Title of Invention	A MEMBRANE SEPARATION PROCESS
Abstract	An Emulsion liquid membrane method for the separation of organic and inorganic species form the various mixtures comprising of the preparation of Emulsion liquid membrane by stirring of internal phase with a surfactant at the suitable pH and rpm then dissolving these emulsion liquid membrane in non-polar solvent and reading the absorbance at UV-visible spectro-photometer.

Full Text

FROM 2 THE PATENT ACT 1970 (39 of 1970) & THE PATENT RULES 2003 COMPLETE SPECIFICATION (see section 10 and rule 13) TITLE OF INVENTION ! A membrane separation process APPLICANT a) NAME OF APPLICANT b) NATIONALITY C) ADDRESS SAIRA B MULLA ; INDIAN OJESH APARTMENT 102-103, NR PATEL NURSING HOME, SECTOR 20, AIOROLI, NAVI-MUMBAI The following specification particularly describes the invention and the manner in which it is to be performed. A Membrane separation process Filed of invention. The field of invention is related to the separation of the of the organic, inorganic species by the membrane separation process, More precisely invention is related to the separation of materials by liquid membrane separation techniques, Prior art. Membranes are the synthetic barriers across which selective permeation of the desired species can be effected by employing appropriate driving force like electrical potential, hydrostatic pressure, concentration gradient etc. Membranes are ubiquitous everywhere in nature. They are responsible for the transport of tremendous quantities of material in animal and plant kingdom. The selectivities of membrane for certain solutes and their rejection for others have fascinated the scientist for centuries.The semi-permrable nature of the membrane is essential to ensure that the separation takes place. Electrodialysis, reverse osomosis, pervaporation, micro and ultra-microfiltration which are membrane integrated processes have proved their potential in varied industrial and biotechnological areas. Membranes may be either: In solid membranes mass transfer of solutes take place due to the different size of the pores and passage of ions or molecules at different rates through the membrane. In the liquid membranes mass transfer takes place due to diffusion process. Different solubilities and diffusion coefficients of the solutes in liquids are responsible for mass transfer. LIQUID MEMBRANE TECHNOLOGY: Profound interest has been focused on LM separation processes both from fundamental and technological view points. The separation of solutes by a LM is based on a well established idea that the two completely miscible liquid phases separated by third liquid immiscible with either of them, can exchange solutes provided there is a difference between their chemical potentials in two phases and provided that the intermediate liquid is able to transport them. In most cases the two miscible liquids are aqueous solutions and the third(membrane) phase is an organic solvent. Here, out of two liquids one is donor and second is acceptor phase. The LM technique is a powerful and novel tool and excels most of the conventional separation techniques because of the following merits: • Low energy requirements. • Compact modular devices. • Simultaneous extraction and stripping in the same device. • Zero pollution. • Flexibility of operation. • Smaller volume of raffinate. Use of mild to moderate process conditions, Some of the advantages of LM over the solid membranes are • Higher fluxes owing to very much higher diffusion coefficients of solutes in the liquids than solids. Liquid membrane technique allow a convection diffusion region instead of molecular one. This also increases the fluxes. • Owing to the availability of great number of substances which when added to the liquid membrane phase increases the In contrast to solvent extraction, in liquid membrane separations amount of solvent used is very small. This permits the use of highly efficient and selective and costly reagents. LM technique is important in the separation science particularly in the instances where solute concentrations are extremely low and large volumes of solution must be processed without generating any secondary waste. US patent 3410794 teaches about a process for separating mixtures in to their components part by means of selective permeation through liquid (surfactant) membranes. The mixture to be separated is first emulsified in aqueous medium contain a selected coated with surfactant are then contacted with a selected true permeate the membrane coating and pass into the solvent phase by conventional means. US patent 4,183,918, this invention relates to the use of liquid membrane technology in preparing medicinals. The medicinal prepared by this invention may be ingested and may be utilized as traps for toxins present in the Gl (gastrointestinal) tract, or as slow-release compositions of drugs, or as reactors. In the trap embodiment, the liquid membrane encapsulated medicinal is an emulsion comprising an exterior phase which is immiscible with the liquids present in the Gl tract and permeable to the toxins therein, and an interior phase which is immiscible with the exterior phase and comprises a reagent capable of converting said toxin into a non-permeable" form, In addition, hydrophilic adsorbents, such as a hydrophilic carbon or a silica gel, may be encapsulated in the emulsions of the instant invention. When the compositions of the instant invention are utilized as slow-release drugs, the interior phase of the emulsion will comprise a drug which is slightly soluble in the exterior phase of the emulsion whereby said drug permeates through said exterior phase of the emulsion over a period of time into the Gl tract. The third method for utilizing the compositions of the instant investment comprise encapsulating a catalyst for a reaction which is in the Gl tract permeate through the exterior phase of the emulsion into an interior phase wherein said catalyst, for example, an enzyme, converts the permeated reactants to reaction products. The reaction products then may permeate through the exterior phase back into the Gl tract. In all cases, the liquid membrane encapsulated medicinals may be administered by either oral ingestion or injection anywhere else into the Gl tract. Object of the Invention. The aim of present work is to develop method for the recovery/ separation of, organic and inorganic species, especially bio-molecules in a very dilute solutions and the comparative study using anionic and cationic reagents such as cyanex-301 and aliquat-336, LA-2, TOMAC, D2EHPA, ALIQUAT-336, CYANEX - 301 or mixture thereof, as the extractants in membrane solvent toluene and span-80 as emulsifying agent, when emulsion liquid membrane technique is applied. Summary of the invention. The scope of the present investigation was to study the effective separation of some of the organic and inorganic species form the various solutions were concentration of the species (analyte) is low, the technique is especially applicable to the bio-molecules such as antibiotics, amino-acids, or bio-molecules possesses the acidic as well as basic groups. Therefore they behave both as anionic as well as cationic species in the solution at different Condition depending upon their pKa values m . Therefore can b© completed by Aliquat-336 and Cyanex --0301 at specified conditions of pH depending upon the. nature of the species. Various anionic and cationic surfactants are studied for the purpose of the separation of the organic and inorganic species form the various solutions, like fermentation broth, Surfactant like Cynaex -301 and Aliquat - 336 are mainly studied are for the purpose of the recovery or separation of the organic or inorganic species form the various media. Cynaex -301 The active components of Cynaex - 301 extractants is bis (2,4,4,-trimethyl pentyl ) dithiophosphoric acid . It is a green mobile liquid having specific gravity, viscosity, and solubility in water 0.95, 78 centipoises, and 7 mg at 24 ° C . Cynaex -301 has been employed as carrier for the recovery of lager number of metal ions by ELM process, Aliquat -336 The active component of aliquat -336 is methyl tricapryl ammonium chloride. Aliquat -336 is yellowish oil. Soluble in chloroform benzene, Aliquat -336 has been employed as carrier in ELM for separation of number of metal ions, bio-molecules, like lactic acid, cephalosphorine-C, amino acid, cephalosphorine . However, there is no literature for the recovery studies of cefadorxil, ampicillin m ciprofloxacin, using this reagent in ELM. General Experimental Procedure adopted for the development of the membrane separation process is as follows, Instruments A UV visible recording spectrophotometer, control dynamics PH meter combined with glass electrode magnetic stirrer, high speed homogenize, (remi India), were used for the absorbance and PH measurements and transport studies respectively chemicals and reagents Standard solution of the material to be separated prepared , of the known concentration, Span -80, Span -80, Paradox- 100, ECA 1152, ECA 4360 or mixture thereof, cynex-301, aliquat -0336 were used as regents. All other chemicals toluene Kerosene, n-heptane decyl alcohol, n-butyl acetate, m-xylene, hexanol, shellol T and n-dodecane, sodium citrate, citric acid, sodium carbonate, sodium chlorides HCI were of the AR grade. Preparation of membrane. Membrane (A) 13 cm 3 Span-80 + 10 CM 3 aliquat -336 diluted to 100 Cm 3 with toluene. Membrane (B) 13 CM3 span -80 + 12 CM 3 Cynaex -301 diluted to 10 Cm3 with toluene. Preparation of Emulsion. Emulsion (A) 10 CM 3 of internal phase citrate buffer of PH 3.0 contain 0.8 mole /dm 3 NaCI was added drop-wise to as 5 cm 3 of membrane (A) stirred vigorously on a magnetic stirrer for 20 minutes to get stable milky white emulsion. Emulsion (B) 10 CM 3 of internal phase carbonate buffer of pH 8.5 containing 1.5 mole /dm 3 of NaCI was added to 5 Cm 3 membrane stirred vigorously on a magnetic stirrer for 20 minuets to get stable milky white solution. Citrate buffer and carbonate buffer are prepared as per the standard procedures. General Extraction procedure for the transport studies of analyte. Using Emulsion (A) 15 CM 3 of Emulsion (A) prepared as above was added into a 100 Cm 3 beaker containing 50 Cm 3 of carbonate buffer with 1.0 mole /dm 3 of Nacl adjusted at pH 8 and 250 mcg of analyte and the concentrate were stirred gently Hi 200 rpm on magnetic Stirred for a given transfer time of 25 minutes. The emulsion was separated by using separating funnel and de-emulsified by treating it with 0.5 CM 3 butanol. Analyte so separated in the aqueous phase is estimated spectro-photometrically at 254 nm or at the maximum absorbance at the ultraviolet range. The amount of the analyte remaining the feed phase was calculated by mass balance. The percentage was then calculated using the formula, Concentration of analyte in the internal phase * 100 Concentration of analyte in the feed phase Emulsion (B) Using the Emulsion (B) same procedure is followed as above for the analyte for getting the similar transportation of analyte from the internal phase to the feed phase and same % ratio is obtained. Various parameters were evaluated for the maximum percent of transport of analyte form an aqueous feed solution to inner phase solution through the ELM (A) or ELM (B) containing aliquat - 336 and Cyanex -301 as carrier. Parameters studied are as follows, 1. Effect of pH or H + ion concentration of the feed phase. 2. Effect of pH of internal Phase. 3. Effect of concentration of NaCI in the internal phase. 4. Effect of stirring time. 5. Effect of concentration of carriers. 6. Effect of surfactant in the membrane. 7 Effect of membrane solvents. 