Chemosphere 134 (2015) 395–401

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Chemosphere
journal homepage: www.elsevier.com/locate/chemosphere

Evaluating the degradation, sorption, and negative mass balances

of pharmaceuticals and personal care products during wastewater
treatment q
Benjamin Blair a,b,⇑, Adam Nikolaus b, Curtis Hedman c, Rebecca Klaper b, Timothy Grundl b
a
School of Public Affairs, University of Colorado Denver, 1380 Lawrence St., Ste. 500 Denver, CO 80204, United States
b
School of Freshwater Sciences, University of Wisconsin at Milwaukee, 600 East Greenfield Ave, Milwaukee, WI 53204, United States
c
State Laboratory of Hygiene, University of Wisconsin-Madison, 2601 Agriculture Drive, Madison, WI 53718, United States

h i g h l i g h t s

 Fate of 57 in situ PPCPs were assessed in activated sludge process.

 Forty-eight PPCPs were detected in soluble and twenty-nine were detected in solids.
 Negative mass balances observed for a subset of PPCPs.
 Some biodegradable PPCPs stop being degraded at low, yet notable, concentrations.

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

Article history: Conventional activated sludge (CAS) wastewater treatment processes are insufficient at removing many
Received 12 January 2015 pharmaceutical and personal care products (PPCPs) from wastewater. In addition, negative mass bal-
Received in revised form 24 April 2015 ances, where the effluent concentration is greater than the influent concentration, have been observed
Accepted 26 April 2015
in wastewater treatment studies and a further understanding of these results is needed. In this study,
the fate and occurrence of 57 PPCPs and hormones were evaluated in an activated sludge process and
the mass balances were determined. The goal of the project was to understand the PPCPs biological
Keywords:
degradation and the extent of sorption to solids. The samples containing in situ PPCPs (i.e. samples were
Pharmaceuticals
Hormones
not spiked with additional PPCPs) were evaluated. Forty-eight of the PPCPs were detected in the soluble
PPCPs form and 29 were detected sorbed to solids. Two notable results were found. First, the results of this
Sorption study indicate a subset of the highly biodegradable PPCPs stop being degraded at low, yet notable, con-
Biodegradation centrations. Second, the results revealed that negative mass balances were present for a subset of the
Wastewater PPCPs when evaluating both the soluble and sorbed concentration, for example carbamazepine and oflox-
Kinetics acin. Desorption from solids was not found to attribute to negative mass balances. Overall, the results
from this study provide new insights into the fate of PPCPs during CAS wastewater treatment by evalu-
ating the degradation kinetics and sorption and the results may explain the consistent levels of highly
degradable PPCPs being emitted from WWTPs worldwide.
Ó 2015 Elsevier Ltd. All rights reserved.

1. Introduction
Risk analyses and exposure studies have led to the concern that
the levels of pharmaceuticals and personal care products (PPCPs)
and hormones discharged into the environment from wastewater
treatment plants (WWTPs) may have a negative impact on the
q
Funding provided by the Milwaukee Metropolitan Sewerage District, Milwau-
kee, WI.
⇑ Corresponding author at: School of Public Affairs, University of Colorado
Denver, 1380 Lawrence St., Ste. 500 Denver, CO 80204, United States.
E-mail address: benjamin.blair@ucdenver.edu (B. Blair).
ecosystem (Al Aukidy et al., 2012; Blair et al., 2013a; Brodin
et al., 2013; Niemuth and Klaper, 2015). Furthermore, influent con-
centrations and the removal of PPCPs from wastewater has been
found to vary widely (Miege et al., 2009; Oulton et al., 2010;
Verlicchi et al., 2012). Information regarding the fate of PPCPs in
wastewater is limited; in particular, the understanding of the fate
of PPCPs in an aerobic wastewater treatment process is
incomplete.
WWTPs that utilize conventional activated sludge (CAS) sys-
tems have been found to present a wide range of removal efficien-
cies for different PPCPs (Verlicchi et al., 2012). During a CAS
process, three mechanisms can be used to determine the fate of

