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Combining Liquid Chromatography Time-Of-Flight Mass Spectrometry for Water Screening with Software Tools to Identify Pesticides and Their Metabolites

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Combining Liquid Chromatography Time-Of-Flight Mass Spectrometry for Water Screening with Software Tools to Identify Pesticides and Their Metabolites VOL. 23 NO. 6 (2011) AARTICLERTICLE Combining liquid chromatography time-of-flight mass spectrometry for water screening with software tools to identify pesticides and their metabolites Wolfgang Schulza and André Schreiberb aLaboratory for Operation Control and Research, Landeswasserversorgung, Germany. E-mail: [email protected] bAB SCIEX, Concord, Ontario, Canada Introduction removes the smallest particles of organic chromatography (LC) coupled with The Landeswasserversorgung (LW) is one contaminants. Finally, the water is disin- mass spectrometry, GC-MS and LC-MS, of the largest water supply companies in fected with chlorine dioxide as a security respectively, to monitor the presence of Germany; it is responsible for providing measure to keep the drinking water safe hundreds of organic contaminants that safe drinking water to more than 250 during transportation to the consumer. are routinely found in water, including cities across the south-west of the coun- Drinking water is strictly regulated and pesticides, industrial chemicals, pharma- try. LW supplies up to 450,000 m3 per the German Drinking Water Ordinance ceuticals and their by-products. These day (6500 s–1) to its 3 million custom- requires over 100 individual param- contaminants can find their way into ers, taken from three different raw water eters to be inspected. For the LW, this water sources directly and indirectly, resources: ground water (from the amounts to over 300,000 separate such as through landfill leachates or Danube marshland), spring water (Egau analyses per year, for which the LW runs accidents and via wastewater treatment waterworks) and the River Danube. Raw its own Laboratory for Operation Control plants. water is extracted from these sources and Research (LOCR) at the Langenau The LW tries to identify all organic and pumped to one of LW’s waterworks waterworks site. This laboratory is respon- substances at trace-level concentrations in Langenau or Dischingen, for rigorous sible for chemical, physico–chemical in its water sources. This approach has a treatment and testing in order to produce and microbiological analyses, provided number of challenges: in particular, many the highest quality drinking water. Water by around 30 staff. The analyses include of the compounds are present at only from the River Danube, for example, monitoring pH, water hardness, micro- very low concentrations, meaning the passes through six stages, beginning biological contaminants (for example, analytical methods must be extremely with two pre-treatment plants for the coliform bacteria, E. coli, Enterococcus, sensitive, but also able to detect diverse removal of humic and clouding materi- and colony count) and inorganic and compounds, including those with low or als, followed by the oxidative removal organic parameters. The laboratory high polarity. Another challenge is coping of flavours and odorous substances and uses a range of chromatography and with the vast number of contaminants oxidation and disinfection of the water at spectrometry-based techniques to (hundreds or thousands) that may be an ozone plant. The water then under- monitor and profile levels of contami- present in any given sample. Finally, a goes a dual-layer filtration process (with nants in raw and treated water (see major challenge is the question of how to hydroanthracite and sand) to remove Table 1). The laboratory has particular detect and identify contaminants that are residual suspended substances, followed expertise in organic trace analysis, using not necessarily expected to be present, by activated carbon filtering, which gas chromatography (GC) and liquid both rapidly and cost-effectively. 18 SPECTROSCOPYEUROPE www.spectroscopyeurope.com VOL. 23 NO. 6 (2011) AARTICLERTICLE Table 1. Range of techniques used by the LW to monitor levels of contaminants in raw and Developing non-target treated water. screening for water Class Examples Analysis method quality testing Inorganic compounds Scientists at LW perform NTS using LC-ToF-MS/MS, using software to iden- Anions Sulfate, chloride, nitrate Ion chromatography tify all peaks present in a sample after Phosphate, ammonium, Photometry non-targeted data acquisition. When a silicate, chlorine dioxide, ozone non-targeted, unexpected compound is discovered, it is important to be able Calcium, magnesium, sodium, to subsequently confirm and identify Cations and heavy metals potassium, iron , chromium, ICP-MS copper, uranium that compound, therefore the mass spectrometer must be sufficiently sensi- Organic compounds tive to detect and identify the compound Pesticides & metabolites Carbofuran, directly, while at the same time having methiocarb sulfone LC/MS and GC/MS high mass accuracy for components Organic chlorine compounds PCB, lindane present at very low concentrations. Industrial chemicals Benzotriazoles The LW started develop- Pharmaceuticals Sulfadiazene, carbamazepine, ing LC-ToF-MS/MS-based