International Journal of Chemical and Petrochemical Technology (IJCPT) ISSN 2277-4807 Vol.

2, Issue 3 Dec 2012 9-22 TJPRC Pvt. Ltd.,

STUDY ON THE EFFECTS OF SOIL PH AND ADDITION OF N-P-K FERTILIZER ON DEGRADATION OF PETROLEUM HYDROCARBON PRESENT IN OIL CONTAMINATED SOIL
MUKUT KALITA & ARUNDHUTI DEVI RM&E Division, Institute of Advanced Study in S & T, Guwahati, India

ABSTRACT
Many of the hydrocarbons are resistant to degradation in the natural environment. The overall degradation rate of hydrocarbons biodegradation in soils is strictly limited by a variety of parameters Two of the most important soil factors that affect degradation are soil pH and available nutrients. Soil pH is an important parameter that predominantly affects the biodegradation process. This is because each type of microorganisms has a preferred pH range for optimal growth and activity. The positive effects as well as the negative effects of different N-P-K levels on the biodegradation of hydrocarbons have been reported by different authors.In the present study, role of soil pH and N-P-K fertilizer on the degradation of petroleum hydrocarbons was evaluated. Remediation studies using petroleum hydrocarbon contaminated soil (artificially contaminated with crude oil of Assam) were conducted under different pH values and different N-P-K environments. The set up of the experimental samples along with the test conditions applied to study the effect of pH and N-P-K fertilizer on degradation of petroleum hydrocarbons has been evaluated. The present study shows the effects of pH on degradation of petroleum hydrocarbons in order to determine the optimum soil pH that gives best result for degradation. The effect of different N-P-K levels in soil on the biodegradation of hydrocarbons was also determined in the study.

KEYWORDS: Crude Oil Contaminants, Hydrocarbons, Soil PH, N-P-K Fertilizer, TpH, Biodegradation, Remediation INTRODUCTION
Petroleum or crude oil is a natural product which has very wide range of uses. In the oil producing states like Assam (India), different companies are engaged in exploration, production and transportation of crude oil. A large number of contaminants including hydrocarbons and heavy metals enter into the nearby areas of an oil collecting station (gathering station) through spills, leaks as well as through emissions from gas flaring and from effluents which are likely to pollute the environment. If oil is spilled on the surface of soil during the drilling operation the hydrocarbons gradually penetrate into the soil system. It has been found that the oil hydrocarbons can infiltrate up to a depth of 50-cm. (Ilangovan and Vivekanandan, 1992). It is known that rainfall prior to or during the spills reduces oil infiltration into the soil and washes petroleum components away to runoff waters (Francke and Clark, 1974). The oil concentration in the soil of the contaminated field decreases with time. Initially the oil concentration is high in the upper 1 to 30-cm layer but after six months, the rapid decrease of oil in the upper layer reverses the situation. Biodegradation, evaporation and leaching could be considered as causes for the decrease but leaching has been shown to be not very significant (Raymond et al., 1976; Dibble and Bartha, 1979a and b). Biodegradation is the metabolic activity of microorganisms to transform or mineralize organic contaminants into less harmful, non-hazardous substances, which are then integrated into natural biogeochemical cycles (Margesin and Schinner, 2001). The time required for degradation of petroleum hydrocarbons in soils depends on

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the chemical compositions of crude oil, the climatic conditions and the properties of soil. Unless all the proper conditions are met for a given compound, biodegradation is not likely to occur (Bitton and Gerba, 1985). Oil infiltration into the soil modifies the soil properties and hence may deteriorate the natural environment (Kalita et al., 2007). Considerable effort is being made to remediate soils contaminated with petroleum hydrocarbons, heavy metals and other organic and inorganic compounds that have resulted from industrial activities, accidental spills and improper waste disposal practices (Saldaa et al.,2005). Remediation can lead to quick recovery of the polluted soils Gradi 1985, Alexander 1978). Biodegradation of oil is one of the most important processes for the eventual removal of petroleum from the environment, particularly for the non-volatile components of petroleum (Albert and Xueqing, 2003). Soil conditions are often controlled to increase the rate of contaminant degradation (Odu, 1978, Gradi, 1985). In the biodegradation process, pH of the soil plays a predominant role. The growth and activity of soil microorganisms are very much dependant on the soil pH. For example, fungi predominate under acidic conditions (pH<7) while bacteria and actinomycetes predominate near pH 7.0. A soil pH of 7.8 has been shown to be the optimum pH for microorganisms growth (Dibble and Bartha, 1979a). The soil pH regulates the solubility, mobility, and the availability of the ionized forms of contaminants (JRB Associates, Inc., 1984).] Changes in salinity may affect oil biodegradation through alteration of the microbial population (Ward and Brock, 1978). Microbial activity is significantly controlled by the availability of nutrients, especially nitrogen and phosphorus (Margesin and Schinner, 1997). The additions of nutrients are necessary to enhance the biodegradation of oil pollutants (Choi et al., 2002). The provision of limiting nutrients has been shown to be effective in stimulating hydrocarbon degradation after oil spills. One possible explanation may be that the presence of nutrient stimulated the activity of indigenous bacterial populations sufficiently to enhance the degradation of the crude oil (Pitchard et al., 1992; Swannell et al., 1999). In theory, approximately 150mg of nitrogen and 30mg of phosphorus are utilised in the conversion of 1g of hydrocarbon to cell materials (Rosenberg and Ron, 1996). The results obtained by Coulon et al. (2004). clearly show that fertilizer addition improves the rate of degradation of both n-alkanes and polycyclic aromatic hydrocarbons (PAHs) in sub-antarctic soils contaminated by petroleum hydrocarbons. The use of lipophilic fertilizer to enhance microbial utilization of crude oil has also been investigated by different authors (Olivieri, et al., 1978, Abu and Ogiji, 1996) and it has been found that the efficiency of a fertilizer depends greatly on the environment and design of the treatment protocol (Riser-Roberts, 1998). In a different study Coulon et al. (2005) found significant positive effects of a commercial oleophilic fertilizer containing N and P (and also of temperature) on hydrocarbon-degrading microbial abundance and total petroleum hydrocarbons (TPH) degradation.

