Environmental Research 156 (2017) 434–442

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Environmental Research
journal homepage: www.elsevier.com/locate/envres

Micropollutants in source separated wastewater streams and recovered MARK

resources of source separated sanitation
⁎
A. Butkovskyia,b, , L. Hernandez Lealb, G. Zeemana, H.H.M. Rijnaartsa
a
Sub-department of Environmental Technology, Wageningen University, P.O. Box 17, 6700 AA Wageningen, The Netherlands
b
Wetsus, Centre of Excellence for Sustainable Water Technology, P.O. Box 1113, 8900 CC Leeuwarden, The Netherlands

A R T I C L E I N F O A B S T R A C T

Keywords: The quality of anaerobic sludge and struvite from black water treatment system, aerobic sludge from grey water
Black water treatment system and effluents of both systems was assessed for organic micropollutant content in order to
Grey water ensure safety when reusing these products. Use of anaerobic black water sludge and struvite as soil amendments
Sludge is recommended based on the low micropollutant content. Aerobic grey water sludge is recommended for
Struvite
disposal, because of the relatively high micropollutant concentrations, exceeding those in sewage sludge.
Micropollutants
Effluents of black and grey water treatment systems require post-treatment prior to reuse, because the measured
micropollutant concentrations in the effluents are above ecotoxicological thresholds.

1. Introduction
Source separated sanitation (SSS) is an innovative treatment
strategy for domestic wastewater aimed at minimizing energy and
water use, and maximizing recovery of water, organic matter and
nutrients. It is based on separate collection and treatment of concen-
trated toilet wastewater (black water) and the rest of domestic waste-
water (grey water) (Zeeman et al., 2008). Contrary to conventional
municipal wastewater collection, transport and treatment, where
wastewater is considered to be a problem, SSS regards wastewater as
a resource. Multiple drivers, such as increased water pollution, scarcity
of water in some areas on the Earth, growth of population, development
of novel wastewater treatment and recycling technologies, stricter
legislation and growth of environmentally oriented society, support
development of SSS (Asano et al., 2007).
The difference in characteristics of wastewater streams facilitates
recovery of valuable products. Black water contains organic matter and
nutrients in much higher concentrations than conventional municipal
wastewater or grey water (De Graaff et al., 2010; Verstraete and
Vlaeminck, 2011). Anaerobic digestion can be applied for energy
recovery from this stream, whereas struvite precipitation can be applied
for nutrient recovery, because concentrated wastewater stream is a pre-
requisite for the utilization of both technologies (Verstraete and
Vlaeminck, 2011). Additionally, sludge from up-flow anaerobic sludge
blanket (UASB) reactor can be also applied on soil, returning accumu-
lated nutrients and organic matter to the environment (Kujawa-
Roeleveld et al., 2006). Grey water, as a relatively diluted stream
without toilet wastewater input has a potential for reuse in various non-
potable applications, after appropriate treatment. Potential grey water
reuse applications include toilet flushing, irrigation of non-agricultural
lands, concrete production, car washing, fire protection, etc. (Eriksson
et al., 2002).
SSS products, such as nutrient-rich solids and recovered water,
could contain harmful wastewater constituents, organic micropollu-
tants being one of them. Pharmaceuticals and human hormones are
excreted with faeces and end up mainly in black water (Zeeman et al.,
2008). Separate black water collection and treatment allows concentra-
tion of these compounds in a significantly lower volume, instead of
dispersion in the total volume of municipal wastewater as employed
within conventional sanitation concepts. This may promote more
efficient removal of the pharmaceuticals and hormones (de Graaff
et al., 2011).
Grey water contains constituents of detergents and personal care
products. Only occasional presence of pharmaceuticals and human
hormones in this stream is expected based on the sources of their origin
in wastewater (Hernández Leal et al., 2010).
Some of the organic micropollutants have a proven negative impact
on the environment and human health, including feminization of male
organisms, growth inhibition and carcinogenicity, which are further
enhanced through bioaccumulation (Halling-Sorensen et al., 2000;
Onesios et al., 2009; Sangion and Gramatica, 2016). Reuse of products
from SSS in land applications could lead to contamination of surface
and ground water, soil and agricultural products. Though the discharge
requirements for organic micropollutants in municipal wastewater

⁎
Corresponding author at: Wageningen UR, Environmental Technology, P.O. Box 17, 6700AA, Wageningen, The Netherlands.
E-mail address: Andrii.Butkovskyi@wur.nl (A. Butkovskyi).

