Title of Invention

METHOD OF BIODEGRADATION OF CHLOROBENZENES USING UASB REACTORS.

Abstract

Chlorobenzenes are used mainly as intermediates in the synthesis of pesticides and other chemicals. They are also used as deodorizers, fumigants, degreasers, insecticides and herbicides. The release of chlorobenzenes into the environment occurs primarily during manufacture, and through the dispersive nature of their uses. Chlorobenzenes found in the range of 5-20 jxg/L in ground water by discharging the effluents from large industrial estates in Ankleshwar, Nandesari and Vapi in state of Gujrat, India. They have been found at abandoned waste sites and in leachates. The presence of chlorobenzenes was also observed in low part per billion in stockpile samples collected from Union Carbide plant site in Bhopal, India. An increased level of chlorobenzenes in the water and wastewater is the matter of concern. These compounds are toxic to plants, vertebrates and invertebrates. Chlorobenznes are recalcitrant substances and difficult to degrade using conventional anaerobic method, following invention successfully explores the ability of UASB reactor to degrade the chlorinated organic compounds mainly Monochlorobenzenes (MCB), 1,2-Dichlorobenzene (o-DCB) and 1,4- Dichlorobenzene (/>-DCB) in synthetic medium using mixed culture at bench scale. Because of this new process and condition, the chlorobenzens which are difficult to degrade can be degraded successfully and people can use this low cost technology/ method to degrade the chlorobenzens particularly MCB, 1,2-DCB and 1,4-DCB using UASB reactor. Following invention is explained precisely with the help of figure 1 which shows schematic diagram of experimental set up. Where 1 denotes Synthetic wastewater, 2 denotes Peristaltic pump, 3 denotes Sampling ports, 4 denotes Aciylic column (0.1m x 0.1m), 5 denotes Settler GSS, 6 denotes Effluent and 7 denotes Biogas.

