J Surfact Deterg (2016) 19:209–218
DOI 10.1007/s11743-015-1763-x
ORIGINAL ARTICLE
Analysis of Linear Alkylbenzene Sulfonate in Laundry
Wastewater by HPLC–UV and UV–Vis Spectrophotometry
Terelle Ramcharan1 • Ajay Bissessur1
Received: 4 June 2015 / Accepted: 16 November 2015 / Published online: 22 December 2015
Ó AOCS 2015
Abstract A lack of natural water resources and an increase in
the demand for fresh potable water has shifted focus to the
possible reuse of recycled laundry wastewater water that is
considered to be relatively clean. Organic components such as
linear alkylbenzene sulfonates (LAS) are the major and most
abundant contributing anionic surfactant constituents found in
laundry detergents. The development and reliability of treat-
ment methods targeted at purification of laundry wastewater
necessitates a fast and accurate method for quantification of
LAS. This paper focuses on a comparative study for the
quantification of LAS based on traditional liquid–liquid
extraction (LLE) and HPLC–UV methods. In the case of LLE,
the anionic surfactant LAS complexes via ion association to a
methylene blue (MB) cationic dye resulting in the formation of
an anionic surfactant–methylene blue (AS–MB) complex. The
AS–MB complex extracted with chloroform absorbs at a k max
of 653 nm. Optimized conditions for quantification of a single
eluted LAS peak using HPLC–UV were obtained by isocratic
elution on a C18 column with a 95 % acetonitrile and 5 %
0.7 M acetic acid mobile phase. Both methods displayed per-
centage recoveries [90 % and statistically showed repro-
ducibility and precision in the quantitation of LAS. HPLC–UV
prevailed over UV–Vis as the method of choice for LAS
determinations given the ease of sample preparation and
applicability to a wider range of samples. Typical levels of LAS
in laundry samples assessed in this study ranged between 116
and 454 mg L-1.
& Ajay Bissessur
bissessura@ukzn.ac.za
1
School of Chemistry and Physics, University of KwaZulu-
Natal, Private Bag X54001, Durban 4000, South Africa
Keywords HPLC–UV  Laundry wastewater  Linear
alkylbenzene sulfonate  Liquid–liquid extraction  Solid
phase extraction  UV–Vis
Introduction
Water is an essential resource for the sustainability of life on
earth. With an increase in the population worldwide the demand
for water has escalated in the urban, industrial and agricultural
domains [1]. To conserve the environment and natural water
resources, recycling and re-use of wastewater is often imple-
mented in most countries [2]. Domestic and industrial laundry
wastewater is relatively ‘clean’ in comparison to other indus-
trial effluents [3]. In view of the current efforts to save the
earth’s energy and water resources, not much interest has been
shown in attempting to filter, purify and re-use domestic and
industrial wastewater [4]. Development of efficient treatment
methods for the recycling and re-use of domestic and industrial
wastewater would require fundamental knowledge of the
chemical constituents present within and adequate methods for
quantification thereof. The composition of laundry wastewater
(LWW) consists primarily of the laundry detergent utilized in
the washing process and the soil removed from the textile items.
Typical soil or more commonly known as ‘dirt’ found in LWW
originates from textile items stained primarily from food,
drinks, body soil and atmospheric dust.
These types of soils mostly contain water soluble salts of
sodium, potassium, calcium and magnesium as well as the
hydrophobic components such as cholesterol, triglycerides and
fatty acids. Metal oxides and silicates contribute to a large
percentage of atmospheric dust, hence would be common in the
textile items laundered. Surfactants are the abundant key
ingredients found in all laundry detergents which effectively
remove the above mentioned chemical constituents from the
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textile items. A considerably high concentration of surfactants
is therefore evidently expected in LWW and requires a fast and
accurate method for quantification [5]. Linear alkylbenzene
sulfonates (LAS) are the most common organic anionic sur-
factant used in laundry detergents due to its excellent perfor-
mance and relatively low cost [6–8]. The commercial product
of LAS as a sodium salt, is comprised of a mixture of homo-
logues containing between 10 and 14 linear carbon atoms with a
phenyl group attached to the linear alkyl chain and the sulfonate
anion (Fig. 1), [9, 10].
The current methods implemented in literature for the
quantification of LAS include UV–Vis spectrophotometry [11,
12] and liquid chromatography [13, 14]. The spectrophoto-
metric method for determination of anionic surfactants pre-
