1. ISBM, Manchester UK, 9th ‐ 11th September 2013
HOW CONFIDENT ARE WE?
Alain LeBlanc
Special projects’ manager,
support services and external quality assessment
Centre de toxicologie du Québec (CTQ)
INSPQ, Québec, Canada
Pierre Dumas
Research & Development division

2. The INSPQ is a provincial government body created to improve
the coordination, development and use of expertise in public health
Among its expertise, the Centre de toxicologie du Québec (CTQ)
offers analytical support in clinical, environmental and occupational health
www.inspq.qc.ca/ctq

3. QUEBEC MINISTRY OF HEALTH AND WEALTHFARE (MSSS)
INSTITUT NATIONAL DE SANTÉ PUBLIQUE DU QUÉBEC (INSPQ)
ENVIRONMENTAL HEALTH AND TOXICOLOGY DEPT. (DSET)
CENTRE DE TOXICOLOGIE DU QUÉBEC (CTQ)
Claude Thellen
QUALITY MANAGEMENT
Sergine Lapointe
TOXICOLOGY
LABORATORY BRANCH
Normand Fleury
SPECIAL PROJECTS, SUPPORT
SERVICES and EQAS SECTOR
Alain LeBlanc
RESEARCH
Pierre Ayotte
METHOD
DEVELOPMENT
Patrick Bélanger
CLINICAL
ORGANICS
Michel Lefebvre
Eric Gaudreau
METALS
Ciprian‐Mihai
Cirtiu
PROJECT
MANAGEMENT
and SUPPORT
SERVICES
EXTERNAL
QUALITY
ASSESSMENT
SCHEMES

4. To make scientists, researchers and
epidemiologists aware of day‐to‐day issues that
laboratories encounter when confronted
with testing new environmental contaminants
Triclosan

5. •
•
•
•
•
To develop reliable, robust and accurate analytical methods
To assure tracability of the metrology
To assure tracability of the data
To assure the comparability of the analytical results
To develop and maintain competence of staff

6. Sample collection, integrity and storage
•
•
•
•
•
•
•
•
Time of collection (spot/24 hrs; day‐to‐day variability)
Handling (freeze/thaw)
Stability (+ > 5 yrs)
Homogeneity: free/conjugated species
Contamination (collection process, container, …)
Sampling container (adsorption onto plastics) / Tubes
Are we sure we are measuring the right biomarker? in the right matrix?
Shipping considerations (overnight, deep freeze)
Analytical innovation
•
•
Keep costs down; compromise with multi‐analyte methods
Tend to less invasive matrices, but are they the right ones?

7. Analytical standards
•
•
Availability of conjugated analytes
Accuracy (certificates, multiple sources)
Impact on biomonitoring data (cycle to cycle)
Availability of appropriate internal standards
13C labeled and unlabeled analogs are usually not available at the same time
• Custom synthesis (TRC, CANSYN, CHIRON, CIL, etc)
High cost, time to delivery, lot‐to‐lot, purity (UV, NMR, IR and MS spectra)
•
•
Analytical methods
•
•
•
•
Classical versus experimental protocols
Sensitivity (LODs) and specificity
Reportable limits (no international consensus; study comparisons)
Laboratory contamination (ex: BPA, Triclosan, Parabens, BDEs, …)

8. Analytical instrumentation
•
Comparability
Reference materials
•
•
•
Availability (certified?)
Appropriateness (same matrix?) commutability
Internal RM 2nd source of standard!!
External quality assessment schemes
•
How to compare study results?
•
•
Collaboration among reference labs or research centers
To help reduce vulnerability

9. SAMPLE INTEGRITY AND STORAGE

10. Sample Temperature Tracking Chart
« Freeze/thaw »
30
Temperature °C
20
10
0
-10
-20
-30
Sample handling: Chronological events
(worst case scenario)

11. Freeze/Thaw ‐ Stability of total BPA in urine (14 samples)
125
120
115
+10%
105
% BPA
110
+5%
100
-5%
95
90
-10%
85
80
cycle 0
cycle 1
cycle 2
cycle 3
cycle 4

