Characterization of POME

3.5.1

Biochemical Oxygen Demand (BOD)

Biochemical oxygen demand (BOD) is defined as the amount of oxygen consumed

during microbial utilization of organics (Peavy et al., 1985). It measures the ability of
naturally occurring microorganisms in wastewater to digest organic matter. BOD is
usually measured with 5 days incubation at 293K the amount of oxygen depletion is
determined. BOD is an indirect measure of biodegradable organic compounds in water
and it can be also used in determining the oxygen demand of industrial discharge and
waste effluent.
To measure BOD of POME, dilution water was first prepared by mixing 1 mL each of
phosphate buffers, magnesium sulphate (MgSO4), calcium chloride (CaCl2), and ferric
chloride (FeCl3) solution into 1L volumetric flask. Then, the solution is diluted to 1L by
using distilled water. 10 mL of POME sample was diluted to 300 mL by using dilution
water. The pH of diluted POME was measured and adjusted to the range of 6.5 to 7.5 by
using concentrated sulphuric acid or sodium hydroxide. The sample solution was then
transferred into a 300mL-incubation bottle. The initial dissolved oxygen (DO)
concentration for POME sample was measured using a dissolved oxygen meter. The
flared mouth of bottle was covered with an aluminium foil. The incubation bottle was
placed in a BOD incubator with temperature of 293 K for five days and the final DO
value was measured. BOD value of POME then was calculated according to the Equation
3.11 and 3.12.
BOD (mg/L) = (D1 D2) x Dilution factor

(3.11)

Dilution factor = Bottle volume (300mL) / Sample volume

(3.12)

where D1 and D2 represent the initial and final DO values of POME sample

3.5.2

Total Suspended Solids (TSS)

Solids suspended in water consist of inorganic particles, organic particles and immiscible
liquids. Common constituents of inorganic solids in water are clay, slit and soil particles
while for organic particles, bacteria, plant fibres and algal cells are suspended in water.
Immiscible liquids such as oils and greases are common constituents of wastewater.
Suspended solids are an important parameter to measure quality of wastewater, to
monitor several treatment processes, and to measure the quality of effluent.
The TSS of POME was determined using vacuum filtration through a 47 mm nylon
membrane with a pore size of 0.45 m. Prior to filtration, the nylon membrane was dried
in the oven at 378 K for 1 hour, cooled in a desiccator and weighed. 100 mL of POME
sample which was mixed to ensure homogeneity was filtered onto centre of filter disk in
a Buchner flask by using gentle suction under vacuum. The filter was washed with three
successive 10 mL volumes of distilled water to allow complete drainage between
washings, and continue suction for about 3 min after filtration was completed. The nylon
membrane was dried in the oven at 378 K for 1 hour, cooled in a desiccator and weighed
until a constant weight was obtained. The weight difference of the membrane was
determined as TSS and was expressed in mg/L.

3.5.3

Ammoniacal Nitrogen Analysis

Ammonia is a gas at temperatures and pressures of natural water system. Ammonia exists
in equilibrium with aqueous ions of ammonium (NH 4+) in water. Oxidation of ammonia
and ammonium ions to nitrate by aquatic microbes will cause elevated biochemical
oxygen demand in wastewater (Peavy et al., 1985).
In ammonia-nitrogen test, ammonia compounds present in wastewater combine with
chlorine to form monochloride. Monochloride then reacts with salicylate to form 5aminosalicylate which is further oxidized in the presence of a sodium nitroprusside
catalyst to give a blue color compound. The blue color will then be masked by the yellow
color from the excess reagent. Finally, a green-color solution will form indicating the
present of ammonia-nitrogen in the wastewater.

Firstly, blank sample (DI water) and POME sample were filled into 10 mL round sample
cell. At the same time, the Hach DR2400 instrument was turned on and the program
385N Ammonia Salic was selected. The content of one Ammonia Salicylate Powder
Pillow was added to each cell. The cells were shaken to dissolve the powder. The timer
icon of the Hach DR2400 was touched. A three-minute reaction period would begin.
When the timer beeped, the content of one Ammonia Cyanurate Reagent Powder Pillow
was added to each cell. The cells were shaken to dissolve the reagent. The timer icon of
the DR2400 was touched and a fifteen-minute reaction period would begin. When the
timer beeped, the blank sample was cleaned by fibreless tissue and subsequently inserted
into the cell holder. For blank sample, zero button was pressed to set as a baseline with
0.00 mg/L of NH3-N. Then, the cell holder filled with POME sample was placed into the
cell holder. Read button was pressed for the reading of POME sample. The result was
expressed in mg/L NH3-N.

3.5.4

pH

pH is a measure of the acidity or alkalinity of the solution. To measure pH of POME, 100

mL of POME sample was collected in a beaker. The tip of pH meter was submerged into
the beaker. The read button on the meter was pressed and when the reading was constant,
pH value was recorded.

3.5.5

Oil and Grease Analysis

Oil and grease (O&G) is a measure of the amount of dispersed oil and grease including
fuels, motor oil, lubricating oil, hydraulic oil, cooking oil, and animal-derived fats in the
water. It is an important parameter for water quality and safety as O&G in water can
induce surface films and shoreline deposits, and also interfere with aerobic and anaerobic
biological process in water, resulting in environmental degradation and water pollution.
In current study, hexane extractable gravimetric method (HEM) was used in oil and
grease analysis. Firstly, 350 mL of POME was prepared in a separatory funnel and the pH
of the solution was adjusted to be 2 or less by using hydrochloric acid. Then, 20 mL of nhexane was added into a separatory funnel. The separatory funnel was shaken vigorously

for 2 minutes. Then, it was clamped vertically for 10 min to allow the separation of the
lower water layer and upper solvent layer. The water layer was drained into a volumetric
flask. The solvent layer was then drained into a pre-weighed boiling flask through a
funnel that contains filter paper and 10 g anhydrous sodium sulphate. The extraction
process was repeated 3 times to ensure complete extraction of oil content. After that, the
n-hexane was evaporated at 343 K until all solvent was removed. The flask was put in a
desiccator for 30 minutes until the flask temperature decreased to room temperature. The
weight of the boiling flask was measured and the difference of the weight was recorded
as the oil and grease content in mg/L of HEM.