Science of the Total Environment 710 (2020) 136277

Contents lists available at ScienceDirect

Science of the Total Environment

journal homepage: www.elsevier.com/locate/scitotenv

Toxicity identification and evaluation of palm oil mill effluent and its
effects on the planktonic crustacean Daphnia magna
Yuya Hashiguchi a,b, Mohd Rafein Zakaria b,c,⁎, Toshinari Maeda a, Mohd Zulkhairi Mohd Yusoff b,
Mohd Ali Hassan b, Yoshihito Shirai a
a
Division of Environmental Bio-Adaptation, Department of Biological Functions Engineering, Graduate School of Life Science and Systems Engineering, Kyushu Institute of Technology, 2-4 Hibikino,
Wakamatsu-ku, Kitakyushu, Fukuoka, Japan
b
Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
c
Laboratory of Biopolymer and Derivatives, Institute of Tropical Forestry and Forest Products, University Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• Toxicity tests were evaluated from palm

oil mill effluent (POME) final discharge.
• Toxicity unit (TU) of whole effluent tox-
icity test for effluents were N 1.0.
• Manipulation of pH, filtration, and aera-
tion had reduced TU values of effluents.
• Volatile, and pH sensitive-substances
are possible toxicants presence in
POME.
• Phenol, Cu and Zn were potential toxi-
cants to invertebrate Daphnia magna.

a r t i c l e i n f o a b s t r a c t

Article history: Palm oil mill effluent (POME) contains complex and highly biodegradable organic matters so discharging it with-
Received 30 August 2019 out appropriate treatment may lead to environmental problems. POME final discharge quality is normally deter-
Received in revised form 21 November 2019 mined based on conventional chemical detection such as by biological oxygen demand (BOD) and chemical
Accepted 20 December 2019
oxygen demand (COD). The novelty of the present study is that the toxicity effects of the POME final discharge
Available online 26 December 2019
samples were evaluated based on whole effluent toxicity (WET) and toxicity identification evaluation (TIE)
Editor: Daqiang Yin tests using Daphnia magna. The toxicity unit (TU) values were recorded to be in the range from TU = 1.1–11 ob-
tained from WET, and the TIE manipulation tests suggested that a substantial amount of toxic compounds was
Keywords: contained in the POME final discharge. Phenol, 2,6-bis (1,1-dimethylethyl)- and heavy metals such as Cu and
Whole effluent toxicity Zn were detected in all the effluents and were recognized as being the main toxicants in the POME final discharge.
Toxicity identification evaluation GC/MS analyses also successfully identified cyclic volatile methyl siloxanes; cyclotetrasiloxane, octamethyl- (D4),
Palm oil mill effluent final discharge cyclopentasiloxane, decamethyl- (D5), cyclohexasiloxane, dodecamethyl- (D6). D4 was detected at
Phenol 0.0148–0.0357 mg/L, which could be potentially toxic. The palm oil industry used only water in the form of
Heavy metals
steam to process the fruits, and the presence of these compounds might be derived from the detergents and
grease used in palm oil mill cleaning and maintenance operations. An appropriate treatment process is thus re-
quired to eliminate these toxicants from the POME final discharge. It is recommended that two approaches,

⁎ Corresponding author at: Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
E-mail address: mohdrafein@upm.edu.my (M.R. Zakaria).