http://dx.doi.org/10.1016/j.envres.2017.03.044
Received 1 August 2016; Received in revised form 16 February 2017; Accepted 28 March 2017
Available online 12 April 2017
0013-9351/ © 2017 Elsevier Inc. All rights reserved.
A. Butkovskyi et al.
effluents are not in place, the need for micropollutant mitigation in
wastewater and solid waste is recognized, especially for treatment
systems which products are intended for resource recovery
(Schwarzenbach et al., 2006; Snyder et al., 2003). Pharmaceuticals
and estrogens in black water and effluent of black water treatment
system, as well as personal care products in grey water and effluent of
grey water treatment system were identified and quantified (de Graaff
et al., 2011; Hernández Leal et al., 2010).
This paper addresses the presence of organic micropollutants,
namely pharmaceuticals and constituents of personal care and house-
hold products (PCPs), in the potential products of full scale SSS system.
Consumption rates, human and environmental toxicity and estrogeni-
city as well as physical and chemical properties of selected micro-
pollutants were considered for the choice of the micropollutants for this
study. The paper covers both liquid and solid streams, also addressing
the presence of pharmaceuticals in grey water and the presence of PCPs
in black water. The quality of the recovered water and solid products
(aerobic and anaerobic sludge and struvite) intended for reuse, and
consequences for their reuse strategies are specific objectives of the
study.
2. Materials and methods
2.1. Source Separated Sanitation System
The full scale demonstration SSS project Noorderhoek (Sneek, the
Netherlands) consists of the separated black and grey water collection
and treatment system, which served 62 apartments during the sampling
period (Fig. 1)(STOWA, 2014). Black water is collected together with
shredded kitchen waste by vacuum pumps. The treatment system
consists of an underground equalization tank (HRT =24 h), up-flow
anaerobic sludge blanket (UASB) reactor (T =32 °C, HRT =35 d),
oxygen-limited autotrophic nitrification-denitrification (OLAND) in a
biorotor and magnesium ammonium phosphate (struvite) precipitation.
Grey water is collected to the equalization tank (HRT =46 h) and
treated in an aerobic activated sludge system (HRT=10 h) followed by
a settler. The average black water flow during the sampling period was
1.1 m3/d, and the average grey water flow – 5.0 m3/d.
The project was launched in summer 2011. The effluent black water
was added to the equalization tank of grey water treatment system since
May 2013 for post-treatment in aerobic reactor. The aerobic grey water
sludge has been added to UASB reactor since July 2013 in order to
increase the energy efficiency of the concept.
Fifteen grab influent and effluent samples of both treatment
systems, UASB sludge, struvite and effluent grey water were collected
in autumn 2014 at random weekdays and at random daytime.
Approximately 5 l of liquid were collected from sampling tap into a
UASB
Struvite
OLAND reactor
Black
water Equalization
tank
Sludge Struvite
Grey
water
Equalization Aerobic sludge
tank reactor
Sludge
Fig. 1. Full-scale Source Separated Sanitation system Noorderhoek. Addition of effluent
black water to grey water treatment system and grey water sludge to UASB reactor are
marked with dotted lines.
Environmental Research 156 (2017) 434–442
plastic bucket, of which 30 ml of the sample was taken and stored at
−20 °C until the analysis. UASB sludge was sampled from the lowest
sampling tap of the UASB reactor (h =1 m). Grey water sludge was
sampled from the bottom of the sedimentation tank. Struvite was taken
directly from the precipitation unit. All samples were stored at −20 °C
till the analysis.
2.2. Micropollutant analysis
Analysed pharmaceuticals included acetaminophen, ibuprofen, di-
clofenac, naproxen, trimethoprim ciprofloxacin, metformin, hydro-
chlorothiazide, gemfibrozil, atenolol, propranolol, metoprolol, carba-
mazepine, oxazepam and three transformation products (desmethylna-
proxen, guanyl urea and carbamazepine-diol). Analysed PCPs included
hexylcinnamaldehyde (HCA), methylparaben, ethylparaben, propylpar-
aben, butylparaben, ethylhexyl metoxycinnamate (EHMC), avoben-
zone, phenylbenzimidazole sulfonic acid (PBSA), benzalkonium chlor-
ide (70/30 v/v mixture of benzyldimethyldodecylammonium chloride
and benzyldimethyltetradecylammonium chloride), methyltriclosan,
2,4-dichlorophenol, triclocarban, triclosan and galaxolide (HHCB).
Single standard micropollutant solutions were prepared in methanol.
Calibration solutions were prepared by mixing of single standard
solutions and diluting the mixture with methanol. All standard solution
were stored at −20 °C.
The solid fractions of influent black water, influent grey water and
UASB sludge were separated by centrifugation before the analysis.
Effluent black water and effluent grey water were analysed without
separation of liquid and solid phases. All liquid samples were spiked
with an internal standard solution and filtrated through the 0.45 µm
PTFE filter. The filter was subsequently washed with 5 ml of acetoni-
trile. The 0.9 ml of this acetonitrile was evaporated till dryness under
nitrogen gas and the precipitate was reconstituted in 0.1 ml of methanol
and 0.9 ml of filtrated liquid.
Solid fractions of the influent black water and grey water, UASB
sludge, struvite and grey water sludge were spiked with internal
standard solution, mixed with 5 ml of acetonitrile and sonicated in
the ultrasonic bath during 20 min. The sonicated samples were
centrifuged and the supernatant was evaporated till dryness under
nitrogen gas. The precipitates were reconstituted in 0.1 ml of methanol
and 0.9 ml of distilled water.
Pharmaceuticals and PCPs with low volatility (triclocarban, avo-
benzone, benzalkonium chloride and PBSA) were analysed by direct
injection of liquid samples or liquid extracts of solid samples to LC-MS/
MS (Agilent 6410 triple quadruple mass spectrometer with electrospray
ionization). The chromatographic separation was done on Phenomenex
Kinetex Phenyl-Hexyl column (h =100 mm, d =2.1 mm, 100 Å pore
size). Mass spectrometer was operated in single reaction monitoring
mode (SRM) (Butkovskyi et al., 2015).
Galaxolide, HCA, EHMC, parabens, triclosan, methyltriclosan and
Effluent 2,4-dichlorophenol were extracted from liquid phase and acetonitrile
washings, liquid samples or liquid extracts of solid samples by PDMS-
twister extraction, as described by Hernández Leal et al., (2010). The
extraction was followed by thermal desorption and gas chromatography
with tandem mass-spectrometry (Agilent 6890N gas chromatograph
(GC) and Agilent 5975XL mass spectrometer (MS) modified with a
Chromtech Evolution triple quadrupole).
Effluent
Internal standards were added to all samples for quality assurance.
Recoveries were determined as a ratio between the peak areas of spiked
matrix sample and spiked blank (milliQ) samples (Ternes Joss, 2006).
The limits of detection (LOD) were calculated as a signal/noise ratio
=3 and the limits of quantification (LOQ) – as a signal/noise ratio
=10. Recoveries, limits of detection and limits of quantification are
presented in the Appendices (Appendices 1–5).
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A. Butkovskyi et al. Environmental Research 156 (2017) 434–442

