Reflexivity in relation to anthropologicalethnographic filmmaking Essay

Reflexivity in relation to anthropologicalethnographic filmmaking - Essay Example The Ax Fight was originally created to show students the difficulty in placing a single point-of-view out of a certain field experience. The footages actualize the teaching process and problematize the translation from one cultural experience to another cultural idiom, within which the anthropologist often condenses, analyzes and-makes intellectual models (Ennis, Asch, 1). In the first unedited section of the film all the events are presented and the sound continues even after the film goes dark and the comments of Chagnon, Asch and Johnson are just heard. In the second part, Napoleon Chagnon explains the socio-political significance of the fighter's behavior. The third part, delves more into the socio-cultural complications of the village people. The final section is an edited version. Timothy wanted his students to understand the shortcoming of a film that is has smooth editing and fibula structure like the Nanook of the North. Films like The Ax Fight show how an intellectual inter vention influences an onlooker. The film is also a direct criticism of the inconsistencies of spectatorship and how the desired effect in understanding is achieved by a good filmmaker. Contrary to pre-disposed notions in science and filmmaking, anthropological faithfulness to observation of another culture cannot ever be beyond bias, unless reflexivity is used to both question that objectivity and nurture mental exercise within the spectators. Anthropological filmmaking combines two processes together. One is the filming process and the other is social science. The balance is crucial since science and art clash headlong. The frame within the camera may help to communicate cultural conditions and also further Western knowledge of the "Other", but a culture cannot be completely understood just by introducing a non-fictional narrative as Flaherty does. The limitations are numerous. First, a narrative forces the plot of the actual observation towards a composition of a fiction. Flaherty wanted to manipulate viewers understanding of the biography of Nanook and not further any racist understanding of the Eskimo culture. But with Timothy Asch reflexivity is the first sta nce to question the all-understanding nature of the anthropologist. He wanted to keep certain signs and their nature open to critical interpretation and never risk a comfortable lapse leading to an acceptance from his audience. The conscious effort to keep the audience visually uncomfortable while watching the fight makes the confrontation of inter-cultural experience more critical. Accumulation of data, the process of accumulating it, narration, and little editing of the film may help the nature of critical viewership and gives minimum control to the maker. Timothy and Chagnon relinquish all control of their point-of-view and plunges their own spectatorship with that of the audience in such a way so that it becomes impossible to neglect the cultural and ideological subjectivity of the anthropologists. The students are left to situate the filmed observations within the sphere of public and uniform scientific study. The anthropologists become a part of the study themselves since their psychological perspectives are of profound value in regard to the choice of the subject itself. Human recording of another human behavior is the proper subject of postmodern dialogism. The indefinite and inadequate interplay of

Article analysis Essay Example | Topics and Well Written Essays - 2500 words

Article analysis - Essay Example The oil producers in the U.A.E. are expected to increase production of oil as a response to this fall (Cornwell, 2014). This reaction of the government is contrary to the conventional economic theory which states that if the price falls then the suppliers should reduce the level of their production. But this is not happening in case of the U.A.E. The economic theory of demand and supply fundamentally states that as price of a product increases less and less quantity of the product is demanded and as the price of a product rises then its supply increases. The equilibrium market price is determined by the point where the quantity demanded by the consumers equals to the quantity supplied by the producers (Frank, 2008). The following diagram shows the same. The initial equilibrium point is marked by E1 and the initial equilibrium price is P1 and the quantity demanded is Q1. A fall in the market demand due to exogenous market conditions like recessions or weak demand of the products causes the price to fall and the demand curve shifts to DD2 where the new quantity is Q2 and the price falls to P2 (Frank, 2008). Following this theory it should be expected that a global fall in prices would discourage the suppliers and they should reduce their production. However, according to the news article this is unlikely to happen. U.A.E. is expected to produce 3 million barrels of oil by the end of the current financial year. According to experts factors like weaker growth of China, rise in shale production in the U.S.A. and oversupply of oil are the main factors that are responsible for the global fall in oil process. A fall in the global oil prices could be represented by a downward shift in the demand curve from point DD1 to DD2 where quantity demanded equals quantity supplied. The new equilibrium point is reached at E2. However, the determination of oil production in the global economy does not follow the cardinal

