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High Throughput Analysis of Endocrine Disruptors (Hormones
and Parabens)
Sébastien Sauvé, Liza Viglino, Paul Fayad and
Michèle PrévostDepartment of Chemistry
sebastien.sauve@umontreal.ca
2
Pharmaceuticals & PCPs
Parent compoundsSoluble Metabolites
IncompleteTreatment
Introduction Into the
environment
Estrogens and Progestogens
Natural Excretion
Anovulants (birth control pills)
Hormone remplacement therapy
Anovulants (birth control)
19.3
9.8
7.6
3.6
3.4
1.4
0.8
0.1
0.1
0 5 10 15 20 25
ETHINYLESTRADIOL & LEVONORGESTREL
ETHINYLESTRADIOL & NORGESTIMATE
DESOGESTREL & ETHINYLESTRADIOL
DROSPIRENONE & ETHINYLESTRADIOL
ETHINYLESTRADIOL & NORETHINDRONE
NORETHINDRONE
ETHINYLESTRADIOL & ETHYNODIOL
ETHINYLESTRADIOL & NORGESTREL
LEVONORGESTREL
Daily consumption per 1000 people
Hormone remplacement therapy (menopause)
16.1
6.5
6.0
3.5
3.3
1.6
1.0
0.4
0 5 10 15 20
ESTROGENIC SUB,CONJUGATED
MEDROXYPROGESTERONE
ESTRADIOL-TRANSDERMIC
ESTRADIOL
PROGESTERONE
ESTROGENIC SUB,CONJUGATED & MEDROXYPROGESTERONE
ESTROPIPATE
ETHINYLESTRADIOL & NORETHINDRONE
Daily Consumption per 1000 people
The sum of estrogens sold in pharmacy mean that about 17% of women, of all ages are actually consuming them
Solid PhaseExtraction
Sébastien Sauvé, Département de chimie
SPE: Enrichment
Detection: Tandem Mass Spectrometry
(selected reaction monitoring)
SPE-LC-MS/MS Analysis
ThermoElectron TSQ Quantum Ultra
Results Hormones
River Montreal STP
Compound Mille Iles St Lawrence North
collector South
collector Effluent
Estriol n.d. n.d. 243 ± 24 230 ± 13 Trace
Estradiol 9 ± 3 8 ± 4 125 ± 4 120 ± 6 90 ± 4
17-α-ethinylestradiol n.d. n.d. 75 ± 3 90 ± 4 n.d.
Estrone n.d. n.d. Trace Trace n.d.
Norethindrone n.d. n.d. 205 ± 3 70 ± 4 53 ± 3
Levonorgestrel n.d. n.d. 150 ± 7 170 ± 4 30 ± 6
Medroxyprogesterone Trace n.d. 5 ± 3 n.d. n.d.
Progesterone 3 n.d. Trace n.d. n.d.
Viglino L, Aboulfadl K, Prévost M, Sauvé S*. 2008. Analysis of natural and synthetic estrogenic endocrine disruptors in environmental waters using online preconcentration coupled with LC-APPI-MS/MS. Talanta 76:1088-1096.
Principles of the LDTD-APCI source: technique that combines thermal desorption (laser diode)
and APCI sample is spotted (1-10 μL) into a 96-well plate and air-dried for
2 min uncharged analytes are thermally desorbed into the gas
phase ionization takes place in the corona discharge region by
APCI and the charged molecules will be transferred to the MS inlet
Source: www.chm.bris.ac.uk/ms/theory/apci-ionisation.html
→ e- + N2 → N2+. +
2e-
Primary ion formation
Secondary ion formation
Proton transfer
→ N2+. + H2O → N2 + H2O+.
→ H2O+. + H2O → H3O+ + HO.
