On-line Dilution Research Articles

Development of Online Dilution System for Quantification of 90Sr Using Automatic Solid-phase Extraction Inductively Coupled Plasma Mass Spectrometry.

In this paper, we propose an online dilution system for the rapid quantification of radioactive strontium-90 (90Sr) with inductively coupled plasma mass spectrometry coupled solid-phase extraction and O2 dynamic reaction (cascade ICP-MS). The proposed system automatically provides a higher dilution ratio, which is at most 3.3 ± 0.2-times the ratio obtained by the previous method, without increasing the analysis time (<15 min). A detection limit of 2.7 Bq/kg wet (0.54 pg/kg wet) was achieved. The recovery test results were consistent with two different spiked values.

Improvement of the Peak Shape in 2D-LC Using a Flow Splitting On-line Dilution

Improvement of the Peak Shape in 2D-LC Using a Flow Splitting On-line Dilution

Pushing the limits of resolving power and analysis time in on-line comprehensive hydrophilic interaction x reversed phase liquid chromatography for the analysis of complex peptide samples

In the present work, we have investigated the combination of hydrophilic interaction liquid chromatography (HILIC) and reversed phase liquid chromatography (RPLC) for the separation of peptides in on-line HILIC x RPLC. This combination usually leads to significant solvent strength mismatch, since a weak solvent in HILIC becomes a strong solvent in RPLC. This may result in band broadening, peak distortion, and breakthrough phenomena. Our focus was directed towards the reduction of band broadening and peak distortion. The conditions of the emergence of breakthrough could be investigated with high resolution mass spectrometry (HRMS) detection. The importance of both the injection volume and the difference in composition between injection and elution solvents was highlighted. Reported strategies to avoid bad peak shapes mostly rely either on flow splitting to limit the injection volume, or on on-line dilution. Here, we propose an alternative approach which consists in injecting large volumes in the second dimension. In this case, no flow-splitting nor dilution prior to the second dimension is required. Our results show that above a certain critical injected volume, depending on both the compound and the elution conditions, narrow and symmetrical peaks can be obtained, despite the persistence of breakthrough. As a result, the injected volume in the second dimension must be larger than the largest critical volume. This counter-intuitive approach was applied for the on-line HILIC x RPLC-UV-HRMS analysis of a complex tryptic digest sample. A peak capacity close to 1500 could be achieved in 30 min, which is two-fold higher than in RPLC x RPLC within the same analysis time.

Two-dimensional liquid chromatography with active solvent modulation for studying monomer incorporation in copolymer dispersants.

A pseudo-comprehensive two-dimensional liquid chromatography approach with size exclusion chromatography in the first dimension and gradient reversed-phase liquid chromatography in the second dimension was successfully developed for the characterization of vinyl acetate/acrylic acid copolymers and vinyl acetate/itaconic acid/acrylic acid terpolymers. Active solvent modulation was exploited to prevent the polymer breakthrough in the second dimension separation caused by the strong solvent used in the first dimension. The conditions of the active solvent modulation valve were optimized to achieve sufficient on-line dilution and to completely prevent polymer breakthrough without adding excessive time to the modulation cycle. Using this approach, copolymers made with different monomer ratios and processes were studied. Heterogeneous composition distribution due to insufficient monomer incorporation was detected in some of the copolymer samples. We demonstrated that with active solvent modulation, the two-dimensional liquid chromatography approach is no longer limited to water-soluble polymers and can be used for a broader range of polymers and copolymers.

Comprehensively qualitative and quantitative analysis of ginsenosides in Panax notoginseng leaves by online two-dimensional liquid chromatography coupled to hybrid linear ion trap Orbitrap mass spectrometry with deeply optimized dilution and modulation system

Panax notoginseng leaves (PNL) was considered as a potential medicinal part with abundant protopanaxdiol type ginsenosides. In this study, an integrated system was developed for simultaneously qualitative and quantitative analysis of ginsenosides in PNL using online comprehensive two-dimensional hydrophilic interaction chromatography and reversed-phase liquid chromatography coupled to a hybrid linear ion trap-Orbitrap mass spectrometry (online HILIC × RP-ESI/HRMS/MSn). The system was configured based on the combination of a XBridge amide column (150 mm × 2.1 mm, 2.5 μm) and Accucore phenyl-hexyl (50 mm × 4.6 mm, 2.6 μm) for the first and second dimensions, respectively. An additional water phase was introduced to dilute the eluent from the first dimension to decrease its elution strength in the second dimension. The online dilution, modulation interface and the second-dimension gradient program were deeply optimized to reduce possible sample loss and improve system resolution. Under the optimal conditions, a total of 226 ginsenosides were unambiguously identified or tentatively characterized by aid of high-resolution accurate mass and MSn fragment data in both negative and positive ion modes, and 93 of them were discovered as potentially new ginsenosides in PNL. Besides, the validated online HILIC × RP-LTQ-MS method was applied to determine 24 ginsenosides directly on 2D-EIC contour plots in nine batches of PNL samples. The powerful separation capability acquired by the developed online HILIC × RP system affords not only reliable structural information for identification, but also accurate quantitation. This combined system can also be used to characterize and quantify bioactive ingredients in the samples with complex matrices.

