OFFICIAL LABORATORY REPORT

ENVIRONMENTAL CHEMISTRY

Extraction in Surfactant Analysis (Detergent)

NAME : CAHYANINGRUM AYU A.

NRP : 03211740000028

LECTURER : ARSETO YEKTI B,ST,MT,M. Phil, PhD

LABORATORY ASSISTANT : ANISA FADHILLAH

ENVIRONMENTAL ENGINEERING DEPARTEMENT

FACULTY OF CIVIL ENGINEERING, ENVIRONMENT AND GEOSCIENCE
SEPULUH NOPEMBER INSTITUTE OF TECHNOLGY 2018
 CHAPTER I
PREFACE

1.1 Purpose
The purpose of this laboratory works are:
1. Comprehend basic principle of extraction application for organic analysis in water sample.
2. Able to determine limited reagent and excess reagent, and its functionality in the analysis.

1.2 Principle
A ratio number of distribution (D) in an extraction is a principle in determining interphase of
substance extraction. This D number is specific on each condition. Surfactant, is an example of organic
substance that is highly dissolve in water, thus if it is extracted with organic solvent, will give not too
high results, because of the low value of D. By conducting reaction of surfactant in slighlt alkaline using
(MB) will form complex substance of Methylen Blue Active Surface (MBAS), that has higher D value,
thus becomes effective during extraction. The residue of MB indicates that all surfactant has formed the
intended/targeted complex substance. This method becomes fundamental of analysis, when it is known
which one is the limited reagent and which one is the excess reagent.

1.3 Basic Theory

The synthetic detergents prepared from polymers of ethylene oxides are resistant to biological
attack because of their (C-O-C) bonds. The ions being smaller, remain less affected by the filtering
action of soil. The most frequently applied surfactants are sodium salts of high molecular weight such as
alkyl sulphates or sulphonates (LAS or linear C12 alkyl- ben- zene sulphonate). Surfactants generally
have high bio- degradability and have a moderate toxicity for fish. The threshold value which can impair
aquatic life is 3–12 mg/l for anionic detergent, 3–38 mg/l for non-ionic de- tergent, and 20mg/l for
cationic detergent. In addition to affecting aquatic life, surfactants affect oxygen-turnover of water and
reduce sedimentation process, and thus de- laying water purification, particularly during aeration of
waste water. Surfactants are also called methylene-blue- active substances (MBAS) because they
produce colour change in aqueous solution of methylene blue dye.

(Ghose et al., 2009)

Coastal ecosystems receive large quantities of pollutants, which are used as the principal
constitute of commercial detergents. Among these, linear alkylbenzenesulfonate (LAS) is the anionic
surfactant used most in the formulation of detergents, with a consumption rate of 5.5 g/day per person
in developed countries. As a result, LAS can be considered as a good indicator of urban-source
pollution. Anionic detergent is toxic for marine life and its film covers surface water and prevents
penetration of oxygen to water from air. From 1 to 20 ppm surfactant is fatal to some fish. In the
research on Oncorhynchus mykiss in aquarium, the lethal dose is 12,5 mg/L for single dose and 35
mg/L for progressive dose and also affects on plasma parameters. Since LAS is removed very
efficiently in wastewater treatment plants, and subsequently in river waters, LAS concentrations in
estuarine and coastal waters are typically below 50 mg/L where sewage treatment systems are
installed. Higher concentrations, up to 2500 mg/L, have been detected in coastal waters close to
untreated discharge outlets.
(Balcıoğlu, 2014)

Surfactants are among the main group of environmental contaminants due to their increasing
usage and potential toxicity. Anionic surfactants (AS) are found commonly in products of everyday use,
such as detergents or washing agents, and linear alkylbenzenesulfonates are presently the most
popular synthetic AS. After entering wastewater treatment plants, such compounds are partially
degraded in aerobic conditions and partially adsorbed onto the activated sludge. Ultimately, they enter
water or soil and act as a major factor influencing the natural environment. Therefore, controlling the
surfactant content in detergents and surface water samples is necessary.
The methylene blue active substances (MBAS) method is a standard method for determination
of anionic surfactants. This method is based on the emergence of ionic pairs, which consist of anionic
surfactants and a cationic dye (methylene blue), and their transport from water phase to organic phase
(chloroform). The sole dye compounds cannot be transported into the organic phase, hence ionic pairs
emerge. The analytical procedure of the standard MBAS method is carried out with a triple extraction of
the ionic pairs from 100 mL of a previously alkalized sample (15, 10 and 10 mL chloroform) and
measuring the absorbance of the extract at k = 650 nm. The MBAS method is useful, cheap and simple;
however, the procedures are rather troublesome and time-consuming. Using chloroform in high doses is
another flaw, as this compound is toxic and harmful for humans and the environment.
(Wyrwas et al., 2014)

