Resources for Modern Methods
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1 Resources for Modern Methods Experiences and Results Department for Environment and Energy Basel-City (AUE) Steffen Ruppe*, Dorrit Griesshaber*, Ingrid Langlois*, Jan Mazacek*, Matthias Ruff #, Martin Loos #, Heinz Singer # *Department for Environment and Energy Basel-City, Switzerland # Eawag: Swiss Federal Institute of Aquatic Science and Technology, Switzerland Kanton Basel-Stadt
2 Department for Environment and Energy Basel (AUE) The first out of seven Rhine monitoring stations is located near Basel, in Weil am Rhein. operated by AUE funded by the Federal Office for Environment (BAFU), Switzerland and the Ministry of the Environment, Sector Climate and Energy, Germany, Baden- Württemberg All samples from this station were analyzed by the AUE laboratory. Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 2
3 Resources for Modern Methods What do we need? LCMS-method measuring relevant targets at very low concentration ranges and detecting non-target substances How do we get there? In July 2009 a project started with the Eawag as scientific partner. 2011: approximately 300 substances were measured 2012: introduction of software EnviMass 1 (Martin Loos, Eawag) 2014: implementation of software EnviMass 2 method development took almost 4 years, primarily due to the missing software link Detection method and analysis software are critical factors for optimal use of resources. Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 3
4 Daily Analytical Strategy Using LC/MS-Orbitrap SPE (multiphase cartridges), 140 internal standards, measurements with a mass resolution of (+MS/MS-fragments) Target List (310) EnviMass Non-Target Priority Substances 50 Targets Manually integrated Semiquan. procedure Screening for relevant masses Trend detection Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 4
5 Priority Substance - obvious application periods in the spring and fall - daily measurements are important to detect violations of alarm limits and to enable estimations of chemical loads in the Rhine in this example, two days contributed 25 % of the annual Isoproturon load in 2013 Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 5
6 Priority Substance Aliskiren: originates from human usage (background level) and industrial production (peaks) dilution effects due to higher discharge can be observed Tizanidin: origin from industrial discharge was verified Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 6
7 Daily Analytical Strategy Using LC/MS-Orbitrap EnviMass Priority substances 50 Targets Target-List (310) Non-Target x calibration x semi-quantification x automated processing peak picking recalibration/alignment componentization prioritization EnviMass2 Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 7
8 Non-Target Screening: Mass (M+H + ) EnviMass2 computes a time series analysis comparing a current sample with last year s data set prioritization is based on a statistical method highlighting mass traces with increasing intensity EnviMass2 (July February 2014, including incident on ) ChemSpider search resulted in nearly 2000 suggestions for possible molecular formulas Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 8
9 Non-Target Screening: 2-Phenyl-2-(2-piperidin)acetamid The compound was identified knowing who discarded the substance and in cooperation with the company in question. 600 kg of the Ritalin reagent were discharged Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 9
10 Non-Target Screening: Mass (M-H - ) 13 C => 7 Carbon 15 N => 4 Nitrogen Mass resolution provide isotop distribution C7H2N4? mass accuracy < 2 ppm ChemSpider search resulted in only two suggestions due to the unusual elemental composition Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 10
11 Non-Target Screening: Mass (M-H - ) 13 C => 7 Carbon BW- 15 Holidays N => 4 Nitrogen taking the intensity profile and the timing of summer holidays in Switzerland and Germany into account, one potential pollution region is singled out discharging company was identified and provided the AUE laboratory with their reference standard Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 11
12 Non-Target Screening: Tetracarbonitrilpropen - nonessential by-product - not further monitored it poses a permanent pollution - concentration ranges from 0.2 µg/l to 1 µg/l - in December 2014 a load of 820 kg was estimated - Tetracarbonitrilpropen is stable Tetracarbonitrilpropen can be detected in samples from along the entire Rhine samples taken in 2013 at Maas-Luis showed nearly the same load It took 2 months from finding the unknown mass to identifying the compound. Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 12
13 Non-Target Screening: AMPS - initially, the concentration was estimated to be 150 ng/l using the internal standard Bentazon-D6 - the molecular structure was identified within one week!!! - the concentration was underestimated by 30-fold!!! Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 13
14 Non-Target Screening: AMPS - due to its polarity AMPS resides predominantly in the water phase - it cannot be completely removed by drinking water treatment detection of AMPS in drinking water It is necessary to pay special attention to polar substances! Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 14
15 15 LCMS-Rhine Alarms from AUE-BASEL in 2014 Substance Use Alarms 2014 Detection Load 2014 (kg) Paracetamol analgetic Feb 2014/ Nov 14 Semi-Quan 320 Aliskiren beta blocker Feb / March / April 14 Priority list 1400 Indometacin 2-Phenyl-2-(2- piperidin)acetamid analgetic beginning and end March 14 Semi-Quan 170 (in 2 weeks) Industrial chemical Feb - March 14 Nontarget 600 Toluol-4-sulfonic acid Industrial chemical Dez 14 Priority list 1500 Unknown_252 Sep 14 Nontarget 1300* Unknown_325 May 14 Nontarget 5000* Tizanidin Muscle relexant Jun 14 Priority list 150 Mecoprop Pesticide Jun 14 Priority list 320 Unknown_139 Jul 14 Nontarget 70* Tetracarbonitrilpropen Industrial chemical Nov 2014 (Info in Basel) Nontarget 14000* * load-estimation red marked substances = permanent pollutions Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 15
16 Rhine Monitoring in Basel 2010 FOCUS 2014 GCMS shifted LCMS Results Alarms 7 19 Load targets 15 t 90 t Load non-targets ca. 3 t (only GCMS) > 25 t Human resources + one full-time position Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 16
17 Summary new LC/MS technologies greatly enhance the analytical spectrum cooperation with scientific partners is necessary a daily target-analysis of nearly 300 relevant compounds is possible observed target-loads were multiplied by factor six daily non-target screening is important, using LC/MS and GC/MS techniques LC/MS non-target screening delivers relevant compounds with high loads personal statement: the benefit of the information gain greatly outweighs the cost of the additional human resources necessary Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 17
18 THANK YOU for your attention! Thanks also to: Anna Brunnert, Steffi Schreiber, Reto Dolf, Paul Svoboda and the entire AUE-Team Kanton Basel-Stadt Workshop IKSR, Steffen Ruppe, Folie 18
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