8. Effect of stirring rate. 1 Effect of pH or H+ ion concentration of the feed phase. The feed phase used for the transport studies of the analyte is 1.0 mole/dm 3 Nacl in carbonate buffer using Emulsion A where as it was in aqueous solution for Emulsion B, The PH of the feed was varied form 6-10 for Emulsion A and HCI concentration was varied for 1X 10 "1 mole /dm 3 for Emulsion B. It was observed that maximum transport 80 % at 1X 10-2 mole therefore the feed phase pH for transport studies of analyte using Emulsion A and that for emulsion b was set at pH 8 1 x 1 O -2 mole /dm3 HCL consontration NH2 groups, or it is zwitter ions contains both the acidic and basic functional groups it ionic forms of different charges in solution depends upon PH of the media and pKa values. At pH values below 4.0 (pK a 1 = 4.0) it exists predominantly in cationic form where as above where as 6.0 it exist predominantly PK a 2 6.0 in anionic form. 2 Effect of pH of internal Phase. The internal phase of the membrane used for the transport studies of analyte at various pH of these ranging form 2-6 and 6.5-10.5 using emulsion (A) and Emulsion (B) respectively indicate that the percent transport of analyte gave maximum transport at pH 3 for emulsion (A) and pH 8.5 for the Emulsion (B). Therefore PH of the internal phase solution for emulsion (A) was set at 3.0 and for emulsion (B) was set up at the 8.5 as optimum conditions for the further studies. 3 Effect of concentration of NaCI in the internal Phase. The presence of chloride ions in the internal phase for the transport of analyte using Emulsion (A) with aliquat-336 was essential as a source of chloride ions generation, where as in case with emulsion (B) with Cyanex -301 it is required to prevent the membrane swelling, variation in the concentration of chloride as Nacl for the maximum % transport of ampicillin was carried out in both systems , it was found that the 0.8mole/dm 3 and 1.5 mole /dm3 are optimum Condition® for emulsion (A) and emulsion (B) respectively. The stirring time was varied form 5-35*minutes keeping all the experimental parameters for both emulsion (A) and emulsion (B) respectively, it is observed that the percent transport of analyte was increased form 5-25 minutes and then remains constant thereafter in both the cases . Thus stirring time of 25 minuets was used as optimum time 5 Effect of concentration carriers Variation in the concentration of carrier used for the preparation used for the preparation of membranes form 5-20% in toluene, indicates that the percent transport of the analyte with increase in carrier concentration found to be maximum 76.77 % and 79.95% with Emulsion A and Emulsion B respectively. AS the carrier concentration in the membrane increases initial flux increases hence percent transport, however much increase in the carrier concentration increases the viscosity of the membrane thereby decreasing the transport. Therefore it is essential that appropriate quantity of carrier must be present in the membrane. 6 Effect of surfactant in the membrane. Variation of surfactant SAN 80 from 6-16 % v/v was carried out in both the systems using Emulsion A and Emulsion B for the preparation of membranes. It was found that 13 % v/v SPAN 80 was sufficient for the maximum transport of analyte in both the emulsion. 7 Effect of membrane solvents. For the preparation of emulsion membrane, the carriers should be taken in the different organic solvents depending upon their solubility, in the present work, the membrane solvents used were kerosene, toluene m ethyl acetate, dichloromethane, .IT was found that the percent transport of analyte were maximum in toluene using both the emulsion systems, therefore toluene was used in the most of cases, experiments were carried out for the other non-polar solvents. 8 Effect of the stirring time. The stirring rate (speed of agitation) affects the p[performance of ELM process to grate extend. Therefore its effect on the percent transport is very important. Variation of stirring rate form 100-500 rpm in the percent system of the transport studies of analyte with emulsion A and Emulsion B found that percent transport of analyte increases with increases in stirring rate up to 300 rpm and deceased above this rate. Increases in speed should increases their mass transfer, however in the present case the percent transport decreases above 300 rpm on the contrary to the general exceptional be attributed ti hydrodynamic instability of ELM at high speed causing sheer on the emulsion globules resulting in breakage. The results of the various parameters obtained as studied above are as tabulated in the form of table 2.1-2.8 B. Example s Application of the method for the separation of ampicillin form synthetic mixture. Application of the proposed methods for the transport of ampicillin using aliquat - 336 and Cynaex - 301 as carriers as developed above has been applied for it suitability of ampicillin from synthetic of mixture of effluents sample of ampicillin manufacturing site. Effluent sample is associated with phenylglycine hydrochloride. Therefore various amounts of pheneylglycine hydrochloride were taken with the fixed amounts of ampicillin and the transport studies were carried out by the procedure developed as above, it is observed that the present method can be satisfactory applied for the separation of the ampicillin. for the recovery of the ampicillin from the waste mixture using cyanex - 301 . However using aliquat - 336, as carrier the method developed has not suitable applied for this separation. This concludes that CYANEX - 301 can be used better carrier for ELM separation of ampicillin. The results obtained are tabulated in the form of table from NO. 2.9 A and 2.9 B. TABLE 2.1 A Transport as a Function of pH of Outer Feed Solution. Feed Phase : 50 cm" of 1 mole/dm NaCl carbonate buffer pH(varying) containing 250µg ampicillin. Internal Phase : 10 cm"1 of 0.8mol/dm3NaCl in citrate buffer pH=3 . Stirring Time : 25 minutes . Stirring Speed : 300 .pm . Membrane : ELM(A). Temperature : Room Temperature . Initial pH % Transport R.S.D. (n=3) Optimum Condition Used 6 50.19 0.80 7 68.31 0.34 8 76.77 0.25 pH=8 9 76.38 0.32 10 60.83 0.40 TABLE 2.!B Effect of feed phase pH on % Transport of Ampicillin External Phase : 50cm3 HCL l x Lo-2 mole/dm3HClcontaining 250µg Ampicillin (varied). Internal Phase 10 cm3 of 1.5mole3 NaCl in carbonate buffer of pH=8.5 . Stirring Speed 300 rpm . Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature . ConcofHCImol/dm3 % Transport RS.D.(n=3) Optimum Condition Used 1x10-3 54.29 0.43 5x10"3 67.81 0.34 1x10-2 79.95 0.45 1xl0-2 mol/dm3 5x10-2 60.48 0.43 x10-1 35.16 0.47 TABLE 2.2A Transport as a Function of Internal Phase pH Feed Phase 50 cm3of 1 moIe/dm3NaCl carbonate buffer pH= 8 containing 250µg ampicillin. Internal Phase 10 cm3 of O.8mol/dm3NaCl in citrate buffer pH (varying). Stirring Time 25 minutes . Stirring Speed 300rpm. Membrane ELM(A). Temperature Room Temperature . Internal Phase pH % Transport R.S.D. (n=3) Optimum Condition Used 2 66.54 0.35 3 76.77 0.50 4 75.20 0.52 pH = 3 5 60.83 0.44 6 42.32 0.51 TABLE 2.2B Effect of Internal Phase pH on % Transport of Ampicillin External Phase : 50cm3 HC1 1x10"2mole/dm3HCl containing 250µg Ampicillin. Internal Phase : 10 cm3 of 1.5mole/dm" NaCl in carbonate buffer of pH(varied). Stirring Speed 300 rpm. Stirring Time : 25 minutes . Membrane . ELM(B). Temperature : Room Temperature. Internal Phase pH % Transport RS.D.(n=3) Optimum ConditionUsed 6.5 58.30 0.34 7.5 68.84 0.36 8.5 79.95 0.48 pH-8.5 9.5 79.49 0.39 10.5 55.21 0.43 Table 2.3A Effect of NaCl Conc.in the Internal Phase on % Transport External Phase : 50 cm3of l mole/dm3NaCl carbonate buffer pH= 8 containing 250µg ampicillin. Internal Phase : 10 cm of 0.8mol/dm3NaCl in citrate buffer pH=3. Stirring Speed : 300 rpm. Stirring Time : 25 minutes. Membrane : ELM(A). Temperature : Room Temperature . NaCl Conc.mole/dm3 % Transport RS.D. (n=3) Optimum ConditionUsed 0.1 59.43 0.47 0.4 72.26 0.62 0.8 76.77 0.24 0.8mole/Dm 1.2 76.59 0.52 1.6 62.08 0.37 TABLE 2.3B Effect of NaCI Conc.in the Internal Phase on % Transport External Phase : 50cm3 HC1 lxl0-2mole/dm3HCl containing 250µg Ampicillin . Internal Phase : 10 cm3 of 1.5mole/dm3 NaCl(varied) in carbonate buffer of pH=8.5. Stirring Speed : 300 rpm. Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature . NaCI Conc.mole/dm3 % Transport RS.D.(N=3) Optimum Condition Used 1.0 77.43 0.37 1.3 75.26 0.26 1.5 79.84 0.25 1.5 mole/dm3 1.7 45.59 0.30 1.9 32.08 0.35 2.0 20.39 0.30 TABLE 2.4A Transport as a Function of StirringTime peed Phase : 50 cm of 1 mole/dm3 NaCl carbonate buffer pH= 8 containing 250µg ampicillin. Internal Phase : 10 cm3 of O.8mol/dm3NaCl in citrate buffer pH=3. Stirring Time : Varying . Stirring Speed : 300 rpm. Membrane : ELM(A) Temperature : Room Temperature . Co : 250µg Ct : [ 250 - (transport at time t) ] ug Time(minutes) % Transport Ct RS.D.(n=3) Optimum ConditionUsed In Co/Ct 10 44.88 112.20 0.36 0.259 15 56.50 141.25 0.37* 0.562 20 76.57 191.43 0.39 25 minutes 0.630 25 76.77 191.93 0.25 0.634 30 76.77 191.93 0.44 0.634 35 76.57 191.43 0.39 0.630 Table 2.4B Transport as a Function of Stirring Time External Phase 50cm3 HCl 1 x l0"2mole/dm3HCl containing 250µg Ampicillin . Internal Phase : 10 cm3 of 1.5mole/dm3 NaCl in carbonate buffer of pH=8.5 . Stirring Speed : 300 rpm. Stirring Time : Varying.. Membrane : ELM(B). Temperature : Room Temperature. Co : 250ug. Ct : [ 250 - (transport at time t)] fig. Time(minutes) % Transport Ct (n=3) OptimumConditionUsed In Co/Ct 10 47.88 119.70 0.43 0.198 15 65.98 164.95 0.26 0.468 20 77.43 193.57 0.42 25 minutes 0.614 25 79.95 199.87 0.41 0.707 30 79.72 199.30 0.33 0.702 35 79.61 199.25 0.35 0.701 TABLE 2.5A Effect of Carrier Concentration (Aliquat -336) on % Transport of ampicilln Feed Phase 50 cm3of 1mole/dm3NaCl carbonate buffer pH= 8 containing 250µg ampicillin. Internal Phase 10 cm3 of O.8mol/dm3NaCl in citrate buffer pH=3. Stirring Time 25 minutes . Stirring Speed 300 rpm Membrane ELM(A). Temperature Room Temperature . Carrier Concentration.(% v/v) % Transport RS.D.(n=3) OptimumCondition Used 1 30.51 0.39 5 53.54 0.37 10 76.77 0.51 10%v/v 15 70.28 0.51 20 54.33 0.44 TABLE 2.5B Effect of Carrier Concentration (Cyanex-301) on % Transport of ampicilln External Phase : 50cm3 HC1 in 1 x l0-2mole/dm3HC1 containing 250ug Ampicillin . Internal Phase : 10 cm' of 1,5mole/dm: NaCl in carbonate buffer of pH=8.5. Stirring Speed : 300 rpm. Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature. TABLE 2.6A Effect of Surfactant Concentration on % Transport of Ampicillin Feed Phase : 50 cm3 of 1 mole/dm3 NaCl carbonate buffer pH= 8 containing 250µg ampicillin. Internal Phase : 10 cm3 of 0.8mol/dm3NaCl in citrate buffer pH=3. Stirring Time 25 minutes . Stirring Speed 300 rpm. Membrane : ELM(A) Temperature : Room Temperature . SurfactantConc.(% V/V) % Transport RS.D.(n=3) OptimumCondition Used 16 54.53 0.37 13 76.77 0.31 13%v/v 10 70.47 0.44 6 40.16 0.41 TABLE 2.6B Effect of Surfactant Concentration on % Transport of Ampicillin External Phase : 50cm3 HC1 lxl0-2mole/dm3HCl containing250µg Ampicillin. Internal Phase : 10 cm3 of 1.5mole/dm3 NaCl in carbonate buffer of pH=8.5. Stirring Speed : 300 rpm Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature . Span-80 ( % v/v) % Transport RS.D.(IF*) Optimum Condition Used 16 35.50 0.36 13 79.84 0.31 13%(v/v) 10 69.30 0.34 6 45.25 0.22 TABLE 2.7A Effect of Various Solvents on % Transport of Ampicillin Feed Phase : 50 cm3of 1 mole/dm3NaCl carbonate buffer pH= 8 containing 250µg ampicillin. Internal Phase : 10 cm3 of 0.8mol/dm3NaCl in citrate buffer pH=3. Stirring Time : 25 minutes . Stirring Speed : 300 rpm. Membrane : ELM(A) varying solvents, Temperature : Room Temperature . Various Solvents % Transport RS.D. (n=3) Solvent Used Ethyl acetate 45.63 0.33 Toluene 76.77 0.21 Toluene Dichloromethane 20.20 0.31 Kerosene 64.44 0.37 TABLE 2.7B Effect of Various Solvents on % Transport of Ampicillin External Phase : 50cm3 HC1 lx1-2mole/dm3HCl containing 250µg Ampicillin . Internal Phase : 10 cm' of 1.5mole/dm NaCl(varied) in carbonate Buffer of pH=8.5. Stirring Speed : 300 rpm. Stirring Time : 25 minutes . Membrane : ELM(B).. Temperature : Room Temperature . Various Solvents % Transport R.S.D.(n=3) OptimumCondition Used Kerosene 64.26 0.34 Toluene 79.84 0.31 Toluene Ethyl acetate 62.24 0.28 Dichloromethane 20.50 0.34 TABLE 2.8A Effect of Stirring Speed on % Transport of Ampicillin Feed Phase : 50 cm3of lmole/dm3NaCl carbonate buffer pH= 8 containing 250µg ampicillin. Internal Phase : 10 cm3 of O.8mol/dm3NaCl in citrate buffer pH=3. Stirring Time : 25 minutes . Stirring Speed : Varying. Membrane : ELM(A) Temperature : Room Temperature . Rotation / minute % Transport RS.D.