methods sulfamethoxazole, iodinated combined with electrospray ionisation x-ray contrast media (ESI) for non-target screening about Aromatic compounds Benzene, xylene five years ago. The approach has the advantage of providing information Benyzl chloride Volatile halogenated on all organic substances in a sample compounds trichloroethene, chloroform, chloroethene that are detectable with this system; Complexing agents EDTA however, there were also a number of disadvantages that the scientists are in the process of addressing. The main issue was sensitivity, and a sample preparation step—solid phase Non-target and a mass spectral library for identification. extraction (SPE)—had to be included, multi-target screening Comparison of the total ion chroma- which leads to interferences. These In general, there are two different tograms (TIC) of unknown samples to had to be removed by blank subtrac- approaches to MS-based screen- that of the control reveals compounds tion during NTS, which was not straight- ing: non-target screening (NTS, also that are either unique to the sample, forward since, initially, the LOCR didn’t known as general unknown screen- or those that are present at signifi- have software capable of performing the ing) and multi-target screening. For cantly higher concentrations than in required calculations. NTS, a single survey scan is carried out the control. Thus, the aim of NTS is to Other challenges to overcome over a defined mass range to discover detect all substances that are accessi- included the difficulties of data analy- which compounds are present in a ble to the particular analysis technique, sis, the need for multivariate statistical sample, prior to their identification with a focus on unexpected or unknown methods, and the need for standard- and quantification. This is commonly substances. ised procedures, particularly with the used when monitoring environmental Multi-target screening, in contrast, large amount of data that NTS gener- samples, for example, that may contain searches for a predefined list of ates. Finally, improved methods were a large number of diverse, unknown compounds in a multiple reaction moni- needed for identification of detected contaminants. toring (MRM) or single ion monitor- compounds, particularly when inter- The MS survey scan could be a time- ing (SIM) experiment. MRM mode is preting accurate mass MS/MS data. The of-flight (ToF), a quadrupole or an ion generally preferred for targeted screen- LOCR’s researchers have spent several trap scan, although high-resolution ing because of its higher selectivity and years overcoming these problems, and instruments such as ToF are generally sensitivity and high S/N.1,2 This allows have recently published a more sensi- used for NTS procedures, due to their direct quantification of target analytes tive, fast and powerful general screening better signal-to-noise (S/N) ratio in at the ng L–1 range, without the need for method that allows rapid comparison scanning mode. The scan can automati- prior enrichment.3 Once a compound between samples and identification of cally trigger the acquisition of a product is detected above a defined thresh- relevant compounds from the vast data ion spectrum if a detected compound’s old, a product ion scan is collected and generated.4 signal is above a defined threshold. This compared against a mass spectral library As an initial step, the scientists devel- spectrum can then be searched against to enhance confidence in detection. oped a highly sensitive LC-MS/MS www.spectroscopyeurope.com SPECTROSCOPYEUROPE 19 VOL. 23 NO. 6 (2011) AARTICLERTICLE multi-targeted screening method using verification of transformation products. information that could not be obtained a QTRAP 5500 System (AB SCIEX, USA), Samples (100 μL) are analysed using previously. The software then produces that allowed direct injection of the water direct injection ToF mass spectrometry, mass vs retention time scatter plots (see sample into the LC system, without the with the following settings: gas 1 40 psi, Figure 1), which represent all the organic need for pre-concentration.3 Since then, gas 2 50 psi, curtain gas 40 psi, temper- trace substances in the sample that are new ToF technology (such as the Triple ature 550°C, Voltage +5500 V resp. recognised by the method. TOF 5600, AB SCIEX) with higher mass –4500 V; scan range 50–1050 m/z. A critical factor of this new approach is resolution and high scan speeds has The resolution power was >38,000 to be able to differentiate between infor- been used that allowed further method at 922.0097 m/z and >20,000 at mation that is identical across different improvement. Another advance was 121,0508 m/z for ESI+, and >38,000 samples and information that is differ- brought about by new, faster software at 980.0164 and >22,000 at 119.0363 ent, such as the presence of a new and algorithms for peak finding and for ESI–. compound in a sample. This is achieved comparing samples. The higher resolution of new ToF tech- by selecting compounds to be submitted The new approach consists of three nology makes it possible to generate full for further identification on the basis of steps: screening/data evaluation, data- sample scans, very rapidly. Speed is really their likely relevance, based on set theory.
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