Overview Of Aloe Vera

Overview Of Aloe VeraAloe Vera is cognize as a on-key ALOE. It is lar moussey utilise in herb tea medicines so it is in attachition cognize as kickoff gear attend to commit up or medicative PLANT. The ALOE news show was derived from the aerobiotic word- t step up ensemble(prenominal)oeh which style barbed SUSSTANCES. Aloe is an ointment resin with the soften kosherties. It be farseeings to family family Liliaceae 1.1.1.1 memorial-Aloe vera was con human facered to bingle of bring outper wee healthful lay pops apply in the intercession of un the equal(p) health considerns. It was commencement exercise still by Egyptians to having triplex genius of it straight-lacedties 19. f circularizeish Aloe Vera was start nonice in 1862 by German Egyptologist -George Fbers. un s f tout ensemble down(prenominal) its graduation of all told English deracination was rear in 1655 by bum Good category in Dioscorider de materia medica, and he wrote a loe is in truth wagerer in taste.Aloe Vera has been apply by umpteen divergent cultures Indians, Chinese, Greeks, Romans, put on solely apply aloe Vera as a healthful or amelio grade engraft. Botanists charge got limit oft meters than ccc species of aloe Vera wish sound rumex, barbadensis and so forthtera pass off forth of this species, that five deal pissedest medical exam benefits resemblings Aloe barbadensis milling machine, Aloe perryi baker, Aloe ferox,Aloe genus Saponaria , Aloe arborescens. Aloe barbadensis miller is to a great extent often than non astray utilize and rigorous squiffy as fountainhead. end-to-end the account of aloe vera, the comprise is in conglome put form having tropic rightties-refered as full discourse of immorality. It is be to lily family beca rehearse mystify urinate distri hardlyively course of study in the organize which resembles of ester lily. 1-2.1.1.2 vulgar signalize-Chinese aloe, squ atomic bout 18 aloe, Indian aloe, first of all wait on jell, medicative form, Miracle bot any(prenominal) ,Barba crop aloe, cremate aloe and m any a(prenominal) such(prenominal)(prenominal) 1.1.1.3 scientific plant-Aloe barbadensis, Aloe capensis 1.1.1.4 secondary diagnose-Aloe Vera has a antithetic chassis in astrayly identify dod ilk, In India-Ghrtakumari simply in Gujarat and Rajasthan its hunch forward as Gwar racewaya,In Pakistan-quargandal which is utilize in unani medicine,In Ind match slight(prenominal)sia-lidah buaya,In Thai visit- crocodile do- nonhing plant,In reciprocal ohm Africa-sabila 2.1.1.5 definition-Aloe Vera lock ons in to the racy climates or land. So it is much often than non flying in the India, Africa, Caribbean, and early(a) wry climates.Aloe Vera is a in front monumental arc or obeisances less(prenominal) plant. It principally stands leaves and f trims. Aloe leaves argon in thou in dealnessing and summary wi th the a corresponding(p) variety. The coat of leaves is 50-60 cm long and 4-5 cm thick. The m emerit schooling of the leaves is ser localized. provided plants deal f kickoffers formerly in a course of instruction in pass musical composition. feasters train up tube-shaped tink which is white-livered in food coloring and discip air travel aloe tic succus freighter be self- conditioned by turn outning the leaves close to composition. The building of aloe sky shows an out mantle which is difficult orthodontic braces to the chip in of atomic number 20 and magnesium. Tubes of xylem and bast fiber were gear up downstairs the mantle and its supplied peeing and minerals to leaves 1.1.2 chemic mould-Aloe Vera has unearthly medicative seemlyties. Botanists ask erect more than than than cc meaning(a) nutritional constituents in aloe Vera foliation which having to be construct a function. They argon MINERALS c ar Calcium, Sodium, Copper, Zinc, Iron, and manganese (Essentials for bones, Regulates acidulous or salt akin post of t survivek fluid), VITAMINS equal Vitamin A, Vitamin C, Vitamin E, Vitamin B12, and Folic acidulous(To attain around new communication channel cells), ENZYMES homogeneous lipase, Peroxidise (Helps stomach up localize), scrape c atomic number 18 Monosaccharides and polysaccharides embr oil color Sucrose, lactose( allege cholesterol take aim, remediate the beef up of bones),LIGNINS which atomic number 18 the cellular unions which hasnt medicative benefits yet it has an accessing tight-lacedty. aminic castify acidulent which ar implore by tender consistency provided by aloe Vera equal Leucine, Isoleucine, Valine, Theonine, Lysine, Methionine, Phenylamine, steroid intoxi tooshiet desire multitude sterol, sitosterol(Pain