Online Measurement System in Reaction Monitoring for Determination of Structural and Elemental Composition Using Mass Spectrometry.

The monitoring of chemical reactions is an important task in chemical engineering, especially in quality control, pharmaceutical and biological processes, or industrial production. The development of new reactions such as catalyst-based procedures requires detailed knowledge about process steps and reaction kinetics. For qualitative and quantitative analysis of reactants and resulting products, proprietary online measurement systems are used, which were designed for special applications. A mobile online reaction monitoring system was developed for a flexible coupling to different mass selective measurement systems for structural (ESI-MS) and elemental (ICP-MS) analysis to determine chemical precursors, reaction products, and internal standard compounds and their elemental composition at any stage of the reaction. Chemical reactions take place in a tempered continuous-flow microreactor. The flow rate in the microreactor can be varied to adjust the residence times in the reactor. An online dilution module was integrated to adapt the concentration of the reaction solutions to the working range of the analyzers. The performance and limitations of the online reaction system were determined using standard solutions and a real chemical reaction. The control software with a graphical user interface enables the adjustment of reaction, sampling, and measurement parameters as well as the system and process control.

A remote-controlled immunochemical system for nephelometric detection of human serum transferrin

The fully-mechanized Multicommutated Flow Analysis (MCFA) system for determination of transferrin in human serum has been developed. The analytical methodology concerns immunoprecipitation measurements with the use of LED-based nephelometric flow-through detectors with polymeric light guides. To improve the mechanization degree of the MCFA system, the Do-It-Yourself electronic module for mobile phone control has been developed. The design and structure of an Android software (build using open source application) controlling the operation of presented flow system has been presented. To bypass the problem of nonlinearity of calibration curve caused by the nature of antigen-antibody interactions, the on-line dilution module has been integrated into the presented manifold. Under optimized conditions, the developed flow analysis system is characterized by relatively low limit of detection and quantitation (2.0 and 4.9 mg L−1, respectively), good precision (RSD < 4%), low reagent and sample consumption per one measurement (16 µL of undiluted reagent and 20 µL of undiluted sample without further on-line dilution, respectively) and relatively high throughput (approximately 35 signal recordings per hour). The developed MCFA system enabled to analyze 16 serum samples with the transferrin concentration from 90 to almost 350 mg dL−1 with statistical agreement to the reference method.

Green analytical method for simultaneous determination of salinity, carbonate and ammoniacal nitrogen in waters using flow injection coupled dual-channel C4D

A flow injection analysis system (FIA) for the simultaneous determination of salinity, carbonate and ammoniacal nitrogen has been developed and reported in this paper. FIA incorporating membrane units was used, not only for the separation of the gaseous carbon dioxide and ammonia, but also for on-line dilution in the salinity measurement. The sample was injected via a 10-port valve with two sample loops. One loop was used for salinity and carbonate measurements and the second loop for ammoniacal nitrogen determination. A dual-channel capacitively coupled contactless conductivity detector was assembled in a single shielding box. Input voltage from the same AC power supply was fed to the input electrodes of both C4D cells. One channel of the C4D was used to monitor the change in conductivity of an acceptor stream that carried a zone of the water sample that has passed through the on-line dilution unit. Conductivity of this zone relates directly to the salinity of the sample. The same sample zone was next acidified to generate carbon dioxide gas that diffused through a hydrophobic membrane of the first gas diffusion (GD) unit. The zone of dissolved carbon dioxide in acceptor stream of water flowed into the same C4D cell as for the salinity measurement, but arriving at a later time. Concurrently, the second channel of the C4D monitored the change in conductivity of the acceptor stream in the second GD unit due to the diffusion of ammonia gas generated by the reaction of base with the sample injected from the second sample loop. The change in conductivity at this second C4D cell correlates with the concentration of ammoniacal nitrogen present in the sample. The proposed method is low cost, simple, rapid and sensitive. The limit of quantitation for salinity, carbonate and ammoniacal nitrogen are 0.31mmolL−1, 1.85 µmol L−1, respectively. Throughput of 20 samples h−1 for simultaneous analysis can be achieved with RSD of less than 3.8%. The system had been applied to the determination of salinity, carbonate and ammoniacal nitrogen in 15 water samples, with results in agreement with those obtained using comparison methods.

