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Indian Journal of Chemical Technology

Vol. 12, March 2005, pp. 145-148

Spectrophotometric determination of anionic surfactants in wastewater using

acridine orange
Asok Adak, Anjali Pal & Manas Bandyopadhyay
Department of Civil Engineering, Indian Institute of Technology, Kharagpur 721 302, India
Received 29 July 2004; revised received 24 December 2004; accepted 10 January 2005

A simple and rapid spectrophotometric method for the determination of sodium dodecyl sulphate (SDS), an anionic
surfactant (AS) is developed. SDS forms a yellow coloured complex with acridine orange (ACO), a cationic dye. The
complex is extracted in toluene. The absorbance of the complex in toluene layer is measured at maximum wavelength (max)
of 467 nm. The calibration curve is linear in the range of 0 6.0 ppm of SDS concentration. The method is compared with
the standard MBAS method in terms of time, ease and accuracy. It is noticed that the present method is much easier and less
time consuming. The method requires chemicals, which are less toxic than those required for the standard method. The
method is applicable to wastewater analysis.

Keywords: Anionic surfactant, sodium dodecyl sulphate, cationic dye, acridine orange, spectrophotometry.
IPC Code: G01J 3/00; C11D 1/02

Surfactants produce intensive pollution of water methods reported so far involving photometric
reservoirs1. Some surfactants are biodegradable under extraction are time consuming and sometimes they
the aerobic conditions, but many are not. The anionic involve toxic chemicals. For example, in methylene
surfactants (AS) are the main constituent of most of blue active substance (MBAS) method chloroform,
the synthetic detergents. Waste effluents from textile which is carcinogenic, is used as extracting solvent.
and organic synthesis production plants and factories, Adjustment of pH is also a big problem in this
may contain up to 2.5-10 g/L of AS2. The method. Here methylene blue forms a complex with
predominant class of AS used in these days includes anionic surfactant, which is extracted three to four
linear alkylbenzene sulphonates and linear alkyl times in chloroform layer. The total time required in
sulphates. Surfactants are harmful3 to human beings, this method is ~90 min14. Thus, elaborating simple,
fishes and vegetation and they act synergistically with rapid and selective method for surfactant
other toxic chemicals present. Moreover, hydrophobic determination is still a challenge to the chemists.
toxic chemicals are largely accumulated in water- Recently, methods using microbial sensor15,
bodies containing surfactant. Thus, many amperometric biosensor16, ion-pair formation with
environmental and public health regulatory authorities insitu flow injection analysis utilizing dynamic
have fixed stringent limits for anionic detergents as surface tension detection17, ion selective electrode18
standard 0.5 mg/L for drinking water and relaxable up and ion-pairing chromatography with suppressed
to 1.0 mg/L for other purposes. conductivity detection19 are also developed. But all
The methods for the determination of AS involve these methods need sophisticated instrumentation and
various techniques viz., gas chromatography4, high expertise. Water pollution due to anionic surfactants
performance liquid chromatography5,6, extraction (AS) is a matter of grave concern in rural areas,
photometry7-10 and extraction fluorometry11-13 using because the people in rural area drink either surface
various dyes such as methylene blue, ethyl violet, water such as pond water or river water, or well water
rhodamine B, rhodamine 6G, safranine-T, acridine directly without proper treatment. They wash their
orange, etc. The conventional techniques of surfactant clothing etc. directly in the pond water thereby
detection, which are mainly based on causing pollution in the surface water. Therefore, a
chromatography, need expensive instrumentation. The simple method applicable to field itself for the AS
____________________________ determination could be of much help to determine the

For correspondence (E-mail: suitability of the water for drinking purpose.