Table 1
Concentration of the studied micropollutants in influent black water, grey water and effluents of black water and grey water treatment systems.

Compound Concentration in black water, Concentration in black water effluent, Concentration in grey water, Concentration in grey water effluent,
µg/l µg/l µg/l µg/l

Acetaminophen 7027.6 ± 1075.6 < LOD 191.7 ± 51.5 < LOD

Ibuprofen 78.1 ± 28.7 < LOD 5.3 ± 3.3 < LOD
Diclofenac 4.8 ± 3.0 1.6 ± 1.1 1.7 ± 0.9 1.7 ± 0.9
Naproxen 111.4 ± 82.8 < LOD 1.3 ± 0.7 < LOQ
Desmethylnaproxen 20.1 ± 15.5 < LOD < LOQ < LOQ
Ciprofloxacin 127.6 ± 125.5 1.8 ± 0.3 < LOQ < LOQ
Metformin 6303.8 ± 668.2 22.0 ± 10.0 25.8 ± 23.7 8.7 ± 8.6
Guanyl Urea 4347.1 ± 996.4 10.2 ± 2.4 < LOD 7.4 ± 8.5
Hydrochlorothiazide 203.8 ± 39.7 22.4 ± 2.5 < LOD 3.8 ± 0.5
Metoprolol 298.7 ± 54.3 62.6 ± 4.8 < LOQ 7.4 ± 1.2
Atenolol < LOD < LOD < LOD < LOD
Propranolol < LOD < LOD < LOD < LOD
Oxazepam 15.4 ± 6.9 3.0 ± 0.5 < LOD < LOD
Carbamazepine < LOD < LOD < LOD < LOD
Carbamazepine-diol < LOD < LOD < LOD < LOD
Gemfibrozil < LOD < LOD < LOD < LOD
Trimethoprim 22.8 ± 9.9 < LOD < LOD < LOD
Galaxolide 1.7 ± 1.6 < LOD 12.5 ± 4.4 4.3 ± 2.7
EHMC < LOD < LOD 13.4 ± 3.1 < LOD
HCA < LOD < LOD 13.0 ± 4.0 < LOD
Methylparaben 1.5 ± 0.8 < LOD 4.4 ± 1.3 < LOD
Ethylparaben 0.1 ± 0.1 < LOD 1.6 ± 0.5 < LOD
Propylparaben 0.2 ± 0.1 < LOD 2.1 ± 0.7 < LOD
Butylparaben < LOD < LOD < LOQ < LOD
Triclosan 0.2 ± 0.2 < LOD 20.5 ± 7.0 5.8 ± 1.7
Triclocarban 0.8 ± 0.9 < LOD < LOQ 0.9 ± 0.2
2,4-dichlorophenol 0.1 ± 0.1 < LOQ 4.1 ± 0.7 < LOQ
Avobenzone < LOD < LOD 1.4 ± 0.3 0.7 ± 0.5
Benzalkonium chloride 15.5 ± 15.3 < LOQ 10.0 ± 3.9 0.5 ± 0.1
Caffeine < LOD < LOD 1174.5 ± 237.5 < LOD
PBSA < LOD < LOD 1.6 ± 0.9 1.5 ± 0.7

