Analysis of Trace Organic Contaminants in Water by LC-MS/MS. Tarun Anumol Agilent Environmental Workshop March 18 th, 2014
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1 Analysis of Trace Organic Contaminants in Water by LC-MS/MS Tarun Anumol Agilent Environmental Workshop March 18 th, 2014
2 Introduction
3 Automation
4 Au.to.mation the technique of making an apparatus, a process, or a system operate automatically Past Present Future?
5 Automation in the lab Workbench for automated sample preparation Robot for automated pipetting into 96 well plates
6 Traditional Extraction Techniques Analysis of TOrCs in Water
7 C o n v e n t i o n a l M e t h o d Sample collection Surrogate addition Extraction (SPE) Evaporation Analysis
8 Sample Transport
9 Sample Volume
10 Instrument Setup
11 Instrument Setup Agilent Flexcube connected to 6460 MS/MS
12 Configuration Flex cube pump Analytical Column Mass Spectrometer Sample Injection Loop Equilibrating SPE cartridge Binary Pump Loading SPE cartridge 1 Waste Flexcube Pump Flow Path Binary Pump Flow Path
13 Configuration Flex cube pump Analytical Column Mass Spectrometer Sample Injection Loop Loading SPE cartridge Binary Pump Elution SPE cartridge 1 Waste Flexcube Pump Flow Path Binary Pump Flow Path
14 Trace Organic Chemicals Atenolol (ß-blocker) Atrazine (Herbicide) Benzophenone (UV-blocker) Bisphenol A (plasticizer) Caffeine (stimulant) Carbamazepine (Anti-seizure) DEET (Insect-repellant) Dexamethasone (glucocorticoid) Trimethoprim (Antibiotic) Estrone (Hormone) Fluoxetine (Anti-depressant) Gemfibrozil (Anti-cholesterol)
15 Trace Organic Chemicals Ibuprofen (Analgesic) Meprobamate (Anti-anxiety) Naproxen (Pain-reliever) PFBS (Fluoro-surfactant) PFOA (Fluoro-surfactant) PFOS (Fluoro-surfactant) Primidone (Anticonvulsant) Simazine (Herbicide) Sulfamethoxazole (Antibiotic) TCPP (Flame-retardant) Triclocarban (Anti-microbial) Triclosan (Anti-microbial)
16 150 ng on column Analysis of OSPE Cartridges <50% 50-60% 60-70% 70-80% 80-90% % % % % >130% % SB-AQ PLRP-s Phenyl-hexyl Carbon-X Absolute recovery range (%) Number of compounds PLRP-s selected for further studies
17 >130% % PLRP-s Flowrate: 1 ml/min 4 ml loading volume selected for further studies Loading Volume 60-70% 70-80% 80-90% % % % % 50-60% <50% ml 2.5 ml 3.0 ml 3.5 ml 4.0 ml 5.0 ml Absolute recovery range (%) Number of compounds
18 Triclosan Loading Volume Sulfamethoxazole Carbamezapine TCEP Atrazine Testosterone PFOA Benzophenone Ibuprofen PFOS Gemfibrozil Primidone Trimethoprim Caffeine Atenolol ml 2.5 ml 3.0 ml 3.5 ml 4.0 ml 5.0 ml Absolute recovery (%)
19 Triclosan PFOS Triclocarban Loading Rate Simazine Ditiazem Carbamezapine TCEP Fluoxetine Atrazine DEET PFHxA Bisphenol A Propylparaben Testosterone Clofibric Acid Benzophenone Naproxen PFOA TCPP Diclofenac Ibuprofen Gemfibrozil Norgestrel Meprobamate Sulfamethoxazole Benzotriazole Hydrocortisone Diphenhydramine Trimethoprim Primidone ml/min 1.0 ml/min 1.5 ml/min Increasing Retention Time Atenolol Caffeine Abusolute Recovery (%)
20 Analytical Method Simultaneous analysis of 36 TOrCs in positive and negative ESI Diphenhydramine Triclosan Gemfibrozil PFOS Triclocarban Atenolol Benzotriazole Caffeine Trimethoprim Primidone Meprobamate Prednisone Simazine Diltiazem TCEP Atrazine Testosterone Naproxen PFOA TCPP Benzophenone Diclofenac Ibuprofen DEET Injection Volume: 1.7 ml Cycle time (Extraction + Analysis): 14.5 min Analytical Column: Poroshell 120 EC, 2.1x50 mm
21 Trimethoprim Triclosan Method Validation DEET Diphenhydramine Ditiazem Gemfibrozil Hydrochlorothiazide Ibuprofen Meprobamate Naproxen PFOA PFOS Primidone Sulfamethoxazole TCPP Testosterone Carbamezapine Caffeine Bisphenol A Atrazine Benzophenone Ultrapure Water (30 ng/l) Ultrapure Water (100 ng/l) Atenolol Recovery (%)
22 Trimethoprim Triclosan Method Validation DEET Diphenhydramine Ditiazem Fluoxetine Gemfibrozil Hydrochlorothiazide Ibuprofen Meprobamate PFOA PFOS Primidone Sulfamethoxazole TCPP Testosterone Caffeine Carbamezapine Bisphenol A Benzophenone Atrazine Surface Water (30 ng/l) Surface Water (100 ng/l) Atenolol Recovery (%)