Chayneau et al. (2005). have also reported the negative effect of a high N-P-K level on the biodegradation of hydrocarbons. The present study was designed with an aim to investigate the effect of soil pH and addition of N-P-K fertilizer on degradation of hydrocarbons present in crude oil contaminated soil by carrying out detailed laboratory scale study.

MATERIALS AND METHODS

Collection of Soil and Crude Oil Soil for the remediation study was collected from a depth of 0 45 cm from a rice-paddy field situated at an uncontaminated area near the Lakowa oil field (District Sibsagar, Assam, India) run by the Oil and Natural Gas Corporation Limited (ONGCL), India. The soil was air dried and sieved with a 2 mm sieve (2 mm). This soil had an

Study on the Effects of Soil PH and Addition of N-P-K Fertilizer on Degradation of Petroleum Hydrocarbon Present in Oil Contaminated Soil

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original pH of 5.0 (1:5 soil-water suspension) with a total organic carbon (TOC) load of 0.49 %. A sample of crude oil was collected from a group gathering station (Lakowa GGS1) of ONGCL under Lakowa oil field, Assam. Collection of N-P-K Fertilizer A commercial N-P-K mixture (with N, P, and K in the weight ratio of 12:12:12) was used in this study. Important Chemicals For controlling soil pH, either sodium hydroxide (GR MERCK), or hydrochloric acid (minimum 35% GR MERCK) were used. Teepol (Intklean-Teepol Grade, International Chemicals, India) was used as de-emulsifier. The Procedure The samples for the laboratory degradation experiment were prepared in duplicate as follows. Experiments were carried out in 1000 ml glass beakers containing 100 g soil in distilled water at soil-to-water ratio of 1:5 in each beaker. Soil solutions in required number were adjusted to pH of 3.5, 4.5, 5.5, 6.5, 7.5, 8.5 by addition of either 1N HCl or 1N NaOH. Adjustments to desired pH values were made at every alternate day over a period of one month till the pH values of the soils stabilized. Soil solutions at original pH (pH5.0) in required number were also kept prepared without adjusting pH. After pH adjustment, crude oil (with the help of TEEPOL-used as de-emulsifier) was added to soil solutions including normal soil solutions to yield four different concentrations of TPH viz. 0.3%, 1.5%, 3.0%, and 5.0% for each pH value. Here % signifies the amount of oil present, in the unit of gram, per 100g of [soil+oil] system. These initially added TPH concentrations were considered as TPH concentrations at 0 day without any hydrocarbon degradation due to biological, physical or chemical phenomenon. Just after the addition of crude oil, the above factors may come into immediate action and hence the TPH calculation at later part of the initial day may give TPH concentrations different from initially added TPH concentrations. For each adjusted pH, 8 (4 x 2) beakers were used. Hence, total number of beakers used was 8 x 6 plus 8 (for normal soil solutions without adjusting pH), i.e., 56 containing 28 types of samples in duplicate. The experimental samples were set up as shown in Table1 and monitored for a period of six months. Samples were drawn after 30, 60, 90, 120, 150 and 180 days for analysis. Each of the soil samples, after collection, was immediately soaked in dichloromethane to prevent further biodegradation of the hydrocarbons and preserved by refrigeration at 4oC for the calculation of TPH (Janiyani et al. 1992). The experiment on remediation was carried out at room temperature. Similarly, remediation studies using petroleum hydrocarbons contaminated soil (at initial TPH 3.0%) were conducted under pH 7.5, original pH and at different N-P-K environments. Here % signifies the amount of oil present, in the unit of gram, per 100g of [soil+N-P-K+oil] system. The experimental samples were set up as shown in Table2. The petroleum hydrocarbons in the samples (collected as above soaking in dichloromethane) were extracted with dichloromethane by the Soxhlet method, and the concentration of TPH was determined by using the gravimetric estimation as described below. In Soxhlet method, fresh solvent (DCM) is continuously refluxed through the finely divided soil sample (W g) contained in a porous thimble and a siphon system removes the extract back into the refluxing solvent. The process was repeated for many times until the completion of extraction of petroleum hydrocarbons from the sample. The extract containing the petroleum hydrocarbons was transferred in to a weighed glass vial (Wi). After the complete evaporation of the solvent, the final weight (Wf) of the glass vial was recorded. TPH was measured by complete evaporation of the extracting solvent and weighing the residue (Potter, 1999).