2.3. Risk assessment (1700 kg/m3) and PNECwater is the Predicted No Effect Concentration
in water.
To assess toxicity of individual micropollutants detected in the
effluent black water and grey water their risk quotients (RQs) were
3. Results and discussion
calculated dividing maximal reported concentrations by predicted no
effect concentrations (PNECs) for these compounds. Only compounds,
3.1. Micropollutants in the influent black water and grey water
which were quantified in the potential products of the source separated
sanitation were included in the assessment. The PNECs were calculated
Most of the analysed pharmaceuticals (ten out of fourteen) and two
from literature data on micropollutant toxicity multiplying effect
out of three transformation products were detected in black water in
concentrations of micropollutants influencing 50% of the population
low µg/l to low mg/l range (Table 1). The highest concentrations were
(EC50) by assessment factor of 1000 (EC, 2003).
observed for analgesic acetaminophen, antidiabetic metformin and its
Potential toxicity of individual micropollutants from UASB sludge
metabolite guanyl urea. Both acetaminophen and metformin are among
and grey water sludge applied as soil amendment was also assessed. The
the most widely used pharmaceuticals in the Netherlands (Oosterhuis
concentrations in the top soil layer after single sludge application event
et al., 2013). Concentrations of a number of detected pharmaceuticals
were calculated according to the formula:
in black water were higher than reported for conventional sewage. For
Csludge × APPLsludge example, Oosterhuis et al. (2013) reported metformin concentrations of
Csoil =
DEPTHsoil × RHOsoil 64–98 µg/l in the influents of two Dutch wastewater treatment plants
(WWTPs), thus, two orders of magnitude lower than those found in
where Csludge is the maximal reported concentration of micropollutant
black water in this study. Maximal concentrations of acetaminophen
in sludge, APPLsludge is the sludge application rate (0.5 kg/m2·y),
(56.9 µg/l) and metoprolol (1.52 µg/l) in the WWTP influent are also
DEPTHsoil is the mixing depth of soil (0.2 m) and RHOsoil is the bulk
two orders of magnitude lower than those reported in black water (Luo
density of soil (1700 kg/m3)(EC, 2003). The PNECs for soil compart-
et al., 2014). Contrary, maximal concentrations of diclofenac (94.2 µg/
ment were either derived directly from literature or calculated from
l) and ibuprofen (603 µg/l) in the WWTP influent are higher than
literature data on toxicity of micropollutants to terrestrial species. For
concentrations of these compounds in black water (Luo et al., 2014),
compounds with logKow < 4 PNECs were calculated from the aqueous
which is probably related to the pharmaceutical consumption patterns
toxicity data using the equilibrium partitioning method according to
of the served population. It also shows that influent concentrations of
the formula:
these pharmaceuticals at another SSS sites may significantly deviate
Ksoil − water from those observed at Noorderhoek potentially leading to increased
PNECsoil = × PNECwater × 1000
RHOsoil concentrations in the effluents.
Seven out of twelve studied PCPs and one transformation product
where Ksoil-water is the partitioning coefficient soil-water calculated
were also found in black water (Table 1). They included biocides
according to EC JRC (2003), RHOsoil is the bulk density of soil
triclosan and benzalkonium chloride, fragrance galaxolide and preser-

436
 A. Butkovskyi et al.

Table 2
Maximal concentrations, PNECs and RQs of micropollutants detected in effluent black water and grey water.

Compound Maximal concentration in black Maximal concentration in grey EC50, µg/l Toxicity endpoint PNEC, µg/l RQ (BW RQ (GW References
water effluent, µg/l water effluent, µg/l effluent) effluent)

Diclofenac 3.5 3.1 50 (Salmo trutta f. fario, 21 d, monocyte 0.05 70 62 Hoeger et al., 2005
infiltration, accumulation n liver)
Ciprofloxacin 2.4 1.5 5000 (Lemna minor, 7 d, 5 0.48 0.3 Martins et al., 2012
growth inhibition)
Metformin 35.3 27 6E+06 (Daphnia magna, 48 h, behavioural changes) 6400 0.006 0.004 Cleuvers, 2003
Guanyl urea 14.4 23.8 n.d. n.d. n.d. n.d. n.d.
Hydrochlorothiazide 27.4 4.8 100000 (Daphnia magna, 48 h, 100 0.27 0.05 Carlsson et al., 2006
acute toxicity)
Metoprolol 70.9 10 610000 (Daphnia magna, 9 d, 610 0.12 0.02 Dzialowski et al., 2006