→ H3O+ + M → (M+H)+ + H2O
Laser diode thermal desorption (LDTD)
LDTD
Installation
Auto-sampler
960 samples
Corona needle position (APCI)
IR L
ase
r
(980 n
m,
20 W
)
Can ramp up to 3000oC/sec.Laser power is defined in %Normally ~100-150oC
Laser diode thermal desorption
Laser diode thermal desorption
LDTD Process
(1) Infrared laser (980 nm, 20W) (2) LazWell Plate (96 wells): analyte desorption (1-10 µL spotted) (3) Transfer tube transporting the neutrally desorded analytes to the APCI region(4) Corona needle discharge region (APCI) (5) MS inlet
(1)
(2)
(3)
(4)
(5)
Parameters of the LDTD-APCI source are optimized for signal intensity :
solvent choice for analyte deposition in the well cavities
laser power (%) carrier gas flow rate (L/min) mass deposition (deposition volume in µL) into
plate well laser pattern
No need to optimize liquid chromatography - it has been completely eliminated!
A minimum of 2 SRM transitions were selected + their ion ratios
LDTD Optimization
Optimization for MS (precursor) and MS/MS (SRM transitions) conditions in NI and PI mode.
Results for Feminizing Hormones
Ionisation Linearity range ILD MDLMode (μg/L) (μg/L) (ng/L)
E1 NI 9 - 684 0.0421 0.9999 9 0.0129 0.9984 30E2 NI 18 - 588 0.0084 0.9998 10 0.0030 0.9972 36
PI 18 - 588 0.0384 0.9995 5 0.0035 0.9997 13E3 NI 80 - 749 0.0038 0.9986 24 0.0016 0.9977 30EE2 NI 39 - 915 0.0062 0.9993 17 0.0019 0.9946 42
PI 57 - 915 0.0178 0.9990 8 0.0024 0.9988 14LEVO PI 47 - 780 0.0128 0.9990 11 0.0025 0.9965 20MPROG PI 35 - 677 0.0630 0.9990 6 0.0098 0.9964 30NOR PI 41 - 634 0.0144 0.9989 9 0.0031 0.9950 25PROG PI 21 - 670 0.0320 0.9995 6 0.0080 0.9971 18
Sensitivity R2 Sensitivity R2
In matrixCompound
Matrix-free
Fayad P, Prévost M, Sauvé S. Laser Diode Thermal Desorption-Atmospheric Pressure Chemical Ionization Tandem Mass Spectrometry Analysis of Selected Steroid Hormones in Wastewater: Method Optimization and Application. Submitted to Analytical Chemistry
Very polar analytes (conjugated drug metabolites) or higher mass analytes (e.g. proteins easily ionized with ESI) can be tough to ionize by APCI.
Limits of detection are not as good – may sometime need SPE
Solutions to this and other difficulties: 1) No chromatographic
separation (no column is used to separate same masses in time)
2) Complexe matrices can reduce signal intensity due to competitive interfering compounds in the APCI region
Could de resolved by using two ionization modes (PI and NI)
Must utilize a proper washing step in sample preparation or try sample dilution
3) Polar or higher mass analytes usually more suitable for ESI type analyses
Different solvants can be used to improve signal response and derivitization
Disadvantages of LDTD vs LC
Analysis is done in 15 sec opposed to several minutes in LC-MS/MS
Small sample volume (1-10 µL) compared to 1+ mL for on-line SPE LC-MS/MS
Less contamination sources (tubing, column, injection loop or port, divert valve)
Reduction of backround noise resulting from solvant use Reduction of operation costs (solvants, columns, tubing,
seringes, pumps)
Source: www.ldtd-ionsource.com/fra/default.asp
Advantages of LDTD vs LC
Compound MWg mol-1
Log Kow Use
Estriol 288.4 2.81
EstrogensEstrone 270.4 3.43
Estradiol 272.4 3.94
17-alpha-ethinylestradiol 296.4 4.15 Synthetic
estrogen
Progesterone 314.5 3.87 Progestogen
Norethindrone 298.5 2.97
Synthetic progestogensNorgestrel 312.4 3.08
Medroxyprogesterone 344.5 2.69
Methylparaben 152.2 2.00
Preservatives
Ethylparaben 166.2 2.49
Propylparaben 180.2 2.98
Butylparaben 194.2 3.57
Benzylparaben 228.3 not found
Triclocarban 315.6 4.90 Antibacterial agent
For Solids
1g of freeze-dried samples (0.2 g for municipal sudge) suspended in in 6 mL of acetonitrile: ethyl acetate (5:1) in a 20-mL Teflon tube.
