Characterization of a condensed-phase membrane introduction mass spectrometry (CP-MIMS) interface using a methanol acceptor phase coupled with electrospray ionization for the continuous on-line quantitation of polar, low-volatility analytes at trace levels in complex aqueous samples

We report the development and application of a capillary hollow fibre membrane interface using methanol as an acceptor phase to deliver target analytes to an electrospray ionization source and a triple quadrupole mass spectrometer. Superior fluid handling systems lead to greater signal stability and membrane integrity for the continuous on‐line monitoring of polar and charged analytes in complex aqueous samples with detection limits in the parts‐per‐trillion to parts‐per‐billion range. The system can be operated in either a continuous flow or a stopped acceptor flow mode – the latter giving rise to greater sensitivity. We report detection limits, enrichment factors and signal response times for selected analytes with polydimethylsiloxane and Nafion^® polymer membrane interfaces. In addition, we demonstrate the use of this interface to detect pharmaceuticals and other contaminants in natural water and artificial urine. The improved sensitivity and analytical response times of our CP‐MIMS system make it possible to continuously monitor dynamic chemical systems with temporal resolutions on the order of minutes. Presented is a comparison of the performance of CP‐MIMS versus direct infusion electrospray ionization, demonstrating the potential advantages over direct infusion for trace analyte measurements in complex, high ionic strength samples. Furthermore, by continuously flowing a reaction mixture in a closed loop over the interface, we demonstrate the use of the system as an in situ reaction‐monitoring platform for the chlorination of a model organic compound in aqueous solution. Copyright © 2011 John Wiley & Sons, Ltd.     

An enzyme derivatized polydimethylsiloxane (PDMS) membrane for use in membrane introduction mass spectrometry (MIMS)

Membrane introduction mass spectrometry (MIMS) provides direct measurement of volatile and semivolatile analytes in condensed and gas-phase samples without sample preparation steps. Although MIMS has numerous advantages that include direct, on-line, real-time analysis with low detection limits, current applications of MIMS are predominantly limited to volatile and semivolatile analytes that permeate hydrophobic membranes (e.g., polydimethylsiloxane; PDMS). We report the first enzyme modified PDMS membrane for use with MIMS. This was achieved by immobilizing Candida rugosa lipase directly onto the surface of oxidized PDMS. These surface immobilized enzymes catalyze ester hydrolysis, releasing an alcohol product at the membrane interface that is readily detected. We have successfully used an enzyme modified membrane for the analysis and quantification of low-volatility and hydrophilic esters. We report the quantification of several carboxylic acid esters in dilute aqueous solutions, including a phthalate monoester carboxylate that is not readily detected by conventional MIMS. This new interface demonstrates the potential for extending the range and versatility of MIMS.     

Membrane introduction mass spectrometry

An overview of membrane introduction mass spectrometry (MIMS) is presented and comparisons are made with other direct sample introduction techniques. Special attention is given to the unique advantages and the limitations of newer variants on the MIMS technique, including affinity MIMS, reverse-phase and trap MIMS. The salient features of the interfaces used in MIMS are summarized and the various membrane materials commonly used are delineated. The applicability of MIMS is illustrated via discussion of

1. (i) bioreactor monitoring (represented by yeast fermentation),

2. (ii) environmental monitoring (illustrated by analysis of contaminated ground water samples) and

3. (iii) on-line chemical reaction monitoring (exemplified by the photolysis of aryl esters).

The applicability of MIMS to the analysis of environmental samples, including complex mixtures in water, air and soil, is noted.     

A semi‐quantitative approach for the rapid screening and mass profiling of naphthenic acids directly in contaminated aqueous samples

We report the use of a direct sampling, online analytical approach for the determination of acid extractable naphthenic acids in complex aqueous samples, known as condensed phase membrane introduction mass spectrometry (CP‐MIMS). The technique employs a capillary hollow fibre semi‐permeable membrane probe configured for immersion into a pH adjusted sample. A continuously flowing methanol acceptor phase transfers naphthenic acids to an electrospray ionization source, operated in negative ion mode, whereupon they are analysed by mass spectrometry as [M–H]^? ions. High‐resolution mass spectrometry is used to characterize the influence of sample pH on membrane transport of multiple components of complex naphthenic acid mixtures. We demonstrate the use of CP‐MIMS for semi‐quantitative analysis of real‐world samples using selected ion monitoring and full scan mass spectra at unit mass resolution. The technique has also been employed to continuously monitor the temporal evolution in the mass profile and concentrations of individual naphthenic acid isomer classes in heterogeneous solutions during adsorption processes. Copyright © 2015 John Wiley & Sons, Ltd.     