437
growth inhibition)
Oxazepam 3.7 < LOQ 500 (Daphnia magna, 48 h, 0.5 7.4 – Calleja et al., 1994
acute toxicity)
Galaxolide < LOD 9.69 140 (Pimephales promelas, 32 d, 0.14 – 69.3 HERA, 2004
post-hatching)
Triclosan < LOD 8.97 0.7 (Scenedesmus subspicatus, 96 h, decrease of 0.0007 – 12857.1 Orvos et al., 2002
population biomass)
Triclocarban < LOD 1.35 3.1 (Ceriodaphnia dubia, 48 h, intoxication, 0.0031 – 435.5 EPA/OTS, 1992
immobilization)
Avobenzone < LOD 1.9 n.d. n.d. n.d. – N.D.
Benzalkonium chloride < LOQ 0.65 18 (Daphnia magna, 48 h, behavioural changes) 0.018 – 36.1 Canton and Mathijssen-
Spiekman, 1983
PBSA < LOD 3.1 n.d. n.d. n.d. – n.d.

n.d. - no data
Environmental Research 156 (2017) 434–442
A. Butkovskyi et al. Environmental Research 156 (2017) 434–442

vatives parabens. These compounds possibly originate from toilet

disinfectants and cleaning agents used in the toilets. Parabens are also

Hund-Rinke and Simon, 2005

used as food preservatives, therefore, they can enter black water

Canton and Mathijssen-

Dzialowski et al., 2006
through human excretion.

Amorim et al., 2010

Martins et al., 2012

Snyder et al., 2011

All analysed PCPs were detected in grey water in µg/l range

Pino et al., 2015

Spiekman, 1983
(Table 1). The concentrations of these compounds in grey water were
in the same range with their concentrations in WWTP influents (e.g.,

Reference

EC, 2008
0.03–25 µg/l for galaxolide and 0.03–23.9 µg/l). Grey water also
contained diclofenac, naproxen and ibuprofen with 70% of total

n.d.
influent diclofenac load of the SSS system located in grey water stream.

3.5 − 17.6
These compounds belong to the anti-inflammatory pharmaceuticals,
thus, their presence in grey water can be explained by washout of the

0.0026
0.03

0.05
topical applications from human skin in the showers. Additionally,

n.d.

0.2

0.2

0.6
RQ
urination in the showers might explain occurrence of pharmaceuticals

PNEC, µg/kg
in the grey water.

0.8 − 4
5380

3299
3.2. Micropollutants in liquid products of SSS system

280a
740

n.d.

203
2.2

40
The liquid products intended for water reuse or discharge, include
effluents of black water and grey water treatment systems. Effluent

(Microbial ammonium oxidation test ISO/DIS

Battery of soil species belonging to different

(Daphnia magna, 48 h, behavioural changes)

black and grey water contain most of the micropollutants detected in

(Daphnia magna, 9 d, growth inhibition)

(Eisenia fetida,conditions not specified)

the influents, despite the efficient removal, which was above 80% for

(Lemna minor, 7 d, growth inhibition)

majority of the studied micropollutants (Table 2). Thus, effluent black
water contained eight out of ten pharmaceuticals detected in the
influent of the treatment system. Maximal diclofenac concentrations

EC50aqua = 610000 µg/l

LC50 = 3298.63 mg/kg

measured in effluent black water were significantly higher than its

EC50aqua = 5000 µg/l

EC50soil = 2.2 mg/kg

predicted no effect concentration (PNEC) of 0.05 µg/l in the aquatic

EC50aqua = 18 µg/l
(Eisenia fetida,14 d)

LC50 = 40 mg/kg
Toxicity endpoint

taxonomic levels
environment (RQ =70), indicating the need for effluent post-treatment.
Additionally, RQ of sedative pharmaceutical oxazepam for effluent
black water was also above 1.

15685)
Risk quotients of personal care products detected in the effluent

n.d.
grey water ranged from 36 for benzalkonium chloride to 12,857 for
triclosan (Table 2). Also diclofenac was detected in the effluent grey
Calculated concentration in soil, amended with

water with maximal concentration (3.1 µg/l) similar to that in the

effluent black water (3.5 µg/l). Therefore, post-treatment of effluent
black water and grey water is required to prevent negative influence of
micropollutants on the environment and human health in case of the
effluent reuse for most of the agricultural, urban and recreational
purposes. Risk quotients of micropollutants in effluent black water will
allow its safe discharge providing sufficient dilution at the discharge
point. High risk quotients of biocides triclosan and triclocarban in grey
water effluent indicate the need for effluent post-treatment even in case
sludge, µg/kg

of discharge to surface waters with dilution factors below 12,000.

Maximal concentrations, PNECs and RQs of micropollutants detected in UASB sludge.

Combination of ozonation and activated carbon adsorption, mem-

140.5

116.9

brane filtration and/or combination of sand filtration and disinfection

39.2

12.5
14.1
8.7

9.2

0.5

6.9

can be used for effluent post-treatment (Li et al., 2009). Alternatively,

electrochemical oxidation can be used as a post-treatment step for
- PNEC derived from two long-term tests (conditions not specified)
Maximal measured concentration in UASB

mitigation of micropollutants in grey water effluent if combined with

adsorption to activated carbon for removal of the transformation by-
products (Butkovskyi et al., 2014). As efficient disinfection is also
provided by ozonation or electrochemical oxidation, simultaneous
micropollutant removal and disinfection can be achieved by applying
these post-treatment processes (Bischoff et al., 2012).

3.3. Micropollutants in solid products of SSS system

sludge, mg/kg

The solid products, intended for resource recovery, include UASB

95.4

26.7

79.5

sludge, grey water sludge from an aerobic activated sludge system and
5.9

6.3
8.5
9.6

0.3

4.7

struvite.
Benzalkonium chloride

Pharmaceuticals did not tend to sorb to anaerobic sludge, which

2,4-dichlorophenol

contained three out of seventeen studied pharmaceuticals (ciproflox-

acin, metoprolol and propranolol) and five out of twelve studied PCPs
Ciprofloxacin

Triclocarban
Propranolol

n.d. - no data

(benzalkonium chloride, triclosan, galaxolide, triclocarban and 2,4-

Metoprolol

Galaxolide

Triclosan

dichlorophenol) (Table 3). These compounds were also present in the

Table 3

HCA

influent black water. Moreover, they could end up in UASB sludge if

a

grey water sludge is added to UASB for co-digestion with black water.

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A. Butkovskyi et al. Environmental Research 156 (2017) 434–442

Table 4
Average concentrations, PNECs and RQs of micropollutants detected in grey water sludge.

Maximal measured Calculated concentration in soil, Toxicity endpoint PNEC, µg/ RQ Reference
concentration in grey water amended with sludge, µg/kg kg
sludge, mg/kg

Galaxolide 153.9 226.3 EU risk assessment report, 280 0.8 EC, 2008
derived from two long-term
tests
EHMC 217.7 319.6 n.d. n.d. n.d.
HCA 27.4 40.1 n.d. n.d. n.d.
Triclosan 140.6 206.8 Battery of soil species 0.8 − 4 51.7 − Amorim et al., 2010
belonging to different 258.5
taxonomic levels
Methyltriclosan 2.4 3.6 n.d. n.d. n.d.
Triclocarban 105.5 155.1 LC50 = 40 mg/kg 40 3.9 Snyder et al., 2011
(Eisenia fetida, conditions not
specified)
Benzalkonium 102.7 151.1 EC50aqua = 18 µg/l 203 0.7 Canton and Mathijssen-
chloride (Daphnia magna, 48 h, Spiekman, 1983
behavioural changes)
Avobenzone 46.3 68.1 n.d. n.d. n.d.

n.d. - no data

Triclosan was the only compound, which RQ for terrestrial applica-
tion of UASB sludge exceeded 1. Up to 97% of triclosan can be removed
during composting of UASB sludge with garden waste (Butkovskyi
et al., 2016). Additionally, source control, including substitution of
triclosan with other biocides in toilet disinfection products, can be
introduced.
Pharmaceuticals and PCPs were not sorbed to struvite in the current
study, except for 2,4-dichlorophenol, which concentrations in struvite
were 0.5 ± 0.6 µg/g. Soil concentrations of 2,4-dichlorophenol as low
as 0.005–0.0005 µg/g are expected, when struvite is directly applied as
fertilizer (Lozano et al., 2010). According to the European legislation,
soil is considered contaminated with 2,4-dichlorophenol if concentra-
tion of this compound in soil exceeds 10 µg/g (Bello et al., 2013).
Therefore, struvite recovered in black water treatment system, does not
threat soil contamination if applied as fertilizer. Moreover, according to
research of Ronteltap et al. (2007) concentrations of heavy metals in
struvite precipitated from human urine are lower than existing legal
limits for soil applications and typical concentrations in commercial
fertilizers and manure (Ronteltap et al., 2007).
Aerobic grey water sludge contained eight out of twelve studied
PCPs. The pharmaceutical content of grey water sludge was not studied.
The observed concentrations of galaxolide (86.5 ± 37.7 µg/g), triclosan
(84.3 ± 30.5 µg/g) and triclocarban (64.2 ± 30.5 µg/g) are higher than
those reported for sewage sludge by other authors (Clarke and Smith,
2011). Low toxicity endpoints for some of the PCPs, especially biocides
triclosan and triclocarban, result in high RQs (Table 4) and, conse-
quently, raise concerns of greywater sludge application in agriculture.
Apart from PCPs grey water sludge contains heavy metals of non-
dietary origin (Tervahauta et al., 2014). Hence soil application of grey
water sludge is not recommended, as well as addition of grey water
sludge to UASB reactor treating black water, in case if anaerobic sludge
is going to be used as fertilizer.
4. Conclusions
Most of the analysed pharmaceuticals (ten out of fourteen) and all
analysed PCPs were detected in influent black water and influent grey
water respectively. Black water also contained PCP of certain groups
(biocides, fragrances and preservatives), whereas grey water contain
anti-inflammatory and analgesic micropollutants. Despite high removal
observed for most of the compounds, effluents of black water and grey
water treatment system require additional post-treatment prior to reuse
for agricultural, urban and recreational applications or discharge to the
environment. UASB sludge and struvite can be applied as soil amend-
ment, though decrease of triclosan concentrations in UASB sludge, e.g.,
by composting process, is required. Grey water sludge is not advised for
soil applications due to the high risk quotients calculated for biocides.
Funding
This work was performed in the cooperation framework of Wetsus,
European Centre of Excellence for Sustainable Water Technology
(www.wetsus.nl). Wetsus is cofunded by the Dutch Ministry of
Economic Affairs and Ministry of Infrastructure and Environment, the
European Union Regional Development Fund, the Province of Fryslân,
and the Northern Netherlands Provinces.
Acknowledgment
The authors would like to thank the participants of the research
theme “Separation at Source” for the fruitful discussions and their
financial support. Additionally, we appreciate the efforts of Nico
Elzinga and Willem van Smeden in the performing of the sampling
campaign. The authors would also like to thank Lea Sevenou and Ton
van der Zande for assistance in the experimental part of the study.