https://doi.org/10.1016/j.scitotenv.2019.136277
0048-9697/© 2019 Elsevier B.V. All rights reserved.
2 Y. Hashiguchi et al. / Science of the Total Environment 710 (2020) 136277
chemical-based monitoring as well as biological toxicity-based monitoring, should be utilized for achieving an ac-
ceptable quality of POME final discharge in the future.
1. Introduction
Palm oil mill effluent (POME) is brownish in color, has high temper-
ature, acidic pH, and contains residual oil, plant debris, and nutrients
(Ho et al., 1984). Due to the high content of organic substances in the ef-
fluent, a series of treatments such as anaerobic, aerobic, and facultative
ponds and tank systems are necessary before it can be discharged into
rivers (Poh et al., 2010). A permissible standard limit for a final dis-
charge from the palm oil industry was set by the Department of Envi-
ronment (DOE), Malaysia to control the effluent quality and avoid
polluting the water bodies. Raw POME and POME final discharge quali-
ties are characterized conventionally by using chemical analyses such as
biological oxygen demand (BOD), chemical oxygen demand (COD),
total solids (TS), suspended solids (SS), etc. The BOD, COD, TS and SS
of POME were reported to be in the range from 10,250–43,750 mg/L,
15,000–100,000 mg/L, 11,500–79,000 mg/L and 5000–54,000 mg/L, re-
spectively (Madaki and Seng, 2013).
The concentrations of phenol and other phenolic compounds were
found to be high between 200 and 1000 mg/L in POME and in the efflu-
ent from the anaerobic treatment of POME (Alam et al., 2009). Phenolic
compounds are secondary metabolites, which are produced from the
shikimic acid and the pentose phosphate of plants through
phenylpropanoid metabolization (Randhir et al., 2004). Some heavy
metals such as Fe, Cu and Zn are contained in POME at levels of
46.5 mg/L, 0.85 mg/L, and 2.3 mg/L, respectively (Madaki and Seng,
2013). The conventional analyses may be limited in the detection of
toxic chemicals and metals in the POME final discharges. Even though
the WET test has been recognized as one of the parameters to be
adopted for the best management practice of the discharge of industrial
effluent or mixed effluent it has however not been widely used in the
analysis of the POME final discharge (DOE, 2019). The present work
highlighted important findings from comparative analyses of conven-
tional chemical and biological ecotoxicity tests using Daphnia magna.
The whole effluent toxicity (WET) method was established by the
U.S. Environmental Protection Agency (USEPA) to investigate the toxic-
ity of aquatic organisms from the effluent of wastewater discharge
(USEPA, 1991). The toxicity effect of the effluent was evaluated based
on a specific bioassay using algae, invertebrates, and fish in the WET
test. The WET test is used to avoid continuous discharging of wastewa-
ter containing toxicants into rivers. This WET method has been applied
in many countries such as USA, Brazil, Denmark, and Korea (Lassen
et al., 2005; de Melo et al., 2013; Ra et al., 2016). However, the WET
test is not able to identify the causative toxicants in the effluents.
USEPA has developed a method, namely toxicity identification evalua-
tion (TIE) to determine the toxicants present by using bioassay and
physicochemical analyses (USEPA, 1991). There are three phases of
TIE manipulation, which are characterization (Phase I), identification
(Phase II) and confirmation (Phase III) of the causative toxicants in the
effluent (USEPA, 1991). The TIE procedure is highly efficient for identi-
fying toxic compounds from different types of industrial wastewater
(Lassen et al., 2005; Kusui et al., 2014). This test was successfully per-
formed on cosmetic wastewater (de Melo et al., 2013), pharmaceutical
industry effluent (Ra et al., 2016) and wastewater from other related in-
dustries (Kusui et al., 2014) in that organic chemicals, particulates, salts,
and heavy metal were identified as the causative toxicants to algae,
crustaceans and fish. To the best of the authors' knowledge, there is
no report thus far pertaining to WET and TIE tests on POME final dis-
charge and its effects on the aquatic life of the planktonic crustacean
D. magna.
© 2019 Elsevier B.V. All rights reserved.
Hence, the present study aimed to investigate the extent of the
safety for discharging POME final discharge into streams and rivers by
assessing the acute toxicities of discharge samples from different palm
oil mills by using D. magna. WET and TIE tests were conducted with
the aim to classify types of toxicants and to characterize and confirm
the presence of toxicants in the effluent samples from selected mills.
The fundamental information obtained from this study may safe water
bodies from excessive release of pollutants from POME final discharge
and allow appropriate treatment technologies to be developed in the
future.
2. Materials and methods
2.1. Sample collection
POME final discharge samples were collected from three differ-
ent mills to study the presence of toxicants and to understand the
mechanisms involved in the degradation of organic pollutants
since these three mills have different wastewater treatment sys-
tems. Palm oil mill A is located in Negeri Sembilan, and mills B
and C are located in Terengganu, Malaysia. All mills adopted the
open lagoon system in their POME treatments. Only Mill A was
equipped with a bio-polishing plant, an additional treatment unit
after the lagoon treatment which consisted of an extended aeration
basin and clarifier, to further treat the effluent before its discharge
into the river. POME final discharge samples were filled into 1 L
amber bottles (Duran, Germany) and sealed with a stopper. The
samples were kept in a chilled box during the delivery back to the
laboratory and kept at 4 °C before further analyses. All the toxicity
tests and chemical analyses were performed within 36 h after sam-
pling. The first sampling was carried out in March–April 2017, and
the second in October–November 2017.
2.2. POME final discharge characterization
The POME final discharge samples were characterized by quan-
tifying 5 days biochemical oxygen demand (BOD 5 ), pH, total
suspended solids (TSS) and total solids (TS) according to the stan-
dard procedures (APHA et al., 2005). The chemical oxygen demand
(COD) and the ammoniacal nitrogen were determined by using
the chemical digestion method following the HACH protocol
(HACH, 2005). All analyses were conducted with at least two
replications.
2.3. Whole effluent toxicity (WET) tests
The WET test was conducted under light illumination and dark con-
ditions for16 h and 8 h, respectively at 20 ± 1 °C. The test solutions con-
taining distilled water and POME final discharge samples with
concentrations ranging from 0 to 25% (v/v) were prepared according
to the TIE method (USEPA, 1991). Twenty mL test solution mixtures
were placed into 30 mL glass beakers, and 5–10 D. magna were exposed
to the test solutions for 48 h. The results were expressed as LC50 values.
Probit, Spearman-Karber, trimmed Spearman-Karber were used for the
calculation of inhibition concentrations. The resulting values were then
converted to toxic units (TU = 100/LC50).
 Y. Hashiguchi et al. / Science of the Total Environment 710 (2020) 136277
2.4. Toxicity identification evaluation (TIE) test
2.4.1. Phase I manipulation