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The percentage of TPH content is computed using the formula: % TPH = [(Wf - Wi) / W] 100 Wi = Initial weight of the glass vial (in g) Wf = Final weight of the glass vial (in g) W= Weight of the soil sample (in g) The extent of degradation of crude oil can be determined by estimating the concentration of residual TPH as mentioned above. Also, the total percentage loss in TPH was determined by using the following formula: Total percentage loss in TPH= [{Conc. of initial TPH - Conc. of residual TPH} / Conc. of initial TPH] 100%. Net percentage loss in TPH due to treatment is defined as: Net percentage loss in TPH due to treatment = Total percentage loss in TPH due to treatment - Total percentage loss in TPH without treatment

RESULTS AND DISCUSSIONS

Many of the hydrocarbons are resistant to degradation in the natural environment. The overall degradation rate of hydrocarbons biodegradation in soils is strictly limited by a variety of parameters Rockne et.al 2002. Two of the most important soil factors that affect hydrocarbons degradation are soil pH and available nutrients. The results of the present study reveal considerable effects of soil pH and addition of nutrients (in the form of N-P-K fertilizer) on the hydrocarbons degradation of crude oil contaminated soil. The results obtained from the detailed remediation studies carried out with soil spiked with different amounts of crude oil under different pH values are presented in Table 3, Table 4 and Table 5. The results estimate the loss of TPH from the crude oil contaminated soil samples for each treatment options employed to study the effect of pH on remediation. As already mentioned above, remediation experiment was also carried out using petroleum hydrocarbons contaminated soils (only with initial TPH 3.0%) under pH 7.5, original pH and at different N-P-K environments. The results of the study are presented in Table 6, Table 7 and Table 8. The results determine the loss of petroleum hydrocarbons from the crude oil contaminated soil samples for each treatment options employed to study the effect of fertilizer at optimum p on remediation. Soil pH is an important parameter that predominantly affects the biodegradation process. This is because each type of microorganisms has a preferred pH range for optimal growth and activity.[3] Following important observations can be made from the detailed remediation (enhanced degradation measured with loss of TPH) studies carried out with soil spiked with different amounts of crude oil under different pH values. a) The common order of degradation (DpH, measured as total percentage loss in TPH after six months) for each initial TPH (except for initial TPH 0.3 %) according to pH variation is as followsD7.5>D4.5>DORIGINAL > D5.5>D6.5>D8.5>D3.5 For initial TPH 0.3% the order is slightly differentH

additional N-P-K

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D7.5>D4.5> D5.5>D6.5>D8.5> DORIGINAL >D3.5 In the second case the less degradation at original pH may be due to the non-treatment condition i.e., absence of pH controlling reagents which requires further investigations. b) The degradation was very good at pH 4.5 and showed identical TPH degradation (enhanced degradation) pattern similar to the Control soil which had a pH of 5.0(original pH). As the Control soil is acidic in nature these observations may be attributed to the members of the indigenous microbial community. This may be due to the fact that biological activity in the soil is less affected by small pH variation. c) The soil samples whose pH was adjusted to 5.5, 6.5 showed enhanced degradation for lowest initial TPH (0.3%) concentration and decreased degradation for other higher initial TPH concentrations. This may perhaps be due to the fact that low TPH concentration is lost favorably due to evaporation and remaining part is used in physical processes. At other higher TPH concentrations these processes are not predominant due to high initial TPH. The soil sample whose pH was adjusted to 8.5 showed identical TPH degradation as above. The soil pH between pH 5.5 and 8.5 encourage microbial activity. d) The soil sample whose pH was adjusted to 3.5 showed decreased TPH degradation for all initial TPH concentrations. This may be due to the decreased microbial activities at very low pH (3.5) as compared to original pH (5.0). e) The study conducted at different pHs showed that the highest degradation of petroleum hydrocarbons occurred at pH 7.5. This may be due to the fact that microbial activity is greater at or near neutral pH, which enhances degradation processes, mineralization, and nitrogen transformations (e.g., nitrogen fixation and nitrification). The degradation continued to improve with time and it was observed that TPH continued to degrade more and more up to 180 days. Above observations are graphically represented in the Figure 1-Figure 4. Thus, the pH factor affected total petroleum hydrocarbons (TPH) degradation and remediation of hydrocarboncontaminated soil appears to be feasible. Another most important soil factor that affects degradation is nutrient availability. The nutrient status of soil has direct impact on microbial activity and hence biodegradation of hydrocarbons in soil. The positive effects as well as the negative effects of different N-P-K levels on the biodegradation of hydrocarbons have been reported by different authors.
[25,26]