Full Text

4. DESCRIPTION, Technical field of invention This invention relates industrial wastewater treatment. The invention especially relates to methods and apparatuses using UASB reactor to degrade the chlorinated organic compounds 5. Use Following invention successfully explores the ability of UASB reactor to degrade the chlorinated organic compounds mainly Monochlorobenzenes (MCB), 1,2- Dichlorobenzene (o-DCB) and 1,4-Dichlorobenzene (p-DCB) in synthetic medium using mixed culture at bench scale. Because of this new process and condition, the chlorobenzenes which are difficult to degrade can be degraded successfully and people can use this low cost technology/ method to degrade the chlorobenzenes particularly MCB, 1,2-DCB and 1,4-DCB using UASB reactor. 6. Prior art Chlorobenzenes are used mainly as intermediates in the synthesis of pesticides and other chemicals. They are also used as deodorizers, fumigants, degreasers, insecticides and herbicides. The release of chlorobenzenes into the environment occurs primarily during manufacture, and through the dispersive nature of their uses. Chlorobenzenes found in the range of 5-20 |ig/L in ground water by discharging the effluents from large industrial estates in Ankleshwar, Nandesari and Vapi in state of Gujrat, India. They have been found at abandoned waste sites and in leachates. The presence of chlorobenzenes was also observed in low part per billion in stockpile samples collected from Union Carbide plant site in Bhopal, India. An increased level of chlorobenzenes in the water and wastewater is the matter of concern. These compounds are toxic to plants, vertebrates and invertebrates. Many researchers have tired to degrade the similar organic compounds i.e. refractory organics using UASB reactor under different conditions (shown in Table 1). Problem to be solved: Chlorobenznes are recalcitrant substances and difficult to degrade using conventional anaerobic method. Very few authors have specifically used UASB reactor for degradation of chlorinated organic compounds. After an extensive literature review it is observed that microbial degradation of chlorinated benzenes has not been cited using up flow anaerobic sludge reactor (UASB) in ambient condition without controlling ambient temperature. Object: Primary object of this invention is to develop a process with the help of UASB reactor for degradation of chlorinated organic compounds. Further object of this invention is to carry out the process of degradation in ambient condition without controlling ambient temperature. Other objects, features and advantages will become apparent from detail description and appended claims to those skilled in art. STATEMENT:- Following invention provides a process for degradation of chlorinated organic compounds with the help of UASB reactor which is simple to use, cost effective and which can obtain optimum results at low operational and manufacturing cost. The process can also be used for the purpose of degradation of other organic compounds with slight modifications. Further the device used can also be coristructed with variation in size, shape, geometry or rearrangement of components without changing the basic structure and utility. BRIEF DESCRIPTION OF DRAWING The invention is described by way of example with reference to the following drawing Figure 1 shows schematic diagram of experimental set up. Where 1 denotes Synthetic wastewater 2 denotes Peristaltic pump 3 denotes Sampling ports 4 denotes Acrylic column (0.1 m x 0.1 m) 5 denotes Settler GSS 6 denotes Effluent 7 denotes Biogas Detailed description: The reactors are made of transparent acrylic plastic sheet with inner dimensions of about 0.1 m x 0.1 m, and 1.2 m of length which may vary according to the requirement. Working volume of the reactors is 12.5 L for provided experimental set up. The reactor is provided with a hopper bottom. The reactor is provided with the feed inlet pipe of about 2.5 cm diameter to avoid the choking of pipe during operation. Inlet end opens towards the bottom of the reactor so that feed strikes at the bottom and will get evenly distributed and rises in the hopper bottom. An outlet is provided at the top, which is connected through pipe to the effluent tank. The gas-solids separator (GSS) device is of square pyramid with bottom dimension of about 80 mm x 80 mm which may vary according to the requirement. Gas collector is provided at the top by assuming 20% of the reactor volume with 50° inclined walls. Baffles of sufficient overlap is provided below the GSS in order to avoid entry of biogas into the settling compartment. The outlet of the gas is placed at the apex of the pyramid, which has been connected to the gas collection chamber. The reactor is provided with four equidistant ports along its height to facilitate sampling. Best mode of working of the invention: Four identical bench-scale UASB reactors designated as Rl, R2, R3 and R4 (Figure)!), having 12.5 L volume were used for the experimental studies. Reactor Rl represented control (without addition of pollutant) whereas R2, R3 and R4 reactors were fed with monochlorobenzene (MCB), 1, 2-dichlorobenzene (1, 2- DCB) and 1, 4-dichlorobenzene (1, 4-DCB), respectively along with supplemental^ carbon source and nutrients as mentioned in Table 2. All the reactors were maintained at room temperature (27 ± 4°C) throughout the study. During start-up, all the reactors were run on a continuous mode with an influent COD of 1000 mg/L and HRT of 30 h. Reactors were gradually adapted with carbon sources such as methanol, acetone and sodium acetate at desired loadings in equal COD proportion (1:1:1) prior to the addition of the chlorobenzenes. Table 2: Characteristics of the synthetic wastewater Parameters Concentrations (mg/L), except pH Rl (Blank) R2 (MCB) R3 (1,2-DCB) R4 (1,4-DCB) PH 7.9±0.1 7.9±0.1 7.9±0.1 7.9±0.1 Alkalinity as (CaC03) 1020±20 1020±20 1020±20 1020±20 COD 2000±48 2000±48 2000±48 2000±48 NHf-N 44±4.0 44±4.0 44±4.0 44±4.0 P04J"-P 5.0±1.0 5.0±1.0 5.0±1.0 5.0±1.0 MCB 5-100 1,2-DCB 5-100 1,4-DCB 5-100 During acclimation, the organic pollutants like MCB, 1,2-DCB and 1,4-DCB were added along with carbon sources to R2, R3 and R4 reactors respectively as well as nutrient and buffer solution were also added. HRT of 30 h and total flow of 10 L/d was maintained in each reactor. The reactors were started on continuous mode with an influent COD of 1000 mg/L and pollutant concentration of 5 mg/L. On 32nd day the influent COD was increased to 1500 mg/L and thereafter on 93rd day influent COD finally maintained at 2000 mg/L. Influent pollutant concentration (MCB, 1,2-DCB and 1,4-DCB) increased to 10, 20, 30, 35, 40, 45, 50, 55, 60, 65, 70, 80, 90, and 100 mg/L on 46th, 86th, 135th, 169th, 196th, 223rd, 247th, 277th, 304th, 331st, 358th, 385th, 418th and 448,h day respectively. At each increment the reactors were acclimated for 30-46 days to achieve chlorobenzenes removal more that 85%. During acclimation period HRT 30 h and COD 2000 mg/L was maintained which corresponds to sludge loading rate (SLR) of 0.08 kg COD/kg VSS.d and organic loading rate (OLR) of 1.6 kg COD/m3.d. The influent and effluent samples were analyzed for pH, COD, alkalinity, respective chlorobenzene and total biogas production. Bio-degradation of chlorobenznes under different conditions such as hydraulic retention time (HRT) study, substrate co-substrate study, alternative carbon source study, shock loading study, effluent recycle study, and pH study were studied and found out the degradation of each chlorobenzenes compounds using Gas Chromatophy. Influent samples were collected from the influent buckets meant for providing synthetic wastewater to the reactors. Effluent samples were collected from the effluent ports. Sludge samples were collected frorri the sampling ports provided along the length of the reactor and gas samples were collected from gas collection line and analyzed for various parameters. The analytical procedures for all the tests as per Standard Methods (APHA, 1992 & 1998), unless specified otherwise (Table 3). Daily measurements were taken for the rate of gas production, influent and effluent pH, COD, alkalinity. Influent and effluent samples were analyzed for chlorobenzenes on alternate days. Volatile fatty acids (VFAs) and chlorides were analyzed once in a week. The sludge samples were analyzed for suspended solids (SS) and Volatile suspended solids (VSS) on bi-weekly basis. The organic chlorinated compounds MCB, o-DCB (1,2-dichlorobenzene) and p- DCB (1,4 dichlorobenzene) were analysed by injecting 1 |xL sample to Agilent 6390 N (Agilent Technologies, India) model gas chromatograph (GC) equipped with flame ionization detector (FID). A HP-5 capillary column with dimension of 30 m x 0.25 mm x 0.25 nm was used for separation of the compounds. The oven temperature was 280°C to 300°C; injector and detector temperature was 250°C and 300°C respectively. The carrier gas was nitrogen at a flow rate of 30 mL/min. Hydrogen and zero air at the rate of 40 and 400 mL/min, respectively, were used to fuel the flame. The retention times were 3.56, 5.57 and 5.29 minutes for MCB, o-DCB and p-DCB respectively. The retention ti tries for benzene and internal standard 2,4,6-trichlorophenol (TCP) were 2.43 and 8.92 minutes respectively. Total time required for a single GC run was about 17.5 minutes. Volatile fatty acids (VFAs) were measured by injecting 2 ^L of filtered and acidified samples to GC equipped with an FID. The, analysis was done at an oven temperature of 150°C, injector temperature of 180°C and detector temperature of 250°C, using a 10% FFAP on 60/80 Chromosorb WHP/0.1% H3PO4 ss column. The carrier gas nitrogen was applied at a flow rate of 30 mL/min. Hydrogen and zero air was used to fuel the flame. The methane concentration in biogas was determined by GC equipped with thermal conductivity detector (TCD) and fitted with Porapack- Q (80-100), ss column (2.4 m x 3 mm). Oven temperature was maintained at 55°C, injector and detector temperature was kept at 120°C and 150°C respectively. Hydrogen was used as carrier gas at flow rate of 40 mL/min. Certified gas standards were used for the gas analysis. Table 3: The parameters and analytical methods for influent and effluent Influent/Effluent parameters Analytical Methods pH pH meter Temperature Thermometers COD Closed Reflux, Titrimetric Method BOD5 Azide Method SS Suspended Solids Dried at 103-105°C VSS Suspended Solids Dried at 550°C TP Persulfate Digest/Ascorbic Acid TKN Method NO3'-N Kjeldahl Digestion NH4+-N Brucine Method Alkalinity as CaC03 Nesslerization Method Chlorides Titrimetric Method Titrimetric Method I claim:- 1. Method for biodegradation of chlorobenzenes using UASB reactor by utilizing ability of UASB reactor to degrade the chlorinated organic compounds mainly Monochlorobenzenes (MCB), 1,2-Dichlorobenzene (o-DCB) and 1,4- Dichlorobenzene (p-DCB) in synthetic medium using mixed culture at bench scale. 2. The USAB reactor as claimed in claim 1 can be constructed of any shape size geometry or any suitable material without changing the basic structure and utility. 3. The reactors as claimed in claim 2 is made of transparent acrylic plastic sheet but can also be made of any suitable material. 4. The process as claimed in claim 1 can also be used for degradation of other organic compounds along with chlorobenzene.

Documents:

392-MUM-2010-ABSTRACT(18-3-2014).pdf

392-MUM-2010-ABSTRACT(3-3-2014).pdf

392-mum-2010-abstract.doc

392-mum-2010-abstract.pdf

392-MUM-2010-CLAIMS(AMENDED)-(18-3-2014).pdf

392-MUM-2010-CLAIMS(AMENDED)-(3-3-2014).pdf

392-mum-2010-claims.doc

392-mum-2010-claims.pdf

392-MUM-2010-CORRESPONDENCE(13-2-2014).pdf

392-MUM-2010-CORRESPONDENCE(5-3-2014).pdf

392-mum-2010-correspondence.pdf

392-mum-2010-description(complete).pdf

392-mum-2010-drawing.pdf

392-mum-2010-form 1.pdf

392-mum-2010-form 18.pdf

392-MUM-2010-FORM 2(TITLE PAGE)-(18-3-2014).pdf

392-MUM-2010-FORM 2(TITLE PAGE)-(3-3-2014).pdf

392-mum-2010-form 2(title page).pdf

392-mum-2010-form 2.doc

392-mum-2010-form 2.pdf

392-mum-2010-form 26.pdf

392-mum-2010-form 3.pdf

392-mum-2010-form 9.pdf

392-MUM-2010-REPLY TO EXAMINATION REPORT(11-3-2013).pdf

392-MUM-2010-REPLY TO HEARING(18-3-2014).pdf

392-MUM-2010-REPLY TO HEARING(3-3-2014).pdf