3.

MATERIALS AND METHODS

3.1 Materials and Reagents

The practical dye that will be used, methyl orange in powdered form, will be

obtained from Unit Operation Laboratory II, Department of Chemical Engineering

(DChE), CEAT, University of the Philippines Los Baños, Laguna.

Moreover, for the calibration for pH meter such as pH 7 and pH 4 buffer

solutions, reagents will be obtained from the Analytical Laboratory while the commercial

grade sodium chloride will come from the Bio-fuels Laboratory, Department of Chemical

Engineering (DChE), CEAT, University of the Philippines Los Baños, Laguna.

Furthermore, for the determination of the free chlorine content of the electrolyzed

water, glycine and DPD (N,N Diethyl-1,4 phenylenediaminesulfate) tablets will be used

that will be obtained from LARC Laboratory.

3.2 Glassware and Equipment

The glasswares that will be used in the experiment are graduated cylinders,

pipette, beaker, volumetric flask, screw-capped test tubes and Hach vials.

The equipment that will be used in reading the absorbance of the dye solution is

the Hach Spectrophotometer DR-2800 in the analytical laboratory, DChE-UPLB. The

acidic electrolyzed water will be generated from Cawolo Disinfectant Generator CA-5,

while for the characterization of electrolyzed, pH and the free chlorine content of the

electrolyzed water, will be determined using the pH meter and Lovibond tintometer from

the LARC Laboratory.

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Furthermore, for the other equipments such as the analytical balance, which will

be used in the determination of the mass of the sample, the hot plate for the source of heat

for the effect of the temperature set-up, and the magnetic stirrer for the distribution of

heat throughout the system, these will be obtained in the analytical laboratory, DChE-

UPLB.

3.3 Preparation and Characterization of Wastewater

Three hundred milligrams of methyl orange dye powder will be dissolved in a one

(1) liter of distilled water using a 1-L volumetric flask, in order to obtain a 300 mg/L of

aqueous methyl orange solution that will be characterized based on its pH and color.

3.4 Preparation and Characterization of Acidic Electrolyzed Water

Cawolo Disinfectant Generator CA-5 will be used in the preparation of the

electrolyzed water. About 15 g of commercial grade NaCl will be prepared and will be

placed into the cup then 300 mL water will also be added. The salt solution that will be

formed will be stirred continuously until all the salt is completely dissolved. After this,

the salt solution will be poured into the generator tank and 200 mL of water will also be

dispensed into the generation tank together with the cup. Upon closing the cover, the

Cawolo Disinfectant Generator CA-5 will be plugged, a red LED light will be expected

to blink, which is the indication that the Cawolo Disinfectant Generator CA-5 is already

on. Then, the start button will be pressed and 20 minutes of electrolysis will be done until

the red LED light is turned off and the green LED light is started to blink which will be
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the signal that acidic electrolyzed water is generated. The acidic electrolyzed water that

will be produced will be characterized based on its free chlorine content and pH.

3.5 Preliminary Experiment

For the preliminary experiment, a standard curve showing the relationship

between the absorbance and the concentration of methyl orange solution will be

determined first. In order to obtain a standard curve, the sample (methyl orange solution)

must be first diluted to a lower concentration so that Hach Spectrophotometer DR-2800

can read its absorbance. The linear equation of the standard curve that will be obtained

will be used to determine the actual concentration of the treated wastewater.

Additionally, the determination of the free chlorine content that is responsible for

the decolorization of the aqueous methyl orange dye solution will also be obtained. This

will be done by using the method that was presented by Lovibond Water Testing with the

reagents glycine and DPD tablet. Furthermore, the concentration of methyl orange will be

determined in such a manner that it will be completely degraded in a high dosage of

electrolyzed water, about 2 mL in volume, for 8 hours.

3.6 Experimental Design

In the parametric analysis of this experiment, in order to identify the effect of one

or two variables to another, two-level factorial design will be used. Table 3-1 shows the

constructed experimental design matrix for the four factors: concentration of aqueous
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methyl orange dye solution, electrolyzed water to aqueous methyl orange dye solution

ratio, reaction time and reaction temperature.

Table 3.1. Combination of Different Parameters in the Decolorization of

Aqueous Methyl Orange Dye Solution.

Parameters
Volume ratio of
Concentration of
electrolyzed water to Reaction Reaction
Sample aqueous methyl
aqueous methyl Time, Temperature
# orange dye
orange dye hours (°C)
solution (ppm)
(mL/mL)
1 200 0.5: 9.5 2 RT
2 300 0.5: 9.5 2 RT
3 200 2: 8 2 RT
4 300 2: 8 2 RT
5 200 0.5: 9.5 8 RT
6 300 0.5: 9.5 8 RT
7 200 2: 8 8 RT
8 300 2: 8 8 RT
9 200 0.5:9.5 2 50
10 300 0.5:9.5 2 50
11 200 2:8 2 50
12 300 2:8 2 50
13 200 0.5:9.5 8 50
14 300 0.5:9.5 8 50
15 200 2:8 8 50
16 300 2:8 8 50

3.7 Decolorization of Aqueous Methyl

Orange Dye Solution

Determination of the Standard Curve of Methyl Orange Solution

The stock solution will be diluted using different dilution factor which are within

the range of the spectrophotometer readings. A 100-mL of 10 ppm methyl orange dye
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solution will be prepared from the stock solution using the equation C1V1 = C2V2. Then a

100 mL of the target diluted concentrations for the aqueous methyl orange solution (0.1,

0.2, 0.3, 0.4, 0.5, 1, 3, 5, 7, and 9 ppm) will be prepared using the stock solution. The

maximum absorbance of the 10 ppm methyl orange dye solution at varying wave length

will be obtained using the spectrophotometer, and then using the same wavelength where

the maximum wavelength will be obtained, the absorbances at varying concentrations

will be checked and recorded. An absorbance versus concentration standard curve will

then be plotted.