12. Freeze/Thaw ‐ Stability of conjugated BPA in urine (14 samples)
100
%Conjugated BPA
98
96
94
92
90
88
cycle 0
cycle 1
cycle 2
cycle 3
cycle 4

13. % Recovery in urine
•
•
Tip R 50% ACN
Free Triclosan can be
rapidly adsorbed onto
plastic tips according to
type of material and % of
acetonitrile in spike
solution.
Time residence in the
pipette tips can increase
the degree of adsorption.
100
Tip R 5% ACN
97
Tip R 0% ACN
95
Tip B 50% ACN
101
Tip B 5% ACN
91
Tip B 0% ACN
90
Tip Y 50% ACN 1 Min
98
Tip Y 5% ACN 1 Min
Tip Y 0% ACN 1 Min
82
75
Tip Y 50% ACN
103
Tip Y 5% ACN
Tip Y 0% ACN
93
86

14. Internal QC from
unspiked urine.
Endogenous conjugated
form of trichlorophenol
18.7
Concentration (ug/L)
17.7
16.7
15.7
14.7
13.7
Good stability
12.7
49
76
3
8
49
17
2
59
7
00
48
47
48
42
2
7
83
1
25
46
45
46
66
6
1
45
08
5
49
44
43
91
0
11.7
# séquence
7.250
External QC made from
spiked urine using free
trichlorophenol
5.250
4.250
Poor stability
3.250
2.250
# séquence
3
49
76
8
49
17
2
59
48
48
00
7
2
47
42
7
46
83
1
46
25
6
45
66
1
08
45
49
44
91
0
5
1.250
43
Concentration (ug/L)
6.250

15. ANALYTICAL STANDARDS

16. Supplier
MMP
MEP
MiBP
MnBP MCHP
MBzP
MEHP MEHHP MEOHP MECPP
MOP
MCPP
MNP
CIL‐2006
‐25%
‐2%
‐
‐47%
‐1%
‐63%
‐29%
‐11%
‐7%
‐
12%
‐22%
‐39%
CIL‐2009
‐9%
Certified
solutions CIL‐2010
‐11%
‐5%
‐13%
‐12%
‐76%
‐14%
‐21%
4%
3%
2%
‐70%
‐16%
‐3%
‐9%
0%
‐11%
‐7%
3%
‐11%
‐19%
‐9%
‐2%
3%
‐78%
‐10%
‐
‐
‐3%
‐
‐
‐
‐
‐
‐
‐
‐
‐
‐
Accustandard
‐26%
‐31%
‐
‐19%
‐
‐4%
‐27%
‐
‐
‐
‐
‐
‐
Accustandard
8%
‐7%
‐
‐5%
‐
‐2%
‐27%
‐
‐
‐
‐
‐
‐
Aldrich
9%
‐
‐
‐6%
‐
‐5%
‐
‐
‐
‐
‐
‐
‐
Chiron
Chiron
‐
‐21%
‐
0%
‐
‐
‐
‐
‐
‐
‐
‐
‐
Neat
standards Dr Ehrenstorfer
13%
‐
‐
‐2%
‐
‐
‐29%
‐
‐
‐
‐
‐
‐
CanSyn
9%
‐5%
4%
‐1%
‐6%
‐1%
‐8%
‐6%
‐4%
‐1%
10%
‐7%
‐3%
CIL
0%
0%
0%
0%
0%
0%
0%
0%
0%
0%
0%
0%
0%
TRC
7%
‐3%
4%
‐6%
‐10%
‐2%
‐11%
9%
9%
‐70%
12%
‐2%
‐6%
Langlois E., LeBlanc A., Simard Y., Thellen C. Accuracy investigation of phthalate metabolite standards
Journal of Analytical Toxicology, 2012 ; 36 :270-279

17. BPA
Triclosan
compounds synthesized at CHUL
BPA‐13C12 and TCS‐13C12 were obtained from Cambridge Isotope Laboratories

18. ANALYTICAL METHODS

19. •
•
Cadmium in urine by ICP MS
Mo‐O interference
corrected VS uncorrected
Mean concentration can be
biased by 30 % with individual
error greater than 200%
Comparability between studies
is strongly related to laboratory
methodologies
‐ Mo in sample
‐ Mo in calibration curve
30
21
Cd nmol/L
Laboratories CV %
•
•
35
25
32
20
15
10
23
9.5
78
5
130
0
0
50
100
150
200
Ratio conc. Mo/Cd
250
300

20. Cadmium Urine (nmol/L)
QMEQAS 2011-2
Lab B
Lab A

21. X 1.6
Comparing United States and Canadian population exposures from National Biomonitoring Surveys: Bisphenol A intake as a case study
Judy S. LaKind, Johanne Levesque, Pierre Dumas, Shirley Bryan, Janine Clarke and Daniel Q. Naiman
Journal of Exposure Science and Environmental Epidemiology (2012) 22, 219‐226

22. X 1.6

23. QC Low
QC High
Target
value
2.90 ±0.23
10.17 ±0.58
CTQ 1
2.98
10.6
CTQ 2
2.82
10.2

24. NCI
PCB 153 (1µg/L) + PCB 132 (0.3 µg/L)
Complete overlap of both congeners
EI

25. Blank level µg/L
2.00
1.80
1.60
1.40
1.20
1.00
0.80
0.60
0.40
0.20
0.00
35800
36800
37800
38800
Sequence #
39800

26. None Specific Metabolite
Diethyl thiophosphate (DETP)
Specific Metabolite
3,5,6‐trichloro‐2‐pyridinol

28. G‐BPA
SO3‐BPA
(SO3)2‐BPA
Free BPA

29. 40
BPA
35
y = 0.9149x - 0.0207
R² = 0.9946
BPA total µg/L (GC)
30
25
20
15
10
5
0
0
5
10
15
20
25
30
35
40
BPA free + BPA-glucuronide + BPA-sulfate + BPA-disulfate in µg BPA eq./L (LC)
LC‐MS/MS VS GC‐MS/MS

30. 900
Triclosan
800
y = 1.0229x + 0.1818
R² = 0.9954
700
TCS total µg/L (GC)
600
500
400
300
200
100
0
0
100
200
300
400
500
600
700
800
900
TCS free + TCS-glucuronide + TCS-sulfate µg in TCS eq./L (LC)
LC‐MS/MS VS GC‐MS/MS

31. 137.6
Concentration (ug/L)
127.6
117.6
107.6
97.6
87.6
77.6
67.6
Setting up
Single analyst + single instrument
“Rush” multi analysts and instruments
# séquence
New standard batch

32. External Quality Assessment Schemes
and Reference Materials

33. EQAS
Blood lead (17)
AAFP CAP DigitalPt G‐EQUAS LAMP NEQAS NYSDH OELM PCI Penn QMEQAS RCPA SEKK SKZL TEQAS WIV‐ISP WSLH
Urine BPA (1)
G‐EQUAS
Urine Triclosan (0)
Urine Triclocarban (0)
RM
Blood lead (6)
IAEA IRMM NIST RECIPE SERONORM UTAK
Urine BPA (1)
RECIPE [NIST]
Urine Triclosan (0)
[NIST]
Urine Triclocarban (0)

34. External Quality Assessment Schemes
EQAS
Since 1979
More than 30 countries
250 laboratories

35. • 3 continuing programs in blood, urine, serum and hair
Metals: PCI, QMEQAS POPs: AMAP
• Material from human origin; unexposed volunteers
• 2 developing programs: MDA, Organic compounds in urine
• Voluntary participation
• Funded by the participants
• Reference material virtual store

36. ISO/CEI 17043
REFERENCE MATERIALS
www.inspq.qc.ca/ctq/sales
36

37. • Laboratories involved in HBM face challenges at all levels of operation
• Competence of staff must be developped and maintained
• Method validation must be brought to a higher level
(stability, matrix effect, comparability of standards, …)
• Importance of External quality assessment schemes and reference
materials is obvious
adapted QA strategy (work to do)
• A laboratory facility with a scientific and administratrive infrastructure is
mandatory
• The term « new contaminant » alone shouldn’t set the rationale for
biomonitoring
• A reference laboratory must have the capabilities and experience to
tackle these recurring issues (not necessarily compatible with every
laboratory structure)

38. QUESTIONS?