the surface of Plant. 2 gm of accurately weighed air dried powdered drug was taken in silica crucible. This silica crucible with drug material was kept in muffle furnace and ignited at temperature 4500C. The material was heated till the white coloured ash and constant weight is obtained. The procedure was performed in triplicate. Result is given in table No. 9. The total ash was calculated by subtracting the weight of crucible with ash of drug after ignition from weight of crucible with drug powder before ignition. Percentage of total ash was calculated with reference to air-dried drug. Acid insoluble ash Acid insoluble ash gives the idea about the presence of inorganic material such as calcium oxalate present in plant material. The ash obtained in the total ash method was boiled with 25 ml of 2N hydrochloric acid for 5 min. Insoluble matter was collected on ash less filter paper (Whatman paper) and washed with hot water. The material retained on filter paper and along with filter paper, was further ignited and weighed. Percentage of acid insoluble ash was calculated with reference to air dried material. Result is given in table No. 9. Water soluble ash The ash obtained from total ash was boiled with 25 ml water for 5 min. All insoluble matter was collected on ash less filter paper, washed with hot water and ignited for 15 min at the temperature not exceeding 4500C. The percentage of water soluble ash was calculated by subtracting weight of insoluble matter from weight of total ash. The difference between weights represents water soluble ash. Percentage of water soluble ash was calculated with reference to air dried drug. Result is given in table No. 9. Extractive Value Extraction by cold maceration It is the process of extraction of crude drugs with solvents with several daily shakings or stirring at room temperature.1 kg of powdered plant was extracted with 5 lit of methanol by cold maceration method. The extract was concentrated on rotary vacuum evaporator (Roteva Equitron, Mumbai) and further dried in vacuum dryer [73]. Successive extraction by using Soxhlet apparatus Weighed accurately 200gm of dried, powered crude drug and kept in a filter paper cover which was already placed in thimble. Then the solvent was slowly poured onto it. The solvent from thimble goes to lower round bottom flask via siphon tube due to the siphoning or syphon cycle. Such 2-3 cycles of solvent were performed and then drug powder was kept for 12 hours with solvent for imbibitions. After 12 hours imbibitions, solvent from flask heated to form vapors. Due to heat the solvent from RBF gets converted into its vapors, and then these vapors pass via side tube into the condenser where it gets condensed. This solvent dripped again on to drug material, which was placed in thimble. This process was continued till thimble gets filled with solvent and when level of solvent reaches to syphon tube, pulling of whole solvent into the flask is taken place. All this events repeated several times and drug material gets extracted continuously with fresh solvent. This process was performed for 3 days and when syphon solution showed negative test for phytoconstituents, extraction was completed. Then the heating was stopped and the mixture was collected and cooled. Then this mixture was filtered and concentrated by using rotary flash vacuum evaporator. The extract was dried in vacuum dryer and was stored in freeze. Then this marc obtained after pet ether extraction and subjected again to extraction by following solvents (Table 10) [73]. Moisture content by Loss on Drying 2 g of air powdered drug was placed in a silica crucible. Before that, crucible was cleaned and dried and weight of empty crucible was taken. The powder was spread in a thin uniform layer. The crucible was then placed in the oven at 1050C. The powder was dried for 4 h and cooled in a desiccator to room temperature and weight of the cooled crucible plus powder was noted. Result is given in table no. 9. Analysis of inorganic constituents (Elemental analysis) Ash of drug material was prepared and adds 50% v/v HCl or 50% v/v HNO3 to ash. Keep it for 1 hour. Filtered and with the filtrate performed the test as per method reported [74]. The results of analysis of inorganic constituents are given in (Table 11). Test for calcium a) Add dil. NH4OH and saturated ammonium oxalate solution to filtrate. White ppt of calcium oxalate forms which is soluble in HCl. Calcium present. b) Add ammonium carbonate to filtrate. White ppt which is insoluble in NH4Cl. Calcium present. Tests for iron a) Add 2% potassium ferricyanide to filtrate. Dark blue coloration. Iron present. b) To filtrate, add 5% ammonium thiocyanate. Blood red color. Iron present. c) To filtrate, add dil. HCl and sol. of KMnO4. Pink color. Iron present. Tests for magnesium a) To filtrate add NaOH. White ppt. Magnesium present. b) To filtrate add (NH4)2CO3. White ppt, redissolve in NH4Cl. Magnesium present. Tests for potassium a) Add sodium cobalt nitrite to filtrate. Yellow ppt. Potassium present. b) Flame test. Violet color to flame. Potassium present. Tests for sodium a) Add uranyl zinc acetate to filtrate, shake well. Yellow crystalline ppt. Sodium present. Tests for carbonate a) Add HgCl2 to filtrate. Brownish red ppt. Carbonate present. b) Add dil. Acid to the filtrate. Effervescence of CO2 Carbonate present. c) Add MgSO4 to filtrate. White ppt. Carbonate present. Tests for Sulphate a) Add BaCl2 to filtrate. White crystalline ppt Sulphate present. b) Add filtrate to lead acetate sol. White ppt. Sulphate present. Tests for phosphate a) Add HNO3 and ammonium molybdate