http://dx.doi.org/10.1016/j.chemosphere.2015.04.078
0045-6535/Ó 2015 Elsevier Ltd. All rights reserved.
396 B. Blair et al. / Chemosphere 134 (2015) 395–401
PPCPs: biological degradation, sorption, and volatilization (Joss
et al., 2006). Sorption to solids is expected to be significant for
PPCPs with a log octanol–water partitioning coefficient (Kow)
greater than 4.0 (Thompson et al., 2011). Few PPCPs have a
log Kow greater than 4.0 and removal by sorption was concluded
to be a minor removal mechanism for most PPCPs (Gulde et al.,
2014; Khan and Ongerth, 2002; Radjenović et al., 2009).
Volatilization can be considered negligible for the majority of
PPCPs (Joss et al., 2006). The disparity commonly observed in
PPCPs removal during aerobic wastewater treatment is likely due
to the varying operating parameters that can impact the fate such
as the solids retention time, hydraulic retention time, concentra-
tion of suspended solids, fraction of heterotrophic and autotrophic
biomass, pH, and other operating and influent conditions (Alvarino
et al., 2014; Fernandez-Fontaina et al., 2012; Gulde et al., 2014;
Majewsky et al., 2011a,b; Tran et al., 2013; Verlicchi et al., 2012).
Some studies assessing the fate of PPCPs in wastewater have
encountered issues with negative mass balances, where lower con-
centrations were seen in the influent than the effluent. Reasons for
this can include improperly addressing the fluid dynamics of a
WWTP (Majewsky et al., 2011b; Ort et al., 2010; Rodayan et al.,
2014), conjugate compounds that are not detected at the influent
could be retransformed into the original compound due to biolog-
ical processes (Kumar et al., 2012; Monteiro and Boxall, 2010;
Salgado et al., 2012; Verlicchi et al., 2012), desorption from the
return activated sludge may occur during the secondary treatment
process (Salgado et al., 2012), and PPCPs may be released from
fecal particles as the feces are being broken down by microbes
(Göbel et al., 2007).
While many studies have investigated PPCPs during an aerobic
wastewater treatment process, numerous questions remain
regarding the fate of PPCPs within conventional activated sludge
treatment processes. Therefore, the goals of this study will be to:
(1) further examine the occurrence and fate of 57 commonly used
PPCPs in an aeration basin; (2) determine the degradation rate con-
stants and sorption coefficients; and (3) assess the negative mass
balances observed in previous studies. These goals were evaluated
by determining the fate and kinetics of 57 PPCPs in mixed liquor
activated sludge from a wastewater treatment plant in
Milwaukee, Wisconsin, USA. This paper presents a substantial
PPCP data set by evaluating the hourly soluble and sorbed levels
within an aerated batch reactor using in situ PPCPs.
2. Materials and methods
2.1. Sampling site
Mixed liquor activated sludge was used from South Shore Water
Reclamation Facility (SSWRF) in Oak Creek, WI, which is a facility
that services the greater Milwaukee, WI area. SSWRF uses prelimi-
nary treatment (7 bar screens/grit channels), primary treatment
(16 primary clarifiers), conventional activated sludge treatment
(28 plug flow aeration basins and 24 secondary clarifiers) and chlo-
rine disinfection (2 5-pass contact channels). SSWRF has a treatment
capacity of 1,135,000 m3 day1 with an average flow of approxi-
mately 379,000 m3 day1. The base flow is 227,000–284,000 m3 -
day1 in dry conditions. Average flow rates on the sampling dates,
November 5, 6, and 7, 2013, were 246,000 m3 day1, 238,000 m3 -
day1, and 291,000 m3 day1, respectively. The mean hydraulic
retention time in the aeration basin at SSWRF is approximately 10 h.
2.2. Batch reactor
One hundred ninety liters of mixed liquor activated sludge from
SSWRF was transferred to a batch reactor from the head of a plug
flow aeration basin on three separate dates (November 5, 6, and 7,
2013). Eleven samples were collected per day for the mixed liquor
suspended solids (MLSS) and PPCPs analyses, where the samples
were collected initially and then hourly for the next 10 h. For the
liquid PPCPs analysis, the hourly samples were allowed to settle
for 5 min and the supernatant was decanted with a target volume
of 500 mL for each extraction method (acidic, basic, and hormone).
Two 500 mL samples of mixed liquor activated sludge samples
were collected for the solids PPCPs and MLSS analyses. The 10 h
duration was selected because under normal operating conditions,
a CAS aerobic basin has a hydraulic retention time of 6–12 h
(Metcalf and Eddy Inc., 2003). The mixed liquor was not spiked
with PPCPs; only the in situ PPCPs in the mixed liquor were evalu-
ated. Aeration was completed using a Hakko 60 L Air Pump and a
submerged Matala 22.9 cm membrane disk diffuser. Dissolved
