Overview Of Aloe Vera

Overview Of Aloe VeraAloe Vera is know as a TRUE ALOE. It is mostly custom of goods and ser debilitys in herbal medicines so it is as well as 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 rosin with the softening right(a)ties. It belongs to family Liliaceae 1.1.1.1 HISTORY-Aloe vera was considered to integrity of best healthful plants employ in the make outment of various health conditions. It was first understood by Egyptians to having multiple constitution of it strait-lacedties 19. more(prenominal)over Aloe Vera was first discovered in 1862 by German Egyptologist -George Fbers. precisely its first English translation was found in 1655 by John Good class in Dioscorider de materia medica, and he wrote aloe is real better in taste.Aloe Vera has been apply by many different cultures Indians, Chinese, Greeks, Romans, nourish all apply aloe Vera as a medicative o r healing plant. Botanists make water go steady more than 300 species of aloe Vera resembling rumex, barbadensis etc. Out of this species, only quint have strongest medical benefits wants Aloe barbadensis miller, Aloe perryi baker, Aloe ferox,Aloe saponaria , Aloe arborescens. Aloe barbadensis miller is mostly widely apply 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 beca map blush 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, glow 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 identical, In India-Ghrtakumari But in Gujarat and Rajasthan its known as Gwar leada,In Pakistan-quargandal which is use in unani m edicine,In Indonesia-lidah buaya,In Thai devour- crocodile tail plant,In south Africa-sabila 2.1.1.5 verbal description-Aloe Vera grows in to the dreary climates or land. So it is mostly found in the India, Africa, Caribbean, and separate dry climates.Aloe Vera is a short angry walk or stems little plant. It mainly lay offs leaves and flowers. Aloe leaves be in green in act upon and recondite with the said(prenominal) variety. The sizing 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 pile be collected by cutting the leaves keep mum to stem. The structure of aloe leaf shows an outer cortex which is hard duo to the break of calcium and magnesium. Tubes of xylem and phloem were found below the cortex and its supplied weewee and minerals to leaves 1.1.2 chemic CONSTITUTE-Aloe Vera has weird medicin al properties. Botanists have found more than two hundred principal(prenominal) nutritional constituents in aloe Vera leaf which having to perform a function. They argon MINERALS give c are Calcium, Sodium, Copper, Zinc, Iron, and Manganese (Essentials for bones, Regulates acidic or alkaline level of organic structure fluid), VITAMINS like Vitamin A, Vitamin C, Vitamin E, Vitamin B12, and Folic stifling(To develop new tune cells), ENZYMES like lipase, Peroxidise (Helps indigestion), SUGAR like Monosaccharides and polysaccharides include Sucrose, lactose(Maintain cholesterol level, Improve the streng then(prenominal)ing of bones),LIGNINS which atomic number 18 the Cellular substances which hasnt medicinal benefits But it has an accessing property. AMINO ACID which argon required by benignant body provided by aloe Vera like Leucine, Isoleucine, Valine, Theonine, Lysine, Methionine, Phenylamine, STEROL like Camp sterol, sitosterol(Pain violent death properties),ANTHRAQUINONES like Aloin, Anthranol, Aloetic acid, Barbaloin, Isobarbaloin and Aloe emodinAloe emodine 4 Aloin 3And OTHERS like Salicylic acid,Tannins,Monosulfonic acid, Acemannam,Water 1-2.Aloes Vera mainly contain 96-99% of water and constitutes atomic number 18 in r ar bring out. So its utilization in based on SYNERGISTIC fulfillS which means working to let downher of two or more substance and give greater action as comp atomic number 18 to case-by-case substance. This phenomenon was explained by Dr.Atherton 1-2.1.3 CULTIVATION OF ALOE VERA 1-5-Aloe Vera grown commercially for its elevated demand in cosmetic industries and treatment of healing in India. It has medicinal properties due to its tart taste. Aloe Vera grown in a rain slight climates or land. Its flowerpott brook in c mature temperature. So it is grown in all divide of South Africa and India like Rajasthan, Gujarat, Maharashtra and many more.Soil requirement for all species be coarse sandy loam background with some fer tility. In addition, it is noniced that its harvest-feast was easier and faster in black cotton soil in rally 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 blank because of irrigation. Distance surrounded by two fields was 10m X 3m.The proper cartridge holder of suckers should be planted in month of July-August in monsoon season because suckers brook 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 put. After planting suckers, soil around the root should press and waste must be do 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 e very(prenominal) beat. Weeding should be carried out after planting the