Phase I was carried out to characterize groups of toxicants in aque-
ous samples (USEPA, 1991; Norberg-King et al., 1991). The phase I test
performed on the POME final discharge included EDTA and sodium
thiosulfate additions to determine the presence of acute toxic cationic
metals and oxidative agents, respectively. Control tests for pH adjust-
ment/aeration and pH adjustment/filtration were conducted at three
pH ranges. Sample aliquots at pHi (initial pH), pH 3, and pH 11 were
allowed to stand for 1 h for complete aeration and filtration. The initial
pH of the samples was adjusted to pH 3 and pH 11 by the addition of
concentrated HCl and NaOH, respectively. After sample manipulations
(aeration and filtration) at the adjusted pH, the samples were
readjusted to pHi by the addition of concentrated NaOH and HCl, re-
spectively, before the toxicity test was performed. After the pH adjust-
ment, three manipulations were conducted; aeration, filtration, and
solid-phase extraction (SPE). The samples were aerated for 1 h using
an aquarium air pump to remove volatile, oxidizable, sublatable, or
spargeable compounds. The samples were filtered through 0.45 μm cel-
lulose acetate membranes (Millipore, USA) after centrifugation at
10,000 ×g for 30 min.
To detect the presence of non-polar organic compounds that may
cause toxicity, the effluent at pHi, pH 3, and pH 11 were fractionated
using SPE C18 columns. SPE on C18 cartridges (Strata C18-E, 500 mg/
3 mL, Phenomenex, USA) were used to absorb non-polar organic com-
pounds. The column was rinsed with 5 mL methanol followed by 5 mL
of ultra-pure water before use. This solution was used as a control in
the chemical analysis in Phase II. Before the sample went through the
C18 column, pH 11 sample was adjusted to pH 9. Each sample
(100 mL) was passed through the column at a rate of 1 mL/ min. After
that the C18 columns were washed with 5 mL ultra-pure water. Then,
they were eluted with 6 mL of methanol and water solutions at concen-
trations of 25:75, 50:50, 70:30, 80:20, 90:10 and 100:0, (v/v) to recover
the filtered toxicants. Each eluent (5 mL) was dried with nitrogen gas
and concentrated to 1 mL with methanol. These methanol fractions
were subjected to toxicity assay. Another 1 mL eluent was subjected
to chemical analysis (in Phase II). After each manipulation was com-
pleted, the pH of all test solutions was re-adjusted back to the initial
pH existing before the toxicity test using HCl and NaOH. Ethylenedi-
aminetetraacetic acid (EDTA) and sodium thiosulfate (STS) were
added to remove cationic metals and oxidative compounds, respectively
(Hockett and Mount, 1996). Finally, the D. magna lethality assay was
conducted using these test solutions for 48 h under light illumination
and dark conditions as previously described.
2.4.2. Phase II manipulation
Gas chromatography-mass spectrometry (GC–MS) was used to
identify non-polar organic compounds from the methanol-eluted sam-
ples obtained from the SPE preparations. The GC–MS used was
equipped with a 30 m × 0.25 mm inner diameter DB-5 capillary column
(GCQ Finnigan) with helium as the carrier gas. The GC–MS was operated
as follows: linear velocity, 40 cm/s; injector temperature, 275 °C; trans-
fer line temperature, 300 °C; an ion source, 150 °C. For analysis, 1 μL
Table 1
Properties of POME final discharge samples obtained from different palm oil mills.
Parameter (mg/L) Mill A Mill B
First sampling Second sampling First sampling
BOD5 67 ± 11 38 ± 5 69 ± 3
COD 91 ± 9 116 ± 5 220 ± 9
TS 3940 ± 20 3895 ± 40 2512 ± 85
TSS 31 ± 10 30 ± 3 83 ± 14
Ammonia as N 1.1 ± 0.1 2.2 ± 0.2 0.8 ± 0.2
pH 9.1 ± 0.1 8.5 ± 0.1 8.0 ± 0.2
a
Department of Environment, Malaysia. Environmental Quality (Industrial Effluents), 2009.
3
splitless injection was carried out using an autosampler. The GC temper-
ature was programmed as follows: initial temperature 50 °C, heated to
120 °C by a temperature ramp of 7.5 °C/min, then to 275 °C by a temper-
ature ramp of 15 °C/min then finally heated to 300 °C (held for 2 min) by
a temperature ramp of 25 °C/min (Chokwe et al., 2012).
Inductively coupled plasma – mass spectrometry (ICP/MS) was used
to determine the metal elements present in the final discharge samples.
The effluent samples were sent for analysis by an accredited third-party
laboratory (UNIPEQ Sdn Bhd, Malaysia).
2.4.3. Phase III manipulation
Specific peaks obtained from the GC–MS analysis (Phase II) were an-
alyzed and further selected to confirm their toxicity effects in the Phase
III test. All chemicals were GC-grade and purchased from Tokyo Chem-
ical Industry Co., Ltd. The chemical concentrations ranging from 0.001 to
100 mg/L were prepared and used for the toxicity test. A standard curve
of selected chemicals was established. D. magna were exposed to vari-
ous chemical concentrations for 48 h. The results were shown as 50%
of lethal concentration (LC50). All toxicity tests were conducted with
at least three replications.
3. Results and discussion
3.1. POME final discharge characterization
The POME final discharge samples obtained from three mills at dif-
ferent sampling times (1st sampling and 2nd sampling) were character-
ized, as shown in Table 1. The samplings were carried out at different
times to observe quality of the final discharge samples. March to April
was a dry season and October to December was a rainy season, particu-
larly in the east coast region like Terengganu. Climate factors may influ-
ence the quality of the POME generated due to the lower amount of
fresh fruit bunch (FFB) processed particularly in the rainy season. It
was anticipated that the quality of the POME final discharge might
vary and conventional chemical analyses such as BOD, COD, TS, TSS,
and pH were done in compliance to the effluent discharge limit set by
DOE, Malaysia (DOE, 2019). It may be expected that, in the rainy season,
pollutants in POME final discharge should be more diluted than in the
dry one. Nevertheless, in Table 1 it is clearly observed that often the op-
posite occurred, in particular for BOD and COD. Hence, it was hypothe-
sized that the only possible cause of this observed phenomenon was the
run-off chemical compounds or contaminants that entered the drainage
systems or treatment lagoons. Thus, a detailed investigation on factors
affecting pollution levels in the rainy season should be carried out in
the future.
Only mill A successfully met the minimum permissible effluent dis-
charge standard (Standard B) for BOD5 and COD. Mills B and C failed to
meet the effluent discharge standard, probably due to incomplete bio-
degradation of the organic compounds caused by improper aeration
and mixing at the lagoon. Mill A was equipped with a bio-polishing
plant whereby an extended aeration basin and a clarifier were installed
to treat the POME further before discharge into the river. The TS of efflu-
ent from mill A was around 4000 mg/L, which was higher than the efflu-
ent from mill B (around 3000 mg/L) and mill C around 2000 mg/L,
Mill C Effluent standard limitsa
Second sampling First sampling Second sampling Standard A Standard B
73 ± 17 97 ± 7 196 ± 4 20 40
236 ± 18 118 ± 4 330 ± 3 80 200
2827 ± 12 1730 ± 30 2013 ± 18 – –
72 ± 6 25 ± 5 56 ± 8 50 100
0.9 ± 0.1 0.8 ± 0.1 2.8 ± 0.2 10 20
8.0 ± 0.1 8.2 ± 0.1 8.5 ± 0.1 5.5–9 5–9
4 Y. Hashiguchi et al. / Science of the Total Environment 710 (2020) 136277

