Column-based comprehensive two-dimensional liquid chromatography (LC×LC) is maturing. Very good hardware is available from several manufacturers and dedicated software, while still imperfect, is also increasingly available. One of the main challenges is now to apply these great tool in an adequate or, ideally, optimal matter. Because many more variables affect an LC×LC separation than a conventional one-dimensional LC separation, this is quite a steep challenge. While tools for computer-aided method development are an increasingly welcome option in 1D-LC, they are a virtual necessity in LC×LC. Hence, this will be the focus of a discussion on progress in LC×LC. A major issue for the future is the development of high-performance “spatial” two- and threedimensional LC separations. In theory, this is a superior approach, because all seconddimension (and eventually third-dimension) separations can be performed in parallel instead of sequentially, greatly inhancing the separation power per unit time. However, progressing from column-based (“temporal”) LC×LC to spatial LC×LC implies a paradigm change, requiring new hardware and new methodology. Spatial multi-dimensional LC is being explored from fundamental (optimum gradients), engineering (computer-aided design, computational fluid dynamics, 3D-printing) and application perspectives. Progress along these lines will be discussed.

Column-based comprehensive two-dimensional liquid chromatography (LC×LC) is maturing. Very good hardware is available from several manufacturers and dedicated software, while still imperfect, is also increasingly available. One of the main challenges is now to apply these great tool in an adequate or, ideally, optimal matter. Because many more variables affect an LC×LC separation than a conventional one-dimensional LC separation, this is quite a steep challenge. While tools for computer-aided method development are an increasingly welcome option in 1D-LC, they are a virtual necessity in LC×LC. Hence, this will be the focus of a discussion on progress in LC×LC. A major issue for the future is the development of high-performance “spatial” two- and threedimensional LC separations. In theory, this is a superior approach, because all seconddimension (and eventually third-dimension) separations can be performed in parallel instead of sequentially, greatly inhancing the separation power per unit time. However, progressing from column-based (“temporal”) LC×LC to spatial LC×LC implies a paradigm change, requiring new hardware and new methodology. Spatial multi-dimensional LC is being explored from fundamental (optimum gradients), engineering (computer-aided design, computational fluid dynamics, 3D-printing) and application perspectives. Progress along these lines will be discussed.

Column-based comprehensive two-dimensional liquid chromatography (LC×LC) is maturing. Very good hardware is available from several manufacturers and dedicated software, while still imperfect, is also increasingly available. One of the main challenges is now to apply these great tool in an adequate or, ideally, optimal matter. Because many more variables affect an LC×LC separation than a conventional one-dimensional LC separation, this is quite a steep challenge. While tools for computer-aided method development are an increasingly welcome option in 1D-LC, they are a virtual necessity in LC×LC. Hence, this will be the focus of a discussion on progress in LC×LC. A major issue for the future is the development of high-performance “spatial” two- and threedimensional LC separations. In theory, this is a superior approach, because all seconddimension (and eventually third-dimension) separations can be performed in parallel instead of sequentially, greatly inhancing the separation power per unit time. However, progressing from column-based (“temporal”) LC×LC to spatial LC×LC implies a paradigm change, requiring new hardware and new methodology. Spatial multi-dimensional LC is being explored from fundamental (optimum gradients), engineering (computer-aided design, computational fluid dynamics, 3D-printing) and application perspectives. Progress along these lines will be discussed.

Nowadays, life science is becoming a very important part of natural science. Cell is the basic unit of organism, from which we start to crack the code of life entity. Microfluidic technology, which is being developed fast currently, is widely used in the cell research field, due to its micro-scale channels and flexible design. In this work, we focused on the determination of molecular structure and contents of the cell secretion and signaling factor. Cells were manipulated and captured on the microfluidic devices, and mass spectrometry (MS) was employed for qualitative and quantitative detection. Our key issue is connecting the microchips with MS, realizing the cell culture and drug stimulation in microchannels, and purifying the cell secretion before detected by MS.

Nowadays, life science is becoming a very important part of natural science. Cell is the basic unit of organism, from which we start to crack the code of life entity. Microfluidic technology, which is being developed fast currently, is widely used in the cell research field, due to its micro-scale channels and flexible design. In this work, we focused on the determination of molecular structure and contents of the cell secretion and signaling factor. Cells were manipulated and captured on the microfluidic devices, and mass spectrometry (MS) was employed for qualitative and quantitative detection. Our key issue is connecting the microchips with MS, realizing the cell culture and drug stimulation in microchannels, and purifying the cell secretion before detected by MS.

Nowadays, life science is becoming a very important part of natural science. Cell is the basic unit of organism, from which we start to crack the code of life entity. Microfluidic technology, which is being developed fast currently, is widely used in the cell research field, due to its micro-scale channels and flexible design. In this work, we focused on the determination of molecular structure and contents of the cell secretion and signaling factor. Cells were manipulated and captured on the microfluidic devices, and mass spectrometry (MS) was employed for qualitative and quantitative detection. Our key issue is connecting the microchips with MS, realizing the cell culture and drug stimulation in microchannels, and purifying the cell secretion before detected by MS.