(n=3) Optium ConditionUsed 100 45.08 0.45 200 65.51 0.42 300 76.77 0.31 300 RPM 400 76.57 0.41 500 75.98 0.55 TABLE 2.8B Effect of Stirring Speed on % Transport of Ampicillin External Phase : 50cm3 HC1 lxl0-2mole/dm3HCl containing 250µg Ampicillin. Internal Phase : 10 cm3 of 1.5mole/dm3 NaCl in carbonate buffer of pH=8.5. Stirring Speed : Varied. Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature . Rotation / minute % Transport RS.D.(n=3) OptimumCondition Used 100 56.81 0.19 200 60.59 0.36 300 79.83 0.27 300 rpm 400 79.83 0.34 500 79.38 0.30 TABLE 2.9A Recovery of Ampicillin from Synthetic mixture: Feed Phase : 50 cm3of 1 mole/dm3NaCl carbonate buffer pH= 8 containing 250µg ampicillin+ phenylglycine HC1 (varying cone). Internal Phase : 10 cm3 of 0.8mol/dm3NaCl in citrate buffer pH=3. Stirring Time : 25 minutes . Stirring Speed : 300 rpm. Membrane : ELM(A). Temperature : Room Temperature . Synthetic mixture µ.g/50cm3 Recovery( %) RS.D.(n=3) 10µg Phenylglycine HC1 250ug Ampicillin 76.77 0.30 50µg Phenylglycine HC1 250ug Ampicillin 77.78 0.44 15µg Phenylglycine HC1 250µg Ampicillin 76.79 0.37 250µg Phenylglycine HC1 250µg Ampicillin 78.67 0.33 350µg Phenylglycine HC1 250µ.g Ampicillin 82.78 0.23 TABLE 2.9B Recovery of Ampicillin from Synthetic mixture: External Phase 50cm3 HCI l x lO-2mole/dm3HCl containing 250µg Ampicillin + phenylglycine HCI (Varying cone.) Internal Phase 10 cm3 of 1.5mole/dm3 NaCl in carbonate buffer of pH=8.5 . Stirring Speed 300 rpm. Stirring Time 25 minutes. Membrane ELM(B). Temperature : Room Temperature. Synthetic mixture jig/50cm3 Recovery( %) RS.D.(n=3) 10µg Phenylglycine HCI 250µg Ampicillin 79.72 0.42 50µg Phenylglycine HCI 250µg Ampicillin 79.68 0.5 4 150µg Phenylglycine HCI 250µg Ampicillin 7949 0.48 250µg Phenylglycine HCI 250µg Ampicillin 78.97 0.35 350µg Phenylglycine HCI 250µg Ampicillin 80.78 0.43 ? Removal of toxins from the blood(35) ? Treatment of drug disorders(36.37) ? Slow release of enzymes and drugs(38,39) ? Treatment of chronic uremia(40,41) A large number of separations have been carried out using ELM technique for bio-molecules.Literature survey of these have been summarized in Table l.l.From this survey it has been revealed as follows:- TABLE 1.1 1. Various amino acids,antibiotics and organic acids have been separated by ELM technique. 2. Amberlite LA-2, TOMAC, D2EHPA, Aliquat-336 and Alanine-336 have been used as mobile carriers in ELM. 3. Various surfactants used are span-80, paranox-100, ECA 1152 and ECA-4360. 4. Kerosene, n-Heptane, decyl alcohol, n-butyl acetate, m-xylene, hexanol, shellol T and n-dodecane are used as membrane solvents. 5. Most of the solutes are separated from fermentation broths. I Claim 1 An Emulsion liquid membrane method for the separation of organic and inorganic species form the various mixtures comprising of the preparation of Emulsion liquid membrane by stirring of internal phase with a surfactant at the suitable pH and rpm then dissolving these emulsion liquid membrane in non-polar solvent and reading the absorbance at UV-visible spectro-photometer. 2 An Emulsion liquid membrane method as claimed in claim 1 where as the internal phase is selected form LA-2, TOMAC, D2EHPA, ALIQUAT-336, CYANEX - 301 or mixture thereof, 3 An Emulsion liquid membrane method as claimed in claim 1 where as surfactant is Span -80, Paradox-100, ECA 1152, ECA 4360 or mixture thereof, 4 An Emulsion liquid membrane method as claimed in claim 1 where as the non-polar solvents is Kerosene, n-heptane decyl alcohol, n-butyl acetate, m-xylene, hexanol, shellol T and n-dodecane. 5 An Emulsion liquid membrane method as claimed in claim 1 where as pH is in the range of 3-8. 6 An Emulsion liquid membrane method as claimed in claim 1 stirring is carried in the range of 200-500 rpm. 6 An Emulsion liquid membrane method as claimed in claim 1 where as organic species are bio-molecules comprising of the zwitter ions. 7 An Emulsion liquid membrane method as claimed in claim 1 where as the inorganic species are transition metals of the first and second transition series, and inner transition elements and also transuranium elements. 8 Liquid membrane emulsion method as described in the description of application with reference to example. FROM 2 THE PATENT ACT 1970 (39 of 1970) & THE PATENT RULES 2003 COMPLETE SPECFIIFCATION (see section 10 and rule 13) TITLE OF INVENTION A membrane separation process APPLICANT a) NAME OF APPLICANT b) NATIONALITY C) ADDRESS SAIRA B MULLA ; INDIAN OJESH APARTMENT 102-103, NR PATEL NURSING HOME, SECTOR 20, AIOROLI, NAVI-MUMBAI. The following specification particularly describes the invention and the manner in which it is to be performed. A Membrane separation process Filed of invention. The field of invention is related to the separation of the of the organic, inorganic species by the membrane separation process, More precisely invention is related to the separation of materials by liquid membrane separation techniques, Prior art. Membranes are the synthetic barriers across which selective permeation of the desired species can be effected by employing appropriate driving force like electrical potential, hydrostatic pressure, concentration gradient etc. Membranes are ubiquitous everywhere in nature. They are responsible for the transport of tremendous quantities of material in animal and plant kingdom. The selectivities of membrane for certain solutes and their rejection for others have fascinated the scientist for centuries.The semi-permrable nature of the membrane is essential to ensure that the separation takes place. Electrodialysis, reverse osomosis, pervaporation, micro and ultra-microfiltration which are membrane integrated processes have proved their potential in varied industrial and biotechnological areas. Membranes may be either: i. Solid Membranes (Polymeric) or ii Liquid Membranes (Thin films). In Solid Membranes mass transfer of solutes take place due to the different size of the pores and passage of irons or molecules at different rates through the membrane. In the liquid membranes mass transfer takes place due to diffusion process. Different solubilities and diffusion coefficients of the solutes in liquids are responsible for mass transfer LIQUID MEMBRANE TECHNOLOGY: Profound interest has been focused on LM separation processes both from fundamental and technological view points. The separation of solutes by a LM is based on a well established idea that the two completely miscible liquid phases separated by third liquid immiscible with either of them, can exchange solutes provided there is a difference between their chemical potentials in two phases and provided that the intermediate liquid is able to transport them. In most cases the two miscible liquids are aqueous solutions and the third(membrane) phase is an organic solvent. Here, out of two liquids one is donor and second is acceptor phase. The LM technique is a powerful and novel tool and excels most of the conventional separation techniques because of the following merits: • Low energy requirements. • Compact modular devices. • Simultaneous extraction and stripping in the same device. • Zero pollution. • Flexibility of operation. • Smaller volume of raffinate. • Use of mild to moderate process conditions, Some of the advantages of LM over the solid Membranes are • Higher fluxes owing to very much higher diffusion coefficients of solutes in the liquids than solids. Liquid membrane technique allow a convection diffusion region instead of molecular one. This also increases the fluxes. • Owing to the availability of great number of substances which when added to the liquid membrane phase increases the In contrast to solvent extraction, in liquid membrane separations amount of solvent used is very small. This permits the use of highly efficient and selective and costly reagents. LM technique is important in the separation science particularly in the instances where solute concentrations are extremely low and large volumes of solution must be processed without generating any secondary waste. US patent 3410794 teaches about a process for separating mixtures in to their components part by means of selective permeation through liquid (surfactant) membranes. The mixture to be separated is first emulsified in aqueous medium contain a selected coated with surfactant are then contacted with a selected true permeate the membrane coating and pass into the solvent phase by conventional means. US patent 4,183,918, this invention relates to the use of liquid membrane technology in preparing medicinal. The medicinal prepared by this invention may be ingested and may be utilized as traps for toxins present in the Gl (gastrointestinal) tract, or as slow-release compositions of drugs, or as reactors. In the trap embodiment, the liquid membrane encapsulated medicinal is an emulsion comprising an exterior phase which is immiscible with the liquids present in the Gl tract and permeable to the toxins therein, and an interior phase which is omissible with the exterior phase and comprises a reagent capable of converting said toxin into a non-permeable" form, In addition, hydrophilic adsorbents, such as a hydrophilic carbon or a silica gel, may be encapsulated in the emulsions of the instant invention. When the compositions of the instant invention are utilized as slow-release drugs, the interior phase of the emulsion will comprise a drug which is slightly soluble in the exterior phase of the emulsion whereby said drug permeates through said exterior phase of the emulsion over a period of time into the Gl tract. The third method for utilizing the compositions of the instant invention comprises encansulation a catalyst for a reaction which is in the Gl tract permeate through the exterior phase of the emulsion into an interior phase wherein said catalyst, for example, an enzyme, converts the permeated reactants to reaction products. The reaction products then may permeate through the exterior phase back into the Gl tract. In all cases, the liquid membrane encapsulated medicinals may be administered by either oral ingestion or injection anywhere else into the Gl tract. Object of the Invention. The aim of present work is to develop method for the recovery/ separation of, organic and inorganic species, especially bio-molecules in a very dilute solutions and the comparative study using anionic and cationic reagents such as cyanex-301 and aliquat-336, LA-2, TOMAC, D2EHPA, ALIQUAT-336, CYANEX - 301 or mixture thereof, as the extractants in membrane solvent toluene and span-80 as emulsifying agent, when emulsion liquid membrane technique is applied. Summary of the invention. The scope of the present investigation was to study the effective separation of some of the organic and inorganic species form the various solutions were concentration of the species (analyte) is low, the technique is especially applicable to the bio-molecules such as antibiotics, amino-acids, or bio-molecules possesses the acidic as well as basic groups. Therefore they behave both as anionic as well as cationic species in the solution at different condition depending upon their pka values m. Therefore can b© completed by Alfe|U9t-336 @nd Cpw --0301 at specified conditions of pH depending upon the. nature of the species. Various anionic and cationic surfactants are studied for the purpose of the separation of the organic and inorganic species form the various solutions, like fermentation broth, Surfactant like Cynaex -301 and Aliquat - 336 are mainly studied are for the purpose of the recovery or separation of the organic or inorganic species form the various media. Cynaex -301 The active components of Cynaex - 301 extractants is bis (2,4,4,-trimethyl pentyl ) dithiophosphoric acid . It is a green mobile liquid having specific gravity, viscosity, and solubility in water 0.95, 78 centipoises, and 7 mg at 24 ° C . Cynaex -301 has been employed as carrier for the recovery of lager number of metal ions by ELM process, Aliquat -336 The active component of