to filtrate, heat 10 min. cool. b) Add silver ammonium- nitrate to filtrate Yellow crystalline ppt. Light yellow ppt Phosphate present. Phosphate present. Tests for chloride a) Add AgNO3 to filtrate. b) To filtrate, add manganese dioxide and H2SO4 White curd ppt, soluble in dil. NH3. Odour of chlorine Chloride present. Chloride present. Tests for nitrate a) Add water to filtrate, add H2SO4 from side of test tube. b) Add H2SO4 and copper to filtrate, warm Brown color at junction of two liquid Liberation of red fumes Nitrate present. Nitrate present. Determination of Type of Starch Grains The shape of starch grains present was determined according to the reported method [68]. Size of starch grains were measured with the help of calibrated Photomicroscope using Motic software. Starch grains were identified by staining with Iodine solution. The Motic digital Photomicroscope was calibrated with images obtained with various magnifications (10x, 40x and 100x) by using standard slide in 1.3 software. The images obtained in triplicate and average figures calculated from 20 readings in each parameter (Table no. 12). Crude Fiber Content Pre-weighed dried powder material was extracted with Petroleum ether (b.p. 40- 600C) using soxhlet apparatus for 8 h. The marc obtained after extraction was utilized for determination of Crude Fiber Content. Crude fiber was investigated by acid-base digestion with H2SO4 (1.25%) and of NaOH (1.25%) solution. The marc after extraction was taken into a 500ml beaker and 200ml of boiling H2SO4 added. The content was boiled for 30 minutes, cooled, filtered and the residue washed three times with 50ml of boiling water. The washed residue was further boiled in 200ml of NaOH for 30 minutes. The digest was filtered to obtain residue. This was washed three times with 50ml of boiling water and lastly with 25ml of ethanol. The washed residue was dried in an oven at 1250C to constant weight and cooled in dessicator. The residue was scraped into a pre-weighed porcelain crucible, weighed, ashed at 5500C for 2 hours, cooled in a dessicator and weighed. Crude fiber content was expressed as percentage loss in weight on ignition. Result is given in table No. 13. Phyto-chemical Analysis Extracts Petroleum ether, benzene, chloroform, acetone and methanol extract obtained by successive extraction method and aqueous extract by maceration method [68, 95]. Qualitative analysis All the extracts were subjected to proximate chemical analysis and its result is given in table no. 14. Tests for Acidic compounds: a) To the test solution add sodium bi-carbonate b) Test solution treated with warm water and filter. Test the filtrate with litmus paper. Tests for Alkaloids: a) Dragendorffs Test: Test solution treated with Dragendorffs reagent (potassium bismuth iodide) b) Mayers Test: Test solution treated with Mayers reagent (Potassium mercuric iodide). c) Wagners Test: Test solution treated with Wagners reagent (Iodine in potassium iodide). d) Hagers Test: To the test solution add gives with Hagers reagent (Saturated picric acid solution). e) Tannic acid test: Test solution treated with Tannic acid solution. f) Picrolonic acid test: Test solution treated with Picrolonic acid. Test for amino acids: a) Millions Test: Test solution treated with Millions reagent and heated on a water bath. b) Ninhydrin Test: Test solution boiled with Ninhydrin reagent. Test for Carbohydrates: a) Molischs Test: To the test solution add with few drops of Molischs reagent (Alcoholicà ¯Ã‚ Ã‚ ¡-naphthol) and 2ml of conc. sulphuric acid is added slowly from the sides of the test tube. b) Barfords Test: Test solution heated with Barfords reagent on water bath. c) Selivanoffs test (Test for Ketones): To the test solution add crystals of resorcinol and equal volumes of concentrated hydrochloric acid and heat on a water bath. d) Test for pentose: To the test solution add equal volumes of hydrochloric acid containing small amount of Phloroglucinol and heat. e) Osazone formation test: Heat the test solution with the solution of phenyl hydrazine hydrochloride, sodium acetate, and acetic acid. Test for Flavonoids: a) Shinoda Test: Test solution treated with fragments of magnesium ribbon and conc. Hydrochloric acid. b) Alkaline Reagent Test: Test solution treated with sodium hydroxide solution c) Zinc-Hydrochloride test: Treat test solution with zinc dust and few drops of HCL Test for glycosides: General test: Extract 200 mg of drug with 5 ml of dilute sulphuric acid by warming on a water bath, filter it, and neutralize the acid extract with 5 % solution of sodium hydroxide. Add 0.1 ml of Fehlings solution A and B until it becomes alkaline (test with pH paper) and heat on water bath for 2 minutes. Test B: Repeat Test A procedure by using 5 ml of water instead of dilute sulphuric acid. Note the quantity of red precipitate formed. Chemical tests for specific glycosides: Tests for Anthraquinone glycosides: a) Borntragers test: Boil the test material with 1ml of sulphuric acid for 5minutes. Filter while hot. Cool the filtrate; shake with equal volume of dichloromethane or chloroform. Separate the lower layer of dichloromethane or chloroform; shake it with half of its volume of dilute ammonia. b) Modified Borntragers test: Boil 200 mg of test material with 2ml of sulphuric acid. Treat with 2 ml of 5 % aqueous ferric chloride solution (freshly prepared) for 5 minutes, shake it with equal volume of chloroform and continue the test as above. c) Test for hydroxy anthraquinones: treat the sample with potassium hydroxide solution. Tests for cardiac glycosides: a) Keddes test: Extract the drug with