sented by Chitikela et al. [12] and Jurado et al. [11] involves the
use of methylene blue, a cationic dye which forms a complex
with the anionic surfactant through an ion association.
The anionic surfactant–methylene blue complex (AS–
MB) is extracted by an organic solvent such as chloroform
and quantified by UV–Vis spectrophotometry. Both the
anionic surfactant and cationic dye are insoluble in chloro-
form as individual compounds, however when complexed
through ion association, the AS–MB complex is more sol-
uble in chloroform when compared to the aqueous phase.
½AS  MBaq  ½AS  MBCl ð1Þ
Equation 1 denotes the ionic pair AS–MB at equilibrium
where the subindex ‘‘aq’’ indicates the concentration of the
anionic surfactant LAS in the aqueous phase, and the subindex
‘‘Cl’’ represents the concentration of the surfactant in the
chloroform phase. Methylene blue reacts with the anionic
surfactant LAS in a 1:1 mol ratio. It is noted in the literature that
the partition coefficient for this specific extraction is extremely
favorable allowing for a single extraction with chloroform [15].
To support a favorable partition coefficient the distribution ratio
can be determined experimentally. The distribution ratio is
defined as a ratio between the total concentration of the analyte
Fig. 1 Chemical structure of
linear alkylbenzene sulfonate
C10H 21
(LAS)
SO3 - Na +
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J Surfact Deterg (2016) 19:209–218
in phase 1 and the total concentration of the analyte in phase 2
[16]. The distribution ratio for the AS–MB complex can be
given by Eq. 2, were the numerator denotes the concentration
of AS–MB in the chloroform phase (phase 1) and the denom-
inator denotes the concentration of AS–MB in the aqueous
phase (phase 2). For the analysis of LWW samples, an appro-
priate dilution factor has to be taken into consideration since
LWW samples are known to contain a high concentration of
surfactants. An indication of complete extraction of methylene
blue into the chloroform phase, leaving the aqueous phase
colorless suggests that the sample size was too large and
requires further dilution. The pH of the reaction also plays an
important role in the LLE of AS–MB complexes. At a high pH
([8) the methylene blue cationic dye loses its cationic character
and at a too low pH (\3) the protonation of the anionic sur-
factant becomes a competitive process to ion pair formation.
½AS  MBCl
D¼ ð2Þ
½AS  ½MBaq
Quantification of LAS as an AS–MB complex is less
commonly used due to the numerous chemical interfer-
ences associated with this method. Anionic compounds
other than surfactants in samples analyzed may complex
with the cationic dye thus resulting in a false presentation
of the quantity of LAS in samples.
In contrast, little chemical interference is associated with the
quantification of LAS by liquid chromatography. The sample
preparation, HPLC column and detector utilized isolates the
targeted analyte, hence allowing for specific quantification of
LAS. Solid phase extraction (SPE) is currently the most widely
used technique for isolation of surfactants from aqueous sam-
ples [17]. In the work presented by Matthijs and De Henau, SPE
was used for the efficient isolation of LAS from aqueous
environmental samples [18]. Washing of the SPE cartridges
with a 40 % aqueous MeOH solution prior elution of LAS with
MeOH, was essential for elimination of polar substances [18].
Reversed phase chromatography with a C18 solid phase
extraction (SPE) cartridge and UV detector has been reported
by Wangkarn et al. [14], Akyuz et al. [13], Guo et al. [19] and
Villar et al. [20] for the quantification of LAS [17]. The struc-
ture of LAS comprising of different homologues complicates
the quantification of LAS through chromatographic methods.
Majority of the work presented on quantification of LAS by
HPLC, focused on separation of the homologues of LAS as it
was a necessity for assessment of the environmental fate of
LAS. Many authors have isolated the homologues of LAS as
individual peaks, and quantified LAS as a sum of the homo-
logues [13, 14, 19, 21]. Separation of the homologues requires
the addition of phase modifiers such as sodium chloride,
ammonium acetate or sodium acetate to be added to the mobile
phase [14]. Good peak resolution of LAS and a short retention
time is difficult to obtain due to the similarities between the
homolog structures. As reported by Akyuz et al. [13],
J Surfact Deterg (2016) 19:209–218
Bengoechea et al. [21], Guo et al. [19]. and Wangkarn et al.
[14], the retention time of LAS homologues was 50, 22, 15 and
8 min respectively.
The paper presented focuses on the quantification of
LAS in LWW collected from domestic sources and a
comparison of methods by UV–Vis spectrophotometry and