In the present study, role of N-P-K fertilizer added at optimum pH and original pH on the degradation of petroleum

hydrocarbons was evaluated. There was significant degradation of petroleum hydrocarbons with the addition of N-P-K fertilizer. The common order of degradation according to N-P-K contents (DpH/NPK%, only for initial TPH 3.0%) after six months of experimentation is shown below. The term NPK% means the amount (in the unit of gram) of N-P-K fertilizer added to 100g soil system. D 7.5/90%> D7.5/60%> D7.5/30%>D7.5/0%> DORIGINAL/0% The degradation continued to improve with increase in concentration of additional N-P-K fertilizer. Thus, soil responded most positively to 90% additional N-P-K fertilizer. This is clearly visible from the Figure 5. It is important to mention that the results of the present study may vary from the results obtained by other studies due to the difference in crude oil compositions, climatic conditions, soil characteristics, soil microbial community and many other important factors. Present findings will, ultimately, help to carry out further investigations to prepare a suitable in situ method for the degradation of hydrocarbons in oil field soil.

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CONCLUSIONS
The detailed remediation (degradation measured with net loss of TPH) studies during six months of experimentation show significant degradation of petroleum hydrocarbons at pH 4.5 and 7.5. In general soil responded most positively to pH increase from original pH to pH 7.5. The soil sample whose pH was adjusted to 3.5 showed less TPH degradation at all initial TPH concentrations which may be attributed to the decreased microbial activities at very low pH (pH3.5) as compared to original pH (pH5.0). Also, there was significant degradation of petroleum hydrocarbons with the addition of N-P-K fertilizer. The degradation continued to improve with increase in concentration of N-P-K fertilizer and available results indicate most positive result for 90% additional N-P-K fertilizer. The above observations strongly support that both the factors (that is soil pH and addition of N-P-K fertilizer) in combination enhance the total petroleum hydrocarbons degradation. This will, ultimately, help in preparing a suitable method for remediation of petroleum hydrocarbon contaminated soil.

ACKNOWLEDGEMENTS
Authors are highly thankful to DBT (Govt. of India) for funding the project entitled Assessment of oil field soil (with special reference to polyaromatic hydrocarbons) for their eventual remediation and reclamation. Authors are also thankful to all the members of the institutions, viz., IASST & Gauhati University for providing all the necessary helps to carry out the experiment.

REFERENCES
1. Abu, G. O.& Ogiji, P. A. (1996). Initial test of a bioremediation scheme for the clean up of an oil polluted water body in a rural community in Nigeria, Bioresource Technology, 58:7-12. 2. Albert, D.V. & Xueqing, Z.(2003). Biodegradation of crude oil contaminating marine shorelines and Freshwater Wetlands. Spill Science & Technology Bulletin. 8(2): 163-178. 3. 4. 5. Alexander, M. (1978). An introduction to soil microbiology, 2nd ed., John Wiley & Sons/ New York p223-330. Bitton, G. & Gerba, C. P. (1985). Groundwater Pollution Microbiology. John Wiley & Sons, New York. Chayneau, C.H., Rougeux, G.,Yepremian, C. & Oudot, J.(2005). Effects of nutrient concentration on the biodegradation of crude oil and associated microbial populations in the soil. Soil. Biol. Biochem., 37: 1490-1497. 6. Choi, S. C., Kwon, K. K., Sohn, J.H.& Kim, S. J. (2002). Evaluation of fertilizer additions to stimulate oil biodegradation in sand seashore mescocosms. J. Microbiol. Biotechnol., 12: 431-436. 7. Coulon, F., Pelletier, E., Gourhant, L.& Delille, D. (2005). Effects of nutrient and temperature on degradation of petroleum hydrocarbons in contaminated sub-Antarctic soil. Chemosphere. 58(10):1439-48. 8. Coulon F, Pelletier E, St Louis R, Gourhant L and Delille D. 2004. Degradation of petroleum hydrocarbons in two sub-antarctic soils: influence of an oleophilic fertilizer.Environ Toxicol Chem. 2004 Aug;23(8):1893-901. 9. Dibble, J.T. & Bartha,R.(1979a). Effects of environmental parameters on the biodegradation of oil sludge. Appl. Environ. Microbiol.37, 729-739. 10. Dibble, J. T. & Bartha, R. (1979b). Leaching aspects of oil sludge biodegradation in soil. Soil Science. 127, 365370. 11. Francke, H.C. & Clark, R E. (1974). Disposal of oily wastes by microbial assimilation. Report Y-1934. U.S. Atomic Energy Commission, Washington, D.C.