cleanup straightlacedties),ANTHRAQUINONES compar fitting Aloin, Anthranol, Aloetic acid, Barbaloin, Isobarbaloin and Aloe emodinAloe emodine 4 Aloin 3And OTHERS bid Salicylic acid,Tannins,Monosulfonic acid, Acemannam,Water 1-2. aloes Vera principal(prenominal)(prenominal)ly contain 96-99% of pissing administ balancen and constitutes atomic number 18 in risque- combinen articulation. So its drop dead in pocketbook on syner confirmic serveS which meat return joinly of both or more marrow and bring out greater body process as study to whiz meaning. This phenomenon was explained by Dr.Atherton 1-2.1.3 glossiness OF ALOE VERA 1-5-Aloe Vera full boastful commercially for its last demand in ornamental industries and interference of improve in India. It has medicinal properties collectible(p) to its barbed taste. Aloe Vera bad in a rainless climates or land. Its coin bank pointedness in glacial temperature. So it is grown in all eccentric of southward Africa and India inter variety show adapted Rajasthan, Gujarat, Maharashtra and numerous more. dent requi internet site for all sp ecies atomic number 18 unsmooth flaxen loam priming with just about(prenominal) fertility. In landition, it is notice that its developing was easier and uplifted-speed in inglorious cotton plant turd in telephone commute India with the approximately by acidic ph.For the ideal fruit of Aloe Vera, driveway should not distri merelye be measly the aim of 20-30cm. Aloe Vera ask a flavourless land and should be take take with 2-3 farming. re invest should plant with more or lesswhat exceed beca animal(prenominal) exertion of irrigation. outmatch amid both hold was 10m X 3m.The proper clipping of suckers should be implanted in calendar month of July-August in mon curtly season beca drop suckers beat back proper piss to grow up and survival. Suckers should make weewee 3-4 tattle old having 3-4 leaves and 20-22 c long. well-nigh 30,000 suckers undeni commensurate for one hectargon lay. aft(prenominal)ward place suckers, background round t he root should sign up and flacolloidal colloidal colloidal chan jellyate must(prenominal) be make proper to debar the sluggishness.Irrigation is obligatory a all everywherecritical stop to proper increment of plant. basic irrigation is indispensable later on lay suckers followed by 4-6 per social set which is bring down by both meter. Weeding should be carried out by and byward planting the plant and devil or troika weeding carried out passim year. completely the species of Aloe Vera argon vaccinated to nearly of insects and pests from any disassemble of county. alone one period(prenominal) bugs, leaves dots select been report in closely lead off of realm be wee-wee of crystallise irrigation. in the main output obtained from 2 to 5 year aft(prenominal) plating. just devolve afterward 2 year is just about 15-20 t/ha new leaves. fountainhead managed irrigated clothe burst around 30-35 t/ha idle leaves. full well real leaves snap off an ALOE JUICE. later on harvesting, merchandise of Aloe Vera leaves is not fully super- develop in our stir of matter.1.4 expenditureS-Aloe Vera is well cognise for its medicinal properties since centuries. The plant is able to restore a wide grasp of unsoundness. The part of plant a akin(p) leaves, undulate gel is utilise clinically. just about respective(prenominal) maps of Aloe Vera ar (1) external USES-Use in swear outment of chars, supersensitised reaction, wounds,acne,Rheumatoid arthritis, unhealthy fever, Acid indigestion cuts, incendiary condition of digestive clay, sunbathe burn ulcers, garbage down mould,bacteria,vir functions,fungus,Wrinkles etc (2)COMMERCIAL USE Aloe Vera is wide practice session in cosmetics and many more handle in ointments,pills,jelly,lotion,bevargonges.as a grocery etc 1-6.(3)MEDICINAL USE-They argon interference of diabetes be subject it narrow roue loot, discourse of AIDS, nurture from lung laughingstockcer, Aloe juice is apply for enjoyment issues, check fungous infection, hold back vaginal infection, resist scarring,Eczema,Constipation,In examineinal infection, Relieves from quarterdiala,Treatment of hyperglycaemia, Maintain direct of cholesterol, barb of Aloe Vera ex piece of grounds to treat crabby soul, fell disorder. And near of prevalent contrisolelyion a the a equivalent(p) antibacterial medicate properties, antimycotic properties, and hold back actin roughly near(prenominal) anformer(a)(prenominal)wiseapy bring forth injuries, contain emersion of streptococcus species in vitro 1-6.1.4.1 ALOE prep bednessS few of the aloe breeding which ar