The improvements in high resolution mass spectrometry (HRMS) instrumentation and data analysis software in the last decades have popularized screening for water quality assessment. Yet, many chemicals can scape from the screening strategy due to their physico-chemical properties, particularly the most polar analytes which do not perform well on reversed-phase liquid chromatography (RPLC), or those not detected due to an improper sampling strategy [1,2]. Hence, in this work we have investigated the detection rate of a set of ca. 3200 potential pollutants of a wide range of polarities (mean logD at pH 7 was 1.5) by a suspect screening workflow on LC-HRMS by using four differrent retention mechanisms, i.e. RPLC on a polar extended range column, hydrophilic interaction chromatography (HILIC), mixed-mode liquid chromatography (MMLC) and supercritical fluid chromatography (SFC) [2]. In parallel grab sampling was compared to passive sampling with polar organic compounds integrative samplers (POCIS). A first assay was performed by analyzing a wastewater effluent by datadependant LC-HRMS. The results showed that SFC was the mechanism providing a larger set of detections, followed by RPLC, MMLC and HILIC. Furtheremore, POCIS provided more detections than grab sampling. However, if only POCIS and SFC would be employed, about 1/3 of the analytes would not be detected. Therefore, the combination of the two retention mechanisms providing a higher ortogonality (RPLC + SFC) and parallel passive and grab sampling was finally selected for application of the analysis of a set of inland and coastal surface water samples, where 155 chemicals were tentatively identified (level 2). Current ongoing work, in the frame of the Nor-Water project (www.nor-water.eu), is being performed in order to improve the detectability of even more polar chemicals and prioritization of a suspect list of chemicals meeting the PMT criteria [3]. Acknowledgements: this work was funded by AEI (CTM2017-84763-C3-2-R), Xunta de Galicia (ED431C2017/36 and ED481A-2017/156), and co-financed by the European Regional Development Fund through the Interreg V-A Spain-Portugal Programme (POCTEP) 2014-2020 (0725_NOR_WATER_1_P). This presentation reflects only the author´s view, thus Programme authorities are not liable for any use that may be made of the information contained therein.

The improvements in high resolution mass spectrometry (HRMS) instrumentation and data analysis software in the last decades have popularized screening for water quality assessment. Yet, many chemicals can scape from the screening strategy due to their physico-chemical properties, particularly the most polar analytes which do not perform well on reversed-phase liquid chromatography (RPLC), or those not detected due to an improper sampling strategy [1,2]. Hence, in this work we have investigated the detection rate of a set of ca. 3200 potential pollutants of a wide range of polarities (mean logD at pH 7 was 1.5) by a suspect screening workflow on LC-HRMS by using four differrent retention mechanisms, i.e. RPLC on a polar extended range column, hydrophilic interaction chromatography (HILIC), mixed-mode liquid chromatography (MMLC) and supercritical fluid chromatography (SFC) [2]. In parallel grab sampling was compared to passive sampling with polar organic compounds integrative samplers (POCIS). A first assay was performed by analyzing a wastewater effluent by datadependant LC-HRMS. The results showed that SFC was the mechanism providing a larger set of detections, followed by RPLC, MMLC and HILIC. Furtheremore, POCIS provided more detections than grab sampling. However, if only POCIS and SFC would be employed, about 1/3 of the analytes would not be detected. Therefore, the combination of the two retention mechanisms providing a higher ortogonality (RPLC + SFC) and parallel passive and grab sampling was finally selected for application of the analysis of a set of inland and coastal surface water samples, where 155 chemicals were tentatively identified (level 2). Current ongoing work, in the frame of the Nor-Water project (www.nor-water.eu), is being performed in order to improve the detectability of even more polar chemicals and prioritization of a suspect list of chemicals meeting the PMT criteria [3]. Acknowledgements: this work was funded by AEI (CTM2017-84763-C3-2-R), Xunta de Galicia (ED431C2017/36 and ED481A-2017/156), and co-financed by the European Regional Development Fund through the Interreg V-A Spain-Portugal Programme (POCTEP) 2014-2020 (0725_NOR_WATER_1_P). This presentation reflects only the author´s view, thus Programme authorities are not liable for any use that may be made of the information contained therein.

The improvements in high resolution mass spectrometry (HRMS) instrumentation and data analysis software in the last decades have popularized screening for water quality assessment. Yet, many chemicals can scape from the screening strategy due to their physico-chemical properties, particularly the most polar analytes which do not perform well on reversed-phase liquid chromatography (RPLC), or those not detected due to an improper sampling strategy [1,2]. Hence, in this work we have investigated the detection rate of a set of ca. 3200 potential pollutants of a wide range of polarities (mean logD at pH 7 was 1.5) by a suspect screening workflow on LC-HRMS by using four differrent retention mechanisms, i.e. RPLC on a polar extended range column, hydrophilic interaction chromatography (HILIC), mixed-mode liquid chromatography (MMLC) and supercritical fluid chromatography (SFC) [2]. In parallel grab sampling was compared to passive sampling with polar organic compounds integrative samplers (POCIS). A first assay was performed by analyzing a wastewater effluent by datadependant LC-HRMS. The results showed that SFC was the mechanism providing a larger set of detections, followed by RPLC, MMLC and HILIC. Furtheremore, POCIS provided more detections than grab sampling. However, if only POCIS and SFC would be employed, about 1/3 of the analytes would not be detected. Therefore, the combination of the two retention mechanisms providing a higher ortogonality (RPLC + SFC) and parallel passive and grab sampling was finally selected for application of the analysis of a set of inland and coastal surface water samples, where 155 chemicals were tentatively identified (level 2). Current ongoing work, in the frame of the Nor-Water project (www.nor-water.eu), is being performed in order to improve the detectability of even more polar chemicals and prioritization of a suspect list of chemicals meeting the PMT criteria [3]. Acknowledgements: this work was funded by AEI (CTM2017-84763-C3-2-R), Xunta de Galicia (ED431C2017/36 and ED481A-2017/156), and co-financed by the European Regional Development Fund through the Interreg V-A Spain-Portugal Programme (POCTEP) 2014-2020 (0725_NOR_WATER_1_P). This presentation reflects only the author´s view, thus Programme authorities are not liable for any use that may be made of the information contained therein.