Then, [13C2]-ethynylestradiol (50ng/g) was added as surrogate standard and the sample was vigorously shaken for a few seconds.
After a sonication of 30 min at 30ºC using a sequential ultrasonic extraction (USE), samples were stirred for 45 min on a wrist-shaker at room temperature and centrifuged for 10 min at 3500 rpm to recover the organic layer.
Before evaporation, under a gentle stream of nitrogen to a volume of 250 L at 20ºC, [13C2]-estradiol and methylparaben-d4 (100 μL of 0.2 mg/L) were added as internal standards.
Sample Preparation
Extract was evaporated to dryness, reconstituted into 5 mL of a heptane-acetone (65/35, v/v) mixture and passed through a silica cartridge
Eluted with 6 mL of a heptane-acetone (65/35, v/v) mixture. The final solution was reduced to a volume of 250 L under a nitrogen stream at 20ºC.
SPE Clean up for sludge
In soils and sedimentsFeminizing hormones & TCC: 0.9 to 4.0 ng g-1
Parabens: 1.4 to 2.9 ng g-1
In biosolidsFeminizing hormones & TCC: 9.0 to 17 ng g-1
Parabens: 2.8 to 6.4 ng g-1
Detection Limits
Viglino L, Prévost M, Sauvé S High throughput analysis of solid-bound endocrine disruptors by LDTD-APCI-MS/MS . In revision
SEDIMENTS SOILS BIOSOLIDS
(ng/g) I II II III I II
Estriol 18 3 6 3 15 3 28 9 69 3 45 8Estradiol 70 7 22 4 93 12 81 11 57 3 41 417-alpha-ethinylestradiol 30 6 n.d 86 12 34 11 55 3 47 12Estrone 16 6 6 3 7 2 10 2 54 3 32 2Norethindrone 90 9 n.d 93 17 91 22 106 3 105 12Levonorgestrel 19 5 n.d 24 12 52 17 53 3 33 7Medroxyprogesterone 29 3 n.d 28 4 13 3 31 3 22 9Progesterone 12 4 LMD 31 9 19 6 89 3 72 9TCC 16 4 n.d 53 9 13 2 59 3 43 7Methylparaben 56 7 n.d 127 12 107 4 72 2 91 9Ethylparaben 12 2 n.d LMD 15 3 LMD n.dPropylparaben 15 4 n.d 5 2 9 1 8 1 n.dButylparaben 19 3 n.d 20 5 23 4 LMD n.dBenzylparaben n.d n.d n.d LMD LMD n.d
Results
Viglino L, Prévost M, Sauvé S High throughput analysis of solid-bound endocrine disruptors by LDTD-APCI-MS/MS . In revision
LDTD allows for a very fast and simple analysis for some endocrine disruptors in the environment for water and solids.
Conclusions
Acknowledgments
Natural Sciences and Engineering Research Council of Canada (NSERC)
Phytronix Technologies inc.
If you want more detailed information, or reprints of
published papers, just send me an e-mail:
sebastien.sauve@umontreal.ca
Questions?
Tandem Mass Spectrometry
Sulfamethoxazole+H+
m/z= 254.0
NH2 S
O
O
N
NO
HH
S
O
O
NH2
N
ONH2
m/z=156.0
m/z=108.0
m/z=92.0
Argon-induced Fragmentation
Tandem Mass Spectrometry
Quadrupole 1
Collision Cell
Quadrupole 3
Analyte=254.0 m/z
Interference=245.2 m/zFiltre=254.0m/z
156.0 m/z, 108.0 m/z, etc
Filtre=156.0m/z124.2 m/z, etc.