chloroform, evaporate to dryness. Add one drop of 90 % alcohol and 2 drops of 2 % sodium hydroxide solution. b) Keller-Killiani Test: (Test for deoxy sugars) Extract the drug with chloroform and evaporate it to dryness. Add 0.4 ml of glacial acetic acid containing ferric chloride, add carefully 0.5 ml of conc. sulphuric acid by the side of test tube. c) Raymonds number: treat the test solution with hot methanolic alkali. d) Baljets Test: The test solution treated with sodium picrate or picric acid. e) Legals Test: Test solution treated with pyridine [made alkaline by adding sodium nitroprusside solution]. f) Tests for coumarins glycosides: Place small amount of sample in test tube and covered it with a filter paper, moistened with dilute sodium hydroxide solution. Placed the covered test tube on water bath for several minutes. Remove the paper and expose it to ultraviolet (UV) light. Cynogentic glycosides: Place 200 mg of drug in conical flask and moisten with few drops of water.( Flask should be completely dry because hydrogen cyanide produced will dissolve in the water rather than come off as gas to react with paper) moisten a piece of picric acid paper with 5% aqueous sodium carbonate solution and suspended in neck of flask. Warm gently at about 37oC. Observe the change in color. Saponin glycosides: Froth test: Place 2 ml solution of drug in water in a test tube, shake well. Tests for steroids and triterpenoids: a) Liebermann Burchard Test: Treat the extract with few drops of acetic anhydride, boil and cool, add conc. sulphuric acid from the sides of test tube. b) Salkowski test: Treat the extract with few drops of conc. sulphuric acid. c) Sulfur powder test: Add small amount of sulfur powder to the test solution. d) Tests for inulin: To the test solution add the solution of à ¯Ã‚ Ã‚ ¡-naphthol and sulphuric acid. e) Tests for Lignin: Treat the sample with hydrochloric acid and Phloroglucinol. Tests for Mucilage: Treat the sample with thionine solution. After 15 min wash with alcohol Tests for tannins: a) Ferric-Chloride Test: Treat test solution with few drops of ferric chloride solution. b) Gelatin test: To the test solution add 1 % gelatin solution containing 10 % sodium chloride. Tests for proteins: a) Heat test: Heat the test solution in boiling water bath. b) Biuret Test: Test solution treated with Biuret reagent (40% sodium hydroxide and dilute copper sulfate solution). c) Xanthoproteic test: To the test solution, add 1 ml of conc. nitric acid and boil yellow precipitate is formed. After cooling it, add 40 % sodium hydroxide solution. d) Test for starch: To the test solution, add weak aqueous iodine solution. Blue color indicates presence of starch, which disappears on heating and reappears on cooling. Effervescence produces Litmus paper turns blue Gives reddish brown colored precipitate Gives cream colored precipitate Gives reddish brown colored precipitate Gives yellow colored precipitate Gives buff colored precipitate Gives yellow colored precipitate White colored precipitate Gives violet color Purple to violet ring appears at the junction of two liquids If red cupric oxide is formed Rose color is produced Red color produced. Yellow crystals formed. Observe under microscope. Shows pink scarlet, crimson red or occasionally green to blue color after few minutes. Shows increase in the intensity of yellow color on addition of few drops of dilute acid. Shows red color after few minutes. Red Precipitate formed compared with precipitate of test A A rose pink to red color is produced in ammonical layer. A rose pink to red color is produced in ammonical layer. Red color produced Purple color is produced. Acetic acid layer shows blue colour. Violet colour produced Gives yellow to orange color Gives blood red color Paper shows green fluorescence. Reddish purple color Stable froth (foam) formed Brown ring is formed at the junction of two layers, If upper layer turns green If upper layer turns deep red Red color at lower layer Yellow color at lower layer It sinks at the bottom Brownish red color formed Pink color formed Mucilage turns violet red. Gives dark blue color Green color appears Precipitate formed Proteins gets coagulated Gives violet color Orange color formed Blue color, which disappears on heating and reappears on cooling Acidic compounds present Acidic compounds present Alkaloids present Alkaloids present Alkaloids present Alkaloids present Alkaloids present Alkaloids present Amino acids present Amino acids present Carbohydrates present Monosaccharides are present. Carbohydrates present Carbohydrates present Carbohydrates present Flavonoids present Flavonoids present Flavonoids present If the precipitate in Test A is greater than in Test B then glycoside may be present. Anthraquinone glycosides present Anthraquinone glycosides present Hydroxy anthraquinones present Cardiac glycosides present Cardiac glycosides present Cardiac glycosides present Cardiac glycosides present Cardiac glycosides present Coumarins glycosides present Cynogentic glycosides present Saponin glycosides Present Steroids present Triterpenoids present Steroids present Triterpenoids present Steroids present Inulin Present Lignin Present Mucilage present Hydrolysable tannins Condensed tannins Tannins present Proteins present Proteins present Proteins present Starch present Floroscence Analysis of various extracts Petroleum ether, Benzene, Chloroform, Acetone, Methanol and Aqueous extracts were screened for fluorescence characteristic. The observation pertaining to their colour in day light and under ultra-violet light were noticed and represented in table. Many substances