reversed phase HPLC–UV. For the chromatographic
method, quantification of LAS is optimized by elution of
LAS as a single peak with a reduced retention time.
Experimental Section
Chemicals
HPLC grade solvents acetonitrile and methanol as well as
analytical grade methylene blue and LAS (sodium dodecyl-
benzene sulfonate) comprising of a mixture of homologues
was purchased from Sigma AldrichÒ. Sep-pakÒ 360 mg C18
SPE cartridges were purchased from Waters. Analytical grade
acetic acid, phosphoric acid, chloroform, sodium hydroxide
and sodium tetraborate were purchased from Associated
Chemical EnterprisesÒ, MerckÒ, Set Pure ChemicalsÒ, Pro-
mark ChemicalsÒ and SAAR ChemÒ respectively.
Sample Collection and Storage
Replicates of LWW were sampled from a domestic
washing machine. The washing machine program com-
prised of one wash cycle followed by two sequential rinse
cycles. Samples were collected in amber glass bottles after
each wash and rinse cycle, and refrigerated. The samples
referred to as 1, 2 and 3rd rinses correspond to water dis-
posed from the 1st wash cycle, 1st rinse cycle and 2nd rinse
cycle of an automated washing machine respectively.
Qualitative Analysis
LAS standards and LWW samples were analyzed by
FTIR–ATR using a 100 Perkin Elmer spectrometer.
Quantitative Analysis of LAS in LWW by UV–Vis
(Spectrophotometric)
LWW samples were diluted and followed a procedure in
accordance to the protocol reported by Jurado et al. [11]
with minor changes. A 5.0-mL sample aliquot was alkalin-
ized with 200.0 lL of 50 mM sodium tetraborate buffer at
pH 10.5 followed by the addition of 100.0 lL methylene
blue reagent prepared in 10 mM sodium tetraborate buffer at
pH 5.5. The AS–MB complex was extracted with 5.0 mL of
chloroform and its UV–Vis absorbance was measured using
211
a Lambda 35 UV–Vis Perkin Elmer double beam spec-
trometer against a prepared blank.
An external standard calibration was set-up with LAS
standards ranging from 0.25 to 2.5 mg L-1. For the recovery
study, samples were spiked with known concentrations LAS.
The stability of the LAS was evaluated by analyzing samples
over 3 different time intervals (1, 2 and 5 days apart).
Quantitative Analysis of LAS in LWW by
HPLC–UV
HPLC–UV analysis was carried out using a Perkin Elmer
200 series pump with a UV detector set at 225 nm and a
20-lL injector loop. A C18 PhenomenexÒ column with
dimensions of 250 9 4.6 mm and an AgilentÒ C18 eclipse
plus column with dimensions of 2.1 9 100 mm was uti-
lized as the stationary phase for HPLC.
LWW samples were diluted and subjected to SPE with a
C18 cartridge in a vacuum manifold prior to HPLC–UV
analysis. The cartridge was conditioned and equilibrated
with 2 mL each of acetonitrile and Millipore water respec-
tively. A volume of 5.0 mL of the sample was loaded onto
the cartridge followed by 3 mL of a wash solution com-
prising of 70 % Millipore water and 30 % (v/v) methanol.
The C18 cartridge was dried under vacuum for a period of
10 min prior to elution of the analyte with acetonitrile. A
calibration curve was prepared by analyzing LAS standards
ranging from 5 to 30 mg L-1. All standards were prepared
in 100 % acetonitrile. Recovery studies and precision anal-
yses by HPLC–UV were performed analogously to the UV–
Vis method [described in Quantitative analysis of LAS in
the LWW by UV–Vis (spectrophotometric) section].
For sample preservation, LWW samples were collected
(described in the sample collection and storage section) and
stored in 15 % (v/v) MeOH at 4 °C. The LAS was quan-
tified after days 1, 2 and 5 using the HPLC–UV method B.
Limit of Detection and Limit of Quantification
The theoretical LOD (limit of detection) was calculated as:
3r
LOD ¼ ð3Þ
N
where r and N, in the above equation represent the standard
deviation of the y-intercept and slope of the calibration curve
respectively. The LOQ (limit of quantification) was calcu-
lated according to Eq. 3; however 10 r was used instead of
3 r. The MLOD (method limit of detection) and MLOQ
(method limit of quantification) were experimentally deter-
mined by preparation of LAS standards with concentrations
corresponding to the LOD and LOQ respectively and
quantified by UV–Vis spectrophotometry and HPLC–UV.
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Results and Discussion
Qualitative Analysis of LAS
LAS in laundry wastewater samples were qualitatively
assessed by HPLC–UV and Infrared spectroscopy. An
established kmax of 225 nm for LAS was carried out by
scanning a dilute standard solution between 200 and
700 nm. The chromatographic analysis of standard LAS
and LWW produced a single symmetrical peak respec-
tively with a consistent flat baseline (Fig. 2). The reported
sample retention time of 0.73 min (Fig. 2a) compared