Important Models of Justices' Behaviour at the Supreme Court Research Paper

Important Models of Justices' Behaviour at the Supreme Court - Research Paper Example There are three models that are mostly displayed by the justices and the judges of the Supreme Court when deciding cases in the supreme courts. These are strategic, attitudinal and the legal orientations. Strategic behaviour refers to the justices’ actions to maximize their overall benefits in light of their expectations concerning their choices of other actors involved in the decision making process. Others are legal and the attitudinal models that aid explain the legal verdict arrived by the justices at the supreme courts Though the attitudinal models of justice behaviour was initially establish in the US, students and other legal practitioner have found the strategic models of behaviour alluring. This dissent of behaviour focuses on the interpedently nature of judges and justices’ behaviour in their decision making, which does not shun the policy preferences of the justices’ .This a strategic begins with assumption that justices are motivated by their policy p references, but further acknowledges that realization of those preferences is a function of other relevant actors. As notes by (Epstein and Knight) Justices may be primarily seekers of legal policy, but they are not unconstrained actors who make decisions based only on their own ideological attitudes. Rather, justices are strategic actors who realize their ability to achieve their goals depends on a consideration of the preferences of other actors, the choices they expect others to make, and the institutional context in which they act. (10). This implies that whole a justice acting on the basis of his attitudes, another justices acting on the same preferences may be expected to behave differently based on strategic considerations. For instance, for justice behaviour‘s preferences mat depend on the other justice and judges reactions and opinions (08-956 wood v. Allen. (01/20/10) Recent literature provides that strategic concerns of behaviour go beyond attitudinal as they tend t o explain further other reasons that attitudinal approach may not offer.(Wood v. Allen, Wood v, state and Alabama, US, 1998) In recent findings, attitudinal concerns have challenged by judges who tend to adopt strategic orientation towards and it was evidenced in a variety of contexts. First, when setting their agendas (Rice v. Collins, 546 US 333,339), second, when writing majority opinions (Wood v. State, 715 819 (1998)) and lastly issuing separate opinions, even on the same case(s) (Williams v. Taylor, 529 US 362,462). Several studies undertaken by Brace and Hall at the aggregate (Brace and Hall; Hall and Brace, 147-162) and individuals (Brace and Hall; Hall and Brace) provides explanations on the dissenting behaviour. They note that when making judicial decisions, justices and judges decide not to follow their ideological differences in certain circumstances in anticipation of decisions by exogenous factors such as electoral constituency, in order to keep job . Other evidence th at pursue a strategic line of inquiry as is vital in the decision making process. This is because the strategic models of supreme court justices takes into account other factors more

Reflexivity in relation to anthropologicalethnographic filmmaking Essay

Reflexivity in relation to anthropologicalethnographic filmmaking - Essay Example The Ax Fight was originally created to show students the difficulty in placing a single point-of-view out of a certain field experience. The footages actualize the teaching process and problematize the translation from one cultural experience to another cultural idiom, within which the anthropologist often condenses, analyzes and-makes intellectual models (Ennis, Asch, 1). In the first unedited section of the film all the events are presented and the sound continues even after the film goes dark and the comments of Chagnon, Asch and Johnson are just heard. In the second part, Napoleon Chagnon explains the socio-political significance of the fighter's behavior. The third part, delves more into the socio-cultural complications of the village people. The final section is an edited version. Timothy wanted his students to understand the shortcoming of a film that is has smooth editing and fibula structure like the Nanook of the North. Films like The Ax Fight show how an intellectual inter vention influences an onlooker. The film is also a direct criticism of the inconsistencies of spectatorship and how the desired effect in understanding is achieved by a good filmmaker. Contrary to pre-disposed notions in science and filmmaking, anthropological faithfulness to observation of another culture cannot ever be beyond bias, unless reflexivity is used to both question that objectivity and nurture mental exercise within the spectators. Anthropological filmmaking combines two processes together. One is the filming process and the other is social science. The balance is crucial since science and art clash headlong. The frame within the camera may help to