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12. Gradi,P.C.(1985). Biodegradation. Its Management and Microbiology Basis. Biotechnology and Bio-Engineering 27:660-674. 13. Illangovan, K. & Vivekanandan,M.(1992). Effect of soil pollution on soil respiration and growth of vigna mungo(L). Sci. Total Environ. 116, 187-194. 14. Janiyani, K. L., Wate, S. R., Muthal, P. L. & Joshi, S. R.(1992). Hydrocarbons in oil and oil sludge from refinery. Indian J. Environ. Hlth.34, 169-179. 15. JRB Associates, Inc. (1984). Summary report: Remedial Response at Hazardous Waste Municipal Environmental Research Laboratory, Cincinnati, OH. PB 85-124899. 16. Kalita, M., Das, H., Khanikar, N., Bhattacharyya, K .G. & Devi, A.(2007). Assessment of pollution risks generated by Group Gathering Station: A case study in Lakowa Oil Field(GGS-1) of ONGCL. Enviro-Spectra. Vol.2 (1). pp 52-59. 17. Margesin, R. & Schinner, F.(2001). Biodegradation and bioremediation of hydrocarbons in extreme environments. Appl Microbiol Biotechnol 56:650-663. 18. Margesin, R. & Schinner, F.(1997). Bioremediation of diesel-oil-contaminated alpine soils at low temperatures. Appl Microbiol Biotechnol 147:462-468. 19. Odu, C. T .I. (1978). The effects of nutrients application and aeration on oil degradation in soil. Environmental Pollution. 15:235-240. 20. Olivieri, R., Robertiello, A. & Degen, L. (1978). Enhancement of microbial degradation of oil pollutants using lipophilic fertilizers.Mar.Pollut. Bull. 9:217-220. 21. Pitchard, P. H., Mueller, J.G., Rogers, J. C., Kremer, F. V. & Glaser, J .A. (1992). Oil spill bioremediation: exoeriences, lessons and results from the Exxon Valdez oil spill in Alaska. Biodegradation. 3:315-335. 22. Potter, T. L. (1999). Assessment and Remediation of Oil Contaminated Soils. New Age International (P) Ltd., Publishers, New Delhi. pp 13-40. 23. Raymond, R. L., Hudson, J. O. & Jamison, V. W. (1976). Oil degradation in soil. Appl. Environ. Microbiol., 31: 522-535. 24. Riser-Roberts Eve. (1998). Remediation of Petroleum Contaminated Soils Biological, Physical, and Chemical Processes. Lewis Publishers, Boca Raton Boston London New York Washington, DC. 25. Rockne, K. J., Shor, L. M., Young, L. Y., Taghon, G. L. & Kosson, D.S.(2002). Distributed sequestration and release of PAHs in weathered sediment. The role of sediment structure and organic carbon properties. Environ. Sci. Technol., 36: 2636-2644. 26. Rosenberg, E. & Ron, E. Z. (1996). Bioremediation of petroleum contamination. Cambridge University Press, UK. pp100-124. 27. Saldaa, M.D., Nagpal, V. & Guigard, S. E. (2005). Remediation of contaminated soils using supercritical fluid extraction: a review (1994-2004). Environ Technol. 26(9):1013-32. 28. Swannell, R.P.J., Mitchell, D., Lethbridge, G., Jones, D., Heath, D., Hagley, M., Jones, M., Petch, S., Milne, R., Croxford, R.& Lee, K. (1999). A field demonstration of the efficacy of bioremediation to treat an oiled shoreline following the Sea Empress incident. Environ. Technol. 20:863-873. 29. Ward,D.M.& Brock T.D.(1978).Hydrocarbon biodegradation in hypersaline environments. Appl. Environ. Microbiol. 35(2):353-359. Sites.Prepared for

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70 TOTAL TPH(%) LOSS A 60 50 40 30 20 10 0 pH3.5 pH4.5 B

INITIAL TPH 0.3% C D E

pH5.5

pH6.5 pH

pH7.5

pH8.5 Original pH

Figure 1: Total percentage loss in TPH for the samples with initial TPH 0.3% [A=0 day, B=30 days, C=60 days, D=90days, E= 120 days, F= 150 days, G=180 days]