and TU = 8.3, respectively. On the other hand, effluents from mills A

and C in the 1st sampling showed toxicity of TU = 4.9 and TU = 5.0, re-
spectively. The lowest toxicity observed was from mill C from the 2nd
sampling with TU = 3.4. Overall, different effluents showed different
toxicity levels.
Interestingly, the highest toxicity was observed from mill A's 2nd
sampling with TU = 11, which was contradictory to the results obtained
from the conventional chemical analysis (BOD5 and COD). Similarly, the
effluent from the 2nd sampling of mill C had higher BOD5 and COD
while having the lowest TU = 3.4. It was concluded that the effluent
quality could not be evaluated solely by using conventional chemical
parameters. The causative toxicants, although existing at low concen-
trations in the effluents, might still cause substantial toxicities to aquatic
organisms. Thus, the WET test is encouraged to be adopted for the best
management practice of POME discharge.

3.3. TIE tests

Fig. 1. WET test after 48 h with initial POME final discharge from different palm oil mills.
respectively. Other parameters like TSS, ammoniacal-N, and pH had met
the permissible discharge standards.
Although the effluent generated from the mills had met the mini-
mum permissible effluent discharge as determined by the COD
(Table 1), the presence of possible toxicants in the effluent might be det-
rimental to living organisms in the receiving water bodies. Hence, the
following WET and TIE tests were performed on effluents from the dif-
ferent mills as described below. To the best of the authors' knowledge,
this was the first attempt to identify and evaluate the presence of toxi-
cants in POME final discharge samples.
3.2. WET test
Due to the limitations of chemical analyses as a result of the com-
plexity of wastewater, biological toxicity test had been implemented
to assess the potential effects of wastewater discharges on aquatic life
(Sarakinos et al., 2000). The WET test has been recognized as one of
the parameters and best practices to be adopted for the analysis of in-
dustrial effluent or mixed effluent (DOE, 2019). However, the WET
test has not been widely used in POME final discharge analysis. A WET
test was performed in the present work to observe the toxicity potential
of the POME final discharge on an aquatic organism. D. magna was ex-
posed to POME final discharges, and their toxicity levels are shown in
Fig. 1. A substantial toxicity effect was observed from the 2nd and 1st
samplings of effluents from mills A and B, which recorded TU = 11
3.3.1. TIE Phase I
Results of the toxicity tests from the original samples and after TIE
Phase I manipulations are shown in Table 2. The toxicities of all the ef-
fluents were sharply decreased after pH adjustment. The treatment
with acid was more effective than alkaline to reduce toxicity. Thus,
pH-sensitive compounds might be the main toxicants in the POME
final discharge. Similarly, aeration and filtration were effective treat-
ments to reduce toxicity, especially after pH adjustment. Based on the
results obtained, it could be postulated that the toxicity of POME final
discharge was caused by volatile or oxidizable compounds and
suspended solids sensitive to pH variations (USEPA, 1991; Norberg-
King et al., 1991).
Toxic compounds removed at lower pH were possibly from ammo-
nia or deionized compounds under acidic conditions. Ankley and
Burkhard (1992) revealed that acidic treatment affected the decompo-
sition of some organic compounds by hydroxylation. Therefore, major
toxicants in the effluents might be hydrolyzed and degraded to non-
toxic compounds in acidic condition. The addition of EDTA and
Na2S2O3 showed no reduction in toxicity, suggesting that few metals
and oxidants were present (Table 2). The presence of other groups of
toxicants in the final discharge samples was determined from the solu-
tion after manipulation by filtration using C18 SPE. C18 SPE treatment
retained toxic compounds on the column and the toxicity levels re-
corded were lower than those of treatment by filtration. It could be an-
ticipated that non-polar organic compounds contributed to acute
toxicity since the polar organic substances were not adsorbed onto the
C18 cartridge through the SPE procedure. Usually, those adsorbates

Table 2
Toxic units (TUs) to D. magna before and after TIE phase I manipulations of the POME final discharge samples.

Manipulations pHa Toxic units (TUs)b

Mill A Mill B Mill C

First sampling Second sampling First sampling Second sampling First sampling Second sampling

pH adjustment pH i 4.9 ± 0.3 11.0 ± 1.7 8.3 ± 0.8 6.4 ± 0.5 5.0 ± 0.3 3.4 ± 0.2
pH 3 0.0 1.6 ± 1.6 2.3 ± 0.1 1.7 ± 0.0 1.3 ± 0.0 1.3 ± 0.0
pH 11 0.0 1.9 ± 0.1 5.1 ± 0.4 3.1 ± 0.1 2.8 ± 0.1 2.0 ± 0.1
pH adjustment and aeration pH i 5.3 ± 0.3 9.0 ± 0.9 7.0 ± 0.6 7.4 ± 0.6 3.8 ± 0.2 4.2 ± 0.2
pH 3 0.0 1.6 ± 0.0 1.5 ± 0.0 1.8 ± 0.0 0.0 1.3 ± 0.0
pH 11 0.0 1.2 ± 0.0 2.6 ± 0.1 3.0 ± 0.1 1.8 ± 0.0 1.8 ± 0.0
pH adjustment and filtration pH i 6.0 ± 0.8 10.8 ± 1.5 8.7 ± 1.0 8.2 ± 0.8 6.5 ± 0.7 4.8 ± 0.3
pH 3 0.0 1.6 ± 0.0 1.8 ± 0.0 1.7 ± 0.0 0.0 1.2 ± 0.0
pH 11 0.0 1.6 ± 0.0 5.6 ± 0.4 3.1 ± 0.1 1.8 ± 0.0 1.6 ± 0.0
EDTA addition pH i 5.6 ± 0.4 13.6 ± 1.4 5.0 ± 0.3 7.3 ± 0.7 2.9 ± 0.1 6.0 ± 0.5
Sodium thiosulfate addition pH i 6.1 ± 0.6 13.0 ± 1.5 4.6 ± 0.3 7.4 ± 0.7 2.5 ± 0.1 5.4 ± 0.4
pH adjustment, filtration, and C18 SPE pH i 4.5 ± 0.1 8.0 ± 0.7 5.4 ± 0.4 5.1 ± 0.3 3.3 ± 0.1 3.0 ± 0.1
pH 3 0.0 1.4 ± 0.0 2.2 ± 0.1 1.6 ± 0.0 1.1 ± 0.0 1.2 ± 0.0
pH 11 0.0 1.2 ± 0.0 5.2 ± 0.4 2.9 ± 0.1 1.9 ± 0.0 1.6 ± 0.0
a
Initial pH (pHi); First sampling; Mill A (9.1), mill B (8.0) and mill C (8.2). Second sampling; Mill A (8.5), mill B (8.0) and mill C (8.5).
b
TUs = Toxicity units.
 Y. Hashiguchi et al. / Science of the Total Environment 710 (2020) 136277 5

Table 3
Metal concentrations measured by ICP-MS from POME final discharge samples.