Online comprehensive two-dimensional hydrophilic interaction chromatography × reversed-phase liquid chromatography coupled with hybrid linear ion trap Orbitrap mass spectrometry for the analysis of phenolic acids in Salvia miltiorrhiza

In the present study, a system was developed for the analysis of phenolic acids in Salvia miltiorrhiza using online comprehensive two-dimensional hydrophilic interaction chromatography and reversed-phase liquid chromatography coupled to a DAD detector and hybrid linear ion trap-Orbitrap mass spectrometry (HILIC×RP-DAD-ESI/HRMS/MSn). The system was configured based on the combination of an XBridge Amide column (150mm×4.6mm, 3.5μm) and Accucore PFP column (50mm×4.6mm, 2.6μm) for the first and second dimensions, respectively. An additional LC pump was used to dilute the eluent from the first dimension to decrease its elution strength in the second dimension. A back-flush trap column was selected as an interface to make up for the loss of efficiency and resolution due to the online dilution. Under the optimized conditions, a total of 196 peaks of polar compounds were successfully separated and detected in Salvia miltiorrhiza using the developed online HILIC×RP system, which exhibited high orthogonality (73%). The online combination of HILIC and RP provides powerful separation capability for the analysis of polar compounds in samples with complex matrices.

Silver nanoparticles in seawater: A dynamic mass balance at part per trillion silver concentrations

This study investigates the dynamic processes affecting silver (Ag) nanoparticles that have been spiked into seawater at environmentally relevant concentrations (200 and 2000ngAgL−1). Seawater samples were taken at regular time intervals from multiple microcosm tanks and analysed rapidly, without any sample preparation, using a recently developed flow injection on-line dilution single particle inductively coupled plasma mass spectrometry method. Dissolution was found to be the predominant process of Ag nanoparticle transformation, with its rate being influenced by the type and thickness of the nanoparticle organic coating. More specifically the branched poly(ethyleneimine) coating provided additional stability to the 40 and 60nmAg nanoparticles that were tested, compared to the poly(vinylpyrrolidone) coated ones. At high Ag nanoparticle spiking levels and after 24h of exposure an extra Ag-containing nanoparticle peak appeared at the low range of the NP size distribution histogram. This peak corresponds to Ag-containing particles that contain Ag mass equivalent to 25–30nm Ag nanoparticles (assuming spherical shape). However, the composition and the “real” size of these particles remains unknown as the particles may have formed from the in-situ reduction of dissolved silver or they originate from other processes involving nanocrystal formation, as has been shown to occur in sewage sludge, or interaction with natural organic matter. Overall, this study provides additional insight into the physicochemical mechanisms behind Ag nanoparticle behavior in marine media.

Optimization of plasma sampling depth and aerosol gas flow rates for single particle inductively coupled plasma mass spectrometry analysis

We performed experiments to assess the separate and also the combined effect of the sampling depth and the aerosol gas flow rates on the signal formation in single particle inductively coupled plasma mass spectrometry (spICP-MS) measurements by using dispersions containing Ag and Au NPs. It was found that the NP signal can significantly be improved by the optimization of the sampling depth. With respect to the „robust” setting, a signal improvement of nearly 100% could be achieved, which translates into a 25–30% improvement in size detection limits. It was also found that the shape of the spICP-MS signal histograms also change with the change of the plasma sampling depth. It was demonstrated that nanoparticle peak separation can also be significantly enhanced by using sampling depth optimization.The effect of the aerosol dilution gas flow, now standard in most ICP-MS instruments, on the spICP-MS signal formation was also studied for the first time in the literature, as this flow was hoped to make spICP-MS measurements more practical and faster via the on-line dilution of the aerosol generated from nano-dispersions. Our experimental results revealed that the dilution gas flow can only be used for a moderate aerosol dilution in spICP-MS measurements, if the gas flow going to the pneumatic nebulizer is proportionally lowered at the same time. This however was found to cause a significant worsening in the operation of the sample introduction system, which gives rise to a strong NP signal loss. Thus it was concluded that the use of the aerosol dilution gas flow, in its present form, can not be suggested for spICP-MS analysis.