aliquat -336 is methyl tricapryl ammonium chloride. Aliquat -336 is yellowish oil. Soluble in chloroform benzene, Aliquat -336 has been employed as carrier in ELM for separation of number of metal ions, bio-molecules, like lactic acid, cephalosphorine-C, amino acid, cephalosphorine . However, there is no literature for the recovery studies of cefadorxil, Ampicilln m ciprofloxacin, using this reagent in ELM. General Experimental Procedure adopted for the development of the membrane separation process is as follows, Instruments A UV visible recording spectrophotometer, control dynamics PH meter combined with glass electrode magnetic stirrer, high speed homogenize, (remi India), were used for the absorbance and PH measurements and transport studies respectives. Chemicals and reagents concentration, Span -80, Span -80, Paradox- 100, ECA 1152, ECA 4360 or mixture thereof, cynex-301, aliquat -0336 were used as regents. All other chemicals toluene Kerosene, n-heptane decyl alcohol, n-butyl acetate, m-xylene, hexanol, shellol T and n-dodecane, sodium citrate, citric acid, sodium carbonate, sodium chlorides HCI were of the AR grade. Preparation of membrane. Membrane (A) 13 cm 3 Span-80 + 10 CM 3 aliquat -336 diluted to 100 Cm 3 with toluene. Membrane (B) 13 CM3 span -80 + 12 CM 3 Cynaex -301 diluted to 10 Cm3 with toluene. Preparation of Emulsion. Emulsion (A) 10 CM 3 of internal phase citrate buffer of PH 3.0 contain 0.8 mole /dm 3 NaCI was added drop-wise to as 5 cm 3 of membrane (A) stirred vigorously on a magnetic stirrer for 20 minutes to get stable milky white emulsion. Emulsion (B) 10 CM 3 of internal phase carbonate buffer of pH 8.5 containing 1.5 mole /dm 3 of NaCI was added to 5 Cm 3 membrane stirred vigorously on a magnetic stirrer for 20 minuets to get stable milky white solution. Citrate buffer and carbonate buffer are prepared as per the standard procedures. General Extraction procedure for the transport studies of analyte. Using Emulsion (A) 15 CM 3 of Emulsion (A) prepared as above was added into a 100 Cm 3 beaker containing 50 Cm 3 of carbonate buffer with 1.0 mole /dm 3 of Nad idjUSied @t pH 8 and £§Q meg of analyte and the concentrate were stirred gently Hi 20P rpm m m$mp Stirred for a given transfer time of 25 minutes. Th8 emulsion was separated by using separating funnel'afld de-emulsified by treating it with 0.5 CM 3 butanol. Analyte so separated in the aqueous phase is estimated spectro-photometrically at 254 nm or at the maximum absorbance at the ultraviolet range. The amount of the analyte remaining the feed phase was calculated by mass balance. The percentage was then calculated using the formula, Concentration of analyte in the internal phase * 100 Concentration of analyte in the feed phase Emulsion (B) Using the Emulsion (B) same procedure is followed as above for the analyte for getting the similar transportation of analyte from the internal phase to the feed phase and same % ratio is obtained. Various parameters were evaluated for the maximum percent of transport of analyte form an aqueous feed solution to inner phase solution through the ELM (A) or ELM (B) containing aliquat - 336 and Cyanex -301 as carrier. Parameters studied are as follows, 1. Effect of pH or H + ion concentration of the feed phase. 2. Effect of pH of internal Phase. 3. Effect of concentration of NaCI in the internal phase. 4. Effect of stirring time. 5. Effect of concentration of carriers. 6. Effect of surfactant in the membrane. 7. Effect of membrane solvents 8. Effect of stirring rate. 1 Effect of pH or H * ion concentration of the feed phase. The feed phase used for the transport studies of the analyte is 1.0 mole/dm 3 Nacl in carbonate buffer using Emulsion A where as it was in aqueous solution for Emulsion B, The PH of the feed was varied form 6-10 for Emulsion A and HCI concentration was varied for 1X 10 "1 mole /dm 3 for Emulsion B. It was observed that maximum transport 80 % at 1X 10-2 mole therefore the feed phase pH for transport studies of analyte using Emulsion a NH2 groups, or it is zwitter ions contains both the acidic and basic functional groups it ionic forms of different charges in solution depends upon PH of the media and pKa values. At pH values below 4.0 (pK a 1 = 4.0) it exists predominantly in cationic form where as above where as 6.0 it exist predominantly PK a 2 6.0 in anionic form. 2 Effect of pH of internal Phase. The internal phase of the membrane used for the transport studies of analyte at various pH of these ranging form 2-6 and 6.5-10.5 using emulsion (A) and Emulsion (B) respectively indicate that the percent transport of analyte gave maximum transport at pH 3 for emulsion (A) and pH 8.5 for the Emulsion (B). Therefore PH of the internal phase solution for emulsion (A) was set at 3.0 and for emulsion (B) was set up at the 8.5 as optimum conditions for the further studies. 3 Effect of concentration of NaCI in the internal Phase. The presence of chloride ions in the internal phase for the transport of analyte using Emulsion (A) with aliquat-336 was essential as a source of chloride ions generation, where as in case with emulsion (B) with Cyanex -301 it is required to prevent the membrane swelling, variation in the concentration of chloride as Nad for the maximum % transport of Ampicilln was carried out in both systems , it was found that the 0.8mole/dm 3 and 1.5 mole /dm3 are QptpyjTI Condition® for emulsion (A) and efnulsion (B) respectively. The ( stirring time was varied form 5-35'"minutes keeping all the experimental parameters for both emulsion (A) and emulsion (B) respectively, it is observed that the percent transport of analyte was increased form 5-25 minutes and then remains constant thereafter in both the cases . Thus stirring time of 25 minuets was used as optimum time 5 Effect of concentration carriers Variation in the concentration of carrier used for the preparation used for the preparation of membranes form 5-20% in toluene, indicates that the percent transport of the analyte with increase in carrier concentration found to be maximum 76.77 % and 79.95% with Emulsion A and Emulsion B respectively. AS the carrier concentration in the membrane increases initial flux increases hence percent transport, however much increase in the carrier concentration increases the viscosity of the membrane thereby decreasing the transport. Therefore it is essential that appropriate quantity of carrier must be present in the membrane. 6 Effect of surfactant in the membrane. Variation of surfactant SAN 80 from 6-16 % v/v was carried out in both the systems using Emulsion A and Emulsion B for the preparation of membranes. It was found that 13 % v/v SPAN 80 was sufficient for the maximum transport of analyte in both the emulsion. 7 Effect of membrane solvents. For the preparation of emulsion membrane, the carriers should be taken in the different organic solvents depending upon their solubility, in the present work, the membrane solvents used were kerosene, toluene m ethyl acetate, dichloromethane, .IT was found that the percent transport of analyte were maximum in toluene using both the emulsion systems, therefore toluene was used in the most of cases, experiments were carried out for the other non-polar solvents. 8 Effect of the stirring time. The stirring rate (speed of agitation) affects the p[performance of ELM process to grate extend. Therefore its effect on the percent transport is very important. Variation of stirring rate form 100-500 rpm in the percent system of the transport studies of analyte with emulsion A and Emulsion B found that percent transport of analyte increases with increases in stirring rate up to 300 rpm and deceased above this rate. Increases in speed should increases their mass transfer, however in the present case the percent transport decreases above 300 rpm on the contrary to the general exceptional be attributed ti hydrodynamic instability of ELM at high speed causing sheer on the emulsion globules resulting in breakage. The results of the various parameters obtained as studied above are as tabulated in the form of table 2.1-2.8 B. Example s Application of the method for the separation of Ampicilln form synthetic mixture. Application of the proposed methods for the transport of Ampicilln using aliquat - 336 and Cynaex - 301 as carriers as developed above has been applied for it suitability of Ampicilln from synthetic of mixture of effluents sample of Ampicilln manufacturing site. Effluent sample is associated with phenylglycine hydrochloride. Therefore various amounts of pheneylglycine hydrochloride were taken with the fixed amounts of Ampicilln and the transport studies were carried out by the procedure developed as above, it is observed that the present method can be satisfactory applied for the separation of the Ampicilln. for the recovery of the Ampicilln from the waste mixture using cyanex - 301 . However using aliquat - 336, as carrier the method developed has not suitable applied for this separation. This concludes that CYANEX - 301 can be used better carrier for ELM separation of Ampicilln. The results obtained are tabulated in the form of table from NO. 2.9 A and 2.9 B. TABLE 2.1 A Transport as a Function of pH of Outer Feed Solution. Feed Phase : 50 cm" of 1 mole/dm3 NaCl carbonate buffer pH(varying) containing 250ug Ampicilln. Internal Phase : 10 cm"1 of 0.8mol/dm3NaCl in citrate buffer pH=3 . Stirring Time : 25 minutes . Stirring Speed : 300 .pm . Membrane : ELM(A). Temperature : Room Temperature . Initial pH % Transport R.S.D. (n=3) Optimum Condition Used 6 50.19 0.80 7 68.31 0.34 8 76.77 0.25 pH=8 9 76.38 0.32 10 60.83 0.40 TABLE 2. B Effect of feed phase pH on % Transport of Ampicilln External Phase : 50cm3 HCI lxl0-2mole/dm3HClcontaining 250ug Ampicilln (varied). Internal Phase 10 cm3 of 1.5mole/dm3 NaCI in carbonate buffer of pH=8.5 . Stirring Speed 300 rpm . Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature . Optimum ConcofHCI % Transport RS.D. Condition Used mol/dm3 (n=3) lxl0-3 54.29 0.43 5x10-3 67.81 0.34 lxl 0-2 79.95 0.45 lxlO"2 mol/dm3 5x10-2 60.48 0.43 1x10-1 35.16 0.47 TABLE 2.2A Transport as a Function of Internal Phase pH Feed Phase 50 cm3of 1 moIe/dm3NaCl carbonate buffer pH= 8 containing 250ug Ampicilln. Internal Phase 10 cm3 of 0.8mol/dm3NaCl in citrate buffer pH (varying). Stirring Time 25 minutes . Stirring Speed 300rpm. Membrane ELM(A). Temperature Room Temperature . Internal Phase pH % Transport R.S.D. (n=3) Optimum Condition Used 2 66.54 0.35 3 76.77 0.50 4 75.20 0.52 pH = 3 5 60.83 0.44 6 42.32 0.51 TABLE 2.2B Effect of Internal Phase pH on % Transport of Ampicilln External Phase : 50cm3 HC1 1x10-2mole/dm3HCl containing 250ug Ampicilln. Internal Phase : 10 cm3 of 1.5mole/dm" NaCl in carbonate buffer of pH(varied). Stirring Speed 300 rpm. Stirring Time : 25 minutes . Membrane . ELM(B). Temperature : Room Temperature. Internal Phase pH % Transport RS.D.(n=3) Optimum ConditionUsed 6.5 58.30 0.34 7.5 68.84 0.36 8.5 79.95 0.48 pH-8.5 9.5 79.49 0.39 10.5 55.21 0.43 Table 2.3A Effect of NaCl Conc in the Internal Phase on % Transport External Phase : 50 cm3ofl mole/dm VlaCl carbonate buffer pH= 8 containing 250p.g Ampicilln. Internal Phase : 10 cm of 0.8mol/dm'NaCl in citrate buffer pH=3. Stirring Speed : 300 rpm. Stirring Time : 25 minutes. Membrane : ELM(A). Temperature : Room Temperature . NaClConc.mole/dm3 % Transport RS.D.(n=3) Optimum ConditionUsed 0.1 59.43 0.47 0.4 72.26 0.62 0.8 76.77 0.24 0.8mole/dnr 1.2 76.59 0.52 1.6 62.08 0.37 TABLE 2.3B Effect of NaCI Conc.in the Internal Phase on % Transport External Phase : 50cm3 HC1 lxl0~2mole/dm3HCl containing 250ug Ampicilln . Internal Phase : 10 cm3 of 1,5mole/dm3 NaCl(varied) in carbonate buffer of pH=8.5. Stirring Speed : 300 rpm. Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature . NaCI Conc.mole/dm3 % Transport RS.D.