for example quinine in solution in dilute sulphuric acid when suitably illuminated emit light of a different wavelength or colour from that which falls on them. This emitted light (fluorescence) ceases when the exciting light is removed [68].Results given in Table No. 15. HPLC Analysis of sample drug The chromatographic pattern of plant was obtained as per report with some modifications for which the HPLC conditions are as follows. Extract: The methanol extract diluted with HPLC grade methanol and filtered through whatman filter paper and used for analysis Instrument: Shimadzu LC-20AT with UV/visible detector Stationary Phase: Bonda- pack C-18 column with 250ÃÆ'-4mm Mobile Phase: Methanol (80): Water (20) Detection wave length: 350 nm Flow Rate: 2 ml/min. HPLC Chromatogram is given in Fig. 3 and its retention time is given in Table no. 16 HPTLC Analysis of sample drug The chromatographic pattern of plant was obtained as per report with some modifications for which the HPTLC conditions are as follows. Extract: Methanolic Extract Instrument: HPTLC (Camag, Switzerland) Stationary Phase: pre-coated silica gel plates Mobile Phase: Ethyl acetate: Formic acid: Glacial acetic acid: water (100:05:10:20) Spraying Reagent: Natural Product Reagent (NP reagent) Detection: 365 nm. HPTLC Chromatogram is given in Fig. 4 and its retention time is given in Table no. 17. Isolation and characterization of chemical principle Compound I The methanol extract was dissolved in water and partitioned with ethyl acetate and n- butanol. The ethyl acetate fraction was subjected to column chromatography for isolation of compounds. Column chromatography: The separation of extract constituents was done by column chromatography. The clean and dried glass column was used. The silica gel for column chromatography (#60-120) was activated at 1100c.The column was filled with silica gel and mobile phase without formation of any air bubbles. The silica gel was then allowed to stabilize in the column. Mixture of two or three compounds was isolated from the ethyl acetate fraction of methanol extract of the plant with following experimental conditions [73]. Height of column: 20 cm Diameter of column: 3.5 cm. Stationary phase: Silica gel (#60-120). Mobile phase: Benzene† Ã¢â‚¬â„¢ Chloroform † Ã¢â‚¬â„¢ Ethyl acetate† Ã¢â‚¬â„¢ Methanol with variant Proportions Elution: Gradient elution. Fraction quantity: 25 ml Preparative TLC: 20 X 20 glass plates were coated with the thick layer of silica gel or any other adsorbent material. The plates were then activated at 1100c.The sample-containing mixture of two or more compounds were applied in the form of thin band on the plate. The plate was then developed. The different bands separated on the plate were scratched and recovered with methanol. Purity of dried sample was checked by TLC method. One single compound was isolated with the help of preparative chromatography from fractions 54- 58. The compound is given for spectral analysis. FTIR spectra, Mass spectra and 1HNMR are given in fig. no. 5, 6 and 7 respectively. The spectral data of FTIR and 1HNMR are given in Table no. 18 and 19 respectively. The assumed structure of the compound (Quercetin) is given in Fig. No. 8. Compound II Petroleum ether extract obtained is processed for separation of the unsaponifiable and saponifiable matter. Extract is allowed to saponify using alcoholic KOH with reflux and then it is extracted with solvent ether for separation of unsaponifiable matter. The aqueous phase is acidified with concentrated H2SO4 and then again extracted with the solvent ether for separation of the saponifiable matter [73]. Fractionation of unsaponifiable matter Experimental: Height of column: 25 cm Diameter of column: 3.5 cm. Stationary phase: Silica gel for column chromatography (#60-120). Mobile phase: Benzene† Ã¢â‚¬â„¢ Ethyl acetate Elution: Gradient elution. Fraction quantity: 30 ml Fractions No. 24-27 were subjected for thin layer chromatography with following experimental conditions. Stationary phase: Silica gel H Mobile phase: Ethyl acetate: Benzene (1: 9) Detection: Vanilin-sulphuric acid reagent Identification: Whitish Purple colour Fraction was concentrated and single band was applied. After plate development; developed band was scraped (Rf. 0.62). After separation of single compound from the silica, it is dried. This sample was further given for spectroscopic analysis. FTIR spectra, Mass spectra and 1HNMR are given in fig. no. 9, 10 and 11 respectively. The spectral data of FTIR and 1HNMR are given in Table no. 20 and 21 respectively. The assumed structure of the compound (Quercetin) is given in Fig. No. 12. Biochemical Estimations a) Estimation of Total carbohydrate content The estimation of carbohydrate was done using the method acid base digestion. Principle: In hot acidic media glucose is converted to hydroxy methyl furfural by dehydration. This forms a green colour product with phenol. Procedure: 100mg of the aqueous extract was taken and it was hydrolyzed by keeping it on water bath for 3 hours with 5 ml of HCl (2.5N) and cooled at room temperature. Neutralized it with sodium carbonate and volume was made up to 100 ml and from this centrifuge 10 ml of the solution. Then 0.2, 0.4, 0.6, 0.8 and 1ml of working standard was pipetted out into a series of test tube and in separate test tubes 0.1 and 0.2 ml of sample solution was pipetted out and the volume was make up to 1ml with water. The blank was prepared with 1 ml distilled water. Then 1ml phenol solution and 5ml of sulphuric acid (96%) was added to each test tube and shaken well. After 10 min the test tube was placed