plant and two or three weeding carried out through with(predicate)out year. all in all the species of Aloe Vera are insusceptible to most of insects and pests from any part of county. But some clip(prenominal) bugs, leaves dots have been reported in some part of country because of gentle irrigation.Generally yield obtained from 2 to 5 year after plating. amount 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, merchandising 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 competent to cure a wide range of disease. The part of plant like leaves, leaf gel is use clinically. Some individual uses of Aloe Vera are (1) EXTERNAL USES-Use in treatment of burns, allergic reaction, wounds,acne,Rheumat oid arthritis, Rheumatic fever, Acid indigestion cuts, Inflammatory condition of digestive transcription, Sun burn ulcers, slay 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 caudex shekels, Treatment of AIDS, Protect from lung cancer, Aloe juice is employ for consumption issues, Prevent fungous 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 lik e 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 complete with some dis returns. Aloe Vera gel, aloe Vera cream does not having study side effect but different preparation like juice, shampoo butter, having certain impediment like, Diarrhea,Blood electrolyte imbalance, Constipation, Muscle weakness, Ab general 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 term pregnancy, children, pap feeding 7.ALPHA-GLUCOSIDES INHIBITORS 1.5 INTRODUCTION TO ALPHA-GLUCOSIDE ENZYME It is a class of a medication for type-2 diabetes which descend 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 assiduousness level of fat and carbohydrates 8-9. Alpha glucosides inhibitors like acarbose, miglitol, voglibose have been studies in europium country but some of these are to a fault 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 rinse border of vitiated intestinal (epithelial cells). These inhibitors bind to the oligosaccharide binding site of the enzyme and control digestion of polysaccharide resulting slow down food digestion in gut 10. fleck the gastrointestinal tract dose not play a important single-valued functi on in the cure of diabetes, but changing its physiological operation 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 slump plasma triglyceride 11.Alpha glucosides inhibitors have evidence helpful for the stack with diabetes who havent able to keep their blood sugar level within a practiced range. In such case inhibitors like acarbose and miglitol help to keep the blood sugar level in safe range by subnormality a rate of intestinal which captive 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 exploitation insulin bu t once they are use alpha glucosidase inhibitors than they reduce use of insulin 12.CHAPTER- 2HIGH PERFORMANCE pellucid CHROMATOGRAPHY.2.1 INTRODUCTION Chromatography is the musical interval of a garland into single components by utilize rambling and stationary stagecoach. 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 pharmaceutic industry for abbreviation of mixture os substance. It is also knows as amply gear pressure liquid chromatography. It is a high improved as compare to chromatography column chromatography. Smaller speck size cans (3-20) be analysed by hplc and allows much better breakup of a components. Detection method is used in hplc which are highly automatize and extremely sensitive as compare to column chromatography 13-14.Some of the advantage over column chromatography is, (a) small stationary human body 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 precept is base on adsorption. When a mixture of components is introduced in to column, they are turn on according to their affinity. The components which has a more affinity towards adsorbent, travel slower and vice versa 14.NORMAL PHASE HPLC hither, stationary build is frozen in nature and rambling 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 dissolver. Column length is 150-250 mm and less than 4.6 mm diameter 13-14.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 apply long chain of hydrocarbons like C8,C12,C4,octadecyl and mixture of water and alcohol use as a solvent. In this case strong attraction amid polar solvent and molecular(a)(a) 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 deem 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 form chromatography, paper chromatography to bankrupts 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 throwback phases 14.(3) Ion qualify 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 reroot power. The main principle is base on reversible step in of ions between ion present in the resolution and those present in ion commuting resin 15.Cation exchange-Solid-H+ + M+ === solid-M+ + H+(Solution) (Solution)Anion exchange Solid-OH- + A- ==== solid-A- + OH-(Solution) (Solution). 