Surfactants (Surface-active agents) represent a highly essential and beneficial group of

compounds for civilised human communities. Two major types of surfactants are bio-surfactants and
synthetic surfactants. Linear alkylbenzene sulphonate (LAS) is an extensively used anionic surfactant.
LAS are frequently used as the sodium salts as the sole surfactant in a formulation or in conjunction
with other anionic, nonionic or cationic surfactants. Presence of LAS at a concentration 500 mg/L is
reported to inhibit microbial growth and resist biodegradation. Most of the LAS biodegradation studies
were conducted under aerobic condition or in anaerobic condition preceded by a period of aerobic
exposure. Under anaerobic conditions LAS is reported to be non biodegradable due to its long exposure
time and toxicity.
The concentrations of anionic surfactant LAS was determined by a modified version of the
methylene blue active substance assay (MBAS) as described by Hayashi. The MBAS assay is a non
specific analytical method to determine anionic surfactant concentration in a solution using a cationic
dye methylene blue. The anionic surfactant will from a complex with the methylene blue and this
complex (not excess dye) can be extracted in to chloroform. The absorbance of the chloroform layer at
655 nm will be directly proportional to the anionic surfactant in the medium.
(Asok et al., 2015)

Surfactants belong to the class of compounds known as amphiphiles, molecules having both a
hydrophobic and hydrophilic component. The hydrophobic component is generally referred to as the tail
group and hydrophilic group is known as the head. The term surfactant comes from a contraction of
‘‘surface active agent’’ and is defined as a material which when present at low concentrations, adsorb
onto the interface, or surface, of the system and thereby alters the interfacial free energies of the
interface. Due to the fact that surfactants are amphiphilic compounds and have the ability to producing
various micellar phases in aqueous and organic solutions, make them marvelous compound in
extraction of various compounds. According to these abilities they can solved various hydrophilic and
hydrophobic compounds in themselves micellar phases. Phenolics compounds consist of various
hydrophobic and hydrophilic compositions. Extraction methods in the bases of organic solvents cannot
make beneficial extraction efficiency for all of these compounds.
(Hosseinzadeh et al., 2013)
 CHAPTER II
EXPERIMENTAL SCHEME

Standart Solution of LAS

 Prepared a series standart solution of LAS in final concentration of : 0,5M,

1M; 1,5M; and 2M by conducting a dilution as much as 1,25 mL; 2,5mL;
3,75 mL; and 5mL into 25 mL volumetric flask

Sample

 25mL is poured into a volumetric flask

Phenolphthalein

 5 drops added into each volumetric flask

 Shaken well
NaOH

 Dropwise of NaOH is added

 Shaken well until the colour becomes lightly pink
H2SO4

 Dropwise of H2SO4, is added until the solution becomes colorless

0,1 M: 2 drops; 0,5 M: 3 drops; 1 M: 2 drops; 2M: 1 drops; sample: 3 drops
 Transfer all solution into seperate separatory funnel
KH2PO4

 25 mL is poured into the volumetric flask that already been emptied

 Shaken well until dissolves completely

Methylene Blue Solution

 10 mL is added into each solution in the separatory funnel

 Shaken well until dissolves completely

Choloform

 25 mL of Choloform is added, divided into 3 times addition which is 8mL,

8mL, and 9mL
 The funnel is then inverted for 2 minutes and the tap carefully opened to
release excess vapor pressure The bottom layer is drained out into the
volumetric flask containing KH2PO4 so the top layer can be retained in the
separatory funnel
 The stopcock is closed and the upper layer is poured out through the top into
another container
 The former bottom layer within KH2PO4 are returned to the separatory funnel
to be extracted again, drained out to the volumetric flask
 Chloroform is addded until the total volume is 25mL for each flask
 Spectophotometry

 The transmittance of each solution being measured by pouring half into

cuvette and put in the spectrophotometry with the desired wavelength
650nm
 Chloroform used as the blanko solution for calibration
 Notes the reading result and make the calibration curve to determine
each concentration