(N=3) Optimum Condition Used 1.0 77.43 0.37 1.3 75.26 0.26 1.5 79.84 0.25 1.5 mole/dm3 1.7 45.59 0.30 1.9 32.08 0.35 2.0 20.39 0.30 TABLE 2.4A Transport as a Function of StirringTime peed Phase : 50 cm of 1 mole/dm NaCl carbonate buffer pH= 8 containing 250ug Ampicilln. Internal Phase : 10 cm3 of 0.8mol/dm3NaCl in citrate buffer pH=3. Stirring Time : Varying . Stirring Speed : 300 rpm. Membrane : ELM(A) Temperature : Room Temperature . Co : 250pg Ct : [ 250 - (transport at time t) ] ug Optimum Time % Transport Ct RS.D. Condition ■In Co/Ct (minutes) (n=3) Used 10 44.88 112.20 0.36 0.259 15 56.50 141.25 0.37* 0.562 20 76.57 191.43 0.39 25 minutes 0.630 25 76.77 191.93 0.25 0.634 30 76.77 191.93 0.44 0.634 35 76.57 191.43 0.39 0.630 Table 2.4B Transport as a Function of Stirring Time External Phase 50cm'HC1 1xl0-2mole/dm3HCl containing 250fig Ampicilln . Internal Phase : 10 cm3 of 1.5mole/dm3 NaCl in carbonate buffer of pH=8.5 . Stirring Speed : 300 rpm. Stirring Time : Varying.. Membrane : ELM(B). Temperature : Room Temperature. Co : 250ug. Ct : 250 - (transport at time t)] fig. Time(minutes) % Transport Ct RS.D.(n=3) OptimumConditionUsed In Co/Ct 10 47.88 119.70 0.43 0.198 15 65.98 164.95 0.26 0.468 20 77.43 193.57 0.42 25 minutes 0.614 25 79.95 199.87 0.41 0.707 30 79.72 199.30 0.33 0.702 35 79.61 199.25 0.35 0.701 TABLE 2.5A Effect of Carrier Concentration (Aliquat -336) on % Transport of ampicilln Feed Phase 50 cmJof Imole/dm NaCl carbonate buffer pH= 8 containing 250ug Ampicilln. Internal Phase 10 cm3 of O.8mol/dm^NaCI in citrate buffer pH=3. Stirring Time 25 minutes . Stirring Speed 300 rpm Membrane ELM(A). Temperature Room Temperature . Carrier Concentration. % Transport RS.D. Optimum (% v/v) (n=3) Condition Used 1 30.51 0.39 5 53.54 0.37 10 76.77 0.51 10%v/v 15 70.28 0.51 20 54.33 0.44 TABLE 2.5B Effect of Carrier Concentration (Cyanex-301) on % Transport of ampicilln External Phase : 50cm3 HC1 in 1 x10'2mole/dm3HC1 containing 250ug Ampicilln . Internal Phase : 10 cm' of 1,5mole/dm: NaCl in carbonate buffer of pH=8.5. Stirring Speed : 300 rpm. Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature. TABLE 2.6A Effect of Surfactant Concentration on % Transport of Ampicilln Feed Phase : 50 cm of 1 mole/dm NaCl carbonate buffer pH= 8 containing 250ug Ampicilln. Internal Phase : 10 cm' of 0.8mol/dm'NaCl in citrate buffer pH=3. Stirring Time 25 minutes . Stirring Speed 300 rpm. Membrane : ELM(A) Temperature : Room Temperature . SurfactantConc.(% V/V) % Transport RS.D.(n=3) OptimumCondition UsecS 16 54.53 0.37 13 76.77 0.31 13%v/v 10 70.47 0.44 6 40.16 0.41 TABLE 2. 6B Effect of Surfactant Concentration on % Transport of Ampicilln External Phase : 50cm3 HC1 lxl0~2mole/dm3HC! containing250ug Ampicilln. Internal Phase : 10 cm3 of 1.5mole/dm3 NaCl in carbonate buffer of pH=8.5. Stirring Speed : 300 rpm Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature . Span-80 ( % v/v) % Transport RS.D.(IF*) Optimum Condition Used 16 35.50 0.36 13 79.84 0.31 13%(v/v) 10 69.30 0.34 6 45.25 0.22 TABLE 2.7A Effect of Various Solvents on % Transport of Ampicilln Feed Phase : 50 cm3of 1 mole/dm3NaCl carbonate buffer pH= 8 containing 250ng Ampicilln. Internal Phase : 10 cm3 of 0.8mol/dm3NaCl in citrate buffer pH=3. Stirring Time : 25 minutes . Stirring Speed : 300 rpm. Membrane : ELM(A) varying solvents, femperature : Room Temperature . Various Solvents % Transport RS.D. (n=3) Solvent Used Ethyl acetate 45.63 0.33 Toluene 76.77 0.21 Toluene Dichloromethane 20.20 0.31 Kerosene 64.44 0.37 TABLE 2.7B Effect of Various Solvents on % Transport of Ampicilln External Phase : 50cm3 HC1 lx10"2mole/dm3HCl containing 250ug Ampicilln . Internal Phase : 10 cm' of 1.5mole/dm NaCl(varied) in carbonate bufferofpH=8.5. Stirring Speed : 300 rpm. Stirring Time : 25 minutes . Membrane : ELM(B).. Temperature : Room Temperature . Various Solvents % Transport R.S.D. Optimum (n=3) Condition Used Kerosene 64.26 0.34 Toluene 79.84 0.31 Toluene Ethyl acetate 62.24 0.28 Dichloromethane 20.50 0.34 TABLE 2.8A Effect of Stirring Speed on % Transport of Ampicilln Feed Phase : 50 cm3of lmole/dm^aCl carbonate buffer pH= 8 containing 250(g Ampicilln. Internal Phase : 10 cm3 of O.8mol/dnr^NaCl in citrate buffer pH=3. Stirring Time : 25 minutes . Stirring Speed : Varying. Membrane : ELM(A) Temperature : Room Temperature . Rotation / minute % Transport RS.D.(n=3) Optium ConditionUsed 100 45.08 0.45 200 65.51 0.42 300 76.77 0.31 300 RPM 400 76.57 0.41 500 75.98 0.55 TABLE 2.8B Effect of Stirring Speed on % Transport of Ampicilln External Phase : 50cm3 HC1 lx10"2mole/dm3HCl containing 250mg Ampicilln. Internal Phase : 10 cm3 of 1.5mole/dm3 NaCl in carbonate buffer of pH=8.5. Stirring Speed : Varied. Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature . Rotation / minute % Transport RS.D. Optimum (n=3) Condition Used 100 56.81 0.19 200 60.59 0.36 300 79.83 0.27 300 rpm 400 79.83 0.34 500 79.38 0.30 TABLE 2.9A Recovery of Ampicilln from Synthetic mixture: Feed Phase : 50 cm3of 1 mole/dm3NaCl carbonate buffer pH= 8 containing 250mg Ampicilln+ phenylglycine HC1 (varying cone). Internal Phase : 10 cm3 of 0.8mol/dm3NaCl in citrate buffer pH=3. Stirring Time : 25 minutes . Stirring Speed : 300 rpm. Membrane : ELM(A). Temperature : Room Temperature . Synthetic mixture u.g/50cm3 Recovery( %) RS.D.(n=3) 10ug Phenylglycine HC1 250ug Ampicilln 76.77 0.30 50ug Phenylglycine HC1 250ug Ampicilln 77.78 0.44 150(j.g Phenylglycine HC1 250|ag Ampicilln 76.79 0.37 250ug Phenylglycine HC1 250ug Ampicilln 78.67 0.33 350ug Phenylglycine HC1 250|j.g Ampicilln 82.78 0.23 TABLE 2.9B Recovery of Ampicillin from Synthetic mixture: External Phase 50cm HCI lx10-2mole/dm3HCl containing 250^g Ampicilln + phenylglycine HCI (Varying cone.) Internal Phase 10 cm of 1.5mole/dnr NaCl in carbonate buffer of pH=8.5 . Stirring Speed 300 rpm. Stirring Time 25 minutes. Membrane ELM(B). Temperature : Room Temperature. Synthetic mixture mg/50cm3 Recovery( %) RS.D.(n=3) 10mg Phenylglycine HCI 250mg Ampicilln 79.72 0.42 50mg Phenylglycine HCI 250mg Ampicilln 79.68 0.5 4 150mg Phenylglycine HCI 250mg Ampicilln 7949 0.48 250mg Phenylglycine HCI 250mg Ampicilln 78.97 0.35 350mg Phenylglycine HCI 250mg Ampicilln 80.78 0.43 · Removal of toxins from the blood(35) · Treatment of drug disorder(36.37) · Slow release of enzymes and drugs(38.39) · Treatment of chronic uremia(40,41) A large number of separations have been carried out using ELM technique for bio-molecules Literature survey of these have been summarized in Table 1.1 From this survey it has been revealed as follows:- TABLE 1.1 1. Various amino acids antibiotics and organic acids have been separated by ELM technique. 2. Amberlite LA-2, TOMAC, D2EHPA, AJiquat-336 and Alanine-336 have been used as mobile carriers in ELM. 3. Various surfactants used are span-80, paranox-100, ECA 1152 and ECA-4360. 4. Kerosene, n-Heptane, decyl alcohol, n-butyl acetate, m-xylene, hexanol, shellol T and n-dodecane are used as membrane solvents. 5. Most of the solutes are separated from fermentation broths. I Claim 1 An Emulsion liquid membrane method for the separation of organic and inorganic species form the various mixtures comprising of the preparation of Emulsion liquid membrane by stirring of internal phase with a surfactant at the suitable pH and rpm then dissolving these emulsion liquid membrane in non-polar solvent and reading the absorbance at UV-visible spectro-photometer. 2 An Emulsion liquid membrane method as claimed in claim 1 where as the internal phase is selected form LA-2, TOMAC, D2EHPA, ALIQUAT-336, CYANEX - 301 or mixture thereof, 3 An Emulsion liquid membrane method as claimed in claim 1 where as surfactant is Span -80, Paradox-100, ECA 1152, ECA 4360 or mixture thereof, 4 An Emulsion liquid membrane method as claimed in claim 1 where as the non-polar solvents is Kerosene, n-heptane decyl alcohol, n-butyl acetate, m-xylene, hexanol, shellol T and n-dodecane. 5 An Emulsion liquid membrane method as claimed in claim 1 where as pH is in the range of 3-8. 6 An Emulsion liquid membrane method as claimed in claim 1 stirring is carried in the range of 200-500 rpm. 6 An Emulsion liquid membrane method as claimed in claim 1 where as organic species are bio-molecules comprising of the zwitter ions. 7 An Emulsion liquid membrane method as claimed in claim 1 where as the inorganic species are transition metals of the first and second transition series, and inner transition elements and also transuranium elements. Signature and date of applicant. 8 Liquid membrane emulsion method as described in the description of application with reference to example. FROM 2 THE PATENT ACT 1970 (39 of 1970) & THE PATENT RULES 2003 COMPLETE SPECFIIFCATION (see section 10 and rule 13) TITLE OF INVENTION A membrane separation process APPLICANT a) NAME OF APPLICANT b) NATIONALITY C) ADDRESS SAIRA B MULLA ; INDIAN OJESH APARTMENT 102-103, NR PATEL NURSING HOME, SECTOR 20, AIOROLI, NAVI-MUMBAI. The following specification particularly describes the invention and the manner in which it is to be performed. A Membrane separation process Filed of invention. The field of invention is related to the separation of the of the organic, inorganic species by the membrane separation process, More precisely invention is related to the separation of materials by liquid membrane separation techniques, Prior art. Membranes are the synthetic barriers across which selective permeation of the desired species can be effected by employing appropriate driving force like electrical potential, hydrostatic pressure, concentration gradient etc. Membranes are ubiquitous everywhere in nature. They are responsible for the transport of tremendous quantities of material in animal and plant kingdom. The selectivities of membrane for certain solutes and their rejection for others have fascinated the scientist for centuries.The semi-permrable nature of the membrane is essential to ensure that the separation takes place. Electrodialysis, reverse osomosis, pervaporation, micro and ultra-microfiltration which are membrane integrated processes have proved their potential in varied industrial and biotechnological areas. Membranes may be either: i. Solid Membranes (Polymeric) or ii Liquid Membranes (Thin films). In Solid Membranes mass transfer of solutes take place due to the different size of the pores and passage of irons or molecules at different rates through the membrane. In the liquid membranes mass transfer takes place due to diffusion process. Different solubilities and diffusion coefficients of the solutes in liquids are responsible for mass transfer LIQUID MEMBRANE TECHNOLOGY: Profound interest has been focused on LM separation processes both from fundamental and technological view points. The separation of solutes by a LM is based on a well established idea that the two completely miscible liquid phases separated by third liquid immiscible with either of them, can exchange solutes provided there is a difference between their chemical potentials in two phases and provided that the intermediate liquid is able to transport them. In most cases the two miscible liquids are aqueous solutions and the third(membrane) phase is an organic solvent. Here, out of two liquids one is donor and second is acceptor phase. The LM technique is a powerful and novel tool and excels most of the conventional separation techniques because of the following merits: • Low energy requirements. • Compact modular devices. • Simultaneous extraction and stripping in the same device. • Zero pollution. • Flexibility of operation. • Smaller volume of raffinate. • Use of mild to moderate process conditions, Some of the advantages of LM over the solid Membranes are • Higher fluxes owing to very much higher diffusion coefficients of solutes in the liquids than solids. Liquid membrane technique allow a convection diffusion region instead of molecular one. This also increases the fluxes. • Owing to the availability of great number of substances which when added to the liquid membrane phase increases the In contrast to solvent extraction, in liquid membrane separations amount of solvent used is very small. This permits the use of highly efficient and selective and costly reagents. LM technique is important in the separation science particularly in the instances where solute concentrations are extremely low and large volumes of solution must be processed without generating any secondary waste. US patent 3410794 teaches about a process for separating mixtures in to their components