communicate cultural conditions and also further Western knowledge of the "Other", but a culture cannot be completely understood just by introducing a non-fictional narrative as Flaherty does. The limitations are numerous. First, a narrative forces the plot of the actual observation towards a composition of a fiction. Flaherty wanted to manipulate viewers understanding of the biography of Nanook and not further any racist understanding of the Eskimo culture. But with Timothy Asch reflexivity is the first sta nce to question the all-understanding nature of the anthropologist. He wanted to keep certain signs and their nature open to critical interpretation and never risk a comfortable lapse leading to an acceptance from his audience. The conscious effort to keep the audience visually uncomfortable while watching the fight makes the confrontation of inter-cultural experience more critical. Accumulation of data, the process of accumulating it, narration, and little editing of the film may help the nature of critical viewership and gives minimum control to the maker. Timothy and Chagnon relinquish all control of their point-of-view and plunges their own spectatorship with that of the audience in such a way so that it becomes impossible to neglect the cultural and ideological subjectivity of the anthropologists. The students are left to situate the filmed observations within the sphere of public and uniform scientific study. The anthropologists become a part of the study themselves since their psychological perspectives are of profound value in regard to the choice of the subject itself. Human recording of another human behavior is the proper subject of postmodern dialogism. The indefinite and inadequate interplay of

Important Models of Justices' Behaviour at the Supreme Court Research Paper

Important Models of Justices' Behaviour at the Supreme Court - Research Paper Example There are three models that are mostly displayed by the justices and the judges of the Supreme Court when deciding cases in the supreme courts. These are strategic, attitudinal and the legal orientations. Strategic behaviour refers to the justices’ actions to maximize their overall benefits in light of their expectations concerning their choices of other actors involved in the decision making process. Others are legal and the attitudinal models that aid explain the legal verdict arrived by the justices at the supreme courts Though the attitudinal models of justice behaviour was initially establish in the US, students and other legal practitioner have found the strategic models of behaviour alluring. This dissent of behaviour focuses on the interpedently nature of judges and justices’ behaviour in their decision making, which does not shun the policy preferences of the justices’ .This a strategic begins with assumption that justices are motivated by their policy p references, but further acknowledges that realization of those preferences is a function of other relevant actors. As notes by (Epstein and Knight) Justices may be primarily seekers of legal policy, but they are not unconstrained actors who make decisions based only on their own ideological attitudes. Rather, justices are strategic actors who realize their ability to achieve their goals depends on a consideration of the preferences of other actors, the choices they expect others to make, and the institutional context in which they act. (10). This implies that whole a justice acting on the basis of his attitudes, another justices acting on the same preferences may be expected to behave differently based on strategic considerations. For instance, for justice behaviour‘s preferences mat depend on the other justice and judges reactions and opinions (08-956 wood v. Allen. (01/20/10) Recent literature provides that strategic concerns of behaviour go beyond attitudinal as they tend t o explain further other reasons that attitudinal approach may not offer.(Wood v. Allen, Wood v, state and Alabama, US, 1998) In recent findings, attitudinal concerns have challenged by judges who tend to adopt strategic orientation towards and it was evidenced in a variety of contexts. First, when setting their agendas (Rice v. Collins, 546 US 333,339), second, when writing majority opinions (Wood v. State, 715 819 (1998)) and lastly issuing separate opinions, even on the same case(s) (Williams v. Taylor, 529 US 362,462). Several studies undertaken by Brace and Hall at the aggregate (Brace and Hall; Hall and Brace, 147-162) and individuals (Brace and Hall; Hall and Brace) provides explanations on the dissenting behaviour. They note that when making judicial decisions, justices and judges decide not to follow their ideological differences in certain circumstances in anticipation of decisions by exogenous factors such as electoral constituency, in order to keep job . Other evidence th at pursue a strategic line of inquiry as is vital in the decision making process. This is because the strategic models of supreme court justices takes into account other factors more

Complete Course Material Essay Example for Free