oxygen levels were kept between 6 and 9 mg L1 (average of
7.5 mg L1) as to not limit the amount of oxygen available to the
microbes.
2.3. PPCPs analysis
PPCPs were extracted and analyzed based upon US EPA Method
1694 (USEPA, 2007a) for pharmaceuticals and US EPA Method
1698 (USEPA, 2007b) for steroids and hormones by using high per-
formance liquid chromatography combined with tandem mass
spectrometry (HPLC/MS/MS) with modifications as published by
Blair et al. (2013b). The PPCPs were selected for this study based
on the EPA methods. The same 57 PPCPs were assessed for both
the soluble and solid levels.
2.4. Kbiol calculation
To evaluate the degradation of soluble PPCPs, pseudo first-order
kinetics (Joss et al., 2006) was used to determine the biological rate
constant:
dSt
¼ K biol MLSS S0 ð1Þ
dt
where St is the soluble compound concentration at time t (ng L1), t
is hydraulic retention time (h), Kbiol is the intrinsic biological rate
constant (L g 1 h1), MLSS is the concentration of suspended solids
(average daily g L1), and S0 is the initial soluble compound concen-
tration (ng L1). Pseudo-first-order kinetics is used to describe
exponential degradation with the mixed liquor suspended solids
concentration proportionally influencing the rate.
The Kbiol values were found by regression using the negative of
the slope of the natural log of the concentration divided by the ini-
tial concentration over time, with the intercept set at zero. The Kbiol
values are based on the loss of the parent compound. In this study,
no attempt was made to evaluate the reaction products. If a com-
pound approached a degradation plateau (where degradation
appears to stop) or the MDL, the values preceding and ending at
the plateau or MDL values were used. PPCPs with a starting soluble
concentration of less than 3 times the MDL were omitted from the
Kbiol calculations.
2.5. Kd calculation
To evaluate the extent of sorption, the sorption coefficient (Kd)
is typically defined for equilibrium conditions in a batch reactor
(Joss et al., 2006):
X
Kd ¼ ð2Þ
MLSS  S
where Kd is the sorption coefficient of activated sludge (L kg1), X is
the sorbed compound concentration expressed per unit of volume
 B. Blair et al. / Chemosphere 134 (2015) 395–401
(ng L1), S is the soluble compound concentration (ng L1), and
MLSS is the mixed liquor suspended solids concentration (kg L1).
2.6. Detection limits
The minimum detection limit (MDL) and minimum quantifica-
tion limit (MQL) were found using USEPA CFR 40, part 136 (USEPA,
2003). The MDL and MQL were found using ten replicates of a
5.0 ng mL1 standard for each compound. The MDL was found
using the student’s t value for a 99% confidence interval multiplied
by the standard deviation from the ten replicates. The MQL was
defined as the three times the MDL value. Positive and negative
controls were conducted for all PPCPs. Concentrations in the nega-
tive control were below the MDL for the majority of the com-
pounds. Two compounds had concentrations detected in the
negative controls with average values above the MQL: diltiazem
(4.1 ng L1) and triclosan (1.7 ng L1).
3. Results and discussion
3.1. PPCPs detected in liquids
Over the three sampling dates, 48 PPCPs were detected in sol-
uble form above the MQL. The classifications and maximum
detected concentration of the PPCPs detected at SSWRF are shown
in Table 1. The Kbiol values could be calculated for 29 PPCPs and
these values are also shown in Table 1. The daily average for the
MLSS on day 1 was 1.44 g L1 (standard deviation of 0.54), day 2
was 1.99 g L1 (standard deviation of 0.37), and day 3 was
1.76 g L1 (standard deviation of 0.49). In addition, Table 1 includes
the MQL and MDL values for each specific compound and the
removal efficiencies over the 10 h period based on the change in
the soluble concentration. The removal efficiencies used the aver-
age concentration at time zero and the average concentration at
10 h.
A predominant practice in the literature for evaluating PPCP
degradation is reporting PPCPs removal efficiencies; however the
removal efficiencies are subject to many sources of error, notably
variable PPCP influent levels and the analytical uncertainty at
levels that approach the MDL can cause significant errors in the
removal efficiencies. Hereafter, we use the intrinsic biodegradation
rates (Kbiol) as they represent a better understanding of the soluble
degradation of PPCPs within an activated sludge wastewater treat-
ment process. These intrinsic rates can be applied to any treatment
system with known amount of MLSS. For our system, with an over-
all MLSS average of 1.7 g L1, a Kbiol of 0.41 L g1 h1 results in an
observed degradation half-life of 1 h. The fastest observed degrada-
