A. Ratna Kumari et al.

, IJSID 2011, 1 (3), 303-319

ISSN:2249-5347

IJSID
International Journal of Science Innovations and Discoveries
Research Article
An International peer Review Journal for Science

Available online through www.ijsidonline.info

OPTIMIZATION OF LEAD ADSORPTION USING ANIMAL BIOPOLYMERS BY FACTORIAL DESIGN

A. Ratna Kumari1*, U.Kiran Babu2, K. Sobha3
1Department

of Biotechnology, Bapatla Engineering College (autonomous), Bapatla-522 101, AP, India & Centre for Biotechnology, Acharya Nagarjuna University, Nagarjuna Nagar, Guntur-522 510, A.P., India, 2Department of Biotechnology, Bapatla Engineering College, Bapatla-522 101, AP, India, 3Department of Biotechnology, RVR & JC College of Engineering, Chowdavaram, Guntur 522 019, AP, India.

Received: 14.09.2011 Modified: 16.10.2011 Published: 29.12.2011 Key words: Adsorption, Animal biopolymers, Factorial design, lead.
*Corresponding Author

ABSTRACT Heavy metals present in the industrial effluents remain as alarming pollutants due to their nondestructive nature, toxicity, bioaccumulation and subsequent biomagnification. Animal biopolymers viz., chick and duck feathers fibers were treated with 5% tannic acid solution. From the preliminary studies it was found that pH, contact time, and biosorbent dose are significant factors for maximal biosorption of lead by both chick and duck feathers. To study the interactive effects of these three independent parameters (Biosorbent dosage, Contact time, pH) and their optimization for lead biosorption process by response surface methodology (RSM), Box-Behnken design (BBD) was applied. The regression equation coefficients (r2) are 0. 98098 & 0.91785 for chick and duck feathers respectively and the data fitted to a second-order polynomial equation for

Address: Name: A. Ratna Kumari Place: Guntur, AP, India E-mail: ratnamtech@gmail.com

removal of lead (Pb). The critical values of the three parameters obtained for chick feathers, with a maximum biosorption efficiency of 70.73%, are 1.9 g/L of biosorbent, 24.56 hours of contact time, 6.94 pH. For duck feathers with a maximum biosorption efficiency of 58.8%, the critical values obtained are 2.3 g/L of biosorbent, 20.17 hours of contact time and 7.02 pH.

INTRODUCTION

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INTRODUCTION Water bodies are being overwhelmed with bacteria, waste matter and toxic substances
(1).

Among toxic

substances reaching hazardous levels are heavy metals (2). Contamination of water supplies with heavy metals is an area of concern both nationally & internationally where the challenge to remediate hazardous metal containing waste streams from present or past mining operations, industrial sites and ground waters is immense
(3).

Heavy

metals of concern include lead, chromium, mercury, uranium, selenium, zinc, arsenic, cadmium, silver, gold and nickel
(4).

Heavy metal pollution in the aquatic system has become a serious threat today as they are non-

biodegradable and thus persistent (5). Metals are mobilized and carried into food web as a result of leaching from waste dumps, polluted soils and water. The metals increase in concentration at every level of food chain and are passed onto the next higher levela phenomenon called bio-magnification (6) and cause several diseases and health disorders in humans, and other living organisms (7). Thus, removal of heavy metals from industrial wastewater is of prime importance (8). Some of the conventional techniques for removal of metals from industrial wastewater include adsorption
(9),

sedimentation

(10),

electrochemical processes
(10)

(11),

ion exchange

(12),

biological operations
(14)

(13),

cementation

(14)

coagulation / flocculation extraction

(15- 16)

, filtration and membrane processes

, chemical precipitation and solvent

. However, most available methods may show economical and technical disadvantages such as

high capital and operational costs, high sensitivity to operational conditions, significant energy consumption, or sludge generation
(17)