Complete Course Material Essay MGT 426 Complete Course Material MGT 426 Week 1 DQ 1 Why is change necessary? What about change is disruptive? How can meeting the challenge of change become organizational opportunities? MGT 426 Week 1 DQ 2 Change: what is in it for me? What is the cycle of change? What are some examples of continuous change? What are some examples of discontinuous change? What are some differences between continuous and discontinuous change? MGT 426 Week 1 Individual Assignment Article Review MGT 426 Week 2 DQ 1 What is organizational culture? What is the relationship between shared vision and organizational culture? How does organizational culture affect an organization’s ability to cope with change? MGT 426 Week 2 DQ 2 How does modeling a change process facilitate change? What are the strengths and weaknesses of the three change models? What are some applications of the three change models? MGT 426 Week 2 Individual Assignment Roles of Managers and Individuals Paper MGT 426 Week 2 Learning Team Assignment Managing Change Paper Part I MGT 426 Week 3 DQ 1 Why is it important to do a gap analysis? How do you apply gap analysis to the change models? Give an example of when you may use a gap analysis. MGT 426 Week 3 DQ 2 What are the steps in implementing change? What should you consider when selecting a change implementation model? Explain. MGT 426 Week 3 Individual Assignment Resistance to Change Paper MGT 426 Week 3 Learning Team Assignment Managing Change Paper and Presentation Part II MGT 426 Week 4 DQ 1 How do you measure the progress of change? How do you know change has been institutionalized? How do you determine what changes have occurred? MGT 426 Week 4 DQ 2 What organizational systems are impacted by the cycle of change? Explain. What systems are not as easily impacted by change? Why or why not? MGT 426 Week 4 Individual Assignment Ethics During Change paper MGT 426 Week 4 Learning Team Assignment Managing Change Paper Part III MGT 426 Week 5 DQ 1 What characteristics of a learning organization do you see in your organization? What can your organization do to transform into a learning organization? MGT 426 Week 5 DQ 2 How closely does your organization follow Senge’s five disciplines? Is this useful? Why or why not? MGT 426 Week 5 Individual Assignment Learning Organization Paper MGT 426 Week 5 Learning Team Assignment Managing Change Paper and Presentation Part IV

Overview Of Aloe Vera

Overview Of Aloe Vera Aloe Vera is known as a TRUE ALOE. It is mostly used in herbal medicines so it is also known as FIRST AID PLANT or MEDICINAL PLANT. The ALOE word was derived from the aerobic word-alloeh which means BITTER SUSSTANCES. Aloe is an emollient resin with the softening properties. It belongs to family Liliaceae [1]. 1.1.1 HISTORY:- Aloe vera was considered to one of best medicinal plants used in the treatment of various health conditions. It was first understood by Egyptians to having multiple nature of it properties [19]. But Aloe Vera was first discovered in 1862 by German Egyptologist -George Fbers. But its first English translation was found in 1655 by John Goodyear in Dioscorider de materia medica, and he wrote aloe is very better in taste. Aloe Vera has been used by many different cultures: Indians, Chinese, Greeks, Romans, have all used aloe Vera as a medicinal or healing plant. Botanists have find more than 300 species of aloe Vera like rumex, barbadensis etc. Out of this species, only five have strongest medical benefits likes Aloe barbadensis miller, Aloe perryi baker, Aloe ferox,Aloe saponaria , Aloe arborescens. Aloe barbadensis miller is mostly widely used and most potent as well. Throughout the history of aloe vera, the plant is in various form having tropical properties-refered as plant of immorality. It is belonging to lily family because flower produce annually in the spring which resembles of ester lily. [1-2]. 1.1.2 COMMON NAME:- Chinese aloe, True aloe, Indian aloe, First aid plant, Medicinal plant, Miracle plant ,Barbadose aloe, Burn aloe and many more [1]. 1.1.3 SCIENTIFIC NAME:- Aloe barbadensis, Aloe capensis [1]. 1.1.4 ALTERNATIVE NAME:- Aloe Vera has a different name in worldwide like, In India-Ghrtakumari But in Gujarat and Rajasthan its known as Gwarpatha,In Pakistan-quargandal which is used in unani medicine,In Indonesia-lidah buaya,In Thailand- crocodile tail plant,In south Africa-sabila [2]. 1.1.5 DESCRIPTION:- Aloe Vera grows in to the dreary climates or land. So it is mostly found in the India, Africa, Caribbean, and other dry climates. Aloe Vera is a short stem or stems less plant. It mainly contains leaves and flowers. Aloe leaves are in green in colour and thick with the same variety. The size of leaves is 50-60 cm long and 4-5 cm thick. The margin of the leaves is serrated. But plants bear flowers once in a year in summer season. Flowers contain tubular collar which is yellow in colour and contain aloe tic juice can be collected by cutting the leaves close to stem. The structure of aloe leaf shows an outer cortex which is hard duo to the present of calcium and magnesium. Tubes of xylem and phloem were found below the cortex and its supplied water and minerals to leaves [1]. 