156.0 m/z
Laser diode thermal desorption (LDTD): Optimization1) Solvant choice for deposition: example for hormones in
NI and PI mode
0.0E+00
5.0E+05
1.0E+06
1.5E+06
2.0E+06
2.5E+06
3.0E+06
3.5E+06
4.0E+06
4.5E+06
5.0E+06
EE2 E2 MPROG LEVO NOR PROG E1 E3
Pea
k A
rea
ACN at 20% laser power
MeOH at 20% laser power
EtOAc at 30% laser power
*
* *
*
0.0E+00
1.0E+05
2.0E+05
3.0E+05
4.0E+05
5.0E+05
6.0E+05
7.0E+05
8.0E+05
5 10 15 20 25 30 40 50 60 80
Pea
k A
rea
Laser Power (%)
EE2
E2
MPROG
LEVO
NOR
PROG
Laser diode thermal desorption (LDTD): Optimization2) Laser power (%): example for hormones in PI mode
La puissance laser maximale est de 20%.
0.0E+00
2.0E+04
4.0E+04
6.0E+04
8.0E+04
1.0E+05
1.2E+05
1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0
Pea
k A
rea
Gas Flow (L/min)
E3
E1
EE2
E2
Laser diode thermal desorption (LDTD): Optimisation3) Carrier gas flow (L/min): example for hormones in NI
mode
E3 is more polar than other hormones. A lower gas flow allows more time in the corona discharge region for proper ionization.
As with LC methods and other techniques, a compromise must be made to combine multiple compounds
Laser diode thermal desorption (LDTD): Optimisation4) Deposition volume (µL): example for PROG in PI mode
0.0E+00
2.0E+07
4.0E+07
6.0E+07
8.0E+07
1.0E+08
1.2E+08
1.4E+08
0.0E+00
2.0E+05
4.0E+05
6.0E+05
8.0E+05
1.0E+06
1.2E+06
1.4E+06
1 2 5 8 10
Ma
trix F
ree
Pe
ak
Are
a
In M
atr
ix P
ea
k A
rea
Deposition Volume (μL)
in matrix
matrix free
A clean-up step in sample preparation or a sample dilution will allow for higher S/N ratios, improving the overall sensitivity and reducing the LD of the method.
Laser diode thermal desorption (LDTD): Optimisation5) Laser pattern : example for PROG in PI mode
0.0E+00
2.0E+05
4.0E+05
6.0E+05
8.0E+05
1.0E+06
1.2E+06
1.4E+06
Pea
k A
rea
Laser Pattern
1 sec – 20%
0.5 sec
1 sec – 20%
0.5 sec
2 sec – 20%
0.5 sec
2 sec – 20%
0.5 sec
1 sec
1 sec
3 sec – 2%
3 sec – 20%
1 sec
1 sec
3 sec – 2%
3 sec – 20%
1 sec
3 sec – 20%
1 sec
3 sec – 20%
1 sec
3 sec – 20%
1 sec
3 sec – 20%
Use an enrichment method to extract and:
1. Isolate the compounds of interest
2. Increase their concentration
3. Remove interfering compounds
Solid Phase Extraction (SPE)
Filtration
Filter = 0.7μm, 0.45 μm
N
O
n
Extraction
200 mg Strata-X Cartridges (Phenomenex)
PS-DVB modifiéSurface Area 760-820 m2/g
Pore size 8.1-9.1 nm
Pore Volume 1.10-1.30 mL/g
Particule size 28-34 μm
Solid phase extraction
Sample100-500 ml offline1 ml online
SPE cartridge
Solid phase extraction
Sample200 ml
SPE Cartridge
Wash
Solid phase extraction
Elutionoffline
SPE Cartridge
Into ~250 µl
250 μL mobile phase containing internal standard
2 min ultrasonic bath
5 min centrifugation to improve dissolution
Injection of 1-10 µl
Reconstitution
Only a part of the SPE aliquot goes into the MS detector
Solid phase extraction offline
10 µl of250 µl
Chromatography MS/MS
The entire 1.0 ml goes through the MS detector
Solid phase extraction online
1.0 ml Chromatography MS/MSSPE
Waste
Pharmas/pesticidesRT: 0.00 - 15.00