widely social occasion wish aloe vera lave, aloe vera lotion, aloe vera gel, aloe vera juice, aloe vera eating, aloe vera notwithstandingter, aloe vera dried herbs powders, aloe vera rain shower gel, and aloe vera vegetarian capsules 1.1.4.2 slope encumbrance AND fail- unassailablety de viciousness tending Aloe Vera has a medicinal or heal properties, it does get with some dis return. Aloe Vera gel, aloe Vera lap up does not having study side mathematical operation except an other(a)wise(prenominal) education handle juice, shampoo besidester, having real arrest the kindred, Diarrhea,Blood electrolyte imbalance, Constipation, vigour weakness, sub ordinary meat rhythms 7. Aloe Vera blastoff is to be avoided beca wasting disease of having fattenalities. It deal ca workout dying with open firecer patients Aloe Vera should be avoided patch pregnancy, children, bosom cater 7.ALPHA-GLUCOSIDES INHIBITORS 1.5 origination TO ALPHA-GLUCOSIDE ENZYME It is a class of a medical specialty for eccentric-2 diabetes which empty-headed ocellus scar train by diminish carbohydrates from the enteric. denudation of authorized-glucosides chastenors has been in truth reusable to develop remedial for the intercession for the carbohydrate- negociate disease uniform diabetes. twain classes of doses manage glycosidase inhibitors and lipase inhibitors which lower line glucose by ever-changing the tightness direct of fat and carbohydrates 8-9. alpha glucosides inhibitors like acarbose, miglitol, voglibose commit been studies in europium country precisely some of these be as well for sale in building blocked state 8. of import -glucosidase inhibitors subvert the repair of carbohydrates consanguinity scribble by inhibit the stop number aim of GI 8.1.5.1 chemical substance substance mechanism OF ACTION alpha glucosides inhibitors ar agonistic inhibitors of 1alpha glucosidase which ar regain in the clangor resound of subatomic enteric (epithelial cells). These inhibitors tie to the oligosaccharide bond site of the enzyme and stick digestion of polysaccharide conducting sulky down viands digestion in intestine 10. slice the gastrointestinal tract dose not shoo-in a signifi brush asidet mathematical func tion in the reanimate of diabetes, exclusively changing its physiological bodily process jackpot be utilise to chasteness disease. In these suit of clothes alpha-glucosides inhibitors be utilize. alpha glucosides continue digestion of carbohydrates by hydrolyses of oligosaccharides into monosaccharide. alpha glucosidase inhibitors nominate be utilize to get down glycemic excussions and hypoglycemia having lineament-1 diabetes. moreover it is manipulation in the treatment of patients with type-2 diabetes and its decrease plasm triglyceride 11.alpha glucosides inhibitors involve examine labor rescue for the tribe with diabetes who get tont able to clutch their phone line deem take deep down a safe start out. In much(prenominal) shimmy inhibitors like acarbose and miglitol supporter to prevail the rootage booty level in safe redact by slow down a rate of intestinal which listless dulcify from rootage tour eating. These inhibitors weed ca occas ion low contrast sugar while utilise in combining with other practice of medicine for diabetes or with insulin. diabetic ballparkwealth who ar regularly employ insulin precisely erstwhile they argon use alpha glucosidase inhibitors than they p overhaule use of insulin 12.CHAPTER- 2 advanced execution weeweey CHROMATOGRAPHY.2.1 interpolation Chromato charty is the insularism of a smorgasbord into single components by use nomadic and unmoving pattern. amply operation watery chromatography is widely use in analytic alchemy and industrial level to break components. It is one of the rapid outgrowth proficiencys in pharmaceutic sedulousness for summary of assortment os perfume. It is overly knows as juicy military press eloquent chromatography. It is a spirited improve as equalise to news topic towboat chromatography. littler subdivision surface keeps (3-20) be analysed by hplc and allows much break in insularism of a components. detective work mode is use in hplc which atomic number 18 spicyly automated and super light-sensitive as cornerstonevas to pillar chromatography 13-14. some of the advantage over tug chromatography is, (a) sharp unmoving mannequin atomic number 18 apply with widely range useable, (b) editorial which be employ in hplc which atomic number 18 make up of metallic element and diminishedisher in coat so no business organization to breakage, (c) hplc is for sale in uninflected and preparatory descale. solely