& they also are ineffective when metals are present in high concentrations in aqueous

solution (18). With increasing environmental awareness and stringent government policies, it has become necessary to develop new environmental friendly ways to clean up contaminants using low-cost methods and materials (19). In this aspect, the relatively new technology termed biosorption has dominated. The major advantages of biosorption over conventional treatment methods include low cost, the use of inexpensive and never exhausted biosorbent materials, high efficiency of metal removal from dilute solutions, minimization of chemical and/or biological sludge, no additional nutrient requirement, regeneration of biosorbent and the possibility of metal recovery (20). The biosorption process involves a solid phase (sorbent or biosorbent; usually a biological material) and a liquid phase (solvent, normally water) containing a dissolved species to be sorbed (sorbate, a metal ion) (1). An adsorbent material (biosorbent), both living and nonliving, derived from suitable biomass can be used for the effective removal and recovery of heavy metal ions from wastewater streams (21-22). Recently, the use of non-living biomaterials as metal-binding compounds has been gaining advantage as these compounds require minimum care and maintenance and can be obtained more cheaply (23) . Biomaterials of animal origin, generated as waste such as animal bones (24-25), chick feathers (26) and duck feathers (27) have been used for removal of heavy metals. In particular, lead has been classified as a serious hazardous heavy metal with high priority in the context of environmental risk
(17).

This metal is extremely toxic and can damage kidney, liver, brain and reproductive

organs besides other adverse effects to humans (7) . At present, lead pollution is considered a worldwide problem
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because this metal is commonly detected in several industrial wastewaters (28). Examples of these wastewaters are those produced by processes such as mining, smelting, printing, metal plating, explosive manufacture, and dying. Electroplating mainly discharges huge amounts of lead and its ingestion beyond the permissible level causes various types of acute disorders in man such as anemia, alimentary symptoms, wrist and foot drop, renal damage, embryo toxicity resulting in spontaneous abortions and sometimes encephalopathy. In children it causes behavioral effects and intellectual impairment. In this context, local legislations have established rigorous standards for lead concentrations in industrial effluents. Therefore, special attention has been given to develop proper methods for lead removal from water (29). During the last decade, several studies have shown that different synthetic and natural sorbents can be used to remove lead ions from aqueous effluents
(30-48).

Many examples of natural sorbents are available in the

literature, and they include brewery biomass, cactus pulp, olive stone waste, chitosan, modified wool, cotton, nutshells, rice hulls, pine bark, sawdust, sugar cane bagasse, fruit stones, and pyrolyzed coffee, among others (41-48). In removal processes, most of these sorbents generally show Pb uptakes in the range of 1.0 - 100 mg/g. In particular, chicken feathers and duck feathers are among the natural sorbents that can be used for water treatment
(27, 49-54).

The feathers represent four to six percent of the total body weight and, as a consequence, are a waste

product generated in large quantities from commercial poultry industry. As a natural protein material, feather fiber has polar and ionizable groups on the side chain of constant amino acid residues, which are able to bind charged species. The adsorption of metal cations to feather fibers can be attributed to many characteristics such as low solubility, complex physical form, relatively high content of reactive groups that can serve as binding sites or that can be chemically modified, variety and juxtaposition of reactive sites that can allow cooperative reaction etc. To date, the use of chicken feathers for sorption purposes has achieved satisfactory results for the removal of some heavy metals, colorants, and organic toxic compounds heavy metals (27). Biosorption efficiency depends upon many factors, the critical ones being pH, contact time, and biosorbent concentration. This work is, therefore, primarily aimed at evaluating the effects of pH, contact time, and biosorbent dose on the percentage removal of lead by both Chick and Duck feathers. To study the cumulative/interactive effect and optimization of lead biosorption process, a Factorial design was applied, varying the three independent parameters (initial pH, Contact time, Biosorbent dosage). Response surface methodology was applied to the Box and Behnken experimental design(55). To the best of our knowledge, there is no published report on optimization of biosorption process for removal of lead with animal biopolymers (Chick and Duck feather) using response surface methodology. As the BoxBehnken design minimizes the number of factor combinations and maintains good precision of the predicted response (56-58), this matrix has been used for the optimization of biosorption process for removal of lead using animal biopolymers (Chick and Duck feathers).
(39, 49-54)