in water bath at 25-30à ¯Ã¢â‚¬Å¡Ã‚ °C for 20 min. The absorbance was read at 490 nm. And the amount of total carbohydrate present was calculated in the sample using standard graph. Result pertaining to Total carbohydrate content is given in Table no. 22 and Calibration curve of standard glucose dilutions are gi ven in Fig. No. 13. Estimation of Bitterness value The bitterness value of plant material was compared with diluted solution of Quinine hydrochloride. Preparation of Solutions Preparation of Quinine hydrochloride solution The stock solution of 100 µg/ml was prepared from which a series of dilutions 42, 44, 46, 48, 50, 52, 54, 56 and 58  µg/ml were prepared. Preparation of Sample Preparation Form the stack solution of 1000  µg/ml, 100, 200, 300 and 400 µg/ml dilutions were prepared. Method Tasted all the dilutions of sample and Quinine sulphate by taking the solution in mouth and swirled it for 30 secs in mouth mainly near to the tongue. After tasting each dilution the mouth wash rinsed thoroughly with drinking water and taken the interval of 10 mins. Until the bitter sensation of previous dilution was no more remain. Then compared the dilution of sample which produced the same bitterness equivalent to the dilution of Quinine sulphate. Then bitterness value was calculated according to following formula. Bitterness value in units per gram = 2000 ÃÆ'- A B ÃÆ'- C Where A= quantity of Quinine sulphate (mg) having higher bitterness B= the concentration of stock solution (mg/ml) C= Volume of sample in ml having higher bitterness Result pertaining to estimation of bitterness value is given in Table no. 22 Total Phenolic content The total phenolic content of methanol extract of V. capitellatum Smith. (VCM) was estimated using Folin-Ciocalteu reagent. In this method, the blue colour formed due to the polyphenol was measured at 760 nm using UV spectrophotometer. Chemicals Folin- Ciocalteu reagent (Merck Co.) Gallic acid (Sigma Ltd., USA) Sodium carbonate (SISCO Research Laboratory Pvt. Ltd., Mumbai, India) Reagent preparation Folin-Ciocalteu (phenol) reagent The reagent was prepared by diluting 1ml with 5ml of distilled water. Sodium carbonate 15% solution was prepared in distilled water. Gallic acid solution The stock solution was prepared by dissolving 1mg gallic acid in 10ml of water from which different concentrations (20-100 µg/ml) were prepared. Sample preparation Sample solution was prepared by dissolving 10 mg of the extract in 100 ml of methanol to give (100  µg/ml) solution. Procedure 0.1ml of extract was mixed with the 0.2ml of Folin-Ciocalteu reagent, 2 ml water and 1 ml of sodium carbonate solution, and absorbance was measured at 760 nm after 10 min incubation at 50 0C. The total phenolic was expressed as  µg gallic acid equivalent. Result pertaining to Total phenolic content is given in Table no. 22 and Calibration curve of standard gallic acid dilutions are given in Fig. No. 14. Total Flavonoid Content Total flavonoid content of VCM was determined using method reported [79].

Genetically Modified Foods/What You Need to Know

Genetically modified foods (GM foods) have made for big talk in the public lately. Public interest groups have been actively protesting against GM foods for months. In response to the up swelling of public concern, the U.S. Food and Drug Administration (FDA) have held meetings to solicit public opinions and, begin the process of establishing a new regulatory procedure for government approval of GM foods. I would like to research and maybe, explain the reasons why I feel that GM foods are not humanity's solution to our food consumption problem. What are Genetically Modified Foods?Genetically modified foods are foods produced from organisms that have had specific changes introduced into their DNA using the methods of genetic engineering. These techniques have allowed for the introduction of new crop traits as well as a far greater control over a food's genetic structure than previously afforded by methods such as selective breeding and mutation breeding (Wikapedia.com). To date most ge netic modification of foods have primarily focused on cash crops in high demand by farmers such as soybean, corn, canola, and cotton seed oil.These have been engineered for resistance to pathogens and herbicides and better nutrient profiles. GM livestock have also been experimentally developed, although as of November 2013 none are currently on the market. There are many reasons to not be a fan of GM foods but, before I explain the reasons not to consume GM foods, let me tell you some reasons why scientists and so-called, â€Å"experts†, are pushing to have  developers and, manufacturers of GM foods make sure, that they are various advantages of consuming these foods, as well as, persuade the public to purchase these products. Are there advantages to GM Foods?One advantage to GM foods is that they help to control certain diseases that can cause people to have an allergic reaction to certain foods. With GM foods the DNA system is modified to eliminate the properties causing these allergies (http://www.buzzle.com/articles/genetically-modified-foods-pros-and-cons.html).Another advantage to GM foods is that they are said to be high in nutrients and contain more vitamins than traditionally grown food. They also claim to have a longer shelf life than traditionally grown food, which means less waste. Now that we have heard why developers, manufacturers, and scientists want GM foods massively produced, let's hear about some of the reasons why most of the population is not so accepting.ThreatsThe biggest threat caused by genetically modified