Parameter (mg/L) Mill A Mill B Mill C Effluent discharge

standarda

First sampling Second sampling First sampling Second sampling First sampling Second sampling A B

As 0.15 0.02 N.D N.D N.D 0.01 0.05 0.10

Ba 0.19 N.D N.D N.D 0.12 0.03 1.0 2.0
Cd N.D N.D N.D N.D N.D N.D 0.01 0.02
Cu 0.12 0.05 N.D N.D 0.35 0.28 0.20 1.0
Pb 0.03 0.04 N.D N.D 0.01 0.03 0.10 0.50
Mn 0.21 0.28 0.40 0.21 0.20 N.D 0.20 1.0
Hg N.D N.D N.D 0.002 N.D 0.01 0.005 0.05
Se 0.10 N.D N.D N.D N.D N.D 0.02 0.5
Zn 0.15 1.31 0.05 0.09 0.01 N.D 2.0 2.0
Fe 3.25 0.94 1.18 0.70 3.25 5.82 1.0 5.0
Ni 1.46 0.04 N.D 0.11 N.D N.D 0.20 1.0
Na 140.70 29.89 51.28 29.87 43.98 107.54 N.R N.R
Ca 33.22 68.81 33.32 37.22 57.87 40.99 N.R N.R
K 1592.30 1740.65 1578.20 1331.24 1211.32 890.81 N.R N.R
Mg 251.16 242.63 177.19 181.31 166.67 116.78 N.R N.R

N.D = not detected.

N.R = no regulation.
a
Discharge standard A and B: Malaysia Environmental Quality Regulation, 2009.

were from solvents, hydrocarbons, polycyclic aromatic hydrocarbons
(PAHs), fatty acid methyl esters (FAME), and surfactants (Budi et al.,
2016).
It was inferred that the toxicants could be derived from suspended
solids, volatile or sublatable compounds, and organic compounds sensi-
tive to pH variations. The toxicity levels, TU = 1–3 obtained after pH 3,
aeration, filtration, and C18 SPE treatment indicated the presence of an-
other minor toxicant group in the final discharge samples. As a conclu-
sion, aeration, filtration, and SPE were the most effective treatments for
reducing toxicity, especially after pH adjustments.
3.3.2. TIE Phase II
Metals concentration in the POME final discharge collected were de-
termined by ICP-MS, as shown in Table 3. It was reported that heavy
metals such as Cu, Zn, and Ni caused inhibitory effects on the growth
of D. magna (Okamoto et al., 2015; Ra et al., 2016). In the present
study, Ni and Fe detected from mills A and C were slightly over the per-
missible limits of standard B with concentrations of 1.46 mg/L (1st sam-
pling) and 5.82 mg/L (2nd sampling), respectively. EC50 for Ni was
reported to be in the range from 0.1–1 mg/L (Okamoto et al., 2015).
The absence of Ni, from the 1st and 2nd samplings of Mill C was ob-
served in this study. In general, the distribution of metals in each mill
was different. Obviously, there was no detection of the metals, As, Ba,
Cd, Cu and Pb in the 1st and 2nd samplings from Mill B as well. Relevant
data had been published in the literature in recent decades concerning
the occurrence, fate and behaviour of metals in wastewater treatment
plants. However, the information is dispersed and not standardized in
terms of parameters for comparing the results (Cantinho et al., 2016).
Hargreaves et al. (2017) reported that the distributions of the metals
could be in the dissolved, colloidal and particulate forms and their cor-
relations with the total suspended solid removal. He also proposed the
concept of particle size fractionation in wastewater that helped us to
understand potential treatment options to enhance metal removal.
However, the contributions of metals and their reactions in biological
treatment led to the conclusion that the sources of the metals in the
POME final discharge were still poorly understood.
Cu and Zn showed the highest values at 0.35 mg/L (1st sampling)
and 1.31 mg/L (2nd sampling) from mill C and mill A, respectively.
The presence of these metals might inhibit the growth of D. magna. Ra
et al. (2016) found that heavy metals such as Cu and Zn with concentra-
tions of 0.30 mg/L (TU = 3.42) and 0.54 mg/L (TU = 0.79), respectively
contributed to the toxicity. In another study, Fu et al. (2016) reported
that the Cu and Zn in water posed the most considerable ecological
risk of 99.9% and 50.7% to aquatic species of Tai Lake in Eastern China.
In another study, the 48 h LC50 of Cu and Zn reported with values of
0.192 mg/L (Arambasic et al., 1995) and 1.22 mg/L (Magliette et al.,
1995), respectively, showed the toxicities of these metals on D. magna.
Other metals were detected in ranges below the permissible discharge

Table 4
The presence of organic compounds identified by GC–MS in the effluent from mill A, B and C.

No. R.T.a (min) Detected chemical MWb Mill A Mill B Mill C

First sampling Second sampling First sampling Second sampling First sampling Second sampling

1 6.54 Cyclotetrasiloxane, octamethyl- 297 Present N.D Present N.D N.D N.D
(C8H24O4Si4)
2 9.55 Cyclopentasiloxane, decamethyl- 371 Present N.D Present N.D N.D N.D
(C10H30O5Si5)
3 12.06 Cyclohexasiloxane, dodecamethyl- 445 Present N.D Present N.D N.D N.D
(C12H36O6Si6)
4 14.02 Phenol, 2,6-bis(1,1-dimethylethyl)- 206 Present Present Present Present Present Present
(C14H22O)
5 17.18 Hexadecanoic acid, methyl ester- 270 Present Present Present Present Present Present
(C17H34O2)
6 18.48 Octadecanoic acid, methyl ester- 299 Present Present Present Present Present Present
(C19H38O2)

N.D = not detected.

a
R.T. = retention time.
b
MW = molecular weight.
6 Y. Hashiguchi et al. / Science of the Total Environment 710 (2020) 136277

Table 5
Toxicity of detected organic compounds on D. magna.

Detected chemical This study (48 h-LC50)a Other studies (48 h-EC50)b References

Cyclotetrasiloxane, octamethyl- N.D 0.015 mg/L Sousa et al. (1995)

Cyclopentasiloxane, decamethyl- N.D 0.0029 mg/L IUCLID (2005)
Cyclohexasiloxane, dodecamethyl- N.D 0.0046 mg/L Springborn Smithers Laboratories (2006)
Phenol, 2,6-bis(1,1-dimethylethyl)- 32.49 mg/L 0.45 mg/L Surprenant (1989)
Hexadecanoic acid, methyl ester- N100 mg/L N.R N.R
Octadecanoic acid, methyl ester- N100 mg/L N.R N.R

N.R = no report.
N.D = Not determine in this work due to low solubility and high volatility of cVMS.
a
Toxicity, LC50 of D. magna after 48 h.
b
Toxicity, EC50 of D. magna after 48 h.