On-line monitoring of continuous beer fermentation process using automatic membrane inlet mass spectrometric system

A fully automatic membrane inlet mass spectrometric (MIMS) on-line instrumentation for the analysis of aroma compounds in continuous beer fermentation processes was constructed and tested. The instrumentation includes automatic filtration of the sample stream, flushing of all tubing between samples and pH control. The calibration standards can be measured periodically. The instrumentation has also an extra sample line that can be used for off-line sample collection or it can be connected to another on-line method. Detection limits for ethanol, acetic acid and eight organic beer aroma compounds were from μg l⁻¹ to low mg l⁻¹ levels and the standard deviations were less than 3.4%. The method has a good repeatability and linearity in the measurement range. Response times are shorter than or equal to 3 min for all compounds except for ethyl caproate, which has a response time of 8 min. In beer aroma compound analysis a good agreement between MIMS and static headspace gas chromatographic (HSGC) measurements was found. The effects of different matrix compounds commonly present in the fermentation media on the MIMS response to acetaldehyde, ethyl acetate and ethanol were studied. Addition of yeast did not have any effect on the MIMS response of ethanol or ethyl acetate. Sugars, glucose and xylose, increased the MIMS response of all studied analytes only slightly, whereas salts, ammonium chloride, ammonium nitrate and sodium chloride, increased the MIMS response of all three studied compounds prominently. The system was used for on-line monitoring of continuous beer fermentation with immobilised yeast. The results show that with MIMS it is possible to monitor the changes in the continuous process as well as delays in the two-phase process.     

Membrane Introduction Mass Spectrometry (MIMS): A Versatile Tool for Direct,Real‐Time Chemical Measurements

Membrane introduction mass spectrometry (MIMS) is a direct, continuous, on‐line measurement technique. It utilizes a membrane to semi‐selectively transfer analyte mixtures from a sample to a mass spectrometer, rejecting the bulk of the sample matrix, which can be a gas, liquid or solid/slurry. Analyte selectivity and sensitivity are affected by optimizations at the membrane, ionization and the mass spectrometer levels. MIMS can be roughly classified by the acceptor phase that entrains analyte(s) to the mass spectrometer after membrane transport, either a gaseous acceptor phase (GP‐MIMS) or condensed acceptor phase (CP‐MIMS). The aim of this article is to provide an introduction to MIMS as a technique and to explore current variants, recent developments and modern applications, emphasizing examples from our group, the Applied Environmental Research Laboratories as well as selected work from others in this emerging area. Also provided is a synopsis of current and future directions for this versatile analytical technique. Copyright © 2014 John Wiley & Sons, Ltd.     

Biodegradation studies of 4-fluorobenzoic acid and 4-fluorocinnamic acid: an evaluation of membrane inlet mass spectrometry as an alternative to high performance liquid chromatography and ion chromatography

The biodegradation of 4-fluorobenzoic acid (4-FBA) and 4-fluorocinnamic acid (4-FCA) has been monitored by membrane inlet mass spectrometry (MIMS) using a hollow-fibre silicone membrane. A novel in-membrane pre-concentration/thermal desorption (IMP-MIMS) technique was employed for MIMS analysis using an oven temperature profile that allowed semi-volatile organic compounds to be accumulated in the membrane and then released by rapid heating. Air drying of the membrane between the analyte pre-concentration and thermal desorption stages improved mass spectrometric performance by removing residual water from the membrane. The concentrations of 4-FBA and 4-FCA determined by MIMS compare well with data obtained by high performance liquid chromatography (HPLC). Stoichiometric amounts of fluoride were monitored using ion chromatography (IC). Intermediates in the biodegradation pathway were identified by liquid chromatography/mass spectrometry (LC/MS). These data establish the potential of MIMS as an alternative to chromatographic methods for monitoring the biodegradation of semi-volatile organic compounds.     