1.2 CHEMICAL CONSTITUTE:- Aloe Vera has weird medicinal properties. Botanists have found more than 200 important nutritional constituents in aloe Vera leaf which having to perform a function. They are MINERALS like Calcium, Sodium, Copper, Zinc, Iron, and Manganese (Essentials for bones, Regulates acidic or alkaline level of body fluid), VITAMINS like Vitamin A, Vitamin C, Vitamin E, Vitamin B12, and Folic Acid(To develop new blood cells), ENZYMES like lipase, Peroxidise (Helps indigestion), SUGAR like Monosaccharides and polysaccharides include Sucrose, lactose(Maintain cholesterol level, Improve the strengthening of bones),LIGNINS which are the Cellular substances which hasnt medicinal benefits But it has an accessing property. AMINO ACID which are required by human body provided by aloe Vera like Leucine, Isoleucine, Valine, Theonine, Lysine, Methionine, Phenylamine, STEROL like Camp sterol, sitosterol(Pain killing properties),ANTHRAQUINONES like Aloin, Anthranol, Aloetic acid, Barbaloin, Isobarbaloin and Aloe emodin Aloe emodine [4] Aloin [3] And OTHERS like Salicylic acid,Tannins,Monosulfonic acid, Acemannam,Water [1-2]. Aloes Vera mainly contain 96-99% of water and constitutes are in rare part. So its work in based on SYNERGISTIC ACTIONS which means working together of two or more substance and give greater action as compare to single substance. This phenomenon was explained by Dr.Atherton [1-2]. 1.3 CULTIVATION OF ALOE VERA [1-5]:- Aloe Vera grown commercially for its high demand in cosmetic industries and treatment of healing in India. It has medicinal properties due to its bitter taste. Aloe Vera grown in a rainless climates or land. Its cant stay in cold temperature. So it is grown in all part of South Africa and India like Rajasthan, Gujarat, Maharashtra and many more. Soil requirement for all species are coarse sandy loam soil with some fertility. In addition, it is noticed that its growth was easier and faster in black cotton soil in central India with the nearly by acidic ph. For the perfect growth of Aloe Vera, lane should not distribute below the level of 20-30cm. Aloe Vera need a flat land and should be exact level with 2-3 farming. Plant should plant with some distance because of irrigation. Distance between two fields was 10m X 3m. The proper time of suckers should be planted in month of July-August in monsoon season because suckers get proper water to grow up and survival. Suckers should have 3-4 mouth old having 3-4 leaves and 20-22 c long. About 30,000 suckers needed for one hectare planting. After planting suckers, soil around the root should press and waste must be made proper to avoid the sluggishness. Irrigation is necessary a critical stage to proper growth of plant. First irrigation is required after planting suckers followed by 4-6 per year which is reducing by every time. Weeding should be carried out after planting the plant and two or three weeding carried out throughout year. All the species of Aloe Vera are insusceptible to most of insects and pests from any part of county. But sometime bugs, leaves dots have been reported in some part of country because of light irrigation. Generally yield obtained from 2 to 5 year after plating. Average yield after 2 year is around 15-20 t/ha fresh leaves. Well managed irrigated crop give around 30-35 t/ha fresh leaves. Fully well developed leaves give an ALOE JUICE. After harvesting, marketing of Aloe Vera leaves is not fully developed in our country. 1.4 USES:- Aloe Vera is well known for its medicinal properties since centuries. The plant is able to cure a wide range of disease. The part of plant like leaves, leaf gel is used clinically. Some respective uses of Aloe Vera are (1) EXTERNAL USES-Use in treatment of burns, allergic reaction, wounds,acne,Rheumatoid arthritis, Rheumatic fever, Acid indigestion cuts, Inflammatory condition of digestive system, Sun burn ulcers, Kill mould,bacteria,viruses,fungus,Wrinkles etc (2)COMMERCIAL USE: Aloe Vera is widely use in cosmetics and many more like in ointments,pills,jelly,lotion,bevareges.as a foodstuff etc [1-6]. (3)MEDICINAL USE-They are Treatment of diabetes because it reduce blood sugar, Treatment of AIDS, Protect from lung cancer, Aloe juice is used for consumption issues, Prevent fungal infection, Prevent vaginal infection, Prevent scarring,Eczema,Constipation,Intestinal infection, Relieves from candiala,Treatment of hyperglycaemia, Maintain level of cholesterol, Injection of Aloe Vera extracts to treat cancer, Skin disorder. And some of general use like Antibacterial properties, Antifungal properties, and Prevent radiation induced injuries, Inhibit growth of streptococcus species in vitro [1-6]. 1.4.1 ALOE PREPARATIONS: some of the aloe preparation which are widely use like aloe vera shampoo, aloe vera lotion, aloe vera gel, aloe vera juice, aloe vera eating, aloe vera butter, aloe vera dried herbs powders, aloe vera shower gel, and aloe vera vegetarian capsules [1]. 1.4.2 SIDE EFFECT AND SAFETY PRECAUTION: Aloe Vera has a medicinal or healing properties, it does come with some disadvantage. Aloe Vera gel, aloe Vera cream does not having major side effect but other preparation like juice, shampoo butter, having certain impediment like, Diarrhea,Blood electrolyte imbalance, Constipation, Muscle weakness, Abnormal heart rhythms [7]. Aloe Vera injection is to be avoided because of having fatalities. It can cause death with cancer patients Aloe Vera should be avoided while pregnancy, children, breast feeding [7]. ALPHA-GLUCOSIDES INHIBITORS: 1.5 INTRODUCTION TO ALPHA-GLUCOSIDE ENZYME: It is a class of a medication for type-2 diabetes which decrease blood sugar level by decreasing carbohydrates from the intestinal. Discovery of alpha-glucosides inhibitors has been very useful to develop therapeutic for the treatment for the carbohydrate- mediated disease like diabetes. Two classes of drugs like glycosidase inhibitors and lipase inhibitors which lower blood glucose by changing the absorption level of fat and carbohydrates [8-9]. Alpha glucosides inhibitors like acarbose, miglitol, voglibose have been studies in Europe country but some of these are also available in united state [8]. Alpha -glucosidase inhibitors reduce the impact of carbohydrates blood sugar by inhibit the upper level of gastrointestinal [8]. 