limits (DOE, 2019). A high concentration of Mg, Na, and K was detected
in all effluents. Okamoto et al. (2015) reported that the EC50 for Mg, Na,
and K were N100 mg/L indicating the low-level toxicities of these min-
erals on D. magna.
Non-polar organic compounds from the methanol-eluted samples
prepared by C18 SPE are shown in Table 4. GC–MS analyses identified
six possible toxicants from the collected effluents. Cyclotetrasiloxane,
octamethyl- (D4), cyclopentasiloxane, decamethyl- (D5),
cyclohexasiloxane, dodecamethyl- (D6), were the siloxanes detected
from the effluents of mills A and B, especially in the 1st sampling. Mean-
while, hexadecanoic acid, methyl ester- and octadecanoic acid, methyl
ester- were a group of fatty acid methyl esters (FAME) (Sreenivasilu
et al., 2017) detected from all the samples. Łukasik and Łenyk (2009) re-
ported that FAME was derived from vegetable-based oils and fats, and
was readily biodegradable under anaerobic and aerobic conditions,
and high temperature. The present work intrigued us as to what was
happening during the anaerobic/aerobic degradation of POME and
what other factors influenced the presence of siloxanes since the milling
process did not involve any addition of the chemicals. D4, D5, and D6
known as cyclic volatile methyl siloxanes (cVMS), are highly volatile
and have low solubilities in water (Wang et al., 2013a). These com-
pounds are identified as potential toxicants to the environment based
on their persistence and bioaccumulation (Borgå et al., 2013). In gen-
eral, wastewater treatment plants substantially removed cVMS through
volatilization and degradation upon regular operations with removal ef-
ficiencies of D4, D5, and D6 above 90% (Wang et al., 2013b). In another
study, it was found that these compounds could be removed through
aeration because of their vast vapor pressures (Parker et al., 1999),
and that their half-lifes would be decreased by high temperature
under acidic and alkaline conditions (Sousa et al., 1995).
Phenol, 2,6-bis(1,1-dimethylethyl)- was also detected in all samples.
It was reported that phenol, 2,6-bis(1,1-dimethylethyl) being used as
the oxidation inhibitor and stabilizer for biodiesel was highly toxic for
aquatic organisms at 0.45 mg/L of EC50 using D. magna as the test or-
ganism (Surprenant, 1989). Some phenolic antioxidants were detected
in POME (Sambanthamurthi et al., 2008). The brownish color of raw
POME was probably from the tannins and low molecular weight pheno-
lic compounds (Limkhuansuwan and Chaiprasert, 2010). Phenol and
other phenolic compounds were found at high concentrations
(200–1000 mg/L) in POME and effluent from the anaerobic treatment
of POME (Alam et al., 2009). However, there is a limited number of stud-
ies which reported on the presence of phenol, 2,6-bis(1,1-
dimethylethyl) in the POME final discharge and its presence is also con-
firmed in the present study.
3.3.3. Phase III manipulations
The toxicity levels of potential toxicants detected in Phase II are
shown in Table 5. Based on the results obtained, hexadecenoic acid,
methyl ester- and octadecanoic acid, methyl ester- showed the lowest
toxicities on D. magna with LC50 values of N100 mg/L after 48 h of expo-
sure. Meanwhile, the LC50 for phenol, 2,6-bis(1,1-dimethylethyl)- was
at 32.49 mg/L. This phenol compound was identified in all effluent sam-
ples with an estimated pKa = 11.91 (ECHA - REACH Dossier) and was
therefore sensitive to alterations in pH. Besides, this phenol is toxic to
aquatic organisms, and in particular, to daphnids. The D. magna was
the most sensitive trophic level/species in short-term tests having an
EC50 of 0.45 mg/L. Consequently, this species was also used for chronic
testing. The long-term test with D. magna resulted in a 21 day-NOEC
(mortality, reproduction, growth) of 0.035 mg/L (ECHA - REACH Dos-
sier). In another study a 48 h LC50 for phenol of 9.411 mg/L, showed a
toxicity effect on D. magna (Arambasic et al., 1995). Confirmation of
cVMS toxicity was not performed due to the low water solubilities of
D4, D5, and D6 at 0.056 mg/L, 0.017 mg/L, and 0.0051 mg/L, respectively
and these low solubilities combined with their high Henry's Law con-
stants, made them notoriously difficult to test in aquatic systems. This
was also supported by the study of Sousa et al. (1995).
After confirmation of the types of toxicants present and their toxicity
levels in the TIE phase II and phase III studies, the concentrations of tox-
icants in the final discharge samples were determined by GC–MS, as are
shown in Table 6. The cVMS were present in only two of the six effluents
collected, with the greatest toxicity being observed in a sample with no
cVMS present (Fig. 1). The elimination of possible toxicants through the
toxicant identification evaluation (TIE) did not suggest that the cVMS
materials were responsible for the observed toxicity. The toxicity of all
effluents was sharply decreased after pH adjustment suggesting that
pH-sensitive compounds might be the main toxicants in the POME
final discharge (Table 1). The cVMS materials did not contain any acid
groups, and thus possessed no ionization state and could not be influ-
enced by changes to pH. The presence of phenol detected in the range
from 0.03–0.04 mg/L, indicated it might be the main toxicant together

Table 6
Concentrations of organic compounds in effluent from mills A, B and C.

Organic compounds (mg/L) Mill A Mill B Mill C

First sampling Second sampling First sampling Second sampling First sampling Second sampling

Cyclotetrasiloxane, octamethyl- 0.015 ± 0.00 N.D 0.036 ± 0.0 N.D N.D N.D
Cyclopentasiloxane, decamethyl- 0.005 ± 0.00 N.D 0.008 ± 0.00 N.D N.D N.D
Cyclohexasiloxane, dodecamethyl- 0.002 ± 0.00 N.D 0.004 ± 0.00 N.D N.D N.D
Phenol, 2,6-bis(1,1-dimethylethyl)- 0.03 ± 0.00 0.03 ± 0.00 0.03 ± 0.00 0.04 ± 0.00 0.04 ± 0.00 0.03 ± 0.00
Hexadecenoic acid, methyl ester- 0.15 ± 0.01 0.15 ± 0.01 0.12 ± 0.00 0.11 ± 0.02 0.14 ± 0.00 0.15 ± 0.01
Octadecanoic acid, methyl ester- 0.05 ± 0.01 0.05 ± 0.01 0.04 ± 0.00 0.04 ± 0.01 0.04 ± 0.00 0.03 ± 0.01

N.D = Not detected.