Result

CHAPTER III
DISCUSSION

3.1 Data Observation

NO. TREATMENT OBSERVATION REACTION PICTURE
M1 = 10 mg/L
V2 = 25 mL

Prepare a series standart

For M2 = 0,5 M:
solution of LAS in final
V1 x 10 = 25 x 0,5
concentration of : 0,5M, 1M; Volume total for
V1 = 1,25 mL
1,5M; and 2M by conducting each flask is 25 mL
For M2 = 1 M:
a dilution as much as 1,25
1. V1 x 10 = 25 x 1
mL; 2,5mL; 3,75 mL; and Flammable,
V1 = 2.5 mL
5mL in seperate 25 mL relatively non-toxic,
For M2 = 1,5 M
volumetric flask. Use pipet colorless liquid
V1 x 10 = 25 x 1,5
to take the indicated
V1= 3,75 mL
amount
For M2 = 2M
V1 x 10 = 25 x 2
V1 = 5 mL
Take 25mL of sample by
measuring pipette and Colorless and
2. No reaction
pipette bulb into a 25mL odorless liquid
volumetric flask

Transfer all the standart

There is no
solution and the sample into
3. significant changes No reaction
erlenmeyer flask, give label
in appearance
to each of them
 White or yellowish-
white to pale orange
It is a weak acid, which
fine crystalline
can lose H+ ions in
powder. Odorless.
Add 5 drops of solution. it turns
4. Aqueous solution is
phenolphthalein colorless in acidic
acidic. Colorless in
solutions and pink in
presence of large
basic solutions
amounts of alkali.
Tasteless.
highly soluble in
4 standart soution @
water, insoluble in
1 drops, sample
Add dropwise of NaOH by ether and other non-
solution for 3 drops;
5. using measuring pipette and polar solvents,
pipette bulb liquid, white, waxy,
The solution turns to
opaque crystal,
lightly pink coloured
odorless

H2SO4 is clear, 2 drops for 0,1 M LAS;

colorless liquid, 2 drops for 1,5 M LAS;
Add dropwise of H2SO4 by
odorless, miscible, 1 drop for 1 M LAS; 2
6. using measuring pipette and
and exothermic; drops for 2M LAS; 2
pipette bulb
The solution turns to drops for sample
colorless solution

There is no
Pour each solution in
7. significant changes No reaction
seperate separatory funnel
in appearance

Fill the emptied erlenmeyer

White powder
flask with KH2PO4 25mL by
8. deliquescent, No reaction
using measuring pipette and
odorless
pipette bulb

In the separatory
Appearence:
funnel the methylen
Blue coloured
Add 10 mL of MB solution blue doesn’t mix well
solution
9. by using measuring pipette with the standart
Clarity:
and pipette bulb, mix well solution or with the
clear without any
sample even after
particles
being inverted
 The liquids form
distinct physical
Add 25mL chloroform using Appearence: layers, with the less
pipette bulb and measuring Colorless dense dense liquid (blue
10. pipette. Divided into 3 times liquid coloured, organic
which is 8mL, 8mL, and 9mL Odor: solvent) floating and
addition Heavy, ethereal odor more dense sinking
(colorless, aqueous
solution)

While a separatory
funnel is being The top must be
shaken, the two opened while
Slowly swirl the solution in
solvents mix and releasing the lower
the separatory funnel by
share a large surface phase to allow
11. opening the stopper at the
area, which allows pressure equalization
top of the funnel routinely
each solute to between the inside of
while mixing
migrate to the the funnel and the
solvent in which it is atmosphere
more soluble

Drain off the aqueous layer

into the erlenmeyer flask.
Collect all the aqueous layer The aqueous layer
It turns out to lightly
12. of 3 times extraction (8mL, doesn’t mixed well
blue colored solution
8mL, 9mL) in the same with the wash solution
erlenmeyer flask for the
same concentration
Discard the organic solvent
that remains in the The separatory
13. No reaction
separatory funnel in the funnel emptied
seperate vessel
 Pour the collected aqueous There is no
layer into the separatory
14. significant changes No reaction
funnel to be extracted once
in appearance
again

The two layer

Swirl well for approximately doesn’t have a big
2 minutes, and drain off the The two layer begins
15. difference each
bottom layer into a clean to seperate
other, they are
volumetric flask
lightly blue coloured

Add more chloroform until

the total volume in the There is no
16. volumetric flask is 25 mL, significant changes No reaction
give lable for each in appearance
volumetric flask

Calibration the
spectrophotometry by using
17.
chloroform as the blanko
solution, fulfill the cuvette