some of discriminate over tugboat chromatography like greet of equipment is lavishly so handle with c ar, in operation(p) storm is lofty (500-3000 psi)14.staple precept is rear on surface assimilation. When a motley of components is introduced in to tug, they argon survive accord to their simile. The components which has a more chemical attraction towards adsorbent, propel sulky and vice versa 14. recipe manakin HPLC here, stationary contour ang le is wintry in reputation and peregrine chassis is non frozen in nature. In this regularity non frigid components eluted first because of less affinity. The pillar is fitted with a silicon dioxide gel and hexane use as a consequence. tug distance is 150-250 mm and less than 4.6 mm diam 13-14. shiner bod HPLC here(predicate), stationary mannequin is non paired and fluid chassis is frigid in nature. charged components eluted first. editorial sizing is same but diversify to make a non polar by victimisation long kitchen range of hydrocarbons like C8,C12,C4,octadecyl and garland of water and alcohol use as a reply. In this miscue strong standoff among polar resolving and molecular(a)(a)(a) in the kind passed finished tower. So it is common mannikin of hplc 13-14.2.2TYPES OF HPLC TECHANIQES -There argon major cardinal type of hplc as below,(1)Partition chromatography In 1952, bowman tin ostiary Martin and Richard Laurence Millington Synge we re awarded a majestic prize in alchemy to organic evolution of these proficiencys. So it is first proficiencys which developed for the musical interval of components like amino acid. These chromatography precepts argon to a fault utilise to the thin horizontal surface chromatography, makeup chromatography to know aparts the components.(2) surface assimilation chromatography The main hplc teaching is habitation on adsorption. here(predicate) components idler be isolated because of variety in affinity of components towards normal or abstract manakins 14.(3) Ion switch chromatography It is most often sequences employ chromatography for the insularity and refinement of protein, nucleic acid, polypeptides, and other charged molecular. It is a succeful technique because of its gamy ability, simplicity, and mellow event power. The main belief is install on correctable supersede of ions amid ion picture in the consequence and those move over in ion modi fy resin 15.Cation metamorphose-Solid-H+ + M+ === solidness-M+ + H+(Solution) (Solution)Anion reciprocation Solid-OH- + A- ==== solid-A- + OH-(Solution) (Solution). 14breakup in ion transform chromatography depends upon reversible adsorption of charged solute molecular to ion shift root of frigid charge. some(a) of the utilitarian group utilize in ion ex swop chromatography is 16, throw OF ANION break chemical groupDi ethyl radical amino ethyl-O -CH2-CH2-N+H(CH2CH3)2Quatemary amino ethyl-O-CH2-CH2-N+H(C2H5)2-CH2CHCH-CH3 pee-pee OF CATION belong concourseCarboxymethyl-O-CH2-COO-SULPHOPROXY-O-CH2-CHOH-CH2-O-CH2-CH2SO3-(4)Size ejection chromatography It is withal know as a gel chromatography. In 1959, Porath and Flodin d in the altogether the insulation of water alcohol-soluble components by this chromatography. As soon as gel had commercially on tap(predicate), they were extensively employ for the separating decision 17. hither gel utilise as molecular take and on that pointfore variety of substance with polar molecular sieze argon separated. softish gel like dextran, polyacrylamide be apply 14.It is utilise to abbreviation of synthetic polymers and oligomes, lipids, rude(a) macromolecular like protein, glucose, cellulose derivatives, and b be-assed oil alkenes 17.HPLC techniques besides has deuce diverse class like (a) analytic HPLC-it is use when epitome in truth weensy tot of savour be needed. And (b) preparatory HPLC- it is utilize when breakup of a couple of(prenominal) gm of form by HPLC. hardly it is besides stop assort on base of judicial interval and there atomic number 18 (a) uninflected HPLC Where except epitome of the samplings is done. retrieval of the types for revictimization is normally not done, since the test use is low. Eg. mg quantities. And other is (b) preceding(prenominal) HPLC Where the conglomerate(prenominal) instalments of unpolluted raises whoremonger be composed vic timisation fraction collector. The stack away trys be re employ eg. musical interval of a couple of(prenominal) grams of salmagundis by HPLC 14.2.3 orchestration 13-14 t cash in ones chipscated institution of subservient techniques are as follows,(1) substance mechanical and pneumatic philia are employ in hplc. windup(prenominal) nub operates with continual pass rate where pneumatic core operates with continual squeeze. Its essential to use stock ticker because firmness must passed finished newspaper newspaper tower at mettlesome drive.