Procedure

2.
0 METHODOLOGY
2.1 CHEMICAL AND APPARATUS USED
In this experiment, the chemical and reagent that will be used for removal of nitrogen
in anoxic batch reactor are Biochemical Oxygen Demand (BOD) testing reagent, waste water,
mineral stabilizer, nessler reagent and polyvinyl alcohol dispersing for direct nesslerization
(refer appendix 2.1.1). For apparatus and instrument are Oxidation Reduction Potential
(ORP) meter, Dissolved Oxygen (DO), pH meter, temperature meter, Mixed Liquor
Suspended Solid (MLSS) meter, filter paper, DR6000 Spectrophotometer, beaker, measuring
cylinder, pipette, spatula, dropper, test tube, and oven for heating of total suspended solid.
2.2 GENERAL PROCEDURE
According to research paper advanced nitrogen removal without addition of external

carbon source in an anoxic, anaerobic and aerobic sequencing batch reactor that conduct by
[5], the working volume of SBR are 2.5 L operated with A/O/A model was applied as shown
in figure 1 below.
Figure 1: Experimental set up and operational mode of AOA SBR
a) Schematic diagram b) Operational mode
Each cycle time was 24 hours consisting of 4 hours anaerobic reaction, 3 – 4 hours of aerobic
reaction evaluate by valley of ammonia on the pH profile and will be stopped automatically
by programmable logic controller (PLC), 12 – 15 hours of anoxic reaction evaluate by nitrate
knee on the ORP profile and stopped by PLC. Figure 1b shown the process of feeding,
settling, and decanting.
In the feed process, 1.25 L of wastewater was pumped to the reactor and the
corresponding VER was 50%. A magnetic stirrer was used to mixing equally in the reaction
stages. To keep a constant aeration intensity, the air pump was applied during the aerobic
reaction stage and dissolve oxygen (DO) concentration was above 3.0 mg/L. Moreover, no
sludge will be discharge during long term operation. . The sludge retention time (SRT) was
80–90 days, which was estimated based on the effluent suspended solids concentration (Ss).
2.3 EXPERIMENT PROCEDURE
2.3.1 The removal of Carbon/ Nitrogen in Aerobic Batch Reactor
First must make sure the valve was closed. Valve of V14 must open and insert the
wastewater into the anoxic tank of B4 followed by 20L of supernatant wastewater from feed
tank, B1 to make up the volume to 30L by using the peristaltic pump, P1. Next, set the
peristaltic pump speed up to 600 rpm to expedite the filling and stop the pump when desired
volume is reached. Then, turn on the stirrer, M1 and regulate the speed to around 100-150
rpm. Maintain the value of dissolved oxygen, DO01 between 0.2-0.5 mg/L and determine the
initial value of COD, NH3-N and TSS in the reactor by taking 100 ml sample from the anoxic
reactor, B4 (refer appendix 2.3.1.1). The sample then will be stored in the ice container to
hibernate the bacteria and stop the reaction (refer appendix 2.3.1.2). 100 ml of sample was
taken from the aerobic reactor, B2 at appropriate time interval and analyse the value of BOD,
NH3-N and TSS value. Lastly, the process will stopped when the desired retention time is
achieved.
2.3.2 Ammonia Nitrogen Analysis
After 30 minutes the sample were collected initial and final to test the amount of ammonia by
using a chemical reagent such as Mineral Stabilizer, Nessler Reagent and Polyvinyl Alcohol
Dispersing. First step, prepare the small beaker for sample about 25 ml for three sample
blank, initial and final. Prepare the deionized water for the blank sample. Then, added 3 drop
of mineral stabilizer into the sample for each of beaker. Stir the sample until it dissolves
equally. After that, added 3 drops of polyvinyl alcohol dispersing for each beaker and stir
until it dissolves. Lastly, added 1.0 ml of Nessler reagent into the sample for each beaker and
stir until it dissolves (refer appendix 2.3.2.1). Let it be for 1 minutes to make the chemical
reagent react before pour into the vial. Make sure wipe the vial before put it into the
Spectrophotometer (refer appendix 2.3.2.2). After the sample are ready, insert the sample into
DR6000 to read result (refer appendix 2.3.2.3).
2.3.3 Biochemical Oxygen Demand (BOD) Analysis
Prepare 150 mL of 2 sample of wastewater from initial sampling for BOD. Then,
added 150 mL of nutrient BOD into the sample and record the dissolved oxygen (DO) in the
sample (refer appendix 2.3.3.1). Repeat the same procedure for final sampling. Next, the
sample will be freeze in the freezer for 3 days and after 3 days (refer appendix 2.3.3.2), check
the DO for initial and final sampling by using DO indicator.
References
[5] Shi, L., Ma, B., Li, X., Zhang, Q., and Peng, Y., 2019, “Advanced Nitrogen Removal
without Addition of External Carbon Source in an Anaerobic / Aerobic / Anoxic
Sequencing Batch Reactor,” Bioprocess Biosyst. Eng., (100).
Appendix
Appendix 2.2.1
Appendix 2.3.1.1
Appendix 2.3.1.2
Appendix 2.3.2.1
Appendix 2.3.2.2
Appendix 2.3.2.3
Appendix 2.3.3.1
Appendix 2.3.3.2

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2.

2.1 CHEMICAL AND APPARATUS USED

2.2 GENERAL PROCEDURE

According to research paper advanced nitrogen removal without addition of external

Figure 1: Experimental set up and operational mode of AOA SBR

a) Schematic diagram b) Operational mode

2.3 EXPERIMENT PROCEDURE

2.3.1 The removal of Carbon/ Nitrogen in Aerobic Batch Reactor

2.3.2 Ammonia Nitrogen Analysis

2.3.3 Biochemical Oxygen Demand (BOD) Analysis

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