23 Trimethoprim Method Validation Clofibric Acid DEET Diclofenac Diphenhydramine Ditiazem Fluoxetine Gemfibrozil Hydrochlorothiazide Hydrocortisone Ibuprofen Meprobamate Naproxen Norgestrel PFHxA PFOA PFOS Primidone Propranolol Propylparaben Simazine Sulfamethoxazole TCEP TCPP Testosterone Triclocarban Triclosan Caffeine Carbamezapine Benzotriazole Bisphenol A Atenolol Atrazine 0 1:5 Diluted Waste Water (100 ng/l) Benzophenone Recovery (%)
24 Method Validation Benzotriazole Bisphenol A Caffeine Carbamezapine Clofibric Acid DEET Diclofenac Diphenhydramine Ditiazem Fluoxetine Gemfibrozil Hydrochlorothiazide Hydrocortisone Ibuprofen Meprobamate Naproxen Norgestrel PFHxA PFOA PFOS Primidone Propranolol Propylparaben Simazine Sulfamethoxazole TCEP TCPP Testosterone Triclocarban Triclosan Trimethoprim Intra-day variability (n=4) Inter-day variability (n=4) Atenolol Atrazine Benzophenone Relative Standard Deviation (%)
25 Method Detection Limit 8 replicates using Glaser et al. method 16 8 MDL (ng/l) Atenolol Atrazine Benzophenone Benzotriazole Bisphenol A Caffeine Carbamezapine Clofibric Acid DEET Diclofenac Diphenhydramine Ditiazem Fluoxetine Gemfibrozil Hydracortisone Hydrochlorothiazide Ibuprofen Meprobamate Naproxen Norgestrel PFHxA PFOA PFOS Primidone Propranolol Propylparaben Simazine Sulfamethoxazole TCEP TCPP Testosterone Triclocarban Triclosan Trimethoprim 0.5 Concentration (ng/l)
26 Average C12 13 C6 13 Triclosan Trimethoprim d3 Matrix Effects C12 13 C3 Carbamezapine d10 DEET d6 Diclofenac 13 C6 Diphenhydramine d5 Ditiazem d3 Fluoxetine d5 Gemfibrozil d6 Ibuprofen d3 Meprobamate d3 Naproxen 13 C1d3 PFHxA 13 PFOA C2 13 C4 13 PFOS C4 Primidone d5 Propylparaben d4 Sulfamethoxazole 13 C6 TCEP d12 Triclocarban Benzotriazole d4 Bisphenol A 13 Caffeine Atenolol d7 Atrazine d5 Benzophenone d10-30 SW WWE (1:5 dil) WWE Matrix Effect (%)
27 Ion Suppression: Online SPE Carbamazepine_d10 Concentration: 100 ppt Injection Vol: 1.5 ml Mass: 150 pg Ultrapure water Wastewater effluent (0.2 um filtered)
28 Ion Suppression: Conventional SPE Carbamazepine_d10 Concentration: 50 ppb Injection Vol: 3 µl Mass: 150 pg Ultrapure water Wastewater effluent (0.2 um filtered)
29 Ion Suppression: Direct Injection Carbamazepine_d10 Concentration: 3 ppb Injection Vol: 50 µl Mass: 150 pg Ultrapure water Wastewater effluent (0.2 um filtered)
30 Ion Suppression Effects Comparison of 3 methods Conventional SPE (500 fold) Online SPE Direct Injection Surrogate Recovery (%) 0 Atenolol d7 Sulfamethoxazole 13C3 Carbamazepine d10 DEET d6 TCEP d12 Gemfibrozil d6
31 Ion Suppression Effects Meprobamate Sulfamethoxazole Carbamazepine DEET TCEP Gemfibrozil Atenolol CSPE OSPE LVI Method Detection Limit (ng/l)
32 Ion Suppression Effects Meprobamate Sulfamethoxazole Carbamazepine DEET TCEP Gemfibrozil Atenolol CSPE-MRL OSPE-MRL LVI-MRL Method Reporting Limit (ng/l)
33 Analysis of Emerging Contaminants in Water Conventional SPE Method Online SPE Method Direct Injection Method 1 L sample 1.5 ml sample 0.1 ml sample 5 hours 30 min 15 min 36 CECs ng/l 33 CECs ng/l 21 CECs ng/l
34 Conclusions Online SPE allows sensitive analysis of trace organics in water while allowing significant time and labor savings. Method has been proven to be robust in several different water matrixes. Online SPE is significantly less affected by ion suppression compared with conventional offline extraction techniques. LVI is currently not sensitive enough to attain desired MRLs but offers promise with rapid increase in sensitivity of newer mass spectrometers. Use of online SPE with accurate mass detectors (ToF & Q-ToF) could allow for real-time analysis of trace unknowns in water.
35 Acknowledgements Dr. Shane Snyder Snyder Research Lab Dr. Sylvain Merel Dr. Sonia Dagnino Agilent Technologies Joe Weitzel Dr. Sheher Mohsin
36 Questions Contact:
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