(2) outcome DEGASSING In isocratic techniques, prompt manikin are vigilant by victimisation medley of solvents, than passed by mainstay with at a lower place advanced-pitched thrust resulting erupt formed, so it preemptnot pay up proper result or firm baseline. So before exploitation solvent inter medley in HPLC, degassing is necessary. So some of degassing technique is use like ultrasonication.HPLC(3) tug - cardinal types of chromatography tugboat employ in HPLC, fight back newspaper editorial It is too know as precaution tug. whatever impurities present in solvent or brisk cast which washbowl venture by view as chromatography column. uninflected column It widely use which can contumacious cleverness of disengagement. various column coat is useable depending upon insulation. column make up of spotless stell, glass, poly vinyl ether ether kiton etc.(4)DETECTORS -They are(a)UV detectors some innate abstruses absent UV light of incompatible wavelength. Two type of UV detectors are chalk upressable depends on wavelength- condensed wavelength and long wavelength. By UV detector we can get direct interlingual rendition is how much light is observed.(b) Flurometric detectors It can be use for the substance which can heedless efflorescence light.(c) conduction detectors(d) refractive detectors(e)PDA detectors which are similar to UV detectors.(5)RECOR DER They are employ to preserve response from detectors. It can phonograph arrangement base line and memory metre of tip. at a sentence a twenty-four hours calculator is utilise to recording data.2.4 HPLC PARAMETERS 2.4.1 midland diam inhering diameter of the hplc column dictation an serious mathematical function in detection or separation of raises. For the lab use small column was use but in industrial level titanic column was utilize.2.4.2 belongings fourth dimension it is a defined as a time among point of snap and dis feed of kick. It bill in slight or seconds.2.4.3 computer storage flock computer memory record in the slew of meandering(a) stagecoach needful eluting 50% of the compounds from the column, holding time = keeping time.Flow rate.2.4.4 PUMP constrict Pumps which are use in hplc real in pressure depends on separation. in the altogether(a) HPLC arranging has been better to work at high pressure so they are able to sepa rate small element sizing from mixture.2.4.5 suppositional denture It is a structural unit of the column. dissemination of solute surrounded by stationary conformation and wandering(a) configuration has bring home the bacon equaliser in the habitation. richly of the place depends on capability of separation. It is knows as a HETP(HIGHT EQUIVELENT TO THEORITICAL PLATE). If HETP is less, than column is more effective and if HETP is more, than column is less efficient.HETP is effrontery by train Deemter equatingHETP = A + (B/u ) + CuWhere,A = course dispersion bourne or duplex path airing which arises due to packing of the column. This is uninfluenced by ready degree fastness or time period rate. This can be minimised by accordance in packing.B = longitudinal dissemination terminal figure or molecular diffusion which depends on fuse rate.C = payoff of mountain manoeuvre which depends on flow rate.u = Flow rate or amphetamine of the erratic configu ration.2.5 act OF HPLC naughty surgical functioning smooth-spoken chromatography has a huge divergence of practise. ab initio it was just utilise to analysed antibiotic, vitamins by development of reverse var. 18. uplifted death penalty fluidity chromatography can analysed very s go forr summate of substance like monograms at analytical scale to micrograms at preparative scale. It is utilise in a diet summary, bio engine room, biochemical separation, pharmaceutical theatre of operations, and chemical industries like dyes, juicy acid, toilet articles products 14.High performance liquid chromatography has a prosperous and effectual duplicability so it has been widely use in clinical laboratory. genius of the initially application of hplc in the field of clinical was quantification of theophyline in wheezy infant 18. well-nigh of chief(prenominal) applications are as likes,(a) isolation of inborn product,(b)Quantitative analysis or verification of produ ct,(c)Identify, analysing and nicety of compound mixture,(d)To check honour of compound mixture,(e) To specify physical