Fig. 2 Chromatograms showing peaks of LAS in a LWW, b LWW spiked with 5 ppm standard LAS and c LWW spiked with 15 ppm standard LAS

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J Surfact Deterg (2016) 19:209–218 213

Table 1 Chromatographic conditions for HPLC analysis [22]. Sharp, definite absorption bands at 1012 and
Method Column Mobile phase Flow rate
1043 cm-1 for the LAS standard (Fig. 3) corresponds to
length (mm) (mL min-1) the presence of an alkylbenzene sulfonate group according
to the ASTM referenced assigning of 1010 and 1042 cm-1.
A 25 ACN/H2O (19:1) 1
Absorption bands at 1010 and 1042 cm-1 respectively
B 10 ACN/0.7 M Acetic 0.5 relates to the stretching between the sulfur atom and aro-
acid (19:1)
matic carbon, and the stretching between sulfur and oxygen
in LAS. Other identifiable absorption bands of LAS namely
aromatics, sulfonate groups and para-substituted sulfonates
Table 2 ASTM guideline to characteristic FTIR absorption bands of at 1497, 1128 and 830 cm-1 respectively are shown in
LAS
Table 2.
Frequency Band Band Functional group Isolation of the dissolved form of LAS from LWW
(cm-1) shape intensity samples is difficult and requires tedious techniques such as
1493 Shoulder Weak Aromatic bands sublation. Consequently, in order to obtain a suitable and
1235–1176 Broad Strong Sulfonate group representative FTIR spectrum of the LAS standard (Fig. 4)
1136 Sharp Moderate Sulfonate group it was dissolved in AR grade methanol. Characteristic
1042 Sharp Strong Alkylbenzene sulfonate absorption bands observed at 1013.76 cm-1 for LWW
1010 Sharp Strong Alkylbenzene sulfonate sample and 1015.83 cm-1 for LAS standard corresponds to
833 Broad Moderate Para substitution the presence of an alkylbenzene sulfonate group. The
impact of methanol as a solvent system resulted in broad,
favourably to LAS standard retention time of 0.71 min overlapping IR absorption bands at 3258.61 and
(Fig. 2b, c). The retention times of the analyte and standard 3276.88 cm-1, which corresponds to the presence of a
LAS served as a qualitative tool in recognizing LAS in the hydroxyl group. This was not the case for the pure solid
laundry wastewater samples, while spiking of LWW sam- form of LAS.
ples with LAS standard shown in Fig. 2 served to highlight
the purity of LAS. Quantitative Analysis of LAS
The characteristic absorption bands of functional groups
by FTIR–ATR obtained for LAS in both standards and The distribution ratio of the AS–MB complex in the organic
samples were compared to an ASTM guideline which lis- and aqueous phase was calculated to ensure that a single
ted the characteristic absorption bands of LAS (Table 2), extraction was appropriate for quantitative analysis of LAS