part by means of selective permeation through liquid (surfactant) membranes. The mixture to be separated is first emulsified in aqueous medium contain a selected coated with surfactant are then contacted with a selected true permeate the membrane coating and pass into the solvent phase by conventional means. US patent 4,183,918, this invention relates to the use of liquid membrane technology in preparing medicinal. The medicinal prepared by this invention may be ingested and may be utilized as traps for toxins present in the Gl (gastrointestinal) tract, or as slow-release compositions of drugs, or as reactors. In the trap embodiment, the liquid membrane encapsulated medicinal is an emulsion comprising an exterior phase which is immiscible with the liquids present in the Gl tract and permeable to the toxins therein, and an interior phase which is omissible with the exterior phase and comprises a reagent capable of converting said toxin into a non-permeable" form, In addition, hydrophilic adsorbents, such as a hydrophilic carbon or a silica gel, may be encapsulated in the emulsions of the instant invention. When the compositions of the instant invention are utilized as slow-release drugs, the interior phase of the emulsion will comprise a drug which is slightly soluble in the exterior phase of the emulsion whereby said drug permeates through said exterior phase of the emulsion over a period of time into the Gl tract. The third method for utilizing the compositions of the instant invention comprises encansulation a catalyst for a reaction which is in the Gl tract permeate through the exterior phase of the emulsion into an interior phase wherein said catalyst, for example, an enzyme, converts the permeated reactants to reaction products. The reaction products then may permeate through the exterior phase back into the Gl tract. In all cases, the liquid membrane encapsulated medicinals may be administered by either oral ingestion or injection anywhere else into the Gl tract. Object of the Invention. The aim of present work is to develop method for the recovery/ separation of, organic and inorganic species, especially bio-molecules in a very dilute solutions and the comparative study using anionic and cationic reagents such as cyanex-301 and aliquat-336, LA-2, TOMAC, D2EHPA, ALIQUAT-336, CYANEX - 301 or mixture thereof, as the extractants in membrane solvent toluene and span-80 as emulsifying agent, when emulsion liquid membrane technique is applied. Summary of the invention. The scope of the present investigation was to study the effective separation of some of the organic and inorganic species form the various solutions were concentration of the species (analyte) is low, the technique is especially applicable to the bio-molecules such as antibiotics, amino-acids, or bio-molecules possesses the acidic as well as basic groups. Therefore they behave both as anionic as well as cationic species in the solution at different condition depending upon their pka values m. Therefore can b© completed by Alfe|U9t-336 @nd Cpw --0301 at specified conditions of pH depending upon the. nature of the species. Various anionic and cationic surfactants are studied for the purpose of the separation of the organic and inorganic species form the various solutions, like fermentation broth, Surfactant like Cynaex -301 and Aliquat - 336 are mainly studied are for the purpose of the recovery or separation of the organic or inorganic species form the various media. Cynaex -301 The active components of Cynaex - 301 extractants is bis (2,4,4,-trimethyl pentyl ) dithiophosphoric acid . It is a green mobile liquid having specific gravity, viscosity, and solubility in water 0.95, 78 centipoises, and 7 mg at 24 ° C . Cynaex -301 has been employed as carrier for the recovery of lager number of metal ions by ELM process, Aliquat -336 The active component of aliquat -336 is methyl tricapryl ammonium chloride. Aliquat -336 is yellowish oil. Soluble in chloroform benzene, Aliquat -336 has been employed as carrier in ELM for separation of number of metal ions, bio-molecules, like lactic acid, cephalosphorine-C, amino acid, cephalosphorine . However, there is no literature for the recovery studies of cefadorxil, Ampicilln m ciprofloxacin, using this reagent in ELM. General Experimental Procedure adopted for the development of the membrane separation process is as follows, Instruments A UV visible recording spectrophotometer, control dynamics PH meter combined with glass electrode magnetic stirrer, high speed homogenize, (remi India), were used for the absorbance and PH measurements and transport studies respectives. Chemicals and reagents concentration, Span -80, Span -80, Paradox- 100, ECA 1152, ECA 4360 or mixture thereof, cynex-301, aliquat -0336 were used as regents. All other chemicals toluene Kerosene, n-heptane decyl alcohol, n-butyl acetate, m-xylene, hexanol, shellol T and n-dodecane, sodium citrate, citric acid, sodium carbonate, sodium chlorides HCI were of the AR grade. Preparation of membrane. Membrane (A) 13 cm 3 Span-80 + 10 CM 3 aliquat -336 diluted to 100 Cm 3 with toluene. Membrane (B) 13 CM3 span -80 + 12 CM 3 Cynaex -301 diluted to 10 Cm3 with toluene. Preparation of Emulsion. Emulsion (A) 10 CM 3 of internal phase citrate buffer of PH 3.0 contain 0.8 mole /dm 3 NaCI was added drop-wise to as 5 cm 3 of membrane (A) stirred vigorously on a magnetic stirrer for 20 minutes to get stable milky white emulsion. Emulsion (B) 10 CM 3 of internal phase carbonate buffer of pH 8.5 containing 1.5 mole /dm 3 of NaCI was added to 5 Cm 3 membrane stirred vigorously on a magnetic stirrer for 20 minuets to get stable milky white solution. Citrate buffer and carbonate buffer are prepared as per the standard procedures. General Extraction procedure for the transport studies of analyte. Using Emulsion (A) 15 CM 3 of Emulsion (A) prepared as above was added into a 100 Cm 3 beaker containing 50 Cm 3 of carbonate buffer with 1.0 mole /dm 3 of Nad idjUSied @t pH 8 and £§Q meg of analyte and the concentrate were stirred gently Hi 20P rpm m m$mp Stirred for a given transfer time of 25 minutes. Th8 emulsion was separated by using separating funnel'afld de-emulsified by treating it with 0.5 CM 3 butanol. Analyte so separated in the aqueous phase is estimated spectro-photometrically at 254 nm or at the maximum absorbance at the ultraviolet range. The amount of the analyte remaining the feed phase was calculated by mass balance. The percentage was then calculated using the formula, Concentration of analyte in the internal phase * 100 Concentration of analyte in the feed phase Emulsion (B) Using the Emulsion (B) same procedure is followed as above for the analyte for getting the similar transportation of analyte from the internal phase to the feed phase and same % ratio is obtained. Various parameters were evaluated for the maximum percent of transport of analyte form an aqueous feed solution to inner phase solution through the ELM (A) or ELM (B) containing aliquat - 336 and Cyanex -301 as carrier. Parameters studied are as follows, 1. Effect of pH or H + ion concentration of the feed phase. 2. Effect of pH of internal Phase. 3. Effect of concentration of NaCI in the internal phase. 4. Effect of stirring time. 5. Effect of concentration of carriers. 6. Effect of surfactant in the membrane. 7. Effect of membrane solvents 8. Effect of stirring rate. 1 Effect of pH or H * ion concentration of the feed phase. The feed phase used for the transport studies of the analyte is 1.0 mole/dm 3 Nacl in carbonate buffer using Emulsion A where as it was in aqueous solution for Emulsion B, The PH of the feed was varied form 6-10 for Emulsion A and HCI concentration was varied for 1X 10 "1 mole /dm 3 for Emulsion B. It was observed that maximum transport 80 % at 1X 10-2 mole therefore the feed phase pH for transport studies of analyte using Emulsion a NH2 groups, or it is zwitter ions contains both the acidic and basic functional groups it ionic forms of different charges in solution depends upon PH of the media and pKa values. At pH values below 4.0 (pK a 1 = 4.0) it exists predominantly in cationic form where as above where as 6.0 it exist predominantly PK a 2 6.0 in anionic form. 2 Effect of pH of internal Phase. The internal phase of the membrane used for the transport studies of analyte at various pH of these ranging form 2-6 and 6.5-10.5 using emulsion (A) and Emulsion (B) respectively indicate that the percent transport of analyte gave maximum transport at pH 3 for emulsion (A) and pH 8.5 for the Emulsion (B). Therefore PH of the internal phase solution for emulsion (A) was set at 3.0 and for emulsion (B) was set up at the 8.5 as optimum conditions for the further studies. 3 Effect of concentration of NaCI in the internal Phase. The presence of chloride ions in the internal phase for the transport of analyte using Emulsion (A) with aliquat-336 was essential as a source of chloride ions generation, where as in case with emulsion (B) with Cyanex -301 it is required to prevent the membrane swelling, variation in the concentration of chloride as Nad for the maximum % transport of Ampicilln was carried out in both systems , it was found that the 0.8mole/dm 3 and 1.5 mole /dm3 are QptpyjTI Condition® for emulsion (A) and efnulsion (B) respectively. The ( stirring time was varied form 5-35'"minutes keeping all the experimental parameters for both emulsion (A) and emulsion (B) respectively, it is observed that the percent transport of analyte was increased form 5-25 minutes and then remains constant thereafter in both the cases . Thus stirring time of 25 minuets was used as optimum time 5 Effect of concentration carriers Variation in the concentration of carrier used for the preparation used for the preparation of membranes form 5-20% in toluene, indicates that the percent transport of the analyte with increase in carrier concentration found to be maximum 76.77 % and 79.95% with Emulsion A and Emulsion B respectively. AS the carrier concentration in the membrane increases initial flux increases hence percent transport, however much increase in the carrier concentration increases the viscosity of the membrane thereby decreasing the transport. Therefore it is essential that appropriate quantity of carrier must be present in the membrane. 6 Effect of surfactant in the membrane. Variation of surfactant SAN 80 from 6-16 % v/v was carried out in both the systems using Emulsion A and Emulsion B for the preparation of membranes. It was found that 13 % v/v SPAN 80 was sufficient for the maximum transport of analyte in both the emulsion. 7 Effect of membrane solvents. For the preparation of emulsion membrane, the carriers should be taken in the different organic solvents depending upon their solubility, in the present work, the membrane solvents used were kerosene, toluene m ethyl acetate, dichloromethane, .IT was found that the percent transport of analyte were maximum in toluene using both the emulsion systems, therefore toluene was used in the most of cases, experiments were carried out for the other non-polar solvents. 