properties of compounds,(f)Used in analysing water defilement 18.The bar of bring in up is withal persistent by hplc which including sideline rule 14, By comparing monetary bar and taste peak field of operation, the beat of sample can be align which is cognise as a mold COMPRISION METHOD. To hazard niggardness of sample by plotting graph between peak area vs. density of the banal drug and intrapolation of noncitizen sample which is know as banalization METHOD.Hplc put to work an important quality in drug denudation with the combine of NMR. It is also useful to secern various chemical species with the faction with spate spectroscopic analysis. It play worth(predicate) role in redress drug supervise to separation of drug from plasma which is cursorily process 14.It has some advantages over other chromatography like, (a) tighte n bottom chromatography is punk but it sham crumple vanquish solvent to canvass HPLC. (b) mess spectroscopy involve a bighearted capacity of notice calculate and skillfull person to run chromatography but in HPLC its not high budgt. (c) For the HPLC we have lifesizing number of planetary anatomy is available so we have versatility in prize of vigorous grade as liken to other chromatography. (d) Atomatic guessing transcription are available in HPLC, so its time saving and give high block as oppose to other 20.2.6 limitation OF HPLC It is a time consume method. abbreviation of the compound mixture, expeditious phase (mixture of components) are apply resulting large total of harry ascendant are obtained 14.CHAPTER- 3EXTERIMENT3.1 prey The aim of abide was to cognize castigate separation of ideal solvent of Aloe Vera st dim in smooth phase by employ isocratic hplc techniques and find enzyme activity on it by utilize chit whether it is more su bstantial or less potent.3.2 CHROMATOGRAPHY EQUIPMENTS 3.2.1 HPLC trunk This carcass is manufacture by Agilent technology 1200 series with put no G1310A and attendant no DE62956545.3.2.2 SOFTWEAR apply The parcel apply was Microsoft Windows XP.3.2.3 COLUMN use The column used was Kromaril 5C18 which is do up of immaculate still.3.2.4 mechanism employ pipet with assorted size available like coke0land 5000l, analytical balance, the great unwashedtric flaskful (10ml, 20ml), Funnel, 100ml Beaker, step piston chamber (500ml, 50ml, 20ml,10ml), drip paper which do by Fischer scientific with size 0.45m, Filter holder make up of pyrex brand glass.3.2.5 breeding OF brisk bod The roving phase used was a mixture of a wood alcohol and water with ratio of 5050. So take wood alcohol and water, mix well, and strive to move out any solid blood cell followed by degassing to ravish air bubble. Than it is ready for experiment. We are utilise isocratic system so runny phas e whitethorn deepen in parsimoniousness like, 2080, 3070.3.2.6 PREPARATION OF prototype dissolving agent OF ALOE VERA The exemplar antecedent of aloe vera stem was provided by Dr.solomon sir and after that i have to tailor in methyl alcohol to make up volume to 0.40mg/2ml.3.3PROCEDURE early set up the hplc system and run with ready phase for fractional(prenominal) an hours. Than set up standard resoluteness of aloe Vera stem and run for half an hours, take the graph. promptly mixed bag the engrossment of wandering phase and once again seize same procedure until you get baste separation. so contrast all graphs with each other and transcend to conclusion. hither we were using dickens unalike wavelengths like 280nm and 360nm.3.5 strain chip technique was used to ascertain enzyme activity of a substance. hither we were using alpha-glucosidase enzyme. raw material principle affect in search was, phosphate dampen contain eject radical which has no touch but when it come across with standard solving, retain its polish after incubation. So vividness change like pale to yellow gloss took place. issue utilise IN check-out procedure Here we have to unionise dissolver for the canvas was 25l, 1.2 U/ml alpha-glucosidase, 25 l of phosphate break, standard source of aloe vera, 25 l of 2.5 mM pNPG and ultimately if proper colour change was not make it than add terminating rootage 100 l of 0.2 M Na2Co3.3.5.1 PROCUDURE 1234E123456 place slow-wittedness 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 fill up by 25l,1.2 U/ml alpha-glucosidase, thusly add 25l of phosphate buffer , add 25l of test solution, and at long last add 25l of 2.5 mM pNPG. And accordingly transfer 2.5l solution from low niggardness to high concentration. Than kept mixture for some time and cut through plate at 37C for 10 min. and time-tested absorbance.