Noora Perkola Finnish Environment Institute · 11/9/2011 · Noora Perkola Finnish Environment...
Transcript of Noora Perkola Finnish Environment Institute · 11/9/2011 · Noora Perkola Finnish Environment...
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Noora Perkola
Finnish Environment InstituteNordic MS Symposium
November 9, 2011, Båstad, Sweden
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Artificial Sweeteners Used as additives in various food products to replace
sugar
4 non-caloric sweeteners
Acesulfame Potassium (ACS-K)
Saccharin (SAC)
Cyclamate (cyclamic acid) (CYC)
Sucralose (SCL)
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Table 1. Analytes; Properties and origin of the standards
1 TRC = Toronto Research Chemicals Inc. (North York, Ontario, Canada)2 Ehrenstorfer = Dr. Ehrenstorfer-Schäfers (Augsburg, Germany)3 ADI = Acceptable Daily Intake
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Artificial Sweeteners in Environment ACS-K, SAC, CYC and SCL detected in environmental
water samples
Levels up to over 2 µg/L of ACS-K
SAC and CYC 10-150 ng/L
SCL 10-1000 ng/L
Loos et al, Journal of Chromatography A (2009) 1216: 1126-1131
Scheurer et al, Anal Bioanal Chem (2009) 394: 1585-1594
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Artificial Sweeteners in Environment Pass through human body and WWTPs
SAC and CYC eliminated over 90 %
ACS-K up to 41 %, SCL 20 %
ACS-K and SCL – tracers of domestic waste water
Effects to aquatic ecosystems unknown
Scheurer et al, Anal Bioanal Chem (2009) 394: 1585-1594
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Artificial Sweeteners in Environment Pass through human body and WWTPs
SAC and CYC eliminated over 90 %
ACS-K up to 41 %, SCL 20 %
ACS-K and SCL – tracers of domestic waste water
Effects to aquatic ecosystems unknown
Scheurer et al, Anal Bioanal Chem (2009) 394: 1585-1594
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Sample pretreatment Reversed phase SPE (Waters Oasis HLB and J.T. Baker
Bakerbond SDB 1) and ion exchange SPE (Waters Oasis WAX and Oasis MAX) were tested
Oasis WAX chosen – possibility to have SCL in different extract (different concentration factor)
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Sample pretreatment SPE procedure (Oasis WAX cartridges):
Conditioning 10 ml MeOH + 5 ml H2O
Load sample (pH 3.0)
Wash with 2 ml 2 % HCOOH (aq)
1st elution 6 ml MeOH
2nd elution 6 ml 1 % NH3 in MeOH
Recoveries between 70 and 120 % with extractionvolumes 50 – 200 ml
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Noora Perkola - 2nd Nordic MS Symposium 9.11.2011
Sample
• 50-200 mL
• pH 3.0
• Add ISTD (d-ACS-K, d-SCL, d-CYC, 13C-SAC)
SPE (WAX)
• Condition: 10 mL MeOH + 5 mL H2O
• Load Sample: 1 drop sec-1
• Elute: 6 mLMeOH + 6 mL 1 % NH3 (MeOH)
Evaporation
• To dryness
• Redissolve to H2O
LC-MS/MS analysis
Figure 1. Sample pretreatment procedure
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LC separation Acquity UPLC
Flow rate 0.400 mL/min, Column T 40 °C
• LC eluents A: 2 mM NH4OAc (aq); B: 2 mM NH4OAc (MeOH)
Gradient:
Initial: 99.9 % A
0.10 min: 99.9 % A
2.00-2.30 min: 0.1 % A
2.40-3.00 min: 99.9 % A
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LC separation• Comparison of HSS T3 1.8 µm 2.1 x 50 mm
and BEH C18 1.7 µm 2.1 x 50 mm columns
• HSS: better retention for polar analytes –longer retention times but less matrixinteractions?
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Figure 2. Chromatogram of standard solution of ACS-K (1), SAC (2), CYC (3) and SCL (4) with BEH C18 column (1a-4a) and HSS column (1b-4b).
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14-Oct-2011 2 WAX, HSS
Time0.05 0.10 0.15 0.20 0.25 0.30 0.35 0.40 0.45 0.50 0.55 0.60 0.65 0.70 0.75 0.80 0.85 0.90 0.95 1.00 1.05 1.10 1.15 1.20
%
0
100
chromtest_111014_21 Sm (Mn, 2x1) 1: MRM of 6 Channels ES- 162 > 82 (acesulfame)
2.47e50.96
14-Oct-2011 2 WAX, HSS
Time1.12 1.14 1.16 1.18 1.20 1.22 1.24 1.26 1.28 1.30 1.32 1.34 1.36 1.38 1.40 1.42 1.44 1.46 1.48 1.50 1.52 1.54 1.56 1.58
%
0
100
chromtest_111014_21 Sm (Mn, 2x1) 2: MRM of 3 Channels ES- 178 > 80 (cyclamate)
7.70e31.29
1.10
1.15 1.39
Figure 3. Example: Chromatogram of river water sample
ACS-K
CYC
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MS/MS analysis Xevo TQ MS
ESI-
MRM
Table 2. Optimization of MS parameters
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ESI- MS/MS LOQs for ACS-K, SAC, CYC and SCL 25 ng/L, 25 ng/L,
6.5 ng/L and 210 ng/L, respectively
Sample volume 100 mL
Extract volume 1.0 mL
Matrix suppression
70-80 % for ACS-K and SAC
60-90 % for SCL
Less than 20 % for CYC
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Enhancing ionization MS optimization with matrix?
Lower matrix effects in MeOH than in 1 % NH3
(MeOH)
More efficient wash step in SPE or clean-up for extract?
Modificator to enhance ionization in ES?
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IDMS Isotope Dilution Mass Spectrometry
Mass labelled standards for all analytes
d4-ACS-K
d11-CYC
d6-SCL
13C6-SAC
Quantification using labelled standards
Reliable results despite low recoveries
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AcknowledgementsI wish to thank
Waters
Association of Maa- ja Vesitekniikan Tuki ry for financial support
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Thank You!
Noora Perkola - 2nd Nordic MS Symposium 9.11.2011