Quantitative method for analysis of tobacco-specific N-nitrosamines in mainstream cigarette smoke by using heart-cutting two-dimensional liquid chromatography with tandem mass spectrometry.

A heart-cutting two dimensional liquid chromatography coupled with tandem mass spectrometry method was developed for the analysis of tobacco-specific N-nitrosamines (TSNAs) at low concentration level in Virginia-type cigarette smoke. A strong cation exchange column was utilized for the first dimensional separation, which effectively removed acidic and neutral components in the smoke, followed by a reversed phase liquid chromatography coupled with tandem mass spectrometric analysis. To capture components of the TSNAs in the effluent on the trapping column, a compensating pump was applied for online dilution and pH adjustment during the period of the TSNAs fraction transferring and enrichment. Highly sensitive determination of the TSNAs in mainstream cigarette smoke was achieved by isotope deuterated internal standards under the multiple reaction monitoring mode. Compared with traditional methodologies, the method was almost no matrix interference. Limits of quantity for the TSNAs were within 0.027-0.094ng/mL, and the results showed good reproducibility and accuracy. Finally, the new method was applied for analysis of the Kentucky reference cigarettes and the results agreed well with joint experiments of Cooperation Centre for Scientific Research Relative to Tobacco.

Direct Determination of Arsenic and Chromium in Seawater Samples Using On-Line Dilution and ICP-MS Analysis

Direct Determination of Arsenic and Chromium in Seawater Samples Using On-Line Dilution and ICP-MS Analysis

New approach of a transient ICP-MS measurement method for samples with high salinity

In the near future it is necessary to establish a disposal for high level nuclear waste (HLW) in deep and stable geological formations. In Germany typical host rocks are salt or claystone. Suitable clay formations exist in the south and in the north of Germany. The geochemical conditions of these clay formations show a strong difference. In the northern ionic strengths of the pore water up to 5M are observed. The determination of parameters like Kd values during sorption experiments of metal ions like uranium or europium as homologues for trivalent actinides onto clay stones are very important for long term safety analysis. The measurement of the low concentrated, not sorbed analytes commonly takes place by inductively coupled plasma mass spectrometry (ICP-MS). A direct measurement of high saline samples like seawater with more than 1% total dissolved salt content is not possible. Alternatives like sample clean up, preconcentration or strong dilution have more disadvantages than advantages for example more preparation steps or additional and expensive components. With a small modification of the ICP-MS sample introduction system and a home-made reprogramming of the autosampler a transient analysing method was developed which is suitable for measuring metal ions like europium and uranium in high saline sample matrices up to 5M (NaCl). Comparisons at low ionic strength between the default and the transient measurement show the latter performs similarly well to the default measurement. Additionally no time consuming sample clean-up or expensive online dilution or matrix removal systems are necessary and the analysation shows a high sensitivity due to the data processing based on the peak area.

Production of trace elements in coastal sea water certified reference material NMIA MX014.

A certified reference material (CRM) for trace elements in acidified sea water, NMIA MX014, has been produced by the National Measurement Institute Australia (NMIA). The CRM consists of natural coastal sea water with 12 elements (As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Se and V) fortified to levels relevant to environmental regulatory testing in Australia ranging from 0.4 to 22μg/kg. Certified values for these 12 elements were assigned using reference methods developed at NMIA, using either isotope dilution or standard addition with ICP-MS measurement. Specialised sample preparation (coprecipitation) and ICP-MS optimisation (online dilution, collision/reaction chemistry, high mass resolution) were used to negate the effect of the high level of dissolved solids. Multiple confirmatory experiments were performed in order to verify that ICP-MS spectral interferences were eliminated and to estimate the measurement uncertainty contribution from method precision and method trueness. Extensive homogeneity and stability testing was performed and the measurement uncertainty of certified values includes contributions from between-bottle homogeneity, short-term stability, medium-term stability and long-term stability. Special attention was paid to the stability of Hg due to well-known preservation problems. Acidified sea water matrix was satisfactory for stabilising Hg at 0.4μg/kg for at least 4years. Relative expanded uncertainties (k = 2) for the 12 certified values were between 1 and 11%. NMIA MX014 is intended for use as a reference material for analytical method validation and quality control for quantification of trace elements in saline water and other similar sample types.

Nanomaterial size distribution analysis via liquid nebulization coupled with ion mobility spectrometry (LN-IMS).