tion rate (acetaminophen, Kbiol = 3.05 L g1 h1) yields a half-life of
8 min.
3.2. PPCPs detection in solids
Twenty-nine PPCPs were detected above the minimum detec-
tion limit sorbed to the mixed liquor activated sludge. The Kd val-
ues were calculated using Eq. (2) and the values are presented in
Table 1 along with the standard deviation and count of samples
used in the Kd calculations.
Sorption equilibrium is often assumed for PPCPs in an aeration
basin, for example equilibrium was observed after 0.5 h for 4 g
MLSS L1 (Ternes et al., 2004). However, a system at sorptive equi-
librium would exhibit sorbed concentrations that diminish in con-
cert with soluble concentrations. Diminishing aqueous
concentrations drive desorption from the solids in order to main-
tain sorptive equilibrium. For many PPCPs, the level sorbed to
the mixed liquor exhibited little change over the sampling period,
397
although the soluble concentrations were found to decrease over
time. This is exemplified by the anti-diabetic metformin (Fig. 1)
where soluble concentrations drop nearly two orders of magnitude
whereas sorbed concentrations remain nearly constant at
10 ng L1. Metformin is widespread in WWTP influent at concen-
trations as high as 129,000 ng L1 and the removal efficiency in
WWTPs has been found to range from 41% to over 98% (Scheurer
et al., 2009, 2012; Trautwein and Kümmerer, 2011; Blair et al.,
2013b; Oosterhuis et al., 2013). Metformin is indicative of slow
(non-equilibrium) sorption therefore it should be recognized that
the Kd values given in Table 1 are qualitative indicators of sorptive
behavior, not true equilibrium sorption. These values are useful in
estimating the proportion of any given compound that leaves the
aeration basin on the solids. In our case, with an average MLSS of
1.7 g L1, the transport is 50:50 solid:aqueous when the
Kd = 588 L kg1. Only 6 compounds reach this level of solid phase
transport (4-androstene-3,17-dione, fluoxetine, progesterone,
testosterone, triclocarban, triclosan).
3.3. Mass balances
Mass balances were determined by evaluating the degradation
rate and sorption to solids. Twelve compounds had fast degrada-
tion rates that corresponded to more than 0.2 L g1 h1
(half-life 6 2 h), which included 17,20-dihydroxyprogesterone,
acetaminophen, caffeine, cotinine, digoxigenin, diphenhydramine,
estriol, estrone, ibuprofen, metformin, naproxen, and paraxan-
thine. Metformin illustrates a PPCP with fast degradation (Fig. 1)
and the metformin concentration decreased consistently over time
on the three separate sampling dates, as shown in Fig. 1a.
In contrast to metformin, some PPCPs had little degradation
over the 10 h period, such as diltiazem, the calcium channel
blocker used to treat hypertension and angina. Diltiazem had lim-
ited removal through degradation and the Kbiol values were found
to be 0.00, 0.01, and 0.05 L g1 h1 across the three sampling
dates, as displayed in Fig. 2. Other compounds had consistent
detection and incomplete removal, such as the opiate codeine
(Kbiol values of 0.00, 0.01 and 0.05 L g1 h1) and the fibrate gemfi-
brozil (Kbiol values of 0.03, 0.06, and 0.00 L g1 h1).
A subset of the compounds had greater mass in the solids than
in the soluble form, such as the antimicrobials triclocarban and tri-
closan, as displayed in Fig. 3a and b. Triclosan and triclocarban
often partition to sediments or solids and are among the most fre-
quently detected organic contaminants in environmental samples
(Blair et al., 2013b; Carey and McNamara, 2015; Lozano et al.,
2013; Pycke et al., 2014). Triclocarban was found to have an aver-
age Kd of 12,000 L kg1, which classifies it one of the more
hydrophobic PPCPs and the most hydrophobic compound assessed
in this study. For triclocarban, the level measured in solids was
approximately twelve times greater than the level measured in sol-
uble form. As a whole, the mass of triclocarban in solids was not
found to significantly change over time. Likewise, the mass balance
for triclosan, with an average Kd of 2600 L kg1, is shown in Fig. 3b.
Triclocarban and triclosan have log Kow values greater than 4.5,
therefore sorption was expected in an activated sludge treatment
process (Blair et al., 2013b; Lozano et al., 2013).
3.4. Degradation plateaus
Degradation plateaus – where the degradation appears to slow
or stop contrary to the expected results from pseudo-first-order
kinetics – may explain the low levels of PPCPs emitted from
SSWRF in the previous research (Blair et al., 2013b). A visible
degradation plateau was seen for acetaminophen (90 ng L1), caf-
feine (40 ng L1), and metformin (1000 ng L1), which are easily
degradable PPCPs (Kbiol greater than 0.2 L g1 h1). For metformin,
398 B. Blair et al. / Chemosphere 134 (2015) 395–401