Figure 2: Total Percentage Loss in TPH for the Samples with Initial TPH 1.5% [A= 0 Day, B=30 Days, C=60 Days, D=90days, E= 120 Days, F= 150 Days, G= 180 Days] 70 TOTAL TPH(%) LOSS 60 50 40 30 20 10 0 pH3.5 pH4.5 pH5.5 pH6.5 pH pH7.5 pH8.5 Original pH A B INITIAL TPH 3.0% C D

Figure 3: Total Percentage Loss In TPH for the Samples with Initial TPH 3.0% [A= 0 Day, B=30 Days, C=60 Days, D=90days, E= 120 Days, F= 150 Days, G= 180 Days]

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70 TOTAL TPH(%) LOSS 60 50 40 30 20 10 0 pH3.5 pH4.5 A B

INITIAL TPH 5.5% C D

pH5.5 pH

pH6.5

pH7.5

pH8.5 Original pH

Figure 4: Total percentage loss in TPH for the samples with initial TPH 5.5% [A= 0 day, B=30 days, C=60 days, D=90days, E= 120 days, F= 150 days, G= 180 days] N-P-K EFFECT 3

TOTAL TPH(%) LOSS

80 60 40 20 0 A

D E SAMPLING DAYS

Figure 5: Total Percentage Loss in TPH for the Various Samples treated to Study the Effect of N-P-K Fertilizer. the Sample Nos. 1,2,3,4 and 5 Correspond to Samples with Code Nos. PFX1, PFX2, PFX3, PX18 And C02 Respectively [A= 0 Day, B=30 Days, C=60 Days, D=90days, E= 120 Days, F= 150 Days, G= 180 Days] Table 1: A Summary of Sample Treatment and Test Conditions Employed to Study the Effect of Ph on Remediation S.N. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Sample Code PX1 PX2 PX3 PX4 PX5 PX6 PX7 PX8 PX9 PX10 PX11 PX12 PX13 PX14 PX15 PX16 Sample Treatment 100 g 100 g 100 g 100 g 100 g 100 g 100 g 100 g 100 g 100 g 100 g 100 g 100 g 100 g 100 g 100 g soil in solution (pH controlled at 3.5) treated to yield 5.5% TPH soil in solution (pH controlled at 3.5) treated to yield 3.0% TPH soil in solution (pH controlled at 3.5) treated to yield 1.5% TPH soil in solution (pH controlled at 3.5) treated to yield 0.3% TPH soil in solution (pH controlled at 4.5) treated to yield 5.5% TPH soil in solution (pH controlled at 4.5) treated to yield 3.0% TPH soil in solution (pH controlled at 4.5) treated to yield 1.5% TPH soil in solution (pH controlled at 4.5) treated to yield 0.3% TPH soil in solution (pH controlled at 5.5) treated to yield 5.5% TPH soil in solution (pH controlled at 5.5) treated to yield 3.0% TPH soil in solution (pH controlled at 5.5) treated to yield 1.5% TPH soil in solution (pH controlled at 5.5) treated to yield 0.3% TPH soil in solution (pH controlled at 6.5) treated to yield 5.5% TPH soil in solution (pH controlled at 6.5) treated to yield 3.0% TPH soil in solution (pH controlled at 6.5) treated to yield 1.5% TPH soil in solution (pH controlled at 6.5) treated to yield 0.3% TPH Test Condition pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled

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S.N. 17 18 19 20 21 22 23 24 25 26 27 28

Sample Code PX17 PX18 PX19 PX20 PX21 PX22 PX23 PX24 C01 C02 C03 C04

Sample Treatment 100 g 100 g 100 g 100 g 100 g 100 g 100 g 100 g 100 g 100 g 100 g soil in solution (pH controlled at 7.5) treated to yield 5.5% TPH soil in solution (pH controlled at 7.5) treated to yield 3.0% TPH soil in solution (pH controlled at 7.5) treated to yield 1.5% TPH soil in solution (pH controlled at 7.5) treated to yield 0.3% TPH soil in solution (pH controlled at 8.5) treated to yield 5.5% TPH soil in solution (pH controlled at 8.5) treated to yield 3.0% TPH soil in solution (pH controlled at 8.5) treated to yield 1.5% TPH soil in solution (pH controlled at 8.5) treated to yield 0.3% TPH soil in solution (Original pH) treated to yield 5.5% TPH soil in solution (Original pH) treated to yield 3.0% TPH soil in solution (Original pH) treated to yield 1.5% TPH

Test Condition pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled pH Controlled Original pH Original pH Original pH Original pH