1.5.1 MECHANISM OF ACTION: Alpha glucosides inhibitors are competitive inhibitors of 1alpha glucosidase which are located in the brush border of small intestinal (epithelial cells). These inhibitors bind to the oligosaccharide binding site of the enzyme and delay digestion of polysaccharide resulting slow down food digestion in gut [10]. While the gastrointestinal tract dose not play a important role in the cure of diabetes, but changing its physiological activity can be used to control disease. In these case alpha-glucosides inhibitors are used. Alpha glucosides delay digestion of carbohydrates by hydrolyses of oligosaccharides into monosaccharide. Alpha glucosidase inhibitors can be used to reduce glycemic excussions and hypoglycaemia having type-1 diabetes. Moreover it is used in the treatment of patients with type-2 diabetes and its decrease plasma triglyceride [11]. Alpha glucosides inhibitors have evidence helpful for the people with diabetes who havent able to keep their blood sugar level within a safe range. In such case inhibitors like acarbose and miglitol help to keep the blood sugar level in safe range by slowing a rate of intestinal which absorbed sugar from blood while eating. These inhibitors can cause low blood sugar while used in combination with other medication for diabetes or with insulin. Diabetic people who are regularly using insulin but once they are use alpha glucosidase inhibitors than they reduce use of insulin [12]. CHAPTER- 2 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY. 2.1 INTRODUCTION: Chromatography is the separation of a mixture into single components by using mobile and stationary phase. High performance liquid chromatography is widely use in analytical chemistry and industrial level to identify components. It is one of the fastest growing techniques in pharmaceutical industry for analysis of mixture os substance. It is also knows as high pressure liquid chromatography. It is a high improved as compare to column chromatography. Smaller particle size cans (3-20ÂÂ µ) be analysed by hplc and allows much better separation of a components. Detection method is used in hplc which are highly automated and extremely sensitive as compare to column chromatography [13-14]. Some of the advantage over column chromatography is, (a) small stationary phase are used with widely range available, (b) column which are used in hplc which are made up of metal and small in size so no fear to breakage, (c) hplc is available in analytical and preparative scale. But some of disadvantage over column chromatography like cost of equipment is high so handle with care, operating pressure is high (500-3000 psi)[14]. Basic principle is base on adsorption. When a mixture of components is introduced in to column, they are travel according to their affinity. The components which has a more affinity towards adsorbent, travel slower and vice versa [14]. ÂÂ ®NORMAL PHASE HPLC: Here, stationary phase is polar in nature and mobile phase is non polar in nature. In this method non polar components eluted first because of less affinity. The column is fitted with a silica gel and hexane used as a solvent. Column length is 150-250 mm and less than 4.6 mm diameter [13-14]. ÂÂ °REVERSE PHASE HPLC: Here, stationary phase is non polar and mobile phase is polar in nature. Polar components eluted first. Column size is same but modify to make a non polar by using long chain of hydrocarbons like C8,C12,C4,octadecyl and mixture of water and alcohol use as a solvent. In this case strong attraction between polar solvent and molecular in the mixture passed through column. So it is common phase of hplc [13-14]. 2.2TYPES OF HPLC TECHANIQES: -There are major four type of hplc as below, (1)Partition chromatography: In 1952, Archer John Porter Martin and Richard Laurence Millington Synge were awarded a Noble prize in chemistry to development of these techniques. So it is first techniques which developed for the separation of components like amino acid. These chromatography principles are also applied to the thin layer chromatography, paper chromatography to separates the components. (2) Adsorption chromatography: The main hplc principle is based on adsorption. Here components can be separated because of difference in affinity of components towards normal or reverse phases [14]. (3) Ion exchange chromatography: It is most frequently used chromatography for the separation and purification of protein, nucleic acid, polypeptides, and other charged molecular. It is a succeful technique because of its high capacity, simplicity, and high resolution power. The main principle is base on reversible exchange of ions between ion present in the solution and those present in ion exchange resin [15]. ÂÂ ®Cation exchange:- Solid-H+ + M+ === solid-M+ + H+ (Solution) (Solution) ÂÂ ®Anion exchange: Solid-OH- + A- ==== solid-A- + OH- (Solution) (Solution). [14] Separation in ion exchange chromatography depends upon reversible adsorption of charged solute molecular to ion exchange group of opposite charge. Some of the functional group used in ion exchange chromatography is [16], NAME OF ANION FUNCTION GROUP Diethyl amino ethyl -O -CH2-CH2-N+H(CH2CH3)2 Quatemary amino ethyl -O-CH2-CH2-N+H(C2H5)2-CH2CHCH-CH3 NAME OF CATION FUNCTION GROUP Carboxymethyl -O-CH2-COO- SULPHOPROXY -O-CH2-CHOH-CH2-O-CH2-CH2SO3- (4)Size exclusion chromatography: It is also know as a gel chromatography. In 1959, Porath and Flodin described the separation of water soluble components by this chromatography. As soon as gel had commercially available, they were extensively used for the separating purpose [17]. Here gel used as molecular sieve and hence mixture of substance with different molecular sieze are separated. Soft gel like dextran, polyacrylamide are used [14]. It is used to analysis of synthetic polymers and oligomes, lipids, natural macromolecular like protein, glucose, cellulose derivatives, and crude oil alkenes [17]. HPLC techniques also has two different class like (a) analytical HPLC-it is used when analysis very small amount of sample are needed. And (b) preparative HPLC- it is used when separation of few gram of mixture by HPLC. But it is also can classified on base of separation and there are (a) analytical HPLC: Where only analysis of the samples is done. Recovery of the samples for reusing is normally not done, since the sample used is low. Eg. mg quantities. And other is (b) preparative HPLC: Where the individual fractions of pure compounds can be collected using fraction collector. The collected samples are reused eg. Separation of few grams of mixtures by HPLC [14]. 2.3 INSTRUMENTATION [13-14]: Brief introduction of instrumental techniques are as follows, (1)PUMP: Mechanical and pneumatic pump are used in hplc. Mechanical pump operates with constant flow rate where pneumatic pump operates with constant pressure. Its necessary to use pump because solvent must passed through column at high pressure. (2) SOLVENT DEGASSING: In isocratic techniques, mobile phase are prepared by using mixture of solvents, than passed through column with under high pressure resulting bubble formed, so it cannot give proper result or steady baseline. So before using solvent mixture in HPLC, degassing is necessary. So some of degassing technique is use like ultrasonication. HPLC (3)COLUMN: -Two types of column used in HPLC, ÂÂ ® Guard column: It is also known as safety column. Any impurities present in solvent or mobile phase which can predict by guard column. ÂÂ ®Analytical column: It widely use which can decided efficiency of separation. Different column size is available depending upon separation. Column made up of stainless stell, glass, poly ether ether kiton etc. (4)DETECTORS: -They are (a)UV detectors: Many organic compounds absorbed UV light of different wavelength. Two type of UV detectors are available depends on wavelength- short wavelength and long wavelength. By UV detector we can get direct reading is how much light is observed. (b) Flurometric detectors: It can be used for the substance which can absorbed florescence light. (c) Conductivity detectors (d) Refractive detectors (e)PDA detectors which are similar to UV detectors. (5)RECORDER: They are used to recorded response from detectors. It can record base line and retention time of peak. Now a day computer is used to recording data. 2.4 HPLC PARAMETERS: 2.4.1 INTERNAL DIAMETER: Internal diameter of the hplc column play an important role in detection or separation of compounds. For the laboratory use small column was used but in industrial level big column was used. 2.4.2 RETENTION TIME: it is a defined as a time between point of injection and appearance of peak. It measure in minute or seconds. 2.4.3 RETENTION VOLUME: Retention volume in the volume of mobile phase required eluting 50% of the compounds from the column, Retention time = Retention time.Flow rate. 2.4.4 PUMP PRESSURE: Pumps which are used in hplc very in pressure depends on separation. Modern HPLC system has been improved to work at high pressure so they are able to separate small particle size from mixture. 2.4.5 THEORETICAL PLATE: It is a functional unit of the column. Distribution of solute between stationary phase and mobile phase has attained equilibrium in the plate. High of the plate depends on efficiency of separation. It is knows as a HETP(HIGHT EQUIVELENT TO THEORITICAL PLATE). If HETP is less, than column is more efficient and if HETP is more, than column is less efficient. HETP is given by Van Deemter equation HETP = A + (B/u ) + Cu Where, A = Eddy diffusion term or multiple path diffusion which arises due to packing of the column. This is unaffected by mobile phase velocity or flow rate. This can be minimised by uniformity in packing. B = Longitudinal diffusion term or molecular diffusion which depends on flow rate. C = Effect of mass transfer which depends on flow rate. u = Flow rate or velocity of the mobile phase. 