Table 1
Minimum detection limit (MDL), minimum quantification limit (MQL), maximum concentration, average 10 h removal efficiency, degradation rate constants (Kbiol), sorption
coefficient (Kd) with corresponding standard deviation, and count of the number of samples for 57 PPCPs.

Compound MDL MQL Soluble Solids

Maximum Removal Kbiol Day 1 Kbiol Day 2 Kbiol Day 3 Kd Kd Std. Kd
concentration efficiency deviation Count
ng L 1
ng L 1
ng L1 % Lg 1
h 1
Lg1
h 1
Lg1
h1
L kg 1
L kg1 Countb
17,20-Dihydroxyprogesterone 1.4 4.2 Below MQL NA 1.68a 1.74a 1.27a NA NA 0
17-Alpha-ethinylestradiol 1.2 3.5 ND NA NA NA NA NA NA 0
4-Androstene-3,17-dione 0.5 1.4 5.8 15.20% NA NA NA 1800 1600 31
5-Hydroandrostene-3,17-dione 2.3 6.9 8.8 NA NA NA NA NA NA 0
Acetaminophen 2.5 7.5 13,000 97.10% 2.23a 2.28a 3.05a 84 88 28
Ampicillin 3.2 8 160 1.30% 0.07 0.02 0.05 30 17 7
Androsterone 0.5 1.6 4 13.30% NA NA NA NA NA 0
Azithromycin 3.7 11 1300 47.90% 0.01a 0.07a 0.13a 130 280 33
Boldenone 1.3 4 Below MQL NA NA NA NA NA NA 0
Caffeine 3.1 9.3 11,000 99.30% 2.12a 1.65a 2.03a 140 140 31
Carbadox 3.4 10.1 130 367.80% NA NA NA NA NA 0
Carbamazepine 2.7 8.2 220 92.40% 0.06 0.05 0 10 3 10
Ciprofloxacin 3.3 9.9 2200 88.60% 0.08 0.04 0 500 310 33
Clarithromycin 3.2 9.6 8100 72.50% 0.07 0.03 0.02 130 320 27
Codeine 3.6 10.7 370 32.60% 0 0.01 0.05 14 5 2
Cotinine 3.5 10.6 260 54.20% 0.71a 0.69a 0.73a 34 0 1
Digoxigenin 4.4 13.2 240 94.30% 0.57a 0.72a 0.73a 150 84 6
Dihydrotestosterone 1.1 3.3 51 97.20% NA NA NA NA NA 0
Diltiazem 3.5 10.4 260 13.40% 0 0.01 0.05 22 9 23
Diphenhydramine 3.6 10.9 84 90.50% 0.24a 0.24a 0.44a 170 89 12
Enrofloxacin 1.4 4.1 34 43.80% NA NA NA NA NA 0
Estriol 2 6.1 60 66.80% 1.78a 0.38a 0.74a NA NA 0
Estrone 2.2 6.7 57 93.70% 2.29a 1.54a 0.91a NA NA 0
Flumequine 5.2 15.6 Below MQL NA NA NA NA NA NA 0
Fluoxetine 3.5 10.5 50 23.10% NA NA NA 1200 720 31
Gemfibrozil 1.6 4.8 190 50.80% 0.03 0.06 0 9 2 2
Ibuprofen 4.7 14 4500 99.70% 1.68a 1.79a 2.09a 170 250 7
Lincomycin 3.1 9.3 32 50.40% NA NA NA NA NA 0
Melengestrol acetate 0.6 1.7 220 90.80% 2.92a NA NA NA NA 0
Metformin 0.5 1.5 99,000 98.60% 0.54a 0.47a 0.30a 3 5 33
Nandrolone 2 6 ND NA NA NA NA NA NA 0
Naproxen 1 2.9 3000 96.20% 1.07a 1.04a 1.22a 120 180 29
Norfloxacin 5.1 15.3 140 164.90% NA NA NA 190 220 28
Ofloxacin 3.9 11.7 2100 124.20% 0.09 0.06 0.03 250 210 33
Oxacillin 2.5 7.4 Below MQL NA NA NA NA NA NA 0
Paraxanthine 6.1 18.2 5200 99.30% 1.41a 1.56a 2.02a 85 120 3
Penicillin G 3.4 10.3 30 125.30% NA NA NA 310 0 1
Penicillin V 2.7 8 86 1174.00% NA NA NA NA NA 0
Progesterone 0.7 2 6.1 34.60% NA NA NA 1900 1500 32
Ranitidine 0.9 2.6 40 57.20% 0.06a NA NA 430 590 13
Roxithromycin 4.3 13 120 81.00% NA 0.46a 0.47a NA NA 0
Sarafloxacin 5.4 16.3 130 59.60% NA NA NA NA NA 0
Sulfachloropyridazine 4.1 12.3 30 41.70% NA NA NA NA NA 0
Sulfadiazine 2.8 8.5 20 64.10% NA NA NA NA NA 0
Sulfadimethoxine 2.4 7.1 52 149.70% NA NA NA NA NA 0
Sulfamerazine 2.1 6.2 30 9.90% NA NA NA NA NA 0
Sulfamethazine 4 12.1 40 4.60% NA NA NA NA NA 0
Sulfamethizole 4.2 12.7 60 32.30% NA NA NA NA NA 0
Sulfamethoxazole 4.1 12.4 7400 35.80% 0.06 0.04 0.01 10 9 32
Sulfathiazole 2.6 7.8 92 49.40% NA NA NA NA NA 0
Testosterone 1.1 3.2 Below MQL NA NA NA NA 1400 90 2
Thiabendazole 1.8 5.3 17 440.80% NA NA NA 150 50 10
Triclocarban 0.5 1.4 540 11.40% 0.02a 0.05a 0.18a 12,000 8900 33
Triclosan 0.5 1.6 300 55.30% 0.05a 0.05a 0.15a 2500 1500 33
Trimethoprim 3.4 10.7 570 53.10% 0.06 0.04 0.01 14 6 29
Virginiamycin 6.4 19.1 Below MQL NA 0.06a 0.08a 0.01a NA NA 0
Warfarin 0.8 2.5 ND NA NA NA NA NA NA 0

‘‘ND’’ is no detection.
‘‘Below MQL’’ is used when the concentration is below the MQL, but above the MDL.
‘‘NA’’ was used if the maximum concentration was less than the MQL or influent concentration had no peaks at time 0.
a
Kbiol value approached a plateau or the MDL prior to 10 h and less than 11 samples were used in the Kbiol regression.
b
Count of number of samples with soluble and solid levels both above MDL.

the observed concentration reached a plateau at hour 8, where the trend was observed on each of the three sampling dates. The met-
degradation appeared to slow or stop. As shown in Fig. 1b with a formin degradation plateau is visible from time 8 to 10.
log scale concentration axis, this plateau was reached at approxi- Acetaminophen and caffeine degraded quickly and the degradation
mately 1000 ng L1 at time eight hours for metformin and this plateau started at hour 2 or 3 and stayed approximately level for
 B. Blair et al. / Chemosphere 134 (2015) 395–401 399

100000
a 3.5. Negative mass balances
Concentration (ng/L)