Acridine orange (ACO) has the potential for being toluene, chlorobenzene, 1,2-dichloroethane, a mixture
used as an ion-pairing agent with AS. Its fluorescence of benzene and isobutyl methyl ketone, etc. have been
property has been used recently for developing a tested for extraction purpose. Among these, toluene is
fluorometric method for arsenic determination20,21. A selected because of its lower toxicity and high
fluorescence-based method for AS determination has extraction efficiency. It was observed earlier22 that the
also been developed22. Here, a very simple and quick ion-association complex formed between SDS and
spectrophotometric method for AS determination ACO could be extracted in toluene efficiently and the
using ACO is reported. This method can either be recovery was in the range of 98-104%.
used for the qualitative or a yes/no type detection of In this procedure, extraction of the ion-association
AS using a colour chart, or for the quantitative complex is an important step. The extraction should
determination of AS using a spectrophotometer. The be efficient and quick. There are methods, which
method does not involve any toxic/carcinogenic often require 1090 min shaking and/or standing8,9,12.
chemical like chloroform (which is used in the In some cases tedious centrifugation step is also
standard method of AS determination). The method is necessary11. In the present method, however, a
suitable for field and real sample analysis. shaking time of 1 min and a retention time of 5 min
are quite sufficient for the efficient extraction and
Experimental Procedure phase separation. Also there is no need of using the
Acridine orange [(3,6-bis(dimethylamino)] salting-out agent (i.e. Na2SO4)8,9 and washing of the
acridine, LOBA Chemicals) is used for the organic layer8. The procedure is advantageous
preparation of approximately 5 10-3 M solution to because the extraction is quick as compared to other
be used as a stock. Sodium dodecyl sulphate (SDS) is methods.
purchased from Fluka and dried at 50 oC under To test the best efficiency of toluene for the
reduced pressure before use. The critical micelle quantitative recovery of the ion-association complex
concentration (CMC) of SDS is 8.1 10-3 M (~ 2333 in the present method, a study has been performed
ppm). Toluene (BDH) is used as an extractant. All using different volumes of toluene keeping the
chemicals used are of analytical reagent grade and are volume of water constant. The water : toluene (v/v)
used without further purification. Water is double ratio is varied from 1:1 to 1:0.1. It is observed that
distilled. water : toluene (v/v) 1:1 and 1:0.5 show complete
Sample solution (10 mL) containing SDS recovery of the complex. Therefore, water : toluene
(concentration range of 0.1 6.0 ppm) is taken in a (v/v) 1 : 0.5 was used for the detailed study.
separating funnel. Acridine orange and glacial acetic Each time 5 mL of toluene is added to the water
acid (100 L each) are added followed by the addition solution during the extraction of the complex. It is
of 5 mL toluene. The contents are shaken for 1 min important to see whether after extraction and
and then allowed to settle for 5 min. The aqueous collection also, the volume remains the same or not.
layer is discarded. The toluene layer is collected and After the contents are shaken for 1 min and are settled
2.5 mL of this solution is used directly for the for 5 min the toluene layer containing the complex is
absorbance measurement at a wavelength of 467 nm. collected and the volume is measured. For five such
Results and Discussion individual experiments, it is noticed that for four
Anionic surfactants form 1:1 complex with cationic experiments the volume of toluene is 5.0 mL and for
dyes. This complex is called ion-association complex. one experiment it is 4.8 mL. This indicates that there
The main criterion for this method is that the dye is not much change in the volume of toluene before
should be sufficiently soluble in water so that the and after the extraction. The absorbance
excess dye can be disposed off easily and the ion- measurements are thus performed directly with the
association complex should have large extractability toluene layer.
in a suitable organic solvent. At low pH this ion- It is observed that the optimum concentration range
association complex is extracted in an organic layer of ACO for 10 mL of water sample containing 0.1-6.0
and the excess dye remains in the water layer. ppm SDS is 110-5 - 110-4 M. Hence, the final
Another important criterion is to select a suitable concentration of ACO is maintained as 510-5 M in
organic solvent for which the phase separation is all cases. Glacial acetic acid is used to maintain the
quick. Various organic solvents such as benzene, pH during extraction and it is noticed that acetic acid

20-150 L per 10 mL of sample solution is optimal

for the purpose.
The absorption spectra of the toluene layer
corresponding to 3.0 ppm (present in water, 10 mL) of
SDS concentration (curve A) and the blank (curve B)
are shown in Fig. 1. The absorbance has a direct
relation with the SDS concentration. A calibration
curve is drawn by plotting absorbance versus different
concentrations of SDS in the range of 0-6.0 ppm
present in the water solutions. The equation of the
calibration curve is Abs = 0.0629 C (ppm) 0.045.
The correlation coefficient, the limit of detection
(calculated as 3SB/m; where SB and m represent
standard deviation of the blank and slope of the Fig. 1 Absorption spectra in toluene of the ion-association
complex formed between SDS (3.0 ppm) and ACO (curve A) and
calibration curve respectively) and the relative
reagent blank (curve B)
standard deviation is 0.9936, 0.04 ppm and < 5%
Table 1 Effect of coexisting substances on SDS
The method can be used as a quick yes/no type determination
study and also for on-field qualitative analysis of SDS Maximum tolerable
and other AS if the colour chart of the standard Interfering substance Added as
concentration (ppm)
solutions is prepared and compared. The sample Cl- NaCl 1000
solution treated in the same way gives a qualitative SO42- Na2SO4 500
measure of the AS concentration without measuring NO3- NaNO3 1000
the absorbance in the spectrophotometer. As Ca2+ CaCl2 100
mentioned earlier this type of analysis in villages was Fe2+ FeSO4 50
carried out for arsenic detection20 by using colour Fe3+ Fe2(SO4)3 50
charts. The described method can be applied for the As3+ NaAsO2 5
quantitative measurement of AS also using a As5+ NaAsO3 5
spectrophotometer in the laboratory. Hg2+ HgSO4 10
A detailed study on the effects of the coexisting Cr6+ K2Cr2O7 10
ions is also made. The limiting concentrations of the Zn2+ Zn(NO3)2 10
constituents those do not interfere in the Cu2+ CuSO4 10
determination of SDS are shown in Table 1. The ions Ni2+ Ni(NO3)2 10
are added as their salts in the SDS solution of 2.0 ppm Phenol 100
concentration. Absorbance variation up to 5% was Atrazine 10
considered as tolerance limit. 2,4-D 10
The applicability of the method for a practical Endosulfan 10
sample of domestic water has been examined. For that Humic acid 10
purpose the laboratory tap water (Source: Kangsabati
River), which is free from SDS, has been spiked with
known amount of SDS and the SDS concentration is
found out using the described method. The results
(Table 2) show good agreement (<5.0%) between
the measured concentration and the known value.
River or pond water sometimes contains large amount
of colloidal suspensions, which may produce
emulsion between the aqueous and organic phases
resulting in longer separation time.
Laboratory tests have been conducted to make a
comparative study between the present method and