Initial Characterization of Wastewater

The prepared wastewater is only composed of methyl orange dye and distilled

water. This aqueous methyl orange dye solution will be characterized based on its initial

pH and color.

3.8 Parametric Study on the Decolorization of

Methyl Orange Dye Solution

Different combinations of concentration of aqueous methyl orange dye solution,

free chlorine level of acidic electrolyzed water, reaction time and temperature will be

tested in this experiment. Furthermore, the volume of electrolyzed water-to-volume of

wastewater will be varied in the experiment so that the effect of free chlorine level in a

10-mL volume will be determined. Additionally, the response to determine the color

removal of each combination will be known through the TCU removed per amount of

free chlorine used.

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Also, the pH of each treated aqueous methyl orange solution will be measured in

order to conclude if the treated dye solution passed the effluent standards as presented in

DAO 2016-08. Lastly, the best condition for the decolorization of methyl orange dye

solution will obtained at the combinations with highest decolorization per amount of free

chlorine used.

Effect of Concentration of Methyl Orange Dye Solution

In order to determine the effect of the concentration of dye on the decolorization

efficiency, two different concentrations will be considered, this includes 200 ppm and

300 ppm at the specified combination of reaction time and volume of electrolyzed water

in the 10-mL sample that will be placed in a screw-capped test tube. The generated

electrolyzed water from Cawolo Disinfectant Generator CA-5 will then be added to 200

ppm and 300 ppm, respectively, following the specified condition presented in Table 3.1.

The final concentration of the treated sample will be determined using the standard curve

and the absorbance that will be read by the Hach Spectrophotometer DR-2800 located in

the analytical laboratory at the Department of Chemical Engineering, CEAT-UPLB.

Effect of Electrolyzed Water to Aqueous Methyl Orange Dye Solution Ratio

To determine the effect of volume of electrolyzed water on the decolorization

efficiency, different volume of electrolyzed water will be added to the aqueous dye

solution. The volumes of electrolyzed water that will be considered in this experiment are

0.5 mL and 2 mL at the specified combination of reaction time and concentration of

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aqueous methyl orange dye solution in the 10-mL sample that will be placed in a screw-

capped test tube. The generated electrolyzed water from Cawolo Disinfectant Generator

CA-5 will then be added to the aqueous methyl dye orange solution following the

specified condition presented in Table 3.1. The final concentration of the treated sample

will be determined using the standard curve and the absorbance that will be read by the

Hach Spectrophotometer DR-2800 located in the analytical laboratory at the Department

of Chemical Engineering, CEAT-UPLB.

Effect of Reaction Time

In order to determine the effect of reaction time on the decolorization efficiency,

two different reaction times will be considered in this experiment. The reaction times that

will be considered are 2 hours and 8 hours at the specified combination of added volume

of electrolyzed water and concentration of methyl orange dye solution in the 10-mL

sample that will be placed in a screw-capped test tube. The generated electrolyzed water

from Cawolo Disinfectant Generator CA-5 will then be added to the aqueous methyl dye

orange solution following the specified condition presented in Table 3.1. The final

concentration of the treated sample will be determined using the standard curve and the

absorbance that will be read by the Hach Spectrophotometer DR-2800 located in the

analytical laboratory at the Department of Chemical Engineering, CEAT-UPLB.

Effect of Temperature

In order to determine the effect of reaction temperature on the decolorization

efficiency, two different reaction temperatures will be considered in this experiment. The
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reaction temperatures will be considered are 50 °C and room temperature. The generated

electrolyzed water form Cawolo Disinfectant Generator CA-5 will then be added to the

aqueous methyl dye orange solution following the specified condition presented in Table

3.1. For the 50 °C – condition, this set-up will be submerged in a water bath together with

the thermometer and heat source, wherein a magnetic stirrer will be in-charge in

distributing the heat evenly in the system. When the decolorization of the aqueous methyl

orange dye solution is already seen, the final concentration of the treated sample will be

determined using the standard curve and the absorbance that will be read by the Hach

Spectrophotometer DR-2800 located in the analytical laboratory at the Department of

Chemical Engineering, CEAT-UPLB.

3.9 Kinetics Study on the Decolorization of

Methyl Orange Dye Solution

Among the conditions that will be tested, the best condition, the condition

wherein there is the highest removal of methyl orange per amount of free chlorine used,

will be considered. In this condition, the time profile versus the efficiency of

removal/free chlorine will be plotted. Through this, the order and the kinetic parameters

will be calculated by integral method. In integral method, the order of reaction will be

assumed first, among the assumed orders of reaction; the highest value for R2 for the

linear regression that will be obtained will be the final order of the reaction.

3.10 Data Analysis

The decolorization of the reaction will be computed using the formula:

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Where:
Ci is the initial concentration in mg/L

Cf is the final concentration in mg/L

TCUi is the initial TCU of the treated solution

TCUf is the final TCU of the treated solution

V is the volume in mL

HOCl is hypochlorous acid

ew is electrolyzed water

3.11 Statistical Analysis

To analyze the statistical proposition of the result, Design Expert Version 10 will

be used, wherein two-way analysis of variance (ANOVA) at p-value = 0.5 will be

employed for the statistical analysis of the obtained data.