80000 Negative Kbiol values, where the soluble concentration increased

over time, were seen for seven PPCPs (Table 1). Negative removal
60000 efficiencies were also observed in the previous work at SSWRF
(Blair et al., 2013b) and in numerous other WWTP monitoring
40000
studies (e.g. Dai et al., 2014; Oulton et al., 2010; Sun et al., 2014).
However, by eliminating the hydrodynamic variation and evaluat-
20000
ing in situ PPCPs, we can better assess the fate, and potential for
negative mass balances of PPCPs in an aeration basin. To illustrate
0
0 2 4 6 8 10 PPCPs with negative mass balances, the concentration of the
Time (hours) antiepileptic carbamazepine and the antibiotic ofloxacin are shown
Soluble Sorbed in Fig. 4a and b, respectively.
Desorption from solids does not explain the increase in the sol-
100000 b uble concentration for these compounds as the soluble concentra-
tion increases and the sorbed level remains stable, on average.
Concentration (ng/L)

10000 Others have reported adjusting the sampling by the hydraulic

retention time eliminates the negative mass balances (Rodayan
1000 et al., 2014), however, negative mass balances were observed in
the batch reactors when evaluating the in situ PPCPs. Analytical
100 uncertainty is apparent at the low levels of PPCPs evaluated, yet
the overall negative mass balance and the same trend is observed
10
on each of the three sampling dates for these two compounds.
Therefore, two potential theories exist to explain the observed neg-
1
0 2 4 6 8 10 ative mass balances of these PPCPs in an aerobic treatment pro-
Time (hours) cess: (1) the PPCPs are enclosed in fecal particles and are
Soluble Sorbed
released from these particles when the feces is broken down by
the microbes (Göbel et al., 2007); or (2) the undetected PPCPs
Fig. 1. (a) Average metformin soluble and sorbed concentration over time. (b) metabolites are being retransformed into the parent compound
Average metformin soluble and sorbed concentration, log scale y-axis. Solid line is through microbial activity (Salgado et al., 2012; Verlicchi et al.,
the first order degradation rate using average of the Kbiol values listed in Table 1.
2012). We posit the cause of negative mass balances is a combina-
Error bars are the standard deviations from the three sample dates.
tion of these processes, with the driving factor being compound
specific.
the remaining samples. Therefore, it appeared that the degradation
of some degradable PPCPs slows at a compound specific
concentration.
3000
Triclosan and triclocarban also had notable degradation pla-
a
teaus. Triclosan had fast degradation occur at concentrations
Concentration (ng/L)

2500
greater than 100 ng L1, however, a degradation plateau was
observed when the soluble concentration reached 60 ng L1 on 2000
each of the three sampling dates. Triclocarban is also noteworthy 1500
due to a visible degradation plateau at 50 ng L1. The starting
concentration on day 1 was at the degradation plateau 1000
(54 ng L1) and degradation was not observed (Kbiol value of 500
0.02 L g1 h1). Similar to triclosan, on the second and third sam-
pling dates, degradation was observed (Kbiol values of 0.05 and 0
0 1 2 3 4 5 6 7 8 9 10
0.15 L g1 h1) because starting concentrations were higher than
Time (hours)
the degradation plateau (170 and 540 ng L1 respectively).
Soluble Sorbed Soluble+Sorbed

300 1000
b
Concentration (ng/L)
Concentration (ng/L)

250
800
200
600
150
400
100

50 200

0 0
0 1 2 3 4 5 6 7 8 9 10 0 1 2 3 4 5 6 7 8 9 10
Time (hours) Time (hours)
Soluble Sorbed Soluble Sorbed Soluble+Sorbed