14breakup in ion exchange chromatography depends upon reversible adsorption of charged solute molecular to ion exchange pigeonholing of opposite charge. Some of the operating(a) group used in ion exchange chromatography is 16,NAME OF ANIONFUNCTION GROUPDi ethyl group amino ethyl-O -CH2-CH2-N+H(CH2CH3)2Quatemary amino ethyl-O-CH2-CH2-N+H(C2H5)2-CH2CHCH-CH3NAME OF CATIONFUNCTION GROUPCarboxymethyl-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 disintegrable 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 abstract 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 render 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 meters is done. Recovery of the samples for re using is norm ally not done, since the sample used is low. Eg. mg quantities. And other is (b) preparative HPLC Where the individual fractions of pure immixs 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 nitty-gritty 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, Guard column It is also known as safety column. Any impurities present in solvent or mobile phase which can predict by guard column.analytical column It widely use which can decided dexterity of separation. assorted column size is available depending upon separation. Column made up of pure stell, glass, poly ether ether kiton etc.(4)DETECTORS -They are(a)UV sensing elements 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) conduction detectors(d) Refractive detectors(e)PDA detectors which are similar to UV detectors.(5)RECORDER They are used to enter response from detectors. It can record base line and retention succession of peak. Now a day computer is used to recording data.2.4 HPLC PARAMETERS 2.4.1 innate 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 orotund 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 dust 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 HE TP(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 equationHETP = A + (B/u ) + CuWhere,A = Eddy diffusion term or multiple path diffusion which arises due to packing of the column. This is unaffected by mobile phase focal ratio or flow rate. This can be minimised by unison in packing.B = Longitudinal diffusion term or molecular diffusion which depends on flow rate.C = Effect of mass manoeuvre 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 vauntingly 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 child amount of substance like monograms at analytical scale to micrograms at preparative scale. It is used in a food analysis, biotechnology, bi ochemical 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 taste 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 beat of amount is also determined by hplc which including followers method 14, By comparing standard and sample peak area, the quantity of sample can be determined which is known as a DIRECT COMPRISION METHOD. To find submerging of sample by plotting graph between peak area vs. concentration of the standard drug and intra polation 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 rapidly process 14.It has some advantages over other chromatography like, (a) Thin layer chromatography is inexpensive but it cant give castigate resolution to compare HPLC. (b) d birthday suit spectroscopy required a large capacity of maintain cypher 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. Anal ysis of the compound mixture, mobile phase (mixture of components) are used resulting large amount of waste solution are obtained 14.CHAPTER- 3EXTERIMENT3.1 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 sequent no DE62956545.3.2.2 SOFTWEAR employ 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 5000l, Analytical balance, Volumetric flask (10ml, 20ml), Funnel, 100ml Beaker, Measuring cylinder (500ml, 50ml, 20ml,10ml),Filter paper which made by Fischer scientific with size 0.45m, Filter holder made up of pyrex bran d glass.3.2.5 PREPARATION OF roving PHASE The mobile phase used was a mixture of a wood alcohol and water with ratio of 5050. So take wood alcohol 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, 2080, 3070.3.2.6 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 extend to to conclusion. Here we were using two different wavelengths like 280nm and 360nm .3.5 ASSAY hinderance 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 zone 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 25l, 1.2 U/ml alpha-glucosidase, 25 l of phosphate buffer, standard solution of aloe vera, 25 l of 2.5 mM pNPG and finally if proper colour change was not occur than add terminating solution 100 l of 0.2 M Na2Co3.3.5.1 PROCUDURE 1234E123456BLANK assimilation 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 25l,1.2 U/ml alpha-glucosidase, then add 25l of phosphate buffer , add 25l of test solution, and finally add 25l of 2.5 mM pNPG. And then transfer 2.5l solution from low concentration to high concentration. Than kept mixture for some time and incubate plate at 37C for 10 min. and tested absorbance.