& duck feathers are also used for the removal of

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MATERIALS AND METHODS Biosorbent preparation Chick and Duck feathers were collected from poultry processing facilities. The feather fibers used were in the form of barbs which were detached from the shaft of Chicken feathers and Duck feathers. These were washed several times with deionised water. Feather fibers were immersed in 5% (w/v) aqueous solution of the Tannic acid (material-to-liquor ratio 1:100) at 70C for 113 hr. Tannic acid (TA) is a kind of plant polyphenol. It is reported that TA can form insoluble products with keratin by tanning reaction, which can increase the chemical and physical stability of protein. Moreover, TA can form chelates with many metal cations via the ortho dihydroxy (catehol) or trihydroxy-benzene (galloyl) group (59). Feather fibers were removed after the incubation time, washed thoroughly in a porcelain funnel with distilled water, and then dried at room temperature before metal adsorption experiments. Preparation of metal solution All the reagents were Analytical Reagent Grade and were prepared in Double Distilled water. Dissolved 1.5980g of lead nitrate (Pb (NO3)2 ) in 100ml of Double Distilled water, diluted to 1 liter in a volumetric flask with Double Distilled water. This was used as the source of Pb in the synthetic waste water. pH of the solution was adjusted using 0.1N HCl or NaOH. Solutions of varying concentrations were prepared by diluting the stock solution with Double Distilled water. Fresh dilutions were used for each adsorption study. Lead (Pb) ion determination The change in Pb concentration due to adsorption was determined using AAS (Atomic absorption spectrophotometer) in flame at a wavelength of 283nm. Effect of biosorbent concentration on Adsorption The adsorption of lead by Chick and duck feather fibers was studied at increasing concentration of biosorbent 0.05g, 0.10g, 0.15g, 0.20g, 0.25g, 0.30g respectively in 100ml of lead solution in 250 ml Erlenmeyer flask at constant incubation time and pH 7. Final values were projected in g/L. The biosorbent was removed from the solution by centrifugation and the supernatant was analyzed for the residual concentrations of lead ion using Atomic Absorption Spectrophotometer. Each adsorption experiment was carried out twice, and the average was used for adsorption study. Effect of Contact Time on Adsorption Optimum biosorbent concentration obtained for chick and duck feather were taken to monitor the effect of time on adsorption. The adsorption experiments were carried out at different contact times viz., 5, 10, 15, 20, 25, 35, 45, 55, 65min with a fixed adsorbent dose at pH 7. The biosorbent was removed from the solution by centrifugation and the supernatant was analyzed for the residual concentrations of lead ion using Atomic Absorption Spectrophotometer.

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Effect of pH on Adsorption Optimum Biosorbent concentration and optimum contact time were used to monitor the pH effect on adsorption. The adsorption experiments were carried out for different pH values 4-9 with a fixed adsorbent dose concentration at optimum contact time. The biosorbent was removed from the solution by centrifugation and the supernatant was analyzed for the residual concentrations of lead ion using Atomic Absorption Spectrophotometer. Factorial Design The BoxBehnken design (BBD) model, which is the standard RSM, was established using STATISTICA 6.0 for the optimization of biosorption process. The experimental design, three independent variables, i.e. pH (6.0-8.0), time (15-35hrs) and biosorbent concentration (0.1-0.3g/100ml) were taken to effect biosorption of lead ions. The experimental design was applied after selection range of each variable (maximum and minimum) as shown in Table 1. The BoxBehnken design contained a total of 27 experiments. All the biosorption experiments were conducted in 250mL Erlenmeyer flasks and then the filtrate was analyzed for residual lead concentration using Atomic absorption spectrophotometer. Statistical analysis The quadratic equation model for predicting the optimal point was expressed according to the following equation: Y= 0 + i Xi + ii Xi2 + ij Xi Xj where Y is the predicted response, 0 model constant; i is linear coefficient, ii is the quadratic coefficient and ij is the different interaction coefficients of the model; In this study, the removal of lead was processed using the following equation Y= 0 + i Xi + ij Xi2 + ij Xi Xj = A0 + A1x1+ A2x2 + A3x3 + A4x1x2+ A5x1x3 + A6x2x3 + A7x1x1 + A8x2x2 + A9x3x3 in which Y is the response variable, percentage removal of lead and X1, X2 and X3 are the coded values of the independent variables- biosorbent concentration, time, pH respectively. STATISTICA 6.0, was used for regression analysis of the data obtained and to estimate the coefficient of the regression equation. The quality-of-fit of polynomial model was expressed by the coefficient of determination r2 and statistical significance was checked. To visualize the relationship between responses and experimental levels for each of the factors, the fitted polynomial equation was expressed as surface plots. Three dimensional plots demonstrate relationships between the lead ion uptake with the paired factors (when other factor was kept at its optimal level), describing the behavior of biosorption system in a batch process.