foods is that they can have harmful effects on the body (http://www.csa.com/discoveryguides/gmfood/overview.php). It is believed that consumption of these foods can cause the development of diseases which are immune to antibiotics. According to experts, people who consume these foods have high chances of developing cancer (http://www.csa.com/discoveryguides/gmfood/overview.php).Because these are new inventions on food, the re's not much known about the long-term effects that genetically modified foods will have on humans. Foodstuffs made of genetically modified crops that are currently available (mainly maize, soybean, and oilseed rape) have been judged safe to eat, and the methods used to test them have been deemed appropriate. These conclusions represent the consensus of the scientific evidence surveyed by the International Council for Science (ICSU) and are consistent with the views of the World Health Organization (WHO). However, the lack of evidence of negative effects does not mean that new genetically modified foods are without risk.The possibility of long-term effects from genetically modified plants cannot be  excluded and must be examined on a case-by-case basis. New techniques are being developed to address concerns, such as the possibility of the unintended transfer of antibiotic-resistance genes. Earlier, I mentioned an advantage to GM foods that allows them to be modified to eliminate properties within certain foods, so that people do not have an allergic reaction to them. My question is, â€Å"what in the world makes these developers think that we want to eat anything that has been modified†? Not to mention, knowing that the long-term effects are not certain if I consume one of these products. That's not comfortably sitting on my stomach! ResponseThese developers and, manufacturers claim that GM foods contain more vitamins and nutrients, along with a longer shelf life than traditional foods. The way the public sees it is the genetically modified anything, cannot be better for you than the real thing. Throughout my research, I have found that most of society's response to GM foods is, â€Å"why fix something that's not broken†? Most of society throws criticisms towards genetically modified foods, criticizing agribusinesses for pursuing profit without concern for potential hazards, and the government for failing to exercise adequate regulatory oversi ght(http://www.csa.com/discoveryguides/gmfood/overview.php).ConcernsThe most concerns about genetically modified foods falls into three categories: environmental hazards, human health risks, and economic concern. Environmental hazards are causes of unintended harm to other organisms. For example; the pollen blown around by the wind off of a GM plant, stands a chance of becoming involved with the milkweed plants that often grow near these crop fields. The concern is for the monarch butterfly, monarch butterflies eat milkweed plants and, if the butterflies eat the milkweed plant with the pollen from the GM plant on it, the butterflies then stand a chance of dying if they come in contact with this pollen. This could create a huge problem for the monarch butterflies extinction rate.Human health risks contain allergencity, which are allergies to things like peanuts and other foods. The possibility still remains that introducing a gene into a plant may create a new allergen that will caus e someone to have an allergic reaction and, they could possibly die from that. Economic concern is the most talked about category out of all three. Things like, putting farmers out of business and bringing this product to the market is a costly process, as well as the tech companies wishing to turn a profit on their investment. Consumer advocates are worried that patenting new plants will raise the price of seed. When the prices of seed goes up, the farmers in third world countries cannot afford to purchase the seeds each year. No seeds means no crops for the farmers, no crops means no money.ConclusionConsumers may wish to select conventional foods on the basis of several criteria such as methods of production (e.g. organic or fair-trade food), religious principles (e.g. kosher food), or the presence of known allergens (e.g. groundnuts). Labeling of foods as genetically modified or non-genetically modified may enable consumer choice as to the process by which the food is produced. H owever, it conveys no information as to the content of the foods, and what risks or benefits may be associated with particular foods. More informative food labeling, explaining how food has been transformed and what the resulting changes in food composition are, could enable consumers to assess these risks and benefits (http://www.greenfacts.org).Genetically modified foods is a big deal in today's society, many people have a very strong opinion about GM foods. Many people are also unaware of what a genetically modified food is. Most people consume genetically modified foods on a daily bases and do not even realize it. Nor is the government obligated to inform you that you are consuming genetically modified foods. Throughout my research, I have found out why I feel that GM foods are not humanity's solution to our food consumption problem. Throughout my argument, I have tried to explain the good and, the bad things associated with genetically modified foods. I have come to the conclus ion that genetically modified foods are not for me or my family. There are too many health issues and, risks that I am not willing to take because my family's health may lay in the balance.

What are your perceptions on the universal declaration of...