8 Effect of the stirring time. The stirring rate (speed of agitation) affects the p[performance of ELM process to grate extend. Therefore its effect on the percent transport is very important. Variation of stirring rate form 100-500 rpm in the percent system of the transport studies of analyte with emulsion A and Emulsion B found that percent transport of analyte increases with increases in stirring rate up to 300 rpm and deceased above this rate. Increases in speed should increases their mass transfer, however in the present case the percent transport decreases above 300 rpm on the contrary to the general exceptional be attributed ti hydrodynamic instability of ELM at high speed causing sheer on the emulsion globules resulting in breakage. The results of the various parameters obtained as studied above are as tabulated in the form of table 2.1-2.8 B. Example s Application of the method for the separation of Ampicilln form synthetic mixture. Application of the proposed methods for the transport of Ampicilln using aliquat - 336 and Cynaex - 301 as carriers as developed above has been applied for it suitability of Ampicilln from synthetic of mixture of effluents sample of Ampicilln manufacturing site. Effluent sample is associated with phenylglycine hydrochloride. Therefore various amounts of pheneylglycine hydrochloride were taken with the fixed amounts of Ampicilln and the transport studies were carried out by the procedure developed as above, it is observed that the present method can be satisfactory applied for the separation of the Ampicilln. for the recovery of the Ampicilln from the waste mixture using cyanex - 301 . However using aliquat - 336, as carrier the method developed has not suitable applied for this separation. This concludes that CYANEX - 301 can be used better carrier for ELM separation of Ampicilln. The results obtained are tabulated in the form of table from NO. 2.9 A and 2.9 B. TABLE 2.1 A Transport as a Function of pH of Outer Feed Solution. Feed Phase : 50 cm" of 1 mole/dm3 NaCl carbonate buffer pH(varying) containing 250ug Ampicilln. Internal Phase : 10 cm"1 of 0.8mol/dm3NaCl in citrate buffer pH=3 . Stirring Time : 25 minutes . Stirring Speed : 300 .pm . Membrane : ELM(A). Temperature : Room Temperature . Initial pH % Transport R.S.D. (n=3) Optimum Condition Used 6 50.19 0.80 7 68.31 0.34 8 76.77 0.25 pH=8 9 76.38 0.32 10 60.83 0.40 TABLE 2. B Effect of feed phase pH on % Transport of Ampicilln External Phase : 50cm3 HCI lxl0-2mole/dm3HClcontaining 250ug Ampicilln (varied). Internal Phase 10 cm3 of 1.5mole/dm3 NaCI in carbonate buffer of pH=8.5 . Stirring Speed 300 rpm . Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature . Optimum ConcofHCI % Transport RS.D. Condition Used mol/dm3 (n=3) lxl0-3 54.29 0.43 5x10-3 67.81 0.34 lxl 0-2 79.95 0.45 lxlO"2 mol/dm3 5x10-2 60.48 0.43 1x10-1 35.16 0.47 TABLE 2.2A Transport as a Function of Internal Phase pH Feed Phase 50 cm3of 1 moIe/dm3NaCl carbonate buffer pH= 8 containing 250ug Ampicilln. Internal Phase 10 cm3 of 0.8mol/dm3NaCl in citrate buffer pH (varying). Stirring Time 25 minutes . Stirring Speed 300rpm. Membrane ELM(A). Temperature Room Temperature . Internal Phase pH % Transport R.S.D. (n=3) Optimum Condition Used 2 66.54 0.35 3 76.77 0.50 4 75.20 0.52 pH = 3 5 60.83 0.44 6 42.32 0.51 TABLE 2.2B Effect of Internal Phase pH on % Transport of Ampicilln External Phase : 50cm3 HC1 1x10-2mole/dm3HCl containing 250ug Ampicilln. Internal Phase : 10 cm3 of 1.5mole/dm" NaCl in carbonate buffer of pH(varied). Stirring Speed 300 rpm. Stirring Time : 25 minutes . Membrane . ELM(B). Temperature : Room Temperature. Internal Phase pH % Transport RS.D.(n=3) Optimum ConditionUsed 6.5 58.30 0.34 7.5 68.84 0.36 8.5 79.95 0.48 pH-8.5 9.5 79.49 0.39 10.5 55.21 0.43 Table 2.3A Effect of NaCl Conc in the Internal Phase on % Transport External Phase : 50 cm3ofl mole/dm VlaCl carbonate buffer pH= 8 containing 250p.g Ampicilln. Internal Phase : 10 cm of 0.8mol/dm'NaCl in citrate buffer pH=3. Stirring Speed : 300 rpm. Stirring Time : 25 minutes. Membrane : ELM(A). Temperature : Room Temperature . NaClConc.mole/dm3 % Transport RS.D.(n=3) Optimum ConditionUsed 0.1 59.43 0.47 0.4 72.26 0.62 0.8 76.77 0.24 0.8mole/dnr 1.2 76.59 0.52 1.6 62.08 0.37 TABLE 2.3B Effect of NaCI Conc.in the Internal Phase on % Transport External Phase : 50cm3 HC1 lxl0~2mole/dm3HCl containing 250ug Ampicilln . Internal Phase : 10 cm3 of 1,5mole/dm3 NaCl(varied) in carbonate buffer of pH=8.5. Stirring Speed : 300 rpm. Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature . NaCI Conc.mole/dm3 % Transport RS.D.(N=3) Optimum Condition Used 1.0 77.43 0.37 1.3 75.26 0.26 1.5 79.84 0.25 1.5 mole/dm3 1.7 45.59 0.30 1.9 32.08 0.35 2.0 20.39 0.30 TABLE 2.4A Transport as a Function of StirringTime peed Phase : 50 cm of 1 mole/dm NaCl carbonate buffer pH= 8 containing 250ug Ampicilln. Internal Phase : 10 cm3 of 0.8mol/dm3NaCl in citrate buffer pH=3. Stirring Time : Varying . Stirring Speed : 300 rpm. Membrane : ELM(A) Temperature : Room Temperature . Co : 250pg Ct : [ 250 - (transport at time t) ] ug Optimum Time % Transport Ct RS.D. Condition ■In Co/Ct (minutes) (n=3) Used 10 44.88 112.20 0.36 0.259 15 56.50 141.25 0.37* 0.562 20 76.57 191.43 0.39 25 minutes 0.630 25 76.77 191.93 0.25 0.634 30 76.77 191.93 0.44 0.634 35 76.57 191.43 0.39 0.630 Table 2.4B Transport as a Function of Stirring Time External Phase 50cm'HC1 1xl0-2mole/dm3HCl containing 250fig Ampicilln . Internal Phase : 10 cm3 of 1.5mole/dm3 NaCl in carbonate buffer of pH=8.5 . Stirring Speed : 300 rpm. Stirring Time : Varying.. Membrane : ELM(B). Temperature : Room Temperature. Co : 250ug. Ct : 250 - (transport at time t)] fig. Time(minutes) % Transport Ct RS.D.(n=3) OptimumConditionUsed In Co/Ct 10 47.88 119.70 0.43 0.198 15 65.98 164.95 0.26 0.468 20 77.43 193.57 0.42 25 minutes 0.614 25 79.95 199.87 0.41 0.707 30 79.72 199.30 0.33 0.702 35 79.61 199.25 0.35 0.701 TABLE 2.5A Effect of Carrier Concentration (Aliquat -336) on % Transport of ampicilln Feed Phase 50 cmJof Imole/dm NaCl carbonate buffer pH= 8 containing 250ug Ampicilln. Internal Phase 10 cm3 of O.8mol/dm^NaCI in citrate buffer pH=3. Stirring Time 25 minutes . Stirring Speed 300 rpm Membrane ELM(A). Temperature Room Temperature . Carrier Concentration. % Transport RS.D. Optimum (% v/v) (n=3) Condition Used 1 30.51 0.39 5 53.54 0.37 10 76.77 0.51 10%v/v 15 70.28 0.51 20 54.33 0.44 TABLE 2.5B Effect of Carrier Concentration (Cyanex-301) on % Transport of ampicilln External Phase : 50cm3 HC1 in 1 x10'2mole/dm3HC1 containing 250ug Ampicilln . Internal Phase : 10 cm' of 1,5mole/dm: NaCl in carbonate buffer of pH=8.5. Stirring Speed : 300 rpm. Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature. TABLE 2.6A Effect of Surfactant Concentration on % Transport of Ampicilln Feed Phase : 50 cm of 1 mole/dm NaCl carbonate buffer pH= 8 containing 250ug Ampicilln. Internal Phase : 10 cm' of 0.8mol/dm'NaCl in citrate buffer pH=3. Stirring Time 25 minutes . Stirring Speed 300 rpm. Membrane : ELM(A) Temperature : Room Temperature . SurfactantConc.(% V/V) % Transport RS.D.(n=3) OptimumCondition UsecS 16 54.53 0.37 13 76.77 0.31 13%v/v 10 70.47 0.44 6 40.16 0.41 TABLE 2. 6B Effect of Surfactant Concentration on % Transport of Ampicilln External Phase : 50cm3 HC1 lxl0~2mole/dm3HC! containing250ug Ampicilln. Internal Phase : 10 cm3 of 1.5mole/dm3 NaCl in carbonate buffer of pH=8.5. Stirring Speed : 300 rpm Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature . Span-80 ( % v/v) % Transport RS.D.(IF*) Optimum Condition Used 16 35.50 0.36 13 79.84 0.31 13%(v/v) 10 69.30 0.34 6 45.25 0.22 TABLE 2.7A Effect of Various Solvents on % Transport of Ampicilln Feed Phase : 50 cm3of 1 mole/dm3NaCl carbonate buffer pH= 8 containing 250ng Ampicilln. Internal Phase : 10 cm3 of 0.8mol/dm3NaCl in citrate buffer pH=3. Stirring Time : 25 minutes . Stirring Speed : 300 rpm. Membrane : ELM(A) varying solvents, femperature : Room Temperature . Various Solvents % Transport RS.D. (n=3) Solvent Used Ethyl acetate 45.63 0.33 Toluene 76.77 0.21 Toluene Dichloromethane 20.20 0.31 Kerosene 64.44 0.37 TABLE 2.7B Effect of Various Solvents on % Transport of Ampicilln External Phase : 50cm3 HC1 lx10"2mole/dm3HCl containing 250ug Ampicilln . Internal Phase : 10 cm' of 1.5mole/dm NaCl(varied) in carbonate bufferofpH=8.5. Stirring Speed : 300 rpm. Stirring Time : 25 minutes . Membrane : ELM(B).. Temperature : Room Temperature . Various Solvents % Transport R.S.D. Optimum (n=3) Condition Used Kerosene 64.26 0.34 Toluene 79.84 0.31 Toluene Ethyl acetate 62.24 0.28 Dichloromethane 20.50 0.34 TABLE 2.8A Effect of Stirring Speed on % Transport of Ampicilln Feed Phase : 50 cm3of lmole/dm^aCl carbonate buffer pH= 8 containing 250(g Ampicilln. Internal Phase : 10 cm3 of O.8mol/dnr^NaCl in citrate buffer pH=3. Stirring Time : 25 minutes . Stirring Speed : Varying. Membrane : ELM(A) Temperature : Room Temperature . Rotation / minute % Transport RS.D.(n=3) Optium ConditionUsed 100 45.08 0.45 200 65.51 0.42 300 76.77 0.31 300 RPM 400 76.57 0.41 500 75.98 0.55 TABLE 2.8B Effect of Stirring Speed on % Transport of Ampicilln External Phase : 50cm3 HC1 lx10"2mole/dm3HCl containing 250mg Ampicilln. Internal Phase : 10 cm3 of 1.5mole/dm3 NaCl in carbonate buffer of pH=8.5. Stirring Speed : Varied. Stirring Time : 25 minutes . Membrane : ELM(B). Temperature : Room Temperature . Rotation / minute % Transport RS.D. Optimum (n=3) Condition Used 100 56.81 0.19 200 60.59 0.36 300 79.83 0.27 300 rpm 400 79.83 0.34 500 79.38 0.30 TABLE 2.9A Recovery of Ampicilln from Synthetic mixture: Feed Phase : 50 cm3of 1 mole/dm3NaCl carbonate buffer pH= 8 containing 250mg Ampicilln+ phenylglycine HC1 (varying cone). Internal Phase : 10 cm3 of 0.8mol/dm3NaCl in citrate buffer pH=3. Stirring Time : 25 minutes . Stirring Speed : 300 rpm. Membrane : ELM(A). Temperature : Room Temperature . Synthetic mixture u.g/50cm3 Recovery( %) RS.D.(n=3) 10ug Phenylglycine HC1 250ug Ampicilln 76.77 0.30 50ug Phenylglycine HC1 250ug Ampicilln 77.78 0.44 150(j.g Phenylglycine HC1 250|ag Ampicilln 76.79 0.37 250ug Phenylglycine HC1 250ug Ampicilln 78.67 0.33 350ug Phenylglycine HC1 250|j.g Ampicilln 82.78 0.23 TABLE 2.9B Recovery of Ampicillin from Synthetic mixture: External Phase 50cm HCI lx10-2mole/dm3HCl containing 250^g Ampicilln + phenylglycine HCI (Varying cone.) Internal Phase 10 cm of 1.5mole/dnr NaCl in carbonate buffer of pH=8.5 . Stirring Speed 300 rpm. Stirring Time 25 minutes. Membrane ELM(B). Temperature : Room Temperature. Synthetic mixture mg/50cm3 Recovery( %) RS.D.(n=3) 10mg Phenylglycine HCI 250mg Ampicilln 79.72 0.42 50mg Phenylglycine HCI 250mg Ampicilln 79.68 0.5 4 150mg Phenylglycine HCI 250mg Ampicilln 7949 0.48 250mg Phenylglycine HCI 250mg Ampicilln 78.97 0.35 350mg Phenylglycine HCI 250mg Ampicilln 80.78 0.43 · Removal of toxins from the blood(35) · Treatment of drug disorder(36.37) · Slow release of enzymes and drugs(38.39) · Treatment of chronic uremia(40,41) A large number of separations have been carried out using ELM technique for bio-molecules Literature survey of these have been summarized in Table 1.1 From this survey it has been revealed as follows:- TABLE 1.1 1. Various amino acids antibiotics and organic acids have been separated by ELM technique. 2. Amberlite LA-2, TOMAC, D2EHPA, AJiquat-336 and Alanine-336 have been used as mobile carriers in ELM. 3. Various surfactants used are span-80, paranox-100, ECA 1152 and ECA-4360. 4. Kerosene, n-Heptane, decyl alcohol, n-butyl acetate, m-xylene, hexanol, shellol T and n-dodecane are used as membrane solvents. 5. Most of the solutes are separated from fermentation broths. I Claim 1 An Emulsion liquid membrane method for the separation of organic and inorganic species form the various mixtures comprising of the preparation of Emulsion liquid membrane by stirring of internal phase with a surfactant at the suitable pH and rpm then dissolving these emulsion liquid membrane in non-polar solvent and reading the absorbance at UV-visible spectro-photometer. 2 An Emulsion liquid membrane method as claimed in claim 1 where as the internal phase is selected form LA-2, TOMAC, D2EHPA, ALIQUAT-336, CYANEX - 301 or mixture thereof, 3 An Emulsion liquid membrane method as claimed in claim 1 where as surfactant is Span -80, Paradox-100, ECA 1152, ECA 4360 or mixture thereof, 4 An Emulsion liquid membrane method as claimed in claim 1 where as the non-polar solvents is Kerosene, n-heptane decyl alcohol, n-butyl acetate, m-xylene, hexanol, shellol T and n-dodecane. 5 An Emulsion liquid membrane method as claimed in claim 1 where as pH is in the range of 3-8. 6 An Emulsion liquid membrane method as claimed in claim 1 stirring is carried in the range of 200-500 rpm. 6 An Emulsion liquid membrane method as claimed in claim 1 where as organic species are bio-molecules comprising of the zwitter ions. 7 An Emulsion liquid membrane method as claimed in claim 1 where as the inorganic species are transition metals of the first and second transition series, and inner transition elements and also transuranium elements. Signature and date of applicant. 8 Liquid membrane emulsion method as described in the description of application with reference to example.