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A. Ratna Kumari et al., IJSID 2011, 1 (3), 303-319 Table 1: BoxBehnken design for the optimization of Biosorption of Lead (Pb) using Chick Feather
BoxBehnken design Run No 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 Biosorbent Concentration (g/100ml) 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 Time (hrs) 15 15 15 25 25 25 35 35 35 15 15 15 25 25 25 35 35 35 15 15 15 25 25 25 35 35 35 pH Chick feathers % Removal Efficiency (Observed) 41.46 43.90 46.34 48.78 53.65 53.69 56.09 57.33 58.29 60.48 63.41 65.85 68.29 70.73 68.29 65.85 63.41 60.97 56.09 53.65 48.78 47.07 46.34 41.51 36.40 32.01 30.11 % Removal Efficiency (Predicted) 39.2797 43.3036 45.7142 50.6497 53.6811 55.0992 55.7664 57.8053 58.2308 64.4042 65.8322 65.6469 67.4400 67.8756 66.6978 64.2225 63.6656 61.4953 53.6319 52.4642 49.6831 48.3336 46.1733 42.3997 36.7819 33.6292 28.8631 Duck feathers % Removal Efficiency (Observed) 35.7 37.8 38.1 39.0 41.4 42.7 43.8 45.8 46.9 47.7 48.6 50.1 51.5 52.2 54.1 56.5 58.9 54.4 52.8 51.1 50.9 49.0 48.1 47.0 45.3 43.5 41.1 % Removal Efficiency (Predicted) 33.4519 36.0796 37.7963 39.4657 41.5852 42.7935 45.3685 46.9796 47.6796 50.4657 51.6519 51.9269 52.3296 53.0074 52.7741 54.0824 54.2519 53.5102 51.0352 50.7796 49.6130 48.7491 47.9852 46.3102 46.3519 45.0796 42.8963

6 7 8 6 7 8 6 7 8 6 7 8 6 7 8 6 7 8 6 7 8 6 7 8 6 7 8

RESULTS & DISCUSSION On treatment with tannic acid the chick & duck feather protein chemical and physical stability was increased. 1ppm, 2ppm, 3ppm, 4ppm, and 5ppm lead solutions were prepared using stock solution. Absorbance values were taken using atomic absorption spectrophotometer. Standard graph was plotted by taking known concentration on X-axis and absorbance at 283 nm on Y-axis which is shown in Fig-1 and it obeyed Beer-lamberts law. Effect of biosorbent concentration Lead biosorption by chick & duck feathers were studied at various biosorbent concentrations ranging from 0.05g to 0.3 g in 100ml of 5ppm lead solution. The percent removal of lead increased with an increase in biosorbent concentration because of an increasing adsorption surface area. The maximum biosorption efficiency was obtained at 0.2 g of chick feather and 0.15g of duck feather, but further increase in biosorbent concentration

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decreased the maximum removal of metal ions because of saturation of biosorbent surfaces as shown in Fig- 2. The percentages of removal of lead by chick and duck feathers were 41.6% and 37.5% respectively.

Fig- 1 Standard graph of lead

45 40 35 30 25 20 15 10 5 0 0.05 0.1 0.15 0.2 0.25 0.3 % removal by D.F.

% removal

% removal by C.F.

biosorbent concentration(g/100ml)

Fig - 2 Effect of biosorbent concentration (give units correctly) on biosorption using chick feather (C.F.) and duck feather (D.F.) Effect of contact time The contact time was evaluated as one of the important parameters affecting the biosorption efficiency. The adsorption experiments were carried out for different contact times with a fixed adsorbent dose concentration at pH 7. Fig - 3 shows the biosorption efficiency of lead ions by chick and duck feathers as a function of contact time. The lead uptake was found to increase with increase in contact time up to 25hrs for chick feathers & 35hrs for duck feathers and after that, lead uptake slowly decreased. The fast initial metal biosorption rate was attributed to the surface binding and the following slower sorption was attributed to the interior penetration
(60).

Different

kinds of functional groups, with different affinities to the metal ions, are usually present on the surface of feathers.
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The active binding groups with higher affinities are firstly occupied chick and duck feathers were 69.3% and 55.83% respectively. Effect of pH Biosorption of heavy metal ions is dependent on the pH of solution as it affects biosorbent surface charge, degree of ionization etc. The pH of the solution influences both metal binding sites on the feather surface and the chemistry of metal in solution. In order to demonstrate the effect of pH on biosorption capacity, uptake of lead ions onto chick and duck feathers as a function of pH was studied in the pH ranges of 4 to 8 with a fixed adsorbent dose concentration at optimum contact time. The percentages of removal of lead by chick and duck feathers were76.66% and 58.95% respectively at pH7 as shown in Fig-4.
80 70 60
% removal
(61).