On-line monitoring of biotechnological processes by gas chromatographic-mass spectrometric analysis of fermentation suspensions

An on-line monitoring system has been developed for the control of a biorreactor for the anaerobic pretreatment of an industrial waste water. The monitoring system is based on a process mass spectrometer with a temperature controlled membrane inlet. The membrane introduction mass spectrometer (MIMS) is coupled with a resistively heated metal gas chromatography capillary column, which serves as a transfer line between the bioreactor and the MIMS. Sampling and injection is performed by means of a pneumatically driven membrane probe, which enables monitoring of soluted and gaseous substances in the fermentation broth. The system can also be coupled to other processes.     

Automated Solid-Phase Extraction and Liquid Chromatography Tandem Mass Spectrometry Determination of N-methyl-2-pyrrolidone and its Metabolites in Urine and Plasma

A method for the simultaneous determination of N-methyl-2-pyrrolidone (NMP) and its metabolites 5-hydroxyl-N-pyrrolidone (5HNMP), N-methylsuccinimide (MSI) and 2-hydroxy-N-methylsuccinimide (2HMSI) in plasma and urine has been developed. Samples were purified by SPE using an ASPEC XL4. Analysis was performed using LC–MS equipped with an APCI interface. The analysis provided linear responses in the range of 0.125–12 μg mL⁻¹ for all of the analytes and up to 150 μg mL⁻¹ for 5HNMP and 2HMSI. The within day precision was in the range of 0.9–19.1% for plasma samples and 1.9–10.4% for urine samples whereas the between day precisions were 4.5–11.9% and 1.2–17.5%, respectively. The method was deemed to be suitable for monitoring the levels of NMP and its metabolites in the plasma and urine of occupationally exposed persons.     

LC-DAD UV and LC-MS for the Analysis of Isoflavones and Flavones from In Vitro and In Vivo Biomass of Genista tinctoria L.

Conditions were determined for the separation of a complex set of isoflavones and flavones (free aglycones, monoglucosides, diglucosides and esters) by HPLC-DAD UV and MS detection. A good separation of all analytes was obtained and satisfactory peak shapes were achieved by gradient elution on an RP C18 column. The method was then applied to carry out qualitative and quantitative analysis of bioflavonoids present in the herb and in vitro cultures of Genista tinctoria L. Electrospray ionisation mass spectrometry in the negative mode was used to identify individual flavonoids in G. tinctoria plant material. Photodiode array detection was also utilised in flavonoid characterisation. All flavonoid compounds were then quantified using an internal standard. The method developed is useful for monitoring the process of flavonoid biosynthesis in biomass obtained through in vitro cultures.     

A coaxially heated membrane introduction mass spectrometry interface for the rapid and sensitive on-line measurement of volatile and semi-volatile organic contaminants in air and water at parts-per-trillion levels

A coaxially heated membrane introduction mass spectrometry (MIMS) sampling interface is presented that demonstrates improved on-line performance for the direct measurement of semi-volatile organic compounds (SVOCs) in air and water samples at parts-per-trillion levels. The device is based on a polydimethylsiloxane (PDMS) capillary hollow fibre membrane (HFM) in a pneumatically assisted "flow-over" configuration that is resistively heated on the membrane interior via a coaxial nichrome wire, establishing a thermal gradient counter to the analyte concentration gradient. This arrangement allows for continuous and/or pulsed heating modes, affording excellent sensitivity for the on-line measurement of SVOCs while retaining sensitivity for volatile organic compounds (VOCs). In addition, the signal response time for SVOCs is reduced substantially over conventional "flow-over" MIMS interfaces. Separation and quantitation of analytes are achieved using quadrupole ion trap tandem mass spectrometry.     

