3.

MATERIALS AND METHODS

3.1. 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.

3.2. 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

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.
 3.3 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 UV-Vis Spectrometer 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.

3.4. 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

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

Concentration of Reaction
Volume ratio of Time, Reaction
Sample aqueous methyl eletrolyzed water to hours Temperature
# orange dye aqueous methyl (°C)
solution (ppm) orange dye
(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 30
10 300 0.5:9.5 2 30
11 200 2:8 2 30
12 300 2:8 2 30
13 200 0.5:9.5 8 30
14 300 0.5:9.5 8 30
15 200 2:8 8 30
16 300 2:8 8 30

3.5. 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

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.

3.6. 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 efficiency

of each combination will be known through the amount of methyl orange removed (in

mg) per amount of free chlorine used.

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 efficiency of 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 measured using UV-Vis

Spectrophotometer 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

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 measured using UV-Vis Spectrophotometer 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 measured using UV-Vis Spectrophotometer

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

reaction temperatures will be considered are 30 °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 30 °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

measured using UV-Vis Spectrophotometer located in the analytical laboratory at the

Department of Chemical Engineering, CEAT-UPLB.

3.7. 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.

3.8. Data Analysis

The decolorization efficiency of the reaction will be determined by calculating the

amount of methyl orange removed per amount of free chlorine used using the formula

given below:

𝐷𝑒𝑐𝑜𝑙𝑜𝑟𝑖𝑧𝑎𝑡𝑖𝑜𝑛 𝑒𝑓𝑓𝑖𝑐𝑖𝑒𝑛𝑐𝑦

(𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛𝑓𝑖𝑛𝑎𝑙 − 𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛𝑖𝑛𝑖𝑡𝑖𝑎𝑙 )(𝑉𝑜𝑙𝑢𝑚𝑒𝑤𝑎𝑠𝑡𝑒𝑤𝑎𝑡𝑒𝑟 )

= 𝑥 100
(𝑉𝑜𝑙𝑢𝑚𝑒𝑒𝑙𝑒𝑐𝑡𝑟𝑜𝑙𝑦𝑧𝑒𝑑 𝑤𝑎𝑡𝑒𝑟 )(𝑖𝑛𝑖𝑡𝑖𝑎𝑙 𝐶𝑙2 𝑐𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛)

3.9. Statistical Analysis

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

be used, wherein two-way analysis of variance (ANOVA) will be employed for the

statistical analysis of the obtained data.