The percentages of removal of lead by

50 40 30 20 10 0 5 10 15 20 25 Time(hours) 35 45 55 65 % removal by C.F. % removal by D.F.

Fig - 3 Effect of time on biosorption using chick feather (C.F.) and duck feather (D.F.)
90 80 70 60 50 40 30 20 10 0 4 5 6 pH 7 8

% removal

% removal by C.F % removal by D.F

Fig-4 Effect of pH on biosorption using chick feather (C.F.) and duck feather (D.F.)

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Factorial Design The independent variables like biosorbent concentration, time, pH were used to optimize the adsorption by chick and duck feathers and the results of percentage removal of lead in each case are presented in Table 1. The percentage removal of lead depends on the individual effects of combinations of independent variables and the results show a significant variation for each combination. Multiple regression analysis of the experimental data was obtained from the following regression equation for the biosorption of lead Equation-1 Y= 0 + i Xi + ij Xi2 + ij Xi Xj = A0 + A1x1+ A2x2 + A3x3 + A4x1x2+ A5x1x3 + A6x2x3 + A7x1x1 + A8x2x2 + A9x3x3 The coefficients (p) were highly significant for both chick and duck feathers when compared with interactive effects. Multiple regression coefficient (R) was estimated from the second-degree polynomial Eq. (1). The value of r2 = 0. 98098 & 0.91785 for chick and duck feathers respectively which is closer to one indicates that the correlation is best suited for predicting the performance of the biosorption system and the predicted values were found to be very closer to the experimental results. The results obtained from the BBD, the students T distribution, the p values and the parameter estimates for chick and duck feathers are given in Table 2& 3 respectively. The regression equation coefficients were calculated and the data fitted to a second-order polynomial equation using MATLAB for removal of lead with chick & duck feathers. The optimum values of the test variables and the corresponding maximum percentage removal of lead (70.73%) by chick feathers were obtained in coded units as X1 =1.0705, X2 = 0.0038, X3 = 0.0186 & maximum percentage removal of lead (58.8%) by duck feathers were obtained in coded units as X1 =565.5556, X2 = 1.3436, X3 = 10.7542 were shown in the following equations. Final Polynomial Equation for chick feathers: Y= -0.1456 + 1.0705x1 + 0.0038 x2+ 0.0186 x3 -0.0083x1x2-0.0260 x1x3 - 0.0001 x2x3- 1.7948 x1x1 -0.000 x2x3 - 0.0008 x3x3 Final Polynomial Equation for duck feathers: Y= -63.5856+ 565.5556x1 + 1.3436 x2 + 10.7542 x3 -4.1500 x1 x2 -14.4167 x1 x3 -0.0508 x2x3 -822.2222 x1 x1 0.0006 x2 x3 - 0.4556 x3 x3 The maximum lead removal by chick and duck feathers was 70.73% and 58.8 respectively. This experimental value closely agrees with the values obtained from the response surface methodology, confirming that the RSM using the statistical design of experiments could be effectively used to optimize the process parameters and to study the importance of individual, cumulative and interactive effects of the test variables in biosorption. Each contour plot represents a number of combinations of two test variables with the other variable kept at its optimal level. The maximum percentage removal of lead is indicated by the surface confined in the smallest

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curve of the contour plot. The studies of the contour plots also reveal the best optimal values of the process conditions and are given below: For chick feathers: biosorbent concentration 0.19g, time 25hrs, pH7 and for duck feathers biosorbent concentration 0.15g, time 35hrs, pH7 which is shown in Fig - 5 to 8.

Fig - 5 Response surface contour plot showing interactive effect of biosorbent concentration and time on the removal of lead by chick feathers.

Fig - 6 Response surface contour plot showing interactive effect of time and pH on the removal of lead by chick feathers.

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The graph was plotted by taking observed and predicted values which shows that both observed and predicted values are adjacent to the line for both chick and duck feathers which are shown in graphs 1& 2.