We apply liquid nebulization (LN) in series with ion mobility spectrometry (IMS, using a differential mobility analyzer coupled to a condensation particle counter) to measure the size distribution functions (the number concentration per unit log diameter) of gold nanospheres in the 5-30 nm range, 70 nm × 11.7 nm gold nanorods, and albumin proteins originally in aqueous suspensions. In prior studies, IMS measurements have only been carried out for colloidal nanoparticles in this size range using electrosprays for aerosolization, as traditional nebulizers produce supermicrometer droplets which leave residue particles from non-volatile species. Residue particles mask the size distribution of the particles of interest. Uniquely, the LN employed in this study uses both online dilution (with dilution factors of up to 10(4)) with ultra-high purity water and a ball-impactor to remove droplets larger than 500 nm in diameter. This combination enables hydrosol-to-aerosol conversion preserving the size and morphology of particles, and also enables higher non-volatile residue tolerance than electrospray based aerosolization. Through LN-IMS measurements we show that the size distribution functions of narrowly distributed but similarly sized particles can be distinguished from one another, which is not possible with Nanoparticle Tracking Analysis in the sub-30 nm size range. Through comparison to electron microscopy measurements, we find that the size distribution functions inferred via LN-IMS measurements correspond to the particle sizes coated by surfactants, i.e. as they persist in colloidal suspensions. Finally, we show that the gas phase particle concentrations inferred from IMS size distribution functions are functions of only of the liquid phase particle concentration, and are independent of particle size, shape, and chemical composition. Therefore LN-IMS enables characterization of the size, yield, and polydispersity of sub-30 nm particles.

Flow injection with on-line dilution and single particle inductively coupled plasma – mass spectrometry for monitoring silver nanoparticles in seawater and in marine microorganisms

Description of a flow injection method for monitoring silver nanoparticles by single particle ICP-MS in seawater matrix and seawater microorganisms.

Technical Improvements in Quantitative In Vitro Microdialysis Sampling of Carbohydrates

The use of quantitative in vitro microdialysis has far-reaching applications to monitor a wide variety of bioprocesses without complex sample preparation procedures. On-line in vitro microdialysis sampling followed by high performance anion exchange chromatography and pulsed electrochemical detection was employed to monitor carbohydrate enzyme kinetics. A key improvement to the microdialysis perfusate delivery system is reported to enhance reproducibility, accuracy, and recovery. A computer-programmable pump designed for precision fluid delivery, with flow rates in the nano- and microliter per minute range that did not require perfusate reloading, was used in place of the commonly used syringe pump. Peak-to-peak noise levels were reduced by three orders of magnitude and recoveries at low flow rates (less than 1 µL/min) were greater than 90% for glucose. In addition, accurate and reproducible delivery allows for on-line dilution for flow rates across two orders of magnitude.

Flagging and correcting non-spectral matrix interferences with spatial emission profiles and gradient dilution in inductively coupled plasma-atomic emission spectrometry

Matrix interference remains one of the most daunting challenges commonly encountered in inductively coupled plasma-atomic emission spectrometry (ICP-AES). In the present study, a method is described that enables identification and correction of matrix interferences in axial-viewed ICP-AES through a combination of spatial mapping and on-line gradient dilution. Cross-sectional emission maps of the plasma are used to indicate the presence of non-spectral (plasma-related and sample-introduction-related) matrix interferences. In particular, apparent concentrations of an analyte species determined at various radial locations in the plasma differ in the presence of a matrix interference, which allows the interference to be flagged. To correct for the interference, progressive, on-line dilution of the sample, performed by a gradient high-performance liquid-chromatograph pump, is utilized. The spatially dependent intensities of analyte emission are monitored at different levels of sample dilution. As the dilution proceeds, the matrix-induced signal variation is reduced. At a dilution where the determined concentrations become independent of location in the plasma, the matrix interference is minimized.

New Innovations in Testing Sustained-Release Tablets Using an Automated Dissolution System with Online Dilution

This paper describes the development and validation of an automated method for the dissolution testing of metformin hydrochloride for use in the quality assurance program. Automation could provide significant improvements in the consistency of results compared with existing manual methods. Care and consideration were given to the automation of preparation of samples and standards in line with current manual procedures. Reproducibility and sample preparation accuracy were quantified, and results obtained were compared against tests performed manually. The system was validated in accordance with the Chinese (1), European (2), and United States pharmacopeias (3) using salicylic acid. Salicylic acid testing for bath calibration also required dilution, which provided a direct comparison to the automated system used to test metformin hydrochloride tablets.