What are your perceptions on the universal declaration of human rights would you like to amend any of the articles or add a new article to the declaration? In: International Laws [Edit categories] Answer: The Universal Declaration of Human Rights is half a century old, but critics are still asking whether anything in our multicultural, diverse world can be truly universal. Some ask, isnt human rights an essentially Western concept, ignoring the very different cultural, economic and political realities of the South? Can the values of the consumer society be applied to societies that have nothing to consume? Isnt talking about universal rights rather like saying that the rich and the poor both have the same right to fly first-class and to†¦show more content†¦Culture is too often cited as a defence against human rights by authoritarians who crush culture domestically when it suits them. In any case, which country can truly claim to be following its traditional culture in a pure form? None have remained in a pristine state; all have been subject to change and distortion by external influence, both as a result of colonialism and through participation in modern inter-state relations. You cannot follow the model of a modern nation-state cutting across tribal boundaries and conventions, and then argue that tribal traditions should be applied to judge the human-rights conduct of that modern state. There is nothing sacrosanct about culture anyway. Culture is constantly evolving in any living society, responding to both internal and external stimuli, and there is much in every culture that societies quite naturally outgrow and reject. Are we, as Indians, obliged to defend, in the name of our culture, the practices of sati or of untouchability? The fact that slavery was acceptable across the world for at least two thousand years does not make it acceptable to us now. The basic problem with cultural relativism is that it subsumes all members of a society under a framework they may prefer to disavow. If dissenters within each culture are free to opt out and to assert their individual rights - for example, Muslim women in my country, India, have theShow MoreRelatedAssignments: Human Rights Law4555 Words   |  19 PagesASSIGNMENTS SUBJECT- STRATEGIC MANAGEMENT Select an appropriate generic strategy to position your printing business unit in its competitive environment (map the environment primarily as a pattern of competitive pressures from rivals, suppliers, buyers, entrants and substitutes). The steps need to be followed to strategise printing business unit in its competitive environment:- a) Planning for a brighter future starts with analyzing inner strengths, weaknesses, opportunities and threats. 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[10 Marks] Answer: Research simply means a search for facts – answers to questions and solutions to problems.Itis  a  purposiveinvestigation.  It  is  an  organized  inquiry.  It  seeks  tofind  explanations  tounexplained phenomenon to clarify the doubtfulRead MoreStrategic Marketing Management337596 Words   |  1351 PagesUniversity AMSTERDAM †¢ BOSTON †¢ HEIDELBERG †¢ LONDON †¢ NEW YORK †¢ OXFORD PARIS †¢ SAN DIEGO †¢ SAN FRANCISCO †¢ SINGAPORE †¢ SYDNEY †¢ TOKYO Elsevier Butterworth-Heinemann Linacre House, Jordan Hill, Oxford OX2 8DP 200 Wheeler Road, Burlington, MA 01803 First published 1992 Second edition 1997 Reprinted 1998, 1999, 2001, 2003 Third edition 2005 Copyright  © 1992, 1997, 2005, Richard M.S. Wilson and Colin Gilligan. All rights reserved The right of Richard M.S. Wilson and Colin Gilligan to be identified

Overwriting of an external culture with organisation culture - Free Essay Example

Sample details Pages: 6 Words: 1793 Downloads: 8 Date added: 2017/06/26 Category Management Essay Type Argumentative essay Did you like this example? Introduction Throughout the revolution and development of organisations, cultural attributes have applied to the existence of many organisations. From various backgrounds to individual uniqueness, management has brought forward perspectives and ideals that present diverse techniques that contribute to modern organisations. Behind many of these techniques lies persuasive and contradictive control of employees, in order to benefit their organisation. Don’t waste time! Our writers will create an original "Overwriting of an external culture with organisation culture" essay for you Create order In this essay, I argue that management of an organisation pursue individuals perspectives and values for the beneficiation of their organisation, rather than preserving individuals characteristics. The first part of this essay consists of organisational culture, where the core definition of organisation culture is explained, followed by the means to practice such management. The second part portrays the perspectives of management and the role they present to employees. The existence of hierarchy within the organisation implements individual views and values as an asset, which can be controlled by the reward system. Part three of this essay illustrates the attempts to govern employees perspectives and cultures with the intention to reshape the basis of individual culture, reforming to an organisation implemented society. In this part, the focus of managers to step further into individuals social lives suggests a deeper meaning to administer not only within the organisation, but infilt rate outside the walls of an organisation. Although this essay emphasizes on the aspects towards management, acknowledgment of employees views and values are in regards. Part One: organisation culture The first part of my essay is what Organisation Culture is. According to the textbook Managing and Organizations, the definition of Culture is what represents the totality of everyday knowledge that people use habitually to make sense of the world around them through patterns of shared meanings and understandings passed down through language, symbols, and artifacts (Clegg, Kornberger, Pitsis 2008) . Organizational culture is defined as the deep, basic assumptions and beliefs that are shared by organizational members (Schein, 1997). Also according to Schein, culture can be divided into three levels, which are Artifacts(Level 1), Values(Level 2) and Basic Assumptions(Level 3). Nowadays, organisations developed their management through the knowledge to govern employees by psychological knowledge which means to understand what employees desires are and to shape work so that the workers see in work their self-fulfilment (Rose 1990). Therefore, many organisations or companies try to c reate organisation culture and change culture, however, there are only a few companies changed successfully (Ackroyd 1990). And even though Ackroyd believes that cultures are highly distinctive, resilient and resistant to change, he thought it is still possible to manage organisational culture. From my point of view, create and change organisational culture is feasible and it is necessary. A good corporate culture can make employees to work hard and efficiency. As hard work will produce both psychological and physical rewards for employees, those rewards make employees work harder and more efficiently. A good organisational culture will give workers a sense of belonging. Part two: the perspective of management Secondly, I argue that from the perspective of management, which is upper level, they want to manage the views and values of workers. An organisation is just like a human body, it constitutes of head, hands and feet. CEO and senior-managers are the