Fig. 3 FTIR spectrum of LAS standard in its pure and original solid form

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214
Fig. 4 FTIR spectrum of a dissolved LAS standard and b laundry wastewater sample
in LWW samples. The distribution ratio of 0.99 with a RSD
of 0.92 is indicative that a single extraction of the AS–MB
complex into the chloroform phase was sufficient.
The quantification of LAS using chromatographic
methods A and B (Table 1) was favorable as single peak
elutions (Fig. 5) were obtained thereby increasing the
precision of the analysis by simplifying peak integration.
Optimization of quantification for LAS using reversed
phase HPLC with UV detection was accomplished by
varying the length of the HPLC column and the mobile
Fig. 5 Chromatograms of LAS in laundry wastewater samples
analyzed by method A and method B using HPLC–UV
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phase. The shortened column reduced the retention time of
LAS from 2.40 to 0.68 min and decreased the flow rate by
0.5 mL min-1. Although similar peak resolutions were
obtained from both methods usage of method B (Table 1)
was favored due to minimum solvent consumption.
The validity of the research methodologies were asses-
sed by the limit of detection (LOD), limit of quantification
(LOQ), method limit of detection (MLOD), method limit
of quantification (MLOQ), reproducibility and percentage
recovery. The LOD, LOQ, MLOD and MLOQ analyses
(Table 3) of both the UV–Vis and HPLC–UV methods is in
effect a measure of the sensitivity. Although a lower LOD
and LOQ was obtained for UV–Vis analysis (Table 3), the
R2 value for the HPLC–UV method (0.9970) was closer to
1 when compared to the UV–Vis method (0.9808). This
shows that there is a greater standard deviation associated
between the correlation of the concentration of LAS and
the response from the UV–Vis instrument, when compared
to the HPLC–UV method.
Optimization of Storage of LWW Samples
and Reproducibility of Results
LWW samples for rinse one stored at 4 °C for a period of
5 days with no added preservative showed a decrease in
concentration of LAS by 31.84 % while a 9.07 % decrease
was observed for samples stored in 15 % MeOH at the
same temperature (Fig. 6).
Under aerobic conditions it has been found that LAS
degrades into products such as those listed in Fig. 7, [23].
J Surfact Deterg (2016) 19:209–218 215

Table 3 Experimental and

LOD (mg L-1) MLOD (mg L-1) LOQ (mg L-1) MLOQ (mg L-1) R2
theoretical LOD and LOQ for
both methods of analysis UV–Vis 0.03260 0.1656 0.1090 0.2396 0.9808
HPLC 0.06970 0.07320 0.2320 0.2690 0.9970

500 within 2 days. After 2 days of storage, the average per-

Concentration of LAS (mg L-1)

centage decrease of LAS in LWW samples for both

450
400
350
300
Preserved
250 No added
preservative
200
0 1 2 3
Period of time samples stored (days)
Fig. 6 The effect of methanol as a preservative for storage of LWW
samples
Microbial growth within the LWW further increases the
oxidizing conditions thereby increasing the rate of LAS
biodegradation. The addition of methanol to the LWW
inhibited microbial growth thus illustrating the importance
of the addition of a sample preservative for storage at 4 °C
over a period exceeding 24 h.
The above results correspondingly show the repro-
ducibility for the quantification of LAS in rinse one by
HPLC–UV. Similarly the reproducibility of the UV–Vis
and HPLC–UV methods was statistically assessed by
ANOVA for rinses 1–3 by UV–Vis and rinses 2–3 for
HPLC–UV (Table 4). Although the statistic results shown
in Table 5 indicates no significant difference whereby the
F-calculated \F-critical and p [ 0.05, it is recommended
that LAS quantification in LWW samples are carried out
methods was \5 %, whereas the average percentage
decrease for samples stored for 5 days was [10 %. All
quantitation was performed in triplicate with a relative
standard deviation (RSD) \5 %.
Percentage Recoveries of LAS
The percentage recovery for both methods with the excep-
tion of rinse one analyzed by UV–Vis, was [90 %, thus
4 5
indicating the efficiency of the methods (Table 6). The
LWW disposed after the 1st rinse is heavily polluted hence
hindering the percentage of recovery (74.10 %, Table 6) of
LAS by UV–Vis. In comparison, the percentage recoveries
of the 2nd and 3rd rinses showed much better results (108.35
and 99.89 % respectively). This decrease in percentage
recovery for rinse one is largely due to increased levels of
silicates, and water soluble salts of sodium, potassium, cal-
cium and magnesium that persist in soils which are generally
removed in the first rinse. Consequently an increased amount
of anionic constituents competes with LAS for ion formation
with the methylene blue cationic dye which is more soluble
in the aqueous phase during extraction with chloroform thus
accounting for decreased levels of LAS.
Quantification of LAS in Laundry Powder
and Wastewater Samples
According to specifications by the respective manufactur-
ers of the laundry detergent, LAS contributes 5–30 % of