the standard MBAS method14 used for AS measurement of AS using a colour chart or for the
determination. Same sample (synthetically prepared quantitative spectrophotometric determination of AS.
by adding SDS in tap water) have been tested using The method is very simple, rapid, reliable and
both the methods. The results obtained are applicable to natural water. The method has been
comparable (Table 2). The percentage of error lies compared with the standard method of AS
within 5.0% using both the methods. However, the determination.
present method is more advantageous and convenient
compared to the standard method because of certain References
reasons. It is easy and less time consuming. In the 1 Berth P & Jeschke P, Tenside Surfactant Deterg, 26 (1989)
standard method chloroform, a well-known 75.
carcinogen is used as an extracting solvent. Moreover, 2 Stavskaya S, Udod V, Taranova L & Krivez I,
several washing steps and the maintenance of pH are Microbiological Clean-up of Waters from Surfactants
(Naukova dumka, Kiev), 1988.
needed for the standard method. All these steps 3 Sing B P, Indian J Environ Prot, 14 (1994) 748.
enhance the cost of analysis. 4 Swisher R D, Surfactant Biodegradation, Surfactant Science
All the above experiments have been performed Series 18, 2nd edn, edited by Schick M J & Fowkes F M
with SDS as a representative of AS. Other anionic (Marcel Dekker, New York), 1987.
5 Nakae A, Tsuji K & Yamanaka M, Anal Chem, 52 (1980)
surfactants such as sodium decyl sulphate and sodium 2275.
dodecyl benzene sulphonate have also been examined. 6 Nakae A, Tsuji K & Yamanaka M, Anal Chem, 53 (1981)
They are found to respond well. At the concentration 1818.
level of 1.99 ppm of both the surfactants, the 7 Abbot D C, Analyst, 87 (1962) 287.
measured concentrations have been found to be 2.15 8 Yamamoto K & Motomizu S, Analyst, 112 (1987) 1405.
9 Motomizu S, Fujiwara S, Fujiwara A & Toel K, Anal Chem,
and 2.11 ppm respectively (within 8% error). 54 (1982) 392.
Laboratory tests have been carried out to determine 10 Frigola Canoves A M & Bosch Serrat F, Affinidad, 44 (1987)
the AS concentration in real wastewater collected 483.
from Dhobi Ghat near Kharagpur railway station. The 11 Rubio-Barroso S, Gomez-Rodriguez M & Polo-Diez L M,
Microchem J, 37 (1988) 93.
samples are analyzed using present method, 12 Gao J & Yu F, Fenxi Huaxue, 19 (1991) 1395.
fluorimetric method22, and standard method14. The 13 Rubio-Barroso S, Rodriguez-Gamonal V & Polo-Diez L M,
samples have been diluted to bring the concentration Anal Chim Acta, 206 (1988) 351.
range suitable for the methods. The measured 14 Standard Methods for the Examination of Water and
Wastewater, 17th edn, APHA-AWWA-WPCF, 1989.
concentrations found (in the parent water sample) are
15 Nomura Y, Ikebukuro K, Yokoyama K, Takeuchi T,
116.05, 107.44 and 112.39 ppm for present, Arikawa Y, Ohno S & Karube I, Anal Lett, 27 (1994) 3095.
fluorimetric and MBAS methods respectively. The 16 Reshetilov A N, Semenchuk I N, Iliasov P V & Taranova L
results are comparable. A, Anal Chim Acta, 347 (1997) 19.
17 Young T E & Synovec R E, Talanta, 43 (1996) 889.
Conclusion 18 Alegret S, Alonso J, Bartroli J, Baro-Roma J, Sanchez J &
A simple method is developed for the delValle M, Analyst, 119 (1994) 2319.
determination of anionic surfactant (AS). An ion- 19 Ye M Y, Walkup R G & Hill K D, J Liq Chromatogr, 17
association complex is formed between an anionic (1994) 4087.
20 Pal T, Jana N R & Sau T, Anal Proc, 32 (1995) 369.
surfactant and a cationic dye e.g. acridine orange
21 Pal A, Jana N R, Sau T, Bandyopadhyay M & Pal T, Anal
(ACO). The yellow complex can be easily extracted Commun, 33 (1996) 315.
in toluene. The coloured organic layer has been used 22 Pal A & Bandyopadhyay M, Indian J Chem Technol, 7
either for a yes/no type or for a qualitative (2000) 105.