head of the organisation and employees are the hands and feet. CEO and senior-managers are the core part of organisation, they give orders and instructions to lower level employees ¼Ãƒâ€¦Ã¢â‚¬â„¢and employees just follow their orders. Both managers and employees are important, as without senior managers to give the macro view of the companys short term and long term plans, as well as planning ¼Ãƒâ€¦Ã¢â‚¬â„¢the company will operate disorderly. Moreover, employees are also very important, asemployees are the key to success (Peters and Waterman, 1982). Managers and employees play different roles but both are essential. There is a position which is generally referred to as the leading hand. Leading hand means a position of mana ger which is lower than senior managers but it observes employees. There is such a position because most senior managers have more experience and ability than employees at the element of governing and developing companies, therefore senior managers have leading hands so that employees can simply follow instructions and need not concern themselves over the bigger picture. There is the example of Vulcan. Vulcan was created in 1826, at that time; the company was very small and mainly manufactured pans and pots (Martin, 2000). Furthermore, during 1960 and 1980, Vulcan suffered from declining, business contracts. Consequently, they changed their Managing Director to John Whyte. He invested in reward system and incentives and created an organizational culture and belief, which was that the managers of Vulcan wanted the company to feel as a family for the employees and that it have a common language, traditions and a single shared identity (Martin, 2000). The idea of the reward system w as that with harder work you can get both physical and psychological rewards. His policy worked well, with the effect of employees working much harder, and consequently the business had a huge improvement. In 1985, the company expanded a lot, there were around 800 employees in the company, and their products became more plentiful such as cast iron cookers and fires (Martin, 2000). At that time, Vulcan was very powerful; the company occupied a lot of market shares in the manufactured cookware market. The products they produced were of high quality and standards. However, there are also a few possible drawbacks from managers administering the views and values of their workers. If an organisation only has one culture it will lead to limiting the organisation. First of all, it will result in the organisation having difficulty in adapting to external changes owing to a lack of a diversity of perspectives, principles and views in the organisation. Secondly, it will lead to a lack of va riety which is essential for advance and adjustment to contest constant commitment to inappropriate ideologies. Thirdly, there is a possibility of conformist thinking (Lecture). However, I believe it is still superior to have one culture in an organisation. As managers are an organisations head, they will adapt external change and help the company to innovation. Part three: the perspective of employees Lastly, I argue that employees agree with managers trying to direct the perspective of employees. As an employee, work is a part of their life. Despite family, people spend most of their time working; it is not avoidable as everyone wants for a good life we have to work. If employees have the same perspective, views and values as managers it will make work much more enjoyable and satisfying. In the situation of an employee who works at a company, if he does not agree with the views and values of the managers, and thinks the managers views are wrong, then it will be difficult for him to work with the whole group. As work is a part of life, people have to enjoy it, and the best way of doing this is to treat the organisation as family and work mates as friends (Rosen, 1988). If we have the same point of view as managers, then it will be easier to work hard for the family because the employees want it to progress. Nowadays, there are more organisations which want to create a corpora te culture as it can make employees work more efficiently. Many senior managers have tried various ways to seek to manage the views of employees. In Rosens reading, US adverting company Shoenman and Associates had a tradition with their employees which was a Christmas party. The company held such a party in order to make the relationship of managers and employees closer. Through the social event, hierarchically arranged relationships of the office are to a degree stripped and levelled during and through play (Rosen, 1988). Furthermore, through all of these kinds of customs, the company wants to express an idea to their employees that work is your wife and work mates are your friends (Rosen, 1988). In the party every thing is free; most of the employees enjoy leisure time with work mates from the company. During this kind of practice employees can relax, dance and chat with each other, even with those senior managers who are highly respected and hard to contact at working days, and i n the party the employees do not have working pressure because it is just a company outing and it has nothing to do with working, they do not need to be concerned with what to talk about, how to behave, and all the things that are restricted in the corporation. According to Rosens writing, some employees from Shoenman and Associates do enjoy the company social events while others do not due to them believing that the Christmas party is a business meeting, even though the unwritten rule is that this party is just a social event, and that the employees do not need be stressed and follow the rule of company, that everyone should just enjoy the party dance and chat with working mates, they still feel they have to be concerned about their actions as their senior managers and CEO are all attending the party. They may prefer to spend holiday time with their own family rather than with people who from company. But I believe that most of the employees do enjoy events with people from the business and they want to have more chance to be familiar with each other. Conclusion: To conclude, I reason that administering the perception and ideals of employees to a single purpose benefits them as well as the organisation. I outlined this in three parts, in the first part starting by talking about the difficulty of shaping organisational culture, and the benefits of having a singular organisational culture. In the second part I went on to talk about while the organisation management are the head of the company employees are equally as important, and how a rewards and incentives system can greatly improve the output of the labour workforce while also improving the company itself. I also mentioned the possibilities of how having a single organisational culture can lead to limitations on company advancements. In the third section I talked about how organisational social events can reinforce the organisational view in the minds of the workers, and how they can improve the quality of employees work life, but also how if they have differing views from the managerial class it is more difficult for them to adjust to working for a large corporation. Management is the skill to bringing together the tiniest details of a business into harmony, and how when, as in the case of the company Vulcan and its Managing Director, it is done well it can greatly advance an organisation as well as improving the satisfaction of the workers and quality of the workplace for the people who work in it.