Fig - 7 Response surface contour plot showing interactive effect of biosorbent concentration and time on the removal of lead by duck feathers

Fig 8 Response surface contour plot showing interactive effect of time and pH on the removal of lead by duck feathers
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Observed vs. Predicted Values 3 3-level factors, 1 Blocks, 27 Runs; MS Residual=3.648408 DV: % Removal Efficiency 75 70 65 60 Predicted Values 55 50 45 40 35 30 25 20 25 30 35 40 45 50 Observed Values 55 60 65 70 75

Graph - 1 Observed and predicted values for chick feathers

Observed vs. Predicted Values 3 3-level factors, 1 Blocks, 27 Runs; MS Residual=4.512952 DV: % Removal Efficiency 60

55

Predicted Values

50

45

40

35

30 30

35

40

45

50

55

60

65

Observed Values

Graph - 2 Observed and predicted values for duck feathers The critical values obtained for chick feathers are 0.19 grams biosorbent, 24.56 hours, 6.94 pH and for duck feathers are 0.23 grams biosorbent, 20.17hours, 7.02pH as shown in Table - 4. Therefore, it is apparent that

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the response surface methodology not only gives valuable information on interactions between the factors but also leads to identification of feasible optimum values of the studied factors. Table - 2: Effect Estimates ; Var.:% Removal Efficiency; R-sqr=.98098; Adj:.97091 3 3-level factors, 27 Runs;
Mean/Interc. (1)Biosorbant concentration (gm)(L) Biosorbant concentration (gm)(Q) (2)Time(hrs)(L) Time(hrs)(Q) (3)pH(L) pH(Q) 1L by 2L 1L by 3L 2L by 3L Effect 41.3222 28.3889 35.8967 -2.1667 3.1267 -0.7422 0.8067 -16.6683 -5.1917 -1.9850 Effect 41.70370 22.84444 16.44444 2.60000 0.05556 0.44444 0.45556 -8.30000 -2.88333 -1.01667 Std.Err. 0.636694 1.800841 1.559574 0.900421 0.779787 0.900421 0.779787 1.102786 1.102786 1.102786 Std.Err. 0.708124 2.002876 1.734542 1.001438 0.867271 1.001438 0.867271 1.226506 1.226506 1.226506 t(17) 64.9013 15.7642 23.0170 -2.4063 4.0096 -0.8243 1.0345 -15.1148 -4.7078 -1.8000 t(17) 58.89325 11.40582 9.48057 2.59627 0.06406 0.44381 0.52527 -6.76719 -2.35085 -0.82891 p 0.000000 0.000000 0.000000 0.027771 0.000908 0.421181 0.315409 0.000000 0.000203 0.089634 p 0.000000 0.000000 0.000000 0.018825 0.949671 0.662780 0.606176 0.000003 0.031057 0.418641 -95.% -230.44 961.38 -1959.35 2.63 -0.05 -4.76 -2.45 -9.50 -37.59 -0.22 -95.% 40.2097 18.6187 12.7849 0.4872 -1.7742 -1.6684 -1.3742 -10.8877 -5.4710 -3.6044 +95.% -60.67 1179.54 -1630.31 5.00 -0.01 41.95 0.84 -7.17 -14.32 0.02 +95.% 43.19771 27.07014 20.10401 4.71285 1.88534 2.55729 2.28534 -5.71230 -0.29563 1.57104 Coeff. 41.32222 14.19444 17.94833 -1.08333 1.56333 -0.37111 0.40333 -8.33417 -2.59583 -0.99250 Coeff. 41.70370 11.42222 8.22222 1.30000 0.02778 0.22222 0.22778 -4.15000 -1.44167 -0.50833

Table - 3: Effect Estimates; Var.:% Removal Efficiency; R-sqr=.91785; Adj:.87437 3 3-level

Mean/Interc. (1)Biosorbant concentration (gm)(L) Biosorbant concentration (gm)(Q) (2)Time(hrs)(L) Time(hrs)(Q) (3)pH(L) pH(Q) 1L by 2L 1L by 3L 2L by 3L

factors, 27 Runs;

Table - 4 Critical values; Variable: % Removal Efficiency Solution: maximum Predicted value at solution: 68.07936 (Chick feathers), 53.18945 (Duck feathers)
Observed Biosorbant concentration (g) Time(hrs) pH 0.10000 15.00000 6.00000 Chick feathers Critical Observed 0.19098 24.56185 6.94208 0.30000 35.00000 8.00000 Observed 0.10000 15.00000 6.00000 Duck feathers Critical 0.23246 19.77395 7.02192 Observed 0.30000 35.00000 8.00000

CONCLUSION
This work has demonstrated the use of BoxBehnken design for determining the optimum process conditions leading to the maximum percentage removal of lead from aqueous solutions. Using this experimental design and multiple regression, the parameters namely, biosorbent concentration, pH and contact time were studied effectively and optimized with a lesser number of experiments. This methodology could therefore be successfully employed to study the importance of individual, cumulative and interactive effects of the test variables in biosorption.

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