Appendix A

See Tables A1–A5.

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Table A1
Recoveries of the studied pharmaceuticals in different matrices.

Compound Recovery, %

Black water influent Black water influent Struvite Grey water Grey water UASB sludge Struvite
(liquid phase) (solid phase) effluent influent effluent

Paracetamol 99 107 94 79 87 111 17

Ibuprofen 82 98 93 99 83 130 91
Diclofenac 94 91 103 87 94 140 104
Naproxen 101 98 84 85 85 94 10
Desmethylnaproxen 67 86 45 67 62 79 n.d.
Trimethoprim 84 93 94 89 84 107 83
Ciprofloxacin 75 51 78 78 68 71 n.d.
Metformine 116 153 95 88 86 81 88
Guanyl urea 77 43 89 47 95 17 n.d.
Hydrochlorothiazide 74 106 90 106 113 168 66
Gemfibrozil 96 97 91 89 76 138 n.d.
Atenolol 80 97 91 85 82 48 106
Propranolol 80 67 87 86 82 90 81
Metoprolol 86 98 98 90 81 127 101
Carbamazepine 31 341 93 90 86 103 79
Carbamazepine-diol 62 339 69 76 75 116 75
Oxazepam 84 88 68 73 61 106 26

n.d. – not determined

Table A2
Recoveries of the studied personal care and household products in different matrices.

Compound Recovery, %

Grey water influent Grey water effluent Grey water Black water influent Black water effluent UASB Struvite
(liquid phase) (solid phase) sludge (liquid phase) (solid pbase) sludge

Methylparaben 85.9 109.4 97.5 107.4 138.0 90.8 65.3

Ethylparaben 95.8 115.8 80.3 87.6 148.8 92.3 91.4
Propylparaben 109.1 124.8 84.8 97.9 155.5 146.9 85.4
Butylparaben 155.2 145.8 89.0 90.6 152.6 152.2 78.4
Galaxolide 99.6 104.5 102.6 73.6 95.1 75.2 96.4
Hexylcinnamaldehyde 125.0 97.1 101.9 94.7 109.8 99.9 90.6
EHMC 57.4 101.8 133.0 84.1 53.1 105.7 107.3
Avobenzone 105.7 51.1 112.3 123.1 115.1 56.5 100.8
PBSA 101.2 112.2 95.9 83.4 98.7 89.6 19.9
Benzalkonium chloride 143.3 126.2 171.2 114.0 140.7 117.1 76.2
Triclocarban 93.7 103.9 95.3 106.1 75.8 146.0 121.5
Triclosan 119.9 77.8 92.5 100.9 53.3 73.7 90.2
Methyltriclosan n.d. n.d. 27.6 85.2 91.2 101.0 101.2
2,4-dichlorophenol 103.4 109.8 111.6 98.1 92.4 121.6 107.0

n.d – not determined.

Table A3
Limits of detection and quantification of the studied pharmaceuticals in different matrices.

Compound Limits of detection Limits of quantification

Liquid matrices, µg/l UASB sludge, µg/g Struvite, µg/g Liquid matrices, µg/l UASB sludge, µg/g Struvite, µg/g