Fig. 7 Reaction O O
scheme showing degradation of
LAS under oxidative conditions

HO CH 3
O
R COO -
Acetoacetic acid

OH
O

HO

OH
SO3 -
Sulfophenyl Phenyl
carboxylic acid carboxylic acid O
LAS
Fumaric acid

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Table 4 Reproducibility of
Method Period Rinse 1 (mg L-1) Rinse 2 (mg L-1) Rinse 3 (mg L-1)
analysis of LAS over a time
period for UV–Vis and HPLC– UV–Vis Day 1 357.2 ± 0.14a 217.6 ± 2.54a 159.2 ± 5.02a
UV a a
Day 2 353.5 ± 1.13 208.9 ± 4.82 153.0 ± 4.97a
a a
Day 5 339.3 ± 4.20 192.6 ± 4.77 151.8 ± 2.62a
a a
HPLC–UV Day 1 454.5 ± 0.48 200.9 ± 3.89 125.9 ± 2.07a
a a
Day 2 447.6 ± 1.32 198.0 ± 4.95 118.9 ± 3.54a
Day 5 413.2 ± 1.37a 184.8 ± 3.73a 116.1 ± 3.19a
Rinse 1 = wastewater disposed after 1st wash cycle
Rinse 2 = wastewater disposed after 1st rinse cycle
Rinse 3 = wastewater disposed after 2nd rinse cycle
a
Average of triplicate analysis

Table 5 ANOVA results

Statistical parameter Rinse 1 Rinse 2 Rinse 3
measured at a 95 % confidence
interval for precision analysis of UV–Vis HPLC UV–Vis HPLC UV–Vis HPLC
LAS
F-calculated 2.445 0.678 3.941 2.153 0.6260 4.030
F-critical 5.143 5.143 5.143 5.143 5.143 5.143
p value 0.1671 0.5427 0.08073 0.1973 0.5664 0.07771

Table 6 Percentage recoveries of LAS 500

Concentration of LAS (mg L-1)

450
Method Rinse 1 (%) Rinse 2 (%) Rinse 3 (%) HPLC
400 UV-Vis
UV–Vis 74.10 ± 3.88a 108.35 ± 2.36a 99.89 ± 3.37a 350

HPLC–UV 100.49 ± 2.78a 94.85 ± 1.95a 98.98 ± 3.94a 300

250
a
Average of triplicate analysis 200
150
100
Table 7 Concentration of LAS in powdered laundry detergent ana- 50
lyzed by UV–Vis spectrophotometry and HPLC–UV 0
Rinse 1 Rinse 2 Rinse 3
-1
Method Concentration LAS (mg L ) Laundry Wastewater Sample