Reading result:
Discard the blanko solution, 0,5 ppm = 0,068 A
refill the cuvette with all the 1 ppm = 0,258 A
18. extracted solution
1,5 ppm = 0,218 A
respectively, put in the
spectrophotometry 2 ppm = 0,246 A
Sample = 0,295 A
 Make a calibration curve to
19. determine the concentration (Discussed below)
of the sample

3.2 Discussion

On Friday, 20th of April 2018, conducted an experiment in Laboratory Water of Treatment,

about Extraction in Surfactant Analysis. The objective of this experiment was to comprehend basic
principle of extraction application for organic analysis in water sample and able to determine
limited reagent or excess reagent, and its functionality in the analysis. In this experiment, we
used a set of UV-vis spectrophotometer, separatory funnel, volumetric flask 25 mL,
erlenmeyer flask 50 mL, measuring pipette, dropper pipette, and cuvette.

We started the experiment by preparing a series standart solution of LAS in final

concentration of : 0,5M, 1M; 1,5M; and 2M. Here is the calculation using dillution equation :
M1 = 10 mg/L (initial molarity of LAS solution, before being added)
V2 = 25 mL (final volume of LAS solution, after being added)

For M2 = 0,5 M : For M2 = 1,5 M

V1 x 10 = 25 x 0,5 V1 x 10 = 25 x 1,5
V1 = 1,25 mL V1= 3,75 mL
For M2 = 1 M : For M2 = 2M
V1 x 10 = 25 x 1 V1 x 10 = 25 x 2
V1 = 2.5 mL V1 = 5 mL
So, to get the desired concentration, we should add the indicated amount of LAS solution
based on the calculation above. Label each concentration, let them aside. Do the same way in
preparing the sample with unknown concentration. Take 25 mL of sample by measuring pipette and
pipette bulb into the volumetric flask 25 mL. Transfer all the solution from the volumetric flask to
the erlenmeyer flask.
Next step is adding 5 drops of phenolphtalein indicator into each erlenmeyer flask.
Phenolphthalein is an acid-base indicator which is colorless in acid solution, but turns pink to red as
the solution becomes alkaline. Then add dropwise of NaOH into each flask, the amount depends on
the presence of pink coloured solution. Here we added 1 drop for each LAS soution, and 3 drops for
the sample solution.
After that, adding 2 drops of H2SO4 for LAS 0,1 ppm; 2 drops for LAS 1,5 ppm; 1 drop for 1
ppm; 2 drops for LAS 2ppm; and 2 drops for sample solution. Use the measuring pipette and pipette
bulb. NaOH is highly soluble in water, insoluble in ether and other non-polar solvents, liquid, white,
waxy, opaque crystal, odorless. The colour of the solution turns from pink to colorless, which
indicates the solution becomes alkaline. Then adding H2SO4 in several amount of dropwise by
using measuring pipette and pipette bulb: 2 drops for 0,1 ppm LAS; 2 drops for 1,5 ppm LAS; 1 drop
for 1 ppm LAS; 2 drops for 2 ppm LAS, and 2 drops for sample solution. This addition causes the
solution turn to colorless.
Continued by pouring each solution in seperate separatory funnel. Fill the emptied
erlenmeyer flask with KH2PO4 25mL by using measuring pipette and pipette bulb, let it aside. Next,
add 10 mL of Methylen Blue (MB) solution into the separatory funnel. The methylen blue doesn’t
mix well with the standart solution or with the sample even after being inverted and the mixture is
dark blue colored solution.
Add 25mL chloroform using pipette bulb and measuring pipette. Divided into 3 times which
is 8mL, 8mL, and 9mL addition. The funnel is then closed and shaken gently by inverting the funnel
multiple times; if the two solutions are mixed together too vigorously emulsions will form. So the
tap carefully opened to release excess vapor pressure. The separating funnel is set aside to allow for
the complete separation of the phases.
The liquids form distinct physical layers, with the less dense liquid (blue coloured, organic
solvent) floating and more dense sinking (colorless, aqueous solution). The aqueous phase drained
out to the erlenmeyer flask containing KH2PO4 through a valve away from the organic phase,
which remains in the separatory funnel. The stopcock is closed and the upper layer is poured out
through the top into another container.
The emptied separatory funnel is then filled back with the extracted solution for further
extractions with additional batches of KH2PO4 solution. The bottom layer is drained out to a clean
volumetric flask, next add more chloroform until the total volume in the flask is 25 mL. Give lable to
each of them based on its concentration of LAS solution.

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