Paracetamol 0.28 0.78 0.03 1.25 2.6 0.11

Ibuprofen 1.62 0.56 0.22 7.74 1.85 0.73
Diclofenac 0.16 0.42 0.01 0.72 1.39 0.03
Naproxen 0.09 0.66 0.15 0.39 2.2 0.51
Desmethylnaproxen 0.79 1.32 n.d. 3.52 4.41 n.d.
Trimethoprim 1.01 0.45 0.04 4.57 1.52 0.15
Ciprofloxacin 0.07 n.d. n.d. 0.32 n.d. n.d.
Metformine 0.07 0.31 0.01 0.31 1.03 0.02
Guanyl urea 0.14 0.12 n.d. 0.63 0.41 n.d.
Hydrochlorothiazide 0.65 0.96 0.03 2.95 3.21 0.08
Gemfibrozil 0.68 0.61 n.d. 3.06 2.01 n.d.
Atenolol 0.47 0.82 0.01 2.12 2.72 0.03
Propranolol 0.33 0.85 0.01 1.10 2.84 0.03
Metoprolol 0.45 1.13 0.03 2.04 3.76 0.11
Carbamazepine 1.75 0.62 0.08 7.81 2.06 0.26
Carbamazepine-diol 0.84 0.21 0.01 3.72 0.67 0.03
Oxazepam 0.46 0.59 0.03 2.03 1.98 0.09

n.d. – not determined

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Table A4
Limits of detection of the studied micropollutants in different matrices calculated in µg/l for influent and effluent samples and in µg/g for sludge and struvite samples.

Compound Limits of detection (LOD)

Grey water Grey Influent of Effluent of the Sludge of Black Black UASB Struvite Black water
(liquid) water the activated activated sludge the activated water water sludge effluent
(solid) sludge system system sludge system (liquid) (solid)

Methylparaben 0.04 0.19 0.07 0.05 0.93 0.08 0.05 0.58 0.03 0.04
Ethylparaben 0.03 0.16 0.03 0.02 0.03 0.02 0.04 0.32 0.02 0.03
Propylparaben 0.03 0.04 0.03 0.02 0.22 0.03 0.04 0.04 0.03 0.03
Butylparaben 0.02 0.04 0.02 0.02 0.04 0.02 0.02 0.08 0.03 0.02
Galaxolide 0.24 2.99 0.28 0.26 3.09 0.72 0.06 0.29 0.15 0.24
Hexylcinnamaldehyde 0.06 2.95 0.14 0.16 0.71 0.78 0.09 0.52 0.07 0.06
EHMC 0.08 0.41 0.32 0.35 1.78 0.28 0.04 0.23 0.03 0.08
Avobenzone 0.37 0.33 0.32 0.57 1.97 0.36 0.03 0.19 0.17 0.37
PBSA 0.02 0.07 0.02 0.02 0.04 0.02 0.05 0.31 0.04 0.02
Benzalkonium chloride 0.34 1.13 0.17 0.14 0.20 0.04 0.19 0.04 0.06 0.34
Triclocarban 0.02 0.04 0.03 0.02 0.07 0.03 0.04 0.20 0.03 0.02
Triclosan 0.04 0.04 0.05 0.03 0.05 0.71 0.04 0.22 0.04 0.04
Methyltriclosan 0.02 0.04 0.02 0.02 0.03 0.03 0.02 0.02 0.03 0.02
2,4-dichlorophenol 0.03 0.02 0.03 0.02 0.03 0.03 0.02 0.02 0.02 0.03

Table A5
Limits of quantification of the studied micropollutants in different matrices calculated in µg/l for influent and effluent samples and in µg/g for sludge and struvite samples.

Compound Limits of quantification (LOQ)

Grey water Grey Influent of Effluent of the Sludge of Black Black UASB Struvite Black water
(liquid) water the activated activated sludge the activated water water sludge effluent
(solid) sludge system system sludge system (liquid) (solid)

Methylparaben 0.13 0.62 0.22 0.16 3.11 0.27 0.18 1.92 0.11 0.2
Ethylparaben 0.09 0.54 0.10 0.07 0.10 0.08 0.12 1.08 0.08 0.1
Propylparaben 0.09 0.12 0.11 0.07 0.74 0.11 0.14 0.14 0.09 0.1
Butylparaben 0.07 0.14 0.06 0.06 0.12 0.07 0.06 0.26 0.11 0.1
Galaxolide 0.81 9.98 0.92 0.87 10.30 2.41 0.21 0.96 0.51 1.9
Hexylcinnamaldehyde 0.21 9.82 0.48 0.52 2.35 2.59 0.29 1.72 0.24 0.3
EHMC 0.28 1.37 1.06 1.17 5.93 0.93 0.12 0.77 0.09 0.7
Avobenzone 1.24 1.09 1.07 1.89 6.57 1.21 0.11 0.63 0.57 0.5
PBSA 0.06 0.24 0.07 0.07 0.14 0.07 0.17 1.04 0.13 0.1
Benzalkonium chloride 1.14 3.78 0.55 0.47 0.65 0.13 0.64 0.12 0.19 0.7
Triclocarban 0.08 0.13 0.10 0.07 0.24 0.09 0.14 0.66 0.10 0.1
Triclosan 0.12 0.14 0.18 0.09 0.15 2.35 0.14 0.72 0.14 0.3
Methyltriclosan 0.07 0.14 0.06 0.08 0.11 0.09 0.05 0.05 0.11 0.1
2,4-dichlorophenol 0.10 0.07 0.11 0.08 0.09 0.11 0.06 0.05 0.08 0.1

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