 Y. Hashiguchi et al. / Science of the Total Environment 710 (2020) 136277
Fig. 2. General overview of palm oil mill processing using only water in the form of steam to cook the fresh fruit bunch with no addition of chemical. A) POME final discharge generated
under normal condition, B) Generation of POME final discharges and possible contamination with cVMS when the mill performing maintenance and cleaning up works.
with the presence of metals such as Cu and Zn, which might have syn-
ergistic effects in the effluents.
3.3.4. The presence of cVMS in the POME final discharge
The cVMS is mainly used in various personal care products such as
antiperspirants, grooming products, sun protection products and cos-
metic preparations. They are also used in the production of lubricants,
adhesives, paints and solvents, foams, detergents, fabric softeners and
protectors, seals, and elastomers. It is suggested that these ‘rinse-off’
products had leaked into the wastewater. It was reported that about
90% of the cVMS used in personal care products would volatilize during
use while the other 10% was introduced into wastewater streams (Xu
et al., 2014). Palm oil processing only used water in the form of steam
to cook the fresh fruit bunches with no addition of chemicals (Fig. 2).
It is postulated that the presence of cVMS in the POME final discharges
could be derived from silicone grease and detergents from the cleanup
process during mill maintenance work. The wash water containing de-
tergents, grease and lubricants could be mixed with the POME stream-
line and ended up in the wastewater treatment ponds or digesters
where these materials would accumulate (Lassen et al., 2005). From
the inconsistent presence of the cVMS compounds detected from the ef-
fluents of the three mills it could be inferred that the mill maintenance
and cleaning up activities, mills location, seasonal and other factors
might influence the presence of cVMS compounds in the POME final dis-
charge samples.
4. Conclusion
WET and TIE tests on the POME final discharge samples from differ-
ent mills were successfully performed in the present study. The toxicity
unit values (TU≥1) obtained from both the WET and TIE tests suggested
that the POME final discharge samples contained potential toxicants.
Pretreatment of effluents using filtration, aeration, and SPE C-18 column
reduced the TU. The GC–MS results and the toxicity test evaluations
using D. magna confirmed that the toxicities were caused mainly by
acid- sensitive compounds, phenol, and heavy metals such as Cu and
Zn. It is recommended that two approaches, chemical-based monitor-
ing, as well as biological toxicity-based monitoring, should be
7
considered for achieving acceptable POME final discharge in the future.
The current findings need further verification, in terms of sample size,
mill locations, seasonal and other factors that may affect the ecosystem.
Such a study is currently on-going.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgment
The authors gratefully acknowledge Universiti Putra Malaysia
(UPM) and the Kyushu Institute of Technology (KYUTECH) for financial
and technical assistance in this study. The first author is a recipient of a
scholarship from the Japan Student Services Organization (JASSO). Also,
the Ministry of Higher Education, Malaysia is thanked for a FRGS Grant
(02-01-14-1401FR).
References
Alam, M.Z., Ameem, S.E., Muyibi, A.S., Kabbashi, A.N., 2009. The factors affecting the per-
formance of activated carbon prepared from oil palm empty fruit bunches for adsorp-
tion of phenol. Chem. Eng. J. 155, 191–198.
Ankley, G.T., Burkhard, L.P., 1992. Identification of surfactants as toxicants in a primary ef-
fluent. Environ. Toxicol. Chem. 11, 1235–1248.
APHA, AWWA, WEF, 2005. Standard Methods for the Examination of Water and Waste-
water. 21st ed. American Public Health Association, Washington, DC.
Arambasic, M.B., Bjeli, S., Subako, G., 1995. Acute toxicity of heavy metals (copper, lead,
zinc), phenol and sodium on Allium cepa L., Lepidium sativum L. and Daphnia magna
St. comparative investigations and the practical applications. Wat. Res. 29, 497–503.
Borgå, K., Fjeld, E., Kierkegaard, A., McLachlan, M.S., 2013. Consistency in trophic magnifi-
cation factors of cyclic methyl siloxanes in pelagic freshwater food webs leading to
brown trout. Environ. Sci. Technol. 47, 14394–14402.
Budi, S., Suliasih, B.A., Othman, M.S., Heng, L.Y., Surif, S., 2016. Toxicity identification eval-
uation of landfill leachate using fish, prawn and seed plant. Waste Manag. 55,
231–237. https://doi.org/10.1016/j.wasman.2015.09.022.
Cantinho, P., Matos, M., Trancoso, M.A., Correia dos Santos, M.M., 2016. Behaviour and fate
of metals in urban wastewater treatment plants: a review. Int. J. Environ. Sci. Technol.
13, 359–386.
Chokwe, T.B., Okonkwo, J.O., Sibali, L.L., Ncube, E.J., 2012. Optimization and simultaneous
determination of alkyl phenol ethoxylates and brominated flame retardants in water
8 Y. Hashiguchi et al. / Science of the Total Environment 710 (2020) 136277
after SPE and heptafluorobutyric anhydride derivatization followed by GC/MS.
Chromatographia 75, 1165–1176. https://doi.org/10.1007/s10337-012-2293-6.
de Melo, E.D., Mounteer, A.H., Henrique, L., Leão, D.S., Cibele, R., Bahia, B., 2013. Toxicity
identification evaluation of cosmetics industry wastewater. J. Haz. Mat. 244–245,
329–334.
Department of Environment (DOE), Malaysia, 2019. https://www.doe.gov.my/portalv1/
wp-content/uploads/2015/01/Environmental_Quality_Industrial_Effluent_Regula-
tions_2009_-_P.U.A_434-2009.pdf.