Determination of ultraviolet filter compounds in environmental water samples using membrane‐protected micro‐solid‐phase extraction

A method based on membrane‐protected micro‐solid‐phase extraction coupled with gas chromatography and mass spectrometry was developed for the determination of six ultraviolet filter compounds in various aqueous media. Multiwalled carbon nanotubes as the sorbent were encapsulated in a sealed polypropylene membrane packet and immersed in the sample to extract the analytes, and then dichloromethane was used for desorption purpose. The method was sensitive enough for quantitative analysis of the target analytes, with limits of quantification between 0.01 and 0.06 μg/L, and produced a linear response (R² > 0.991) over the calibration range (0.05–6 μg/L). The obtained reproducibility was practically suitable with relative standard deviation values of less than 14% in pure water (spiked at 0.20/μg L) and less than 15% in real samples. The optimized method was applied for the analysis of real water samples with varying matrix complexity: tap, river, and dam water; geothermal spa; and sewage treatment plant effluent. Various levels and patterns of contamination were observed in the examined samples, while the sample from spa was the most contaminated, regarding the target analytes. Matrix spikes and matrix spike replicates were also analyzed to validate the technique for analysis of real aqueous samples, and satisfactory results were achieved.     

Applications of Raman-based techniques to on-site and in-vivo analysis

Raman-based technologies have proved to be excellent tools for on-site and in-vivo analysis, due to the non-invasive nature of their detection, their capability of providing structure information, their high tolerance to aqueous samples, the ultra-sensitivity of surface-enhanced Raman scattering (SERS) and resonance Raman scattering (RRS), the high spatial resolution of tip-enhanced Raman scattering (TERS), and the ultrashort spectra-acquisition time for coherent anti-Stokes Raman scattering (CARS) and stimulated Raman scattering (SRS).In this review, we highlight the recent success of Raman-based technologies in various applications, including:

• (1) 

  on-site surface analysis and chemical-reaction monitoring;

• (2) 

  on-site identification of cultural objects, archeological studies and planetary science;

• (3) 

  in-vivo analysis of cells and microorganisms;

• (4) 

  in-vivo diagnosis inside human and animal bodies;

• (5) 

  in-vivo fast Raman imaging and mapping;

• (6) 

  the study of SERS processes; and,

• (7) 

  assessment of nanomaterial safety.

     

Simultaneous Determination of Isoniazid and Acetylisoniazid in Human Plasma by Hydrophilic Interaction Liquid Chromatography Tandem Mass Spectrometry and Its Application to a Pharmacokinetics Related to N-Acetyltransferase2 Genetic Polymorphism

A rapid, sensitive, and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with hydrophilic interaction chromatography has been developed and validated for the simultaneous determination of isoniazid and acetylisoniazidin human plasma. Following precipitation of the protein, the analytes were extracted from human plasma, with high extraction recovery (>70%) for both Isoniazid and acetylisoniazid. The analytes were then separated using a hydrophilic interaction chromatography (HILIC) column and detected by electrospray ionization (ESI) mass spectrometry performed with a triple-quadrupole mass spectrometry. The quantification of the analytes was realized by low-energy collision dissociation tandem mass spectrometry (CID-MS/MS) using the multiple reaction monitoring (MRM) mode at m/z of 138.1→121.1 for isoniazid and m/z 180.1 → 138.1 for acetylisoniazid, respectively. The method was linear over the concentration range of 5–50,000 ng/mL for both. The intra-day and inter-day relative standard deviations (RSD) were less than 15% and the relative errors (RE) were all within 15%. The validated method has been successfully used to analyze human plasma samples for application in pharmacokinetic studies of isoniazid related to NAT2 genetic polymorphism in healthy Chinese subjects. The results showed that there were significant differences in the pharmacokinetic parameters of isoniazid and acetylisoniazid between subjects with and without mutations in the NAT2 gene.     

Simple and rapid determination of phthalates using microextraction by packed sorbent and gas chromatography with mass spectrometry quantification in cold drink and cosmetic samples

