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Quantitative and Semi-Quantitative Determination of PPCPs and Their By-products in Wastewater Treatment Plants Samples Using UHPLC-Orbitrap MS
and Data Mining Technologies
Overview
Purpose:
Develop an analytical method to (1) determine PPCP concentrations in
wastewater samples, and (2) examine the transformation of selected PPCPs during
treatment processes.
Methods:
Samples prepared by solid phase extraction (SPE) and analyzed by high
performance liquid chromatography-Orbitrap mass spectrometry (HPLC-Orbitrap MS).
Results:
Quantitative results of selected pharmaceuticals and personal care products
(PPCPs) like DEET, Triclosan (TCS), Triclocarban (TCC), Diclofenac (DCL),
Carbamazepine (CBZ) and semi-quantitative of their degradation products were
obtained.
Introduction
Results obtained from a simple and powerful workflow that can readily determine
PPCPs and their by-products in wastewater treatment plant (WWTPs) samples will be
presented. This workflow was applied in a survey of 43 permeate samples obtained
from a pilot anaerobic membrane bioreactor (AnMBR). Quantitative results show the
prevalence of various PPCPs in wastewater, particularly for compounds with high
usage and/or poor elimination (e.g., caffeine, carbamazepine (CBZ), DEET, lidocaine,
lincomycin, ketoprofen, and bezafibrate). For PPCP by-products, we identified that in-
situ microbial degradation was the dominant pathway for triclocarban (TCC) removal;
whilst triclosan (TCS), diclofenac (DCF) and CBZ were eliminated via a combination of
photodegradation and metabolism. Thirty by-products were detected in this pilot
survey, including the toxic compounds chlorophenol and acridone.
Methods
Sample Preparation
For this study, permeate samples were chosen due to their complex matrix which
poses as a challenge for conventional analytical method. These samples were
collected from a pilot anaerobic membrane bioreactor (AnMBR) pilot plant located at
the Wastewater Technology Centre (Environment Canada, Burlington, Ontario). A total
of 35 permeate samples permeate tank from January 2012 to March 2013. During this
time, the reactor were operated at different temperatures at 20, 35 and 55 °C using
samples collected, respectively, in summer, winter and winter, to investigate the effect
on the removal of PPCPs in permeates. Grab samples were contained in 1L-amber
bottles without headspace and stored in dark, cold storage (4°C) until analysis.
Neat standards of native target compounds were purchased from Sigma-Aldrich
(Oakville, ON, Canada). Deuterium (D) and
13
C-labelled standards were purchased
from CDN Isotopes (Pointe-Claire, QC, Canada) and Cambridge isotope Laboratories
(Andover, MA, US). Five levels of analytical standard solutions were prepared by
diluting intermediate solutions with CH
3
OH HPLC grade acetonitrile (CH
3
CN) and
methanol (CH
3
OH) were purchased from Thermo Fisher Scientific (Ottawa, ON,
Canada). High purity water used for aqueous mobile phases and sample preparation
was produced by passing reverse osmosis water through a Thermo
Scientific™
Barnstead™
Nanopure
™
water purification system (Mississauga, ON, Canada).
Laboratory Services NBranch (LaSB) method E3454
1
was used to prepare samples for
targeted compound analysis and non-targeted compound screening. Waters OASIS®
(Mississauga, ON, Canada) HLB solid phase extraction (SPE) cartridge (6 cc, 500 mg)
was used in the extraction. Method E3454 has been accredited by the Canadian
Association for Laboratory Accreditation (CALA) since 2004.
Liquid Chromatography (or more generically Separations)
Sample analysis was achieved on a Thermo
Scientific™
Dionex™
UltiMate
™
3000
HPLC consisting of a HRG-3400RS binary pump, WPS-3000 autosampler, and a TCC-
3400 column compartment. Separation was made by injecting 5
m
L extracts into a
Thermo
Scientific™
Betasil
™
and a Thermo
Scientific™
Hypersil
™
Gold, 2.1x100 mm
columns, respectively, for positive and negative mode Orbitrap MS analysis.
One positive mode HPLC a
analysis of PPCPs and their
TABLE 1. HPLC mobile ph
Mass Spectrometry
The HPLC
was interfaced t
a heated electrospray ioniz
and calibrated in positive an
MSCAL5 and MSCAL6. Hig
L/min). Spray voltages used
Mass spectrometric data w
width-at-half-maximum pea
1.5 scans/sec when using a
time of 100 msec.
Data Analysis
Thermo
Scientific™
Trac
analysis for 56 PPCPs. Th
screening along with a dat
active compounds and t
perfluorohydrocarbons. Tr
(M+H)
+
, (M+NH
4
)
+
and (M+
negative mode for compou
extracted ion chromatogra
Analytes were automatica
(approximately 25
–
50 pg/m
for the mono-isotopic mass
a relative intensity of 90%
was about 65 sec/sample
TraceFinder software were
carry out ChemSpider
™
se
the SIEVE software
too.
Results
Quantitative Analytical Re
Quantitative analysis deter
antibiotics, non-steroidal a
products such as insect re
ciprofloxacin and sulfa drug
to other therapeutic classes
is reported for the antide
representative (i.e., CBZ), it
Column oven
Mobile phase (Positive)
Mobile phase (Negative I)
Mobile phase (Negative II)
HPLC Gradient