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Time (min)
0
1000
1000
1000
1000
100
Relati
ve Abu
ndance
0
1000
1000
100 NL: 3.20E4TIC F: + c APCI sid=10.00 SRM ms2 315.220@cid28.00 [108.650-109.650] MS 2000ppt2NL: 9.97E3TIC F: + c APCI sid=10.00 SRM ms2 255.160@cid18.00 [158.560-159.560] MS 2000ppt2NL: 6.70E5TIC F: + c APCI sid=10.00 SRM ms2 345.190@cid25.00 [122.580-123.580] MS 2000ppt2NL: 1.14E5TIC F: + c APCI sid=10.00 SRM ms2 271.140@cid12.00 [252.600-253.600] MS 2000ppt2NL: 8.55E4TIC F: + c APCI sid=10.00 SRM ms2 271.160@cid13.00 [252.610-253.610] MS 2000ppt2NL: 6.48E4TIC F: + c APCI sid=10.00 SRM ms2 279.140@cid16.00 [132.550-133.550] MS 2000ppt2NL: 9.86E4TIC F: + c APCI sid=10.00 SRM ms2 299.170@cid30.00 [108.570-109.570] MS 2000ppt2NL: 5.88E4TIC F: + c APCI sid=10.00 SRM ms2 313.180@cid17.00 [244.630-245.630] MS 2000ppt2
Progesterone
Estradiol
Medroxyprogesterone
Estriol
Estrone
17-α-ethinylestradiol
19-Northindrone
D(-) Norgestrel
Chromatograms obtained with the complete method:
SPE-LC-MS/MS (ESI/APPI) in milli-Q water doped @ 100 ng/l
Steroid hormones
Compounds with their LODs (using 1ml injections)
Pharmaceuticals2 - 24 ng/L
Pesticides2 - 17 ng/L
HormonesEstrogens
3 - 25 ng/LProgestogens
2 - 7 ng/L
Viglino L, Aboulfadl K, Daneshvar Mahvelat A, Prévost M, Sauvé S*. 2008. On-line solid phase extraction and liquid chromatography/tandem mass spectrometry to quantify pharmaceuticals, pesticides and some metabolites in wastewaters, drinking, and surface waters. Journal of Environmental Monitoring 10:482-489
Viglino L, Aboulfadl K, Prévost M, Sauvé S*. 2008. Analysis of natural and synthetic estrogenic endocrine disruptors in environmental waters using online preconcentration coupled with LC-APPI-MS/MS. Talanta 76:1088-1096. (doi:10.1016/j.talanta.2008.05.008)
0
50
100
150
Carb
a
Nap
roxe
n
Caffei
ne
Trim
etho
prim
Gem
fibroz
il
MOY entrée Lac St Louis
MOYFleuve St Laurent
Pharmas in the St-Lawrence
Hormones
River Montreal STP
Compound Mille Iles St Lawrence North
collector South
collector Effluent
Estriol n.d. n.d. 243 ± 24 230 ± 13 Trace
Estradiol 9 ± 3 8 ± 4 125 ± 4 120 ± 6 90 ± 4
17-α-ethinylestradiol n.d. n.d. 75 ± 3 90 ± 4 n.d.
Estrone n.d. n.d. Trace Trace n.d.
Norethindrone n.d. n.d. 205 ± 3 70 ± 4 53 ± 3
Levonorgestrel n.d. n.d. 150 ± 7 170 ± 4 30 ± 6
Medroxyprogesterone Trace n.d. 5 ± 3 n.d. n.d.
Progesterone 3 n.d. Trace n.d. n.d.
Viglino L, Aboulfadl K, Prévost M, Sauvé S*. 2008. Analysis of natural and synthetic estrogenic endocrine disruptors in environmental waters using online preconcentration coupled with LC-APPI-MS/MS. Talanta 76:1088-1096.
0
50
100
150
200
250
300
SMX TRI CLA AZI
Co
nce
ntr
atio
n / n
gL
-1
Analyzed Anti-infectives
City of Montréal Wastewater Treatment Plant
North Influent
South Influent
Effluent
Montréal April 2006
Segura PA, Garcia Ac A, Lajeunesse A, Ghosh D, Gagnon C, Sauvé S*. 2007. Determination of six anti-infectives in wastewater using tandem solid phase extraction and LC/MS/MS. Journal of Environmental Monitoring 9:307-313.
