LABORATORY MANUAL FOR DETERMINING OIL AND GREASE FOR

RAW AND TREATED EFFLUENT

Revision No: 1/2021 | Date: 08/09/2021

1.0 Introduction
Oil and grease present in waste water samples extracted using liquid-liquid extraction method.

2.0 Apparatus
Beaker

Measuring cylinder

1 L separatory funnel

Distillation set

Desiccator

Oven

3.0 Reagents
Concentrated sulfuric acid (1.84 SG/93%), diluted 1:1 with distilled water

Anhydrous sodium sulfate

Commercial grade hexane

Isopropyl alcohol

Whatman No. 40 filter paper (or filter paper with pore size 8 micron)

4.0 Procedure
1- Collect a representative waste water sample in a clean bottle. The bottle should be free from
detergent film build-up on the inside wall.
2- Transfer 100-ml aliquot of the sample water into a beaker to allow for addition of sulfuric
acid. Acidify the sample with 2.5 ml sulfuric acid (1:1) if the sample is not yet acidified with
sulfuric acid. Then, transfer the acidified sample water into a separatory funnel. Rinse the
beaker with 30 ml hexane and transfer the hexane rinsing into the separatory funnel. Shake
the separatory funnel for 2 minutes and vent out the pressure build-up every 5-10 shakes.
Break the emulsion layer with 4 ml of isopropyl alcohol if needed.
3- Allow the organic phase to fully separate and drain the aqueous phase into the used beaker.
Repeat the extraction with another three portions of 30 ml hexane rinsing beaker each time
with the hexane before transferring into the separatory funnel.
4- Drain the organic phase into a conical flask containing some anhydrous sodium sulfate. Add
more anhydrous sodium sulfate if there is still water present in the organic phase. Filter off
 the water-less organic phase into a distilling flask (previously dried and weighed) using
Whatman No. 40 filter paper. Distill off the hexane on the water bath. Complete the drying
in the oven at 100 - 105°C for 3 hours. Cool the distilling flask in a desiccator and record the
final mass.

5.0 Calculations
The oil and grease content is calculated using the following formula

mg (m ¿ ¿ distilled−mempty )( g) 1000 mg 1000 ml

Oil∧Grease content ( )L
=
V sample (ml)
×
1g
×
1L
¿

where,

mdistilled = mass of flask after distillation

mempty = mass of empty flask
Vsample = volume of sample used

6.0 Disposal
 The oil and grease contained in the flask after distillation can be washed away using the
distilled used hexane. Discard the used hexane into an organic waste container. UNDER NO
CIRCUMSTANCES should the used hexane disposed into drainage.
 The acidified oil-depleted water sample can be poured down the drain with a lot of water to
lower the acidity of the water.
 The filter paper and sodium sulfate used to dry the organic phase can be discarded into solid
waste container.

7.0 Emergency
 In case of spill, follow spill cleanup procedure.
 Contact immediately in case of emergency: Azri (Lab Operator), Anas (Lab Officer), Aziz (Lab
Exec), Ramzan (Principal Chemist)

8.0 Reference
Oil and Grease for Treated Effluent Reference Method – DOE, 2008

Prepared by Verified by Certified by

Muhammad Anas Yuzairi bin Muhammad Abdul Aziz Ramzan Razali
Mohd Yusri Laboratory Executive Principal Chemist, CEO
Laboratory Officer 08 September 2021 08 September 2021
08 September 2021