Fig. 2. Average soluble and sorbed concentration of diltiazem over time. Solid line Fig. 3. Mass balance for (a) triclocarban and (b) triclosan. Error bars on the standard
is the first order degradation rate using average of the Kbiol values listed in Table 1. deviation on the soluble + solids. Solid line is the linear trend line of the
Error bars are the standard deviations from the three sample dates. soluble + sorbed concentration.
400
250
Concentration (ng/L)
200
150
100
50
0 back regarding the research design.
0 1 2 3 4 5
Time (hours)
Soluble Sorbed
2500
Concentration (ng/L)
2000
1500
1000
500
0 care products, and hormones from wastewater. Sci. Total Environ. 444, 515–
0 1 2 3 4 5
Time (hours)
Soluble Sorbed
2
Fig. 4. Mass balance for (a) carbamazepine (R = 0.66, p < .05) and (b) ofloxacin
(R2 = 0.42, p < .05). Error bars on the soluble + solids represent the standard
deviation. Solid line is the linear trend line of the soluble + sorbed concentration.
4. Conclusion
The presence of PPCPs in treated wastewater and surface waters
is a significant environmental concern. This work presented the
sorption and Kbiol values of 57 PPCPs in a mixed liquor activated
sludge batch reactor and two notable advances in the field of
PPCPs in wastewater were found: (1) demonstrated that the low
concentration of highly degradable PPCPs detected in the effluent
may be due to degradation stopping or slowing at a compound
specific concentration; and (2) a further evaluation of the negative
mass balances observed for a subset of in situ soluble and sorbed
PPCPs within an aerobic batch reactor.
The results demonstrated that a subset of highly degradable
PPCPs, such as acetaminophen, caffeine, and metformin, stopped
being degraded at notable levels within an activated sludge
wastewater treatment process. We found degradation plateaus
levels to be compound specific, with some the degradation of some
compounds slowing or stopping at concentrations as high as
1000 ng L1. Understanding this phenomenon is of critical impor-
tance as it may explain the continuous low levels of degradable
PPCPs being emitted from SSWRF and WWTPs worldwide that uti-
lize an activated sludge wastewater treatment process.
To our knowledge, this is the first study demonstrating in situ
PPCPs negative mass balances – the sum of the soluble and sorbed
concentrations increased over time – within an aerobic batch reac-
tor. By using a batch reactor and evaluating the soluble and sorbed
levels, the potential that the metabolites are retransformed into
the parent compound through biological activity still exists.
Likewise, we posit these PPCPs were potentially enclosed in fecal
particles and released when the particles were disturbed or
degraded or metabolites were retransformed to the parent com-
pound. The results indicate that the use of updated sampling pro-
tocols, such as eliminating variability by using the hydraulic
B. Blair et al. / Chemosphere 134 (2015) 395–401
a retention time, will not eliminate the negative mass balances for
a subset of PPCPs. Overall, by evaluating mass balances through
the degradation and sorption of PPCPs in an aeration basin, a better
understanding of the fate of PPCPs in an aeration basin can be
developed.
Acknowledgments
We thank Patrick McNamara and Daniel Zitomer for their feed-
6 7 8 9 10
References
Soluble+Sorbed
b Al Aukidy, M., Verlicchi, P., Jelic, A., Petrovic, M., Barcelò, D., 2012. Monitoring
release of pharmaceutical compounds: occurrence and environmental risk
assessment of two WWTP effluents and their receiving bodies in the Po Valley,
Italy. Sci. Total Environ. 438, 15–25.
Alvarino, T., Suarez, S., Lema, J.M., Omil, F., 2014. Understanding the removal
mechanisms of PPCPs and the influence of main technological parameters in
anaerobic UASB and aerobic CAS reactors. J. Hazard. Mater. 278, 506–513.
Blair, B.D., Crago, J.P., Hedman, C.J., Klaper, R.D., 2013a. Pharmaceuticals and
personal care products found in the Great Lakes above concentrations of
environmental concern. Chemosphere 93, 2116–2123.
Blair, B.D., Crago, J.P., Hedman, C.J., Treguer, R.J., Magruder, C., Royer, L.S., Klaper,
R.D., 2013b. Evaluation of a model for the removal of pharmaceuticals, personal
6 7 8 9 10 521.
Brodin, T., Fick, J., Jonsson, M., Klaminder, J., 2013. Dilute concentrations of a
psychiatric drug alter behavior of fish from natural populations. Science 339,
Soluble+Sorbed 814–815.
Carey, D.E., McNamara, P.J., 2015. The impact of triclosan on the spread of antibiotic
resistance in the environment. Front. Microbiol. 5, 780.
Dai, G., Huang, J., Chen, W., Wang, B., Yu, G., Deng, S., 2014. Major pharmaceuticals
and personal care products (PPCPs) in wastewater treatment plant and
receiving water in Beijing, China, and associated ecological risks. Bull.
Environ. Contam. Toxicol. 92, 655–661.
Fernandez-Fontaina, E., Omil, F., Lema, J.M., Carballa, M., 2012. Influence of
nitrifying conditions on the biodegradation and sorption of emerging
micropollutants. Water Res. 46, 5434–5444.
Göbel, A., McArdell, C.S., Joss, A., Siegrist, H., Giger, W., 2007. Fate of sulfonamides,
macrolides, and trimethoprim in different wastewater treatment technologies.
Sci. Total Environ. 372, 361–371.
Gulde, R., Helbling, D.E., Scheidegger, A., Fenner, K., 2014. PH-dependent
biotransformation of ionizable organic micropollutants in activated sludge.
Environ. Sci. Technol. 48, 13760–13768.
Joss, A., Zabczynski, S., Göbel, A., Hoffmann, B., Löffler, D., McArdell, C.S., Ternes, T.A.,
Thomsen, A., Siegrist, H., 2006. Biological degradation of pharmaceuticals in