Efficiency of wastewater treatment (Montréal - May
2006),amongst the world’s largest ~ 2 500 000
m3/dayCompound Removal efficiency (%)
Mass flows in the St-Lawrence River
(g day-1) Sulfamethoxazole 12 ± 1 340 ± 30
Trimethroprim N.S. 310 ± 20
Ciprofloxacin 29 ± 2 320 ± 10
Levofloxacin 31 ± 2 118 ± 2
Clarithromycin N.S. 790 ± 60
Azithromycin N.S. 320 ± 20
N.S. – non significatif
1 ton of antibiotic molecules released every year
Pharmaceuticals & PCPs
•Reproductive issues in fish
•Multiple or distorted legs in frogs and other amphibians
•Drug resistance in humans
Possible Effects on Humans and Animals in the Environment
Develop a simple method, sensitive enough to analyze trace contaminants in waste, raw and drinking waters and eventually solids (soil, sediments and biosolids)
Online solid phase extraction preconcentration coupled to liquid chromatography and tandem mass spectrometry (SPE-LC-MS/MS) and
Laser Diode Thermal Desorption (LDTD) – APCI (Atmospheric Pressure Chemical Ionization) -MS/MS
Pharmaceuticals & PCPs
Use an enrichment method to extract and:
1. Isolate the compounds of interest
2. Increase their concentration
3. Remove interfering compounds
SPE (solid phase extraction)
Solution
Filtration
Filter = 0.7μm, 0.45 μm
Sébastien Sauvé, Département de chimie
Solid Phase
Extraction
SPE: Enrichment
LC-MS/MS Analysis
Detection: Tandem Mass Spectrometry
(selected reaction monitoring)
ThermoElectron TSQ Quantum EQuan MAX System
RT: 0.00 - 15.00
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Time (min)
0
1000
1000
1000
1000
100
Rel
ativ
e Ab
unda
nce 0
1000
1000
100 NL: 3.20E4TIC F: + c APCI sid=10.00 SRM ms2 315.220@cid28.00 [108.650-109.650] MS 2000ppt2NL: 9.97E3TIC F: + c APCI sid=10.00 SRM ms2 255.160@cid18.00 [158.560-159.560] MS 2000ppt2NL: 6.70E5TIC F: + c APCI sid=10.00 SRM ms2 345.190@cid25.00 [122.580-123.580] MS 2000ppt2NL: 1.14E5TIC F: + c APCI sid=10.00 SRM ms2 271.140@cid12.00 [252.600-253.600] MS 2000ppt2NL: 8.55E4TIC F: + c APCI sid=10.00 SRM ms2 271.160@cid13.00 [252.610-253.610] MS 2000ppt2NL: 6.48E4TIC F: + c APCI sid=10.00 SRM ms2 279.140@cid16.00 [132.550-133.550] MS 2000ppt2NL: 9.86E4TIC F: + c APCI sid=10.00 SRM ms2 299.170@cid30.00 [108.570-109.570] MS 2000ppt2NL: 5.88E4TIC F: + c APCI sid=10.00 SRM ms2 313.180@cid17.00 [244.630-245.630] MS 2000ppt2
Progesterone
Estradiol
Medroxyprogesterone
Estriol
Estrone
17-α-ethinylestradiol
19-Northindrone
D(-) Norgestrel
Ovarian hormones
SPE-LC-MS/MS (ESI/APPI) in milli-Q water doped @ 100 ng/l
Compounds with their LODsInjection Volume?
Pharmaceuticals2 - 24 ng/L
Pesticides2 - 17 ng/L
HormonesEstrogens
3 - 25 ng/LProgestogens
2 - 7 ng/L
Viglino L, Aboulfadl K, Daneshvar Mahvelat A, Prévost M, Sauvé S*. 2008. On-line solid phase extraction and liquid chromatography/tandem mass spectrometry to quantify pharmaceuticals, pesticides and some metabolites in wastewaters, drinking, and surface waters. Journal of Environmental Monitoring 10:482-489
Viglino L, Aboulfadl K, Prévost M, Sauvé S*. 2008. Analysis of natural and synthetic estrogenic endocrine disruptors in environmental waters using online preconcentration coupled with LC-APPI-MS/MS. Talanta 76:1088-1096. (doi:10.1016/j.talanta.2008.05.008)
Montréal April 2006
0
50
100
150
200
250
300
SMX TRI CLA AZI
Co
nce
ntr
atio
n / n
gL
-1
Analyzed Anti-infectives
City of Montréal Wastewater Treatment Plant
North Influent
South Influent
Effluent
Segura PA, Garcia Ac A, Lajeunesse A, Ghosh D, Gagnon C, Sauvé S*. 2007. Determination of six anti-infectives in wastewater using tandem solid phase extraction and LC/MS/MS. Journal of Environmental Monitoring 9:307-313.
Results – Granby April 2006
City of Granby Wastewater Treatement Plant
0
50100
150
200
250300
350
SMX TRI CLA AZI
Analyzed Anti-infectives
Co
nce
ntr
atio
n /
ng
L-1
Inf luent
Eff luent
Efficiency of Montréal wastewater treatment - May 2006(amongst the world’s largest ~ 2 500 000 m3/day)
Compound Removal efficiency (%)
Mass flows in the St-Lawrence River
(g day-1) Sulfamethoxazole 12 ± 1 340 ± 30
Trimethroprim N.S. 310 ± 20
Ciprofloxacin 29 ± 2 320 ± 10
Levofloxacin 31 ± 2 118 ± 2
Clarithromycin N.S. 790 ± 60
Azithromycin N.S. 320 ± 20
N.S. – non significant
1 ton of antibiotic molecules released every year
What does that mean?
A few nanograms per liter represent the equivalent of a sugar cube diluted in the Olympic stadium – really just traces!
The concentrations observed at the ‘worst case’ sites are far from the levels expected to cause impacts on human health (from 0.02 to 3% of the daily therapeutic dose after the consumption of 2 l/d for 70 years)
•We can certainly detect pharmaceuticals in the environment
•Concentrations of pharmaceuticals in the environment can be of concern for some issues
• Especially antibiotic resistance, endocrine disruptors
Pharmaceuticals in the Environment
What are the effects of chronic exposure to low levels of a mixture of contaminants?
What is the contribution of traces of ovarian hormones as endocrine disruptors to human?
High seasonal variability – not yet considered for risk assessment
What are the risks from chemotherapy agents, and other compounds (surfactants, ionic liquids)?
Some other questions:
Unreasonable to try and regulate consumption• We can improve wastewater treatment to reduce releases – an
engineering option
We can look at pharmacopeia's redundancy• When equivalent drugs are available, we could promote those
with the least impact
Can we think up « degradable » drugs?• More susceptible to oxydation, hydrolysis, UV, microbial
degradation, etc. in the natural environment.
• Tricky considering that they must still remain effective!
And Green Chemistry?
HRMS (High Resolution Mass Spectrometry)
• Time of Flight
• Magnetic Sectors
• Orbitrap Mass Spec.
Accurate mass measurement useful for positive ID when fragmentation is limited or for the screening of unknows
Other MS Techniques for PCPPs
Most prescribed anti-infectives in Québec in 2004
(IMS Health Canada compilation – 8 million people in Quebec).
0 100 200 300 400 500 600 700 800
Thousands of retail prescriptions dispensed
Amoxicillin
Ciprofloxacin
Azithromycin
Clarithromycin
Penicillin V
Cefprozil
Fluconazole
Sulfamethoxazole + Thrimethoprim
Mesazaline
Minocycline
Moxifloxacin
Clindamycin
Cephalexin
Levofloxacin
Nystatin
Nitrofurantoin
Terbinafine
Cloxacillin
Pivampicillin
Gatifloxacin
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