Theoretical value 5–15
UV–Vis spectrophotometry 11.11 ± 2.74a
HPLC–UV 12.61 ± 1.88a
a
Average of triplicate analysis
the total detergent composition. The concentrations of LAS
determined experimentally using both methods (Table 7)
falls within the acceptable range specified for laundry
powder.
For both methods of analysis a marked decrease in
concentrations of LAS ranging from rinse 1 to 3 (Fig. 8)
has been observed. A 55.80 % decrease was observed from
rinse 1 to rinse 2, as expected due to the larger amount of
LAS needed to remove sediments from soiled items during
the washing cycle. However a lower percentage decrease
was observed (37.28 %) for rinse 2 to 3, due to the rinsing
cycle of the washing machine which primarily removes the
surfactants adsorbed by the fibre. Similar percentage
Fig. 8 Concentration of LAS in laundry wastewater samples ana-
lyzed by UV–Vis and HPLC–UV
decreases from rinse 1 to 2, and rinse 2 to 3 were noted for
UV–Vis analysis of LAS (Fig. 8).
Statistically assessed results of rinses 2 and 3 by
ANOVA (Table 8) indicate no significant difference in the
concentrations of LAS quantified using both the UV–Vis
and HPLC–UV method (F-calculated \ F-critical and
p [ 0.05). This was not the case for rinse 1, as the F-
calculated [ F-critical and p \ 0.05 (Table 8). The most
contributing factor to the varying concentration of LAS in
rinse 1 is possibly attributed to the increased levels of
pollutants which negatively influence the LLE of LAS
especially in the case for the UV–Vis method. The pres-
ence of anionic compounds such as chlorides, nitrates and
sulfates etc., competes with LAS in complexing to
methylene blue thus increasing the percentage error of the

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Table 8 ANOVA data of LAS quantified by UV–Vis spectropho-
tometry and HPLC–UV at a 95 % confidence level
Rinse 1 Rinse 2 Rinse 3
F-calculated 990.1 6.108 7.193
F-critical 7.709 7.709 7.709
-6
p value 6.080 9 10 0.06884 0.05511
analysis. Solubility of these anionic methylene blue com-
plexes in the chloroform phase depend on the organic or
inorganic nature of the interfering anionic compounds.
It is therefore evident that the complexation between
methylene blue and the interfering anionic compounds
prevented ion pair formation between LAS and methylene
blue; hence the quantity of LAS extracted decreased
(74.10 % recovery, Table 6). For quantification of LAS in
heavily polluted water by UV–Vis spectrophotometry, it is
suggested that techniques such as sublation is utilized. This
technique isolates the surfactant from the water sample and
yields a dried residue relatively free of non-surfactant
substances. This reduces the error associated with the
presence of other anionic substances in the wastewater
thereby increasing the efficiency of the method.
Few errors are associated with chromatographic analysis
of LAS when compared to the UV–Vis method. The sample
preparation, HPLC column and detector, decreases the
sample matrix interferences isolating LAS. The presence of
different homologues of LAS is the major problem associ-
ated with the quantification of LAS. This affects the chro-
matographic peak quality resulting in either peak fronting or
tailing and split peaks if incorrect mobile phase is used.
Alternatively many authors have separated the homologues
of LAS and quantified total LAS in samples based on the
sum of the peak areas of the homologues [13, 19].
These homologues are structurally closely related; hence
the difficulty arises when attempting to resolutely separate
these compounds by chromatographic methodologies. The
chromatographic elution of LAS as a single resolute peak
increases the accuracy and precision of the analysis thus
increasing accuracy of the peak integration. The sample
preparation using SPE prior to the analysis by HPLC–UV
allows for pre-concentration of LAS thus enabling HPLC–
UV as a suitable and preferred method for quantification of
LAS in environmental samples. In addition the method is
less error prone and can be applied to a wider range of
samples in comparison to the UV–Vis method.
The drawbacks suffered by the implementation of UV–
Vis as a suitable method for LAS determination are sev-
eral. The sample preparation for analysis of LAS by UV–
Vis spectrophotometry is a long and tedious one. Quan-
tification of LAS in heavily polluted samples results in
217
gross errors, and furthermore requires pre-treatment such
as sublation.
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Terelle Ramcharan Terelle Ramcharan received her B.Sc. degree in
Applied Chemistry in 2011, her B.Sc. honors degree in Chemistry in
2012 and her M.Sc degree in Analytical Chemistry in 2015 from the
University of KwaZulu-Natal Westville campus. Her current research
includes the chemical analysis and treatment of wastewater.
Ajay Bissessur Ajay Bissessur obtained his B.Sc. (Hons) and M.Sc.
degrees from the University of Durban-Westville. He is employed at
the University of KwaZulu-Natal as a lecturer in Analytical and
Applied Chemistry. His research interests include renewable energy,
antioxidants, insulation materials and sulfur chemistry.