2.5 APPLICATION OF HPLC: High performance liquid chromatography has a large divergence of application. Initially it was just used to analysed antibiotic, vitamins by development of reverse phase [18]. High performance liquid chromatography can analysed very minor amount of substance like monograms at analytical scale to micrograms at preparative scale. It is used in a food analysis, biotechnology, biochemical separation, pharmaceutical field, and chemical industries like dyes, fatty acid, toiletry products [14].High performance liquid chromatography has a easy and good reproducibility so it has been widely use in clinical laboratory. One of the initially application of hplc in the field of clinical was quantification of theophyline in asthmatic infant [18]. Some of important applications are as likes,(a) Isolation of natural product,(b)Quantitative analysis or assay of product,(c)Identify, analysing and purification of compound mixture,(d)To check purity of compound mixture,(e) To find physical properties of compounds,(f)Used in analysing water pollution [18]. The quantity of amount is also determined by hplc which including following method [14], ÂÂ ® By comparing standard and sample peak area, the quantity of sample can be determined which is known as a DIRECT COMPRISION METHOD. ÂÂ ® To find concentration of sample by plotting graph between peak area vs. concentration of the standard drug and intrapolation of unknown sample which is known as CALIBRATION METHOD. Hplc play an important role in drug discovery with the combination of NMR. It is also useful to identify various chemical species with the combination with mass spectroscopy. It play valuable role in therapeutic drug monitoring to separation of drug from plasma which is quick process [14]. It has some advantages over other chromatography like, (a) Thin layer chromatography is inexpensive but it cant give batter resolution to compare HPLC. (b) Mass spectroscopy required a large capacity of maintain budget and skillfull person to run chromatography but in HPLC its not high budgt. (c) For the HPLC we have large number of mobile phase is available so we have versatility in choice of mobile phase as compare to other chromatography. (d) Atomatic injection system are available in HPLC, so its time saving and give high resolution as compare to other [20]. 2.6 LIMITATION OF HPLC: It is a time consuming method. Analysis of the compound mixture, mobile phase (mixture of components) are used resulting large amount of waste solution are obtained [14]. CHAPTER- 3 EXTERIMENT 3.1 AIM: The aim of project was to known batter separation of standard solution of Aloe Vera stem in mobile phase by using isocratic hplc techniques and find enzyme activity on it by using assay whether it is more potent or less potent. 3.2 CHROMATOGRAPHY EQUIPMENTS: 3.2.1 HPLC SYSTEM: This system is manufactured by Agilent technology 1200 series with model no G1310A and serial no DE62956545. 3.2.2 SOFTWEAR USED: The software used was Microsoft Windows XP. 3.2.3 COLUMN USED: The column used was Kromaril 5C18 which is made up of stainless still. 3.2.4 APPARATUS USED: Pipette with different size available like 1000land 5000ÂÂ µl, Analytical balance, Volumetric flask (10ml, 20ml), Funnel, 100ml Beaker, Measuring cylinder (500ml, 50ml, 20ml,10ml),Filter paper which made by Fischer scientific with size 0.45ÂÂ µm, Filter holder made up of pyrex brand glass. 3.2.5 PREPARATION OF MOBILE PHASE: The mobile phase used was a mixture of a methanol and water with ratio of 50:50. So take methanol and water, mix well, and filter to remove any solid particle followed by degassing to remove air bubble. Than it is ready for experiment. We are using isocratic system so mobile phase may vary in concentration like, 20:80, 30:70. 3.2.6 PREPARATION OF STANDARD SOLUTION OF ALOE VERA: The standard solution of aloe vera stem was provided by Dr.solomon sir and after that i have to dilute in methanol to make up volume to 0.40mg/2ml. 3.3PROCEDURE: First set up the hplc system and run with mobile phase for half an hours. Than inject standard solution of aloe Vera stem and run for half an hours, take the graph. Now change the concentration of mobile phase and again repeat same procedure until you get batter separation. Then compare all graphs with each other and reach to conclusion. Here we were using two different wavelengths like 280nm and 360nm. 3.5 ASSAY: Assay technique was used to determined enzyme activity of a substance. Here we were using alpha-glucosidase enzyme. Basic principle involved in assay was, phosphate buffer contain free radical which has no colour but when it come across with standard solution, retain its colour after incubation. So colour change like colourless to yellow colour took place. SOLUTION USED IN ASSAY: Here we have to prepare solution for the assay was 25ÂÂ µl, 1.2 U/ml alpha-glucosidase, 25 ÂÂ µl of phosphate buffer, standard solution of aloe vera, 25 ÂÂ µl of 2.5 mM pNPG and finally if proper colour change was not occur than add terminating solution 100 ÂÂ µl of 0.2 M Na2Co3. 3.5.1 PROCUDURE: 1 2 3 4 E 1 2 3 4 5 6 BLANK Concentration in raw at no 1-0.50mg/ml, raw no 2- 0.25mg/ml, raw no 3- o.125mg/ml, and raw no 4- 0.0625mg/ml. Here all the column from 1 to 6 filled by 25ÂÂ µl,1.2 U/ml alpha-glucosidase, then add 25ÂÂ µl of phosphate buffer , add 25ÂÂ µl of test solution, and finally add 25ÂÂ µl of 2.5 mM pNPG. And then transfer 2.5ÂÂ µl solution from low concentration to high concentration. Than kept mixture for some time and incubate plate at 37ÂÂ °C for 10 min. and tested absorbance.