Documents:

679-mum-2005-claims (complete).doc

679-MUM-2005-CLAIMS(AMENDED)-(21-9-2012).pdf

679-MUM-2005-CLAIMS(MARKED COPY)-(21-9-2012).pdf

679-mum-2005-claims.pdf

679-MUM-2005-CORRESPONDENCE(5-6-2009).pdf

679-mum-2005-correspondence(6-6-2006).pdf

679-mum-2005-correspondence-received.pdf

679-mum-2005-description (complete).pdf

679-mum-2005-description (provisional).pdf

679-mum-2005-drawings.pdf

679-mum-2005-form 1(6-6-2006).pdf

679-MUM-2005-FORM 18(5-6-2009).pdf

679-mum-2005-form 2(title page)-(complete)-(6-6-2006).pdf

679-mum-2005-form 2(title page)-(provisional)-(7-6-2005).pdf

679-mum-2005-form 3(6-6-2006).pdf

679-mum-2005-form 5(6-6-2006).pdf

679-mum-2005-form-1.pdf

679-mum-2005-form-2 (complete).doc

679-mum-2005-form-2 (provisional).doc

679-mum-2005-form-2 (provisional).pdf

679-mum-2005-form-2.pdf

679-mum-2005-form-3.pdf

679-mum-2005-form-5.pdf

679-MUM-2005-REPLY TO EXAMINATION REPORT(21-9-2012).pdf

abstract1.jpg