municipal wastewater treatment: proposing a classification scheme. Water Res.
40, 1686–1696.
Khan, S.J., Ongerth, J.E., 2002. Estimation of pharmaceutical residues in primary and
secondary sewage sludge based on quantities of use and fugacity modelling.
Water Sci. Technol. 46, 105–113.
Kumar, V., Johnson, A.C., Nakada, N., Yamashita, N., Tanaka, H., 2012. De-
conjugation behavior of conjugated estrogens in the raw sewage, activated
sludge and river water. J. Hazard Mater. 227, 49–54.
Lozano, N., Rice, C.P., Ramirez, M., Torrents, A., 2013. Fate of triclocarban, triclosan
and methyltriclosan during wastewater and biosolids treatment processes.
Water Res. 47, 4519–4527.
Majewsky, M., Gallé, T., Yargeau, V., Fischer, K., 2011a. Active heterotrophic biomass
and sludge retention time (SRT) as determining factors for biodegradation
kinetics of pharmaceuticals in activated sludge. Bioresour. Technol. 102, 7415–
7421.
Majewsky, M., Galle, T., Bayerle, M., Goel, R., Fischer, K., Vanrolleghem, P.A., 2011b.
Xenobiotic mass balances in wastewater treatment plants: residence time
distributions as a guiding principle for sampling strategies. Water Res. 45,
6152–6162.
Metcalf and Eddy Inc., 2003. Wastewater Engineering, fourth ed. Tata McGraw Hill.
Miege, C., Choubert, J.M., Ribeiro, L., Eusèbe, M., Coquery, M., 2009. Fate of
pharmaceuticals and personal care products in wastewater treatment plants–
conception of a database and first results. Environ. Pollut. 157, 1721–1726.
Monteiro, S.C., Boxall, A.B.A., 2010, Occurrence and fate of human pharmaceuticals
in the environment. In: Whitacre, D.M. (Ed.), Reviews of Environ. Contam. and
Toxicol., vol. 202, pp. 53–154.
Niemuth, N.J., Klaper, R.D., 2015. Emerging wastewater contaminant metformin
causes intersex and reduced fecundity in fish. Chemosphere 135, 38–45.
Oosterhuis, M., Sacher, F., ter Laak, T.L., 2013. Prediction of concentration levels of
metformin and other high consumption pharmaceuticals in wastewater and
regional surface water based on sales data. Sci. Total Environ. 442, 380–388.
Ort, C., Lawrence, M.G., Reungoat, J., Mueller, J.F., 2010. Sampling for PPCPs in
wastewater systems: comparison of different sampling modes and optimization
strategies. Environ. Sci. Technol. 44, 6289–6296.
 B. Blair et al. / Chemosphere 134 (2015) 395–401
Oulton, R.L., Kohn, T., Cwiertny, D.M., 2010. Pharmaceuticals and personal care
products in effluent matrices: a survey of transformation and removal during
wastewater treatment and implications for wastewater management. J.
Environ. Monit. 12, 1956–1978.
Pycke, B.F., Roll, I.B., Brownawell, B.J., Kinney, C.A., Furlong, E.T., Kolpin, D.W.,
Halden, R.U., 2014. Transformation products and human metabolites of
triclocarban and triclosan in sewage sludge across the United States. Environ.
Sci. Technol. 48, 7881–7890.
Radjenović, J., Petrović, M., Barceló, D., 2009. Fate and distribution of
pharmaceuticals in wastewater and sewage sludge of the conventional
activated sludge (CAS) and advanced membrane bioreactor (MBR) treatment.
Water Res. 43, 831–841.
Rodayan, A., Majewsky, M., Yargeau, V., 2014. Impact of approach used to determine
removal levels of drugs of abuse during wastewater treatment. Sci. Total
Environ. 487, 731–739.
Salgado, R., Marques, R., Noronha, J.P., Carvalho, G., Oehmen, A., Reis, M.A.M., 2012.
Assessing the removal of pharmaceuticals and personal care products in a full-
scale activated sludge plant. Environ. Sci. Pollut. Res. 19, 1818–1827.
Scheurer, M., Sacher, F., Brauch, H.J., 2009. Occurrence of the antidiabetic drug
metformin in sewage and surface waters in Germany. J. Environ. Monit. 11,
1608–1613.
Scheurer, M., Michel, A., Brauch, H., Ruck, W., Sacher, F., 2012. Occurrence
and fate of the antidiabetic drug metformin and its metabolite guanylurea in
the environment and during drinking water treatment. Water Res. 46, 4790–
4802.
401
Sun, Q., Lv, M., Hu, A., Yang, X., Yu, C.P., 2014. Seasonal variation in the occurrence
and removal of pharmaceuticals and personal care products in a wastewater
treatment plant in Xiamen, China. J. Hazard. Mater. 277, 69–75.
Ternes, T.A., Herrmann, N., Bonerz, M., Knacker, T., Siegrist, H., Joss, A., 2004. A rapid
method to measure the solid–water distribution coefficient (Kd) for
pharmaceuticals and musk fragrances in sewage sludge. Water Res. 38, 4075–4084.
Thompson, K., Zhang, J., Zhang, C., 2011. Use of fugacity model to analyze
temperature-dependent removal of micro-contaminants in sewage treatment
plants. Chemosphere 84, 1066–1071.
Tran, N.H., Urase, T., Ngo, H.H., Hu, J., Ong, S.L., 2013. Insight into metabolic and
cometabolic activities of autotrophic and heterotrophic microorganisms in the
biodegradation of emerging trace organic contaminants. Bioresour. Technol.
146, 721–731.
Trautwein, C., Kümmerer, K., 2011. Incomplete aerobic degradation of the
antidiabetic drug Metformin and identification of the bacterial dead-end
transformation product Guanylurea. Chemosphere 85, 765–773.
USEPA, 2003. CFR 40, Part 136 – Guidelines Establishing Test Procedures for the
Analysis of Pollutants.
USEPA, 2007a. Method 1694: Pharmaceuticals and Personal Care Products in Water,
Soil, Sediment, and Biosolids by HPLC/MS/MS.
USEPA, 2007b. Method 1698: Steroids and Hormones in Water, Soil, Sediment, and
Biosolids by HRGC/HRMS.
Verlicchi, P., Al Aukidy, M., Zambello, E., 2012. Occurrence of pharmaceutical
compounds in urban wastewater: removal, mass load and environmental risk
after a secondary treatment—a review. Sci. Total Environ. 429, 123–155.