100 g soil in solution (Original pH) treated to yield 0.3% TPH

Table 2: A Summary of Sample Treatment and test Conditions Employed in the Effect of N-P-K Fertilizer on Remediation S.N. Sample Sample Treatment Test Condition Code 1 PFX1 100g soil in solution(pH controlled at 7.5) + 30g N-P- N-P-K fertilizer added K fertilizer and treated to yield 3.0% TPH 2 PFX 2 100g soil in solution(pH controlled at 7.5) + 60g N-P- N-P-K fertilizer added K fertilizer and treated to yield 3.0% TPH 3 PFX 3 100g soil in solution(pH controlled at 7.5) + 90g N-P- N-P-K fertilizer added K fertilizer and treated to yield 3.0% TPH 4 PX18 100g soil in solution(pH controlled at 7.5) and treated No N-P-K fertilizer to yield 3.0% TPH added 5 C02 100g soil in solution (Original pH) and treated to yield No N-P-K fertilizer 3.0% TPH added Table 3: Conc. of Residual TPH (%) [A= 0 Day, B=30 Days, C=60 Days, D=90days, E= 120 Days, F= 150 Days, G= 180 Days] S.N. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 Sample Code P X1 P X2 P X3 P X4 P X5 P X6 P X7 P X8 P X9 PX10 PX11 PX12 PX13 PX14 PX15 PX16 PX17 PX18 PX19 PX20 PX21 PX22 A 5.50 3.00 1.50 0.30 5.50 3.00 1.50 0.30 5.50 3.0 1.50 0.30 5.50 3.00 1.50 0.30 5.50 3.00 1.50 0.30 5.50 3.00 B 5.43 2.99 1.44 0.29 4.73 2.97 1.42 0.26 5.22 2.71 1.31 0.25 5.44 2.68 1.47 0.26 4.69 2.87 0.99 0.26 5.38 2.92 C 5.37 2.98 1.42 0.28 4.28 2.76 1.37 0.24 4.99 2.39 1.27 0.23 5.24 2.56 1.42 0.22 4.16 2.86 0.98 0.25 5.32 2.90 D 5.23 2.90 1.41 0.27 3.94 2.71 1.32 0.21 4.55 2.38 1.21 0.22 5.15 2.54 1.41 0.21 3.82 1.71 0.96 0.21 5.28 2.83 E 5.20 2.86 1.30 0.25 3.27 2.65 1.26 0.19 4.54 2.23 1.12 0.19 4.98 2.35 1.38 0.19 3.00 1.45 0.82 0.15 5.19 2.66 F 5.18 2.83 1.28 0.24 2.91 2.05 1.06 0.17 4.50 2.17 1.11 0.18 4.71 2.34 1.23 0.18 2.76 1.40 0.81 0.13 5.03 2.56 G 5.16 2.80 1.26 0.24 2.63 1.47 0.90 0.16 4.47 2.13 1.11 0.17 4.48 2.34 1.14 0.18 2.54 1.34 0.80 0.12 4.89 2.48

Study on the Effects of Soil PH and Addition of N-P-K Fertilizer on Degradation of Petroleum Hydrocarbon Present in Oil Contaminated Soil

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S.N. 23 24 25 26 27 28

Sample Code PX23 PX24 C01 C02 C03 C04

A 1.50 0.30 5.50 3.00 1.50 0.30

B 1.39 0.26 5.40 2.88 0.99 0.29

C 1.35 0.24 4.91 2.80 0.98 0.28

D 1.33 0.22 4.65 2.71 0.94 0.26

E 1.26 0.20 4.43 2.64 0.94 0.24

F 1.25 0.19 4.28 2.23 0.93 0.23

G 1.24 0.19 4.16 1.75 0.93 0.23

Table 4: Total Percentage Loss in TPH (%) [A= 0 day, B=30 days, C=60 days, D=90days, E= 120 days, F= 150 days, G= 180 days] S.N. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 Sample Code PX1 PX2 PX3 PX4 PX5 PX6 PX7 PX8 PX9 PX10 PX11 PX12 PX13 PX14 PX15 PX16 PX17 PX18 PX19 PX20 PX21 PX22 PX23 PX24 C01 C02 C03 C04 A * * * * * * * * * * * * * * * * * * * * * * * * * * * * B 1.27 0.33 0.40 3.33 14.00 1.00 5.33 13.33 5.09 9.67 12.67 16.67 1.09 10.67 2.00 13.33 14.73 4.33 34.00 13.33 2.18 2.67 7.33 13.33 1.82 4.00 34.00 3.33 C 2.36 0.67 5.33 6.67 22.18 8.00 8.67 20.00 9.27 20.33 15.33 23.33 4.73 14.67 5.33 26.67 24.36 4.67 34.67 16.67 3.27 3.33 10.00 20.00 10.73 6.67 34.67 6.67 D 4.91 3.3 6.00 10.00 28.36 9.67 12.00 30.00 17.27 20.67 19.33 26.67 6.36 15.33 6.00 30.00 30.55 43.00 36.00 30.00 4.00 5.67 11.33 26.67 15.45 9.67 36.67 13.33 E 5.45 4.67 13.33 16.67 40.55 11.67 16.00 36.67 17.45 25.67 25.33 36.67 9.45 21.67 8.00 36.67 45.45 51.67 45.33 50.00 5.64 11.33 16.00 33.33 19.45 12.00 37.33 20.00 F 5.82 5.67 14.67 20.00 47.09 31.67 29.33 43.33 18.18 27.67 26.00 40.00 14.36 22.00 18.00 40.00 49.82 53.33 46.00 56.67 8.55 14.67 16.67 36.67 22.18 25.67 38.00 23.33 G 6.18 6.67 16.00 20.00 52.18 51.00 40.00 46.67 18.73 29.00 26.00 43.33 18.55 22.00 24.00 40.00 53.82 55.33 46.67 60.00 11.09 17.33 17.33 36.67 24.36 41.67 38.00 23.33