A simple and rapid method using microextraction by packed sorbent coupled with gas chromatography and mass spectrometry has been developed for the analysis of five phthalates, namely, diethyl phthalate, benzyl‐n‐butyl phthalate, dicyclohexyl phthalate, di‐n‐butyl phthalate, and di‐n‐propyl phthalate, in cold drink and cosmetic samples. The various parameters that influence the microextraction by packed sorbent performance such as extraction cycle (extract–discard), type and amount of solvent, washing solvent, and pH have been studied. The optimal conditions of microextraction using C₁₈ as the packed sorbent were 15 extraction cycles with water as washing solvent and 3 × 10 μL of ethyl acetate as the eluting solvent. Chromatographic separation was also optimized for injection temperature, flow rate, ion source, interface temperature, column temperature gradient and mass spectrometry was evaluated using the scan and selected ion monitoring data acquisition mode. Satisfactory results were obtained in terms of linearity with R² >0.9992 within the established concentration range. The limit of detection was 0.003–0.015 ng/mL, and the limit of quantification was 0.009–0.049 ng/mL. The recoveries were in the range of 92.35–98.90% for cold drink, 88.23–169.20% for perfume, and 88.90–184.40% for cream. Analysis by microextraction by packed sorbent promises to be a rapid method for the determination of these phthalates in cold drink and cosmetic samples, reducing the amount of sample, solvent, time and cost.     

A high-throughput liquid chromatography-tandem mass spectrometry method for simultaneous determination of direct oral anticoagulants in human plasma

Direct oral anticoagulants are widely used in many indications to prevent thromboembolic events. Routine therapeutic monitoring is not required; however, there is increasing evidence suggesting the benefit of plasma level measurement in some situations. In addition, laboratory monitoring might help improve patient and drug non-compliance and thus individualize therapy. In the present study, we developed a sensitive and high throughput ultra-high-performance liquid chromatography-tandem mass spectrometry method for simultaneous quantification of apixaban, dabigatran, edoxaban, and rivaroxaban in human plasma. A one-step extraction procedure in 96-well formate for phospholipid and protein removal was used for sample pre-treatment, and analytes were separated using gradient elution over 4.2 min. Analytes were detected on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring mode. The method was validated according to the European Medicine Agency guideline for the selectivity, linearity, and lower limit of detection, precision and accuracy, matrix effects, extraction recovery, carryover, dilution integrity, and stability over a concentration range of 3.0–1000 ng/ml for all analytes. The validated method was applied to real clinical samples of patients treated with one of the drugs. Therefore, we can conclude that our method is suitable for therapeutic drug monitoring of direct oral anticoagulants.     

Ionization suppression effects with condensed phase membrane introduction mass spectrometry: methods to increase the linear dynamic range and sensitivity

Condensed phase membrane introduction mass spectrometry (CP‐MIMS) is an online analytical method that allows for the direct, trace level measurement of a wide range of analytes in complex samples. The technique employs a semi‐permeable membrane that transfers analytes from a sample into a flowing acceptor solvent, which is directly infused to an atmospheric pressure ionization source, such as electrospray or atmospheric pressure chemical ionization. While CP‐MIMS and variants of the technique have been in the literature for nearly a decade, much of the work has focused on instrument development. Few studies have thoroughly addressed quantitative methods related to detection limits, ionization suppression, or linear dynamic range. We examine ionization suppression in the direct rapid quantitation of analytes by CP‐MIMS and introduce several analytical strategies to mitigate these effects, including the novel implementation of a continuously infused internal standard in the acceptor phase solvent, and modulation of acceptor phase flow rate. Several representative analytes were used to evaluate this approach with spiked, complex sample matrices, including primary wastewater effluent and artificial urine. Also reported are improved measured detection limits in the low part‐per‐trillion range, using a ‘stopped‐flow’ acceptor mode. Copyright © 2015 John Wiley & Sons, Ltd.     

Derivatization of (5R)-hydroxytriptolide from benzylamine to enhance mass spectrometric detection: application to a Phase I pharmacokinetic study in humans

(5R)-Hydroxytriptolide, a semisynthetic structural analog of triptolide, exhibits anti-inflammatory and immunosuppressive effect both in vitro and in vivo. The compound is currently undergoing Phase I clinical trials. This work describes the quantification of (5R)-hydroxytriptolide in human plasma based on chemical derivatization from benzylamine. Analysis through liquid chromatography–tandem mass spectrometry (LC–MS/MS) is performed for characterization. The primary reaction product between (5R)-hydroxytriptolide and benzylamine was identified as a 12,13-epoxide ring adduct. For quantification in plasma, (5R)-hydroxytriptolide and the internal standard (triptolide) were first extracted from diethyl ether–dichloromethane (3:2, