ion chromatography coupled to ms, a powerful approach for polar pesticides determination · 2017....
TRANSCRIPT
-
The world leader in serving science
Dott. Luca Gerardo IC/SP Application Specialist, Thermo Fisher Scientific, Rodano (MI)
Ion Chromatography Coupled to MS, a Powerful Approach for Polar Pesticides Determination
-
What is Glyphosate?
• Glyphosate (N-(phosphonomethyl)glycine) is a broad spectrum systemic herbicide commonly used as weed control.
• It is rapidly degraded to aminomethylphosphonic acid (AMPA) metabolite frequently found in plants, water and soil.
• Very polar, zwitterionic compound • Difficult to retain on C18 LC column
H2N P
O
OHOH
-
• In 2015, the World Health Organization's International Agency for Research on Cancer (IARC) classified glyphosate as "probably carcinogenic to humans." • International scientists found there was a particularly strong connection to non-Hodgkins
lymphoma.
• 2015: The European Food Safety Authority and the European Chemicals Agency have said glyphosate is NOT likely to be carcinogenic to humans. • Under scrutiny for research conducted by chemical companies
• 2016: The World Health Organization/ United Nations Joint Meeting on Pesticide Residues also cleared glyphosate as unlikely to pose a risk to humans, • tarnished by conflicts of interest regarding ties to the International Life Sciences Institute, a
food industry front group
• March, 2017: California Environmental Protection Agency's Office of
Environmental Health Hazard Assessment confirmed that it would add glyphosate to list of cancer causing chemicals.
.
Glyphosate – Health Concerns Timeline
-
• 2016
• The Munich Environmental Institute group found glyphosate in 14 of Germany’s most popular beers (0.46 – 29.74 µg/L).
• Alliance for Natural Health USA tested 24 popular breakfast foods, 10 of 24 goods had detectable levels of glyphosate (86 – 1,327 µg/kg) (www.anh-usa.org).
• FDA found glyphosate in US honey at double the levels allowed in the EU.
• More than 50 lawsuits against glyphosate producers are pending in US District Court in San Francisco.
• Several hundred similar actions are pending in
state courts.
Glyphosate in the News
http://www.anh-usa.org/http://www.anh-usa.org/https://usrtk.org/our-investigations/fda-finds-monsantos-weed-killer-in-u-s-honey/
-
HPLC methods: • Typically require extensive sample preparation
• SPE • QuePP method
• Preparation of buffers/acidic mobile phases
• Non retentive on C18 columns • Pre or post column derivatisation • Speciality columns – poor resolving power • Fluorescence detection
• Matrix effects and interferences
Analysis of Glyphosate
Time and additional source of errors
Robustness issues
-
QuPPe-PO v 9.1-Negative Mode Compounds
• Method lists a total of 42 different (pos and neg mode) analytes.
http://www.crl-pesticides.eu/userfiles/file/EurlSRM/meth_QuPPe-PO_EurlSRM.pdf
http://www.crl-pesticides.eu/userfiles/file/EurlSRM/meth_QuPPe-PO_EurlSRM.pdf
-
Why Couple Ion Chromatography (IC) to MS?
- IC columns provide great specificity and selectivity for ionic compounds.
- Metal-free flow path reduces fouling of ion-exchange columns.
- MS and conductivity detection in series
Possibility to determine high concentration contaminants (Anions / Cations) and trace contaminants (Pesticides/Disinfection byproducts) in the same chromatographic run.
- Eluent suppression to Water
Enable rapid switching from anion to cation analysis sharing MS device.
- Just add water to prepare eluents for anions and cations analysis.
-
Principles of RFIC - Eluent Generation
Vent
Pump
[KOH] α Current
Flow rate
H2
Degas unit
Cation exchange connector
Thermo Scientific™ Dionex ™ CR-ATC anion trap
KOH + H2
KOH electrolytic chamber
Thermo Scientific™ EluGen™ KOH
cartridge
Pt anode
[ + ]
K+
OH–
KOH
K-reservoir
H2O
Thermo Scientific™ RFIC technology - Reagent-free IC without manual eluent preparation
(H2O 2H+ + ½O2 + 2e–)
(2H2O + 2e– 2OH– + H2)
KOH [ - ] Pt cathode
-
NaOH, H2
Waste/Vent Waste/Vent Anode
To Detector
H+ + O2 H2 + OH-
H2O H2O H+ , X- in H2O
H2O H2O
H2O, O2
Cation- Exchange
membranes
OH-
Na+, X- in NaOH eluent Cathode
H2O 2H+ + ½ O2 + 2e
- 2H2O + 2e- 2OH- + H2
X- H+
Na+ OH-
Chemistry and Ion Movement … … in an Thermo Scientific™ Dionex™ ERS™ electrolytically regenerated suppressor
-
HPLC Separation … Thermo Scientific™ Hypercarb™ and Silica C18 columns
-
• 10 µg/kg spike in grapes (Fosetyl & Phosphonic acid @ 100 µg/kg)
IC-MS/MS Pesticide Multi Residue Ion Chromatogram
-
IC Separations - Polar Pesticides …
Glyphosate
Glufosinate
Ethephon
Phosphonic Acid
Fosethyl Al
Maleic Hydrazide
Bromide
AMPA
… on a Thermo Scientific™ Dionex™ IonPac™ AS19 4 µm column
-
AS19 4 um Gradient
Ion Chromatography Selectivity
• 0- 12 minutes - Matrix interferences in conductivity trace can be sent to waste to preserve the ion source.
• 12 minutes +/- analytes of interest directed to MS
-
Polar_mix_20ppb_AS19_161006104310 10/06/16 10:43:10
RT: 0.00 - 31.93
0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30Time (min)
0
1000
1000
1000
100R
ela
tive
Ab
un
da
nce
0
1000
10025.36
25.67 26.91 30.7128.5424.3920.47 22.2310.916.0914.92
15.1915.94 16.91 25.7518.07 26.1620.56 23.72 29.64 31.4321.2213.433.54 9.50 11.832.53 4.9916.63
17.12 18.036.27 20.44 22.24 24.56 25.388.15 28.43 30.273.81 5.51 10.55 11.558.80 15.7314.902.6411.92
12.64 13.47 15.25 16.89 18.34 23.48 31.5028.04 30.1424.27 27.1520.85 22.355.61 9.336.64 8.21 11.494.753.422.681.7024.69
25.0317.03 25.8016.78 27.8217.90 28.98 30.6015.166.42 13.56 18.595.53 23.2412.29 21.0310.787.63 9.314.412.2331.08
30.5724.87 29.0027.7222.5419.85 21.3817.25 19.1013.625.54 16.4812.686.78 8.15 8.73 10.863.682.12
NL: 8.15E5Base Peak m/z= 168.0059-168.0075 F: FTMS - p ESI Full ms [50.0000-200.0000] MS Polar_mix_20ppb_AS19_161006104310
NL: 2.60E5Base Peak m/z= 110.0006-110.0018 F: FTMS - p ESI Full ms [50.0000-200.0000] MS Polar_mix_20ppb_AS19_161006104310
NL: 3.75E6Base Peak m/z= 80.9743-80.9751 F: FTMS - p ESI Full ms [50.0000-200.0000] MS Polar_mix_20ppb_AS19_161006104310
NL: 4.04E6Base Peak m/z= 82.9537-82.9545 F: FTMS - p ESI Full ms [50.0000-200.0000] MS Polar_mix_20ppb_AS19_161006104310
NL: 1.85E5Base Peak m/z= 128.0096-128.0108 F: FTMS - p ESI Full ms [50.0000-200.0000] MS Polar_mix_20ppb_AS19_161006104310
NL: 1.81E6Base Peak m/z= 98.9486-98.9496 F: FTMS - p ESI Full ms [50.0000-200.0000] MS Polar_mix_20ppb_AS19_161006104310
Polar_mix_20ppb_AS19_161006104310 #4117 RT: 18.35 AV: 1 NL: 1.89E7T: FTMS - p ESI Full ms [50.0000-200.0000]
50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200m/z
0
10
20
30
40
50
60
70
80
90
100
Re
lative
Ab
un
da
nce
59.9850
94.9809
60.992896.9602
76.9880
61.9881 183.0123
79.9574 194.9277184.991659.0135 98.9560 172.9915116.928780.9652 140.9673111.947389.0245180.9967
155.988963.9621 75.0087 134.8949 147.0300129.0559 161.0457
um Matrix Interferences Removed
TIC showing 20ppb: *Glyphosate *AMPA *Phosphonic acid *Chlorate *Cyanuric acid *Perchlorate
-
Flow Diagram— IC-HR/AM MS System
Conductivity Detector
EG IC column Suppressor IC pump 25 µL/min
H2O
Methanol
Low dead volume
mixing tee
Waste AXP-MS pump 10 µL/min
High pressure ion chromatography system
Injection from autosampler
To assist desolvation
Thermo Scientific™ Q Exactive™ Orbitrap™
mass spectrometer
Grounding union
H2O
AXP pump 40 µL/min
Trap Guard
40 µL/min
Degas
Wang, J., Christison, T., et al. Submitted to Analytical Chemistry
Data Management
-
Ion Chromatography Coupled with Orbitrap HRMS (Q Exactive Orbitrap MS)
-
HRMS for IC and HPLC
Comprehensive solution for pesticide analysis
-
AMPA Linearity from 20 to 2000 ppt in Drinking Water
-
AMPA Repeatability at 30 ppt in Drinking Water
Picco Area
1 112109
2 110436
3 111219
4 114398
5 111643
Media 111961
StDev 1494.84
CV% 1.3
-
μS
Time (min)
1. Artificial matrix 2. Drinking water 3. Surface water 4. Bottled water 5. Deionized water
1.
2.
5. 4.
3.
Fosetyl-Al Glyphosate Clopyralid AMPA Glufosinate
Conductivity Traces of Anions Present in Different Matrices
-
20 ppt AMPA Spiked in Water Containing Cl- (5 to 50 ppm)
-
Glyphosate Lineatity from 20 to 2000 ppt
-
Glyphosate Repeatability at 10 ppt in Drinking Water
Picco Area 1 199496
2 190406
3 197228
4 195575
5 183081
Media 193157.2
StDev 6551.6
CV% 3.4
-
2 – 5 – 10 ppt of Glyphosate in Drinking Water
Grafico1
35886
105416
195575
Sheet1
PiccoArea
1199496
2190406
3197228
4195575
5183081
Media193157.2
StDev6551.6498456493
CV%3.3918745176
235886
5105416
10195575
Sheet1
-
Repeatability of Glyphosate 500 ppt on Lettuce Extract
Picco Area
1 565481
2 609915
3 543858
4 595630
5 609373
Media 584851.4
StDev 29160.51
CV% 4.99
-
Cation Polar Pesticides
Note: Triazole-lactic acid missing Courtesy of Dr. Richard Fussell
-
Cation Polar Pesticides on Dionex IonPac CS17 Column
Courtesy of Dr. Richard Fussell
-
Paraquat – Mepiquat – Chlormequat - Diquat
Clormequat
Mepiquat
Paraquat
Diquat
Dionex IonPac CS17 column full scan determination of cation polar pesticides
-
• Determination of several analytes in a single anionic run: • Glyphosate, AMPA, Glufosinate, anionic polar pesticides • Haloacetic Acid HAA9 • Bromate (at ppt level) • Perclorate (at ppt level) • Organic ccids • Anions
• Determination of several analytes in a single cationic run: • Diquat, Paraquat, Chlormequat, cationic polar pesticides • Amines • Cations
IC-MS Advantages
-
Do you have any questions?
Do you have additional questions or do you want to talk to an expert from Thermo Fisher Scientific? Please send an E-Mail to [email protected] and we will get back to you.
mailto:[email protected]
Ion Chromatography Coupled to MS, a Powerful Approach for Polar Pesticides DeterminationWhat is Glyphosate?Glyphosate – Health Concerns TimelineGlyphosate in the NewsAnalysis of GlyphosateQuPPe-PO v 9.1-Negative Mode CompoundsWhy Couple Ion Chromatography (IC) to MS?Principles of RFIC - Eluent GenerationChemistry and Ion Movement …HPLC SeparationIC-MS/MS Pesticide Multi Residue Ion ChromatogramIC Separations - Polar Pesticides …Ion Chromatography SelectivityMatrix Interferences Removed Flow Diagram— IC-HR/AM MS SystemIon Chromatography Coupled�with Orbitrap HRMS �(Q Exactive Orbitrap MS)HRMS for IC and HPLCAMPA Linearity from 20 to 2000 ppt in Drinking WaterAMPA Repeatability at 30 ppt in Drinking WaterConductivity Traces of Anions Present in Different Matrices20 ppt AMPA Spiked in Water Containing Cl- (5 to 50 ppm)Glyphosate Lineatity from 20 to 2000 pptGlyphosate Repeatability at 10 ppt in Drinking Water2 – 5 – 10 ppt of Glyphosate in Drinking WaterRepeatability of Glyphosate 500 ppt on Lettuce ExtractCation Polar PesticidesCation Polar Pesticides on Dionex IonPac CS17 ColumnParaquat – Mepiquat – Chlormequat - DiquatIC-MS AdvantagesDo you have any questions?