ECHA-REACH Dossier, d. https://echa.europa.eu/information-on-chemicals/registered-
substances.
Fu, Z., Wu, F., Chen, L., Xu, B., Feng, C., Bai, Y., Liao, H., Sun, S., Giesy, J.P., Guo, W., 2016.
Copper and Zn but not other priority toxic metals pose risks to native aquatic species
in a lake in Eastern China. Env. Pollut. 219, 1069–1076.
HACH, 2005. Hach Water Analysis DR5000 Spectrohotometer Procedures Manual. HACH
Company, USA.
Hargreaves, A.J., Vale, P., Whelan, J., Constantino, C., Dotro, G., Campo, P., Cartmell, E.,
2017. Distribution of trace metals (Cu, Pb, Ni, Zn) between particulate, colloidal and
truly dissolved fractions in wastewater treatment. Chemosphere 175, 239–246.
Ho, C.C., Tan, Y.K., Wang, C.W., 1984. The distribution of chemical constituents between
the soluble and the particulate fractions of palm oil mill effluent and its significance
on its utilization/treatment. Agric. Wastes. 11, 61–71.
Hockett, J.R., Mount, D.R., 1996. Use of metal chelating agents to differentiate among
sources of acute aquatic toxicity. Environ. Toxicol. Chem. 15, 1687–1693.
IUCLID, 2005. IUCLID Dataset for Octamethylcyclotetrasiloxane. Epona Associates LLC,
Willington, CT.
Kusui, T., Yakata, Y., Itatsu, Y., Zha, J., 2014. Whole effluent toxicity assessment of
indystrial effluents in Toyama, Japan with a battery of short-term chronic bioassays.
J. Industr. Environ. Technol. 12, 55–63.
Lassen, C., Hansen, C.L., Mikkelsen, S.H., Maag, J., 2005. Siloxanes – consumption, toxicity
and alternatives. Environmental Protection Agency. Danish Ministry of the
Environmenthttps://www2.mst.dk/udgiv/publications/2005/87-7614-756-8/pdf/87-
7614-757-6.pdf.
Limkhuansuwan, V., Chaiprasert, P., 2010. Decolorization of molasses melanoidins and
palm oil mill effluent phenolic compounds by fermentative lactic acid bacteria.
J. Environ. Sci. 22, 1209–1217.
Łukasik, Z., Łenyk, M., 2009. Fame oxidation and thermal stability. Problemy Eksploatacji
2, 149–158.
Madaki, Y.S., Seng, L., 2013. Palm oil mill effluent (POME) from Malaysia palm oil mills:
waste or resource. Int. J. Sci. Environ. Technol. 2, 1138–1155.
Magliette, R., Doherty, F.G., McKinney, D., et al., 1995. Bull. Environ. Contam. Toxicol. 54,
626.
Norberg-King, T.J., Durhan, E.J., Ankley, G.T., Robert, E., 1991. Application of toxicity iden-
tification evaluation procedures to the ambient waters of the Colusa Basin Drain. En-
viron. Toxicol. Chem. 10, 891–901.
Okamoto, A., Yamamuro, M., Tatarazako, N., 2015. Acute toxicity of 50 metals to Daphnia
magna. A. Appl. Toxic. 35, 824–830.
Parker, W.J., Shi, J., Fendinger, N.J., Monteith, H.D., Chandra, G., 1999. Pilot plant study to
assess the fate of the two volatile methyl siloxane compounds during municipal
wastewater treatment. Environ. Toxicol. Chem. 18 (2), 172–181.
Poh, P.E., Yong, W.J., Chong, M.F., 2010. Palm oil mill effluent (POME) characteristic in
high crop season and the applicability of high-rate anaerobic bioreactors for the treat-
ment of pome. Ind. Eng. Chem. Res. 49, 11732–11740.
Ra, J.S., Jeong, T.Y., Lee, S.H., Kim, S.D., 2016. Application of toxicity identification evalua-
tion procedure to toxic industrial effluent in South Korea. Chemosphere 143, 71–77.
Randhir, R., Lin, Y.T., Shetty, K., 2004. Stimulation of phenolics, antioxidant and antimicro-
bial activities in the dark germinated mung bean sprouts in response to peptide and
phytochemical elicitors. Process Biochem. 39, 637–646.
Sambanthamurthi, R., Tan, Y.A., Manickam, K.S., 2008. Treatment of vegetation liquors de-
rived from oil-bearing fruits. PatentStorm. http://www.freepatentsonline.com/
y2009/0053333. html.
Sarakinos, H.C., Bermingham, N., Paul, A., White, J.B., Rasmussen, J.B., 2000. Correspon-
dence between whole effluent toxicity and the presence of priority substances in
complex industrial effluents. Environ. Toxicol. Chem. 19, 63–71.
Sousa, J.V., McNamara, P.C., Putt, A.E., Machado, M.W., Surprenant, D.C., Hamelink, J.L.,
Kent, D.J., Silberhorn, E.M., Hobson, J.F., 1995. Effects of octamethylcyclo-
tetrasiloxane (OMCTS) on freshwater and marine organisms. Environ. Toxicol.
Chem. 14, 1639–1647.
Springborn Smithers Laboratories, 2006. Dodecamethylcyclohexasiloxane (D6): Full Life-
Cycle Toxicity Test With Water Fleas, Daphnia magna, Under Static Renewal Condi-
tions. Silicones Environmental, Health and Safety Council (Study No. 1203-6149).
Sreenivasilu, B., Paramageetham, C., Sreenivasulu, D., Suman, B., Umamahesh, K., Babu,
G.P., 2017. Analysis of chemical signatures of alkaliphiles using fatty acid methyl
ester analysis. J. Pharmacy Bioallied Sci. 9, 106–114.
Surprenant, D.C., 1989. Acute Toxicity of Diflubenzuron to Quahogs (Mercenaria
mercenaria) Embryo-Larvae under Static Conditions. Duphar B. V. Internal Document
No. 56635/10/89 (25 pp.).
USEPA, 1991. Methods for Aquatic Toxicity Identification Evaluations: Phase I Toxicity
Characterization Procedures (EPA/600/6-91/003).
Wang, D.G., Norwood, W., Alaee, M., Byer, J.D., Brimble, S., 2013a. Review of recent ad-
vances in research on the toxicity, detection, occurrence and fate of cyclic volatile
methyl siloxanes in the environment. Chemosphere 93, 711–725.
Wang, D.G., Steer, H., Tait, T., Williams, Z., Pacepavicius, G., Young, T., Ng, T., Smyth, S.A.,
Kinsman, L., Alaee, M., 2013b. Concentrations of cyclic volatile methylsiloxanes in
biosolid amended soil, influent, effluent, receiving water, and sediment of wastewa-
ter treatment plants in Canada. Chemosphere 93, 766–773.
Xu, S., Kozerski, G., Mackay, D., 2014. Critical review and interpretation of environmental
data for volatile methylsiloxanes: partition properties. Environ. Sci. Technol. 48,
11748–11759.