*Not Applicable

20

Mukut Kalita and Arundhuti Devi

Table 5: Net Percentage Loss in TPH (%) Due to Treatment [A= 0 Day, B=30 Days, C=60 Days, D=90days, E= 120 Days, F= 150 Days, G= 180 Days] S.N. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 Sample Code PX1 PX2 PX3 PX4 PX5 PX6 PX7 PX8 PX9 PX10 PX11 PX12 PX13 PX14 PX15 PX16 PX17 PX18 PX19 PX20 PX21 PX22 PX23 PX24 C01 C02 C03 C04 A * * * * * * * * * * * * * * * * * * * * * * * * * * * * B -0.55 -3.67 -33 0 12.18 -3 -28 10 3.27 5.67 -21 13 -0.73 6.67 -32 10 12.91 0.33 0 10 0.36 -1.33 -26 10 * * * * C -8.37 -6 -29 0 11.45 1.33 -26 13.33 -1.46 13.66 -19 16 -6 8 -29 20 13.63 -2 0 10 -7.46 -3.34 -24 13.33 * * * * D -10 -6.37 -30 -3.33 12.91 0 -24 16.67 1.82 11 -17 13 -9.09 5.66 -30 16.67 15.1 33.33 -0.67 16.67 11.45 -4.00 -25 13.34 * * * * E -14 -7.33 -24 -3.33 21.1 -0.33 -21 16.67 -2 13.67 -12 16 -10 9.67 -29 16.67 26 39.67 8 30 -13.81 -0.67 -21 13.33 * * * * F -16.36 -20.00 -23.33 -3.33 24.91 6.00 -8.67 20.00 -4.00 2.00 -12 16.67 -7.82 -3.67 -20.00 16.67 27.64 27.66 8.00 33.34 -13.63 -11.00 -21.33 13.34 * * * * G -18 -35 -22 -3.33 27.82 9.33 2 23.34 -5.63 -12 -12 20 -5.81 -19 -14 16.67 29.46 13.66 8.67 36.67 -13.27 -24.34 -20 13.34 * * * *

*Not Applicable

Study on the Effects of Soil PH and Addition of N-P-K Fertilizer on Degradation of Petroleum Hydrocarbon Present in Oil Contaminated Soil

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Table 6: Conc. of Residual TPH (%) [A= 0 Day, B=30 Days, C=60 Days, D=90days, E= 120 Days, F= 150 Days, G= 180 Days] S.N. 1 2 3 4 5 Sample Code PFX1 PFX 2 PFX 3 PX18 C02 A 3.00 3.00 3.00 3.00 3.00 B 1.65 2.28 2.02 2.87 2.88 C 1.59 1.29 1.53 2.86 2.80 D 1.20 0.98 0.98 1.71 2.71 E 1.14 0.95 0.94 1.45 2.64 F 1.03 0.92 0.85 1.40 2.23 G 0.94 0.81 0.73 1.34 1.75

Table 7: Total Percentage Loss in TPH (%) [A= 0 Day, B=30 Days, C=60 Days, D=90days, E= 120 Days, F= 150 Days, G= 180 Days] S.N 1 2 3 4 5 *Not Applicable Table 8: Net Percentage Loss In TPH (%) Due To Treatment [A= 0 Day, B=30 Days, C=60 Days, D=90days, E= 120 Days, F= 150 Days, G= 180 Days] S.N 1 2 3 4 5 *Not Applicable Sample Code PFX1 PFX 2 PFX 3 PX18 C02 A * * * * * B 41.00 20.00 28.70 0.33 * C 40.33 50.33 42.33 -2 * D 50.33 57.66 57.66 33.33 * E 50.00 56.33 56.67 39.67 * F 40.00 43.66 46.00 27.66 * G 27.00 31.33 34.00 13.66 * Sample Code PFX1 PFX 2 PFX 3 PX18 C02 A * * * * * B 45.00 24.00 32.67 4.33 4.00 C 47.00 57.00 49.00 4.67 6.67 D 60.00 67.33 67.33 43.00 9.67 E 62.00 68.33 68.67 51.67 12.00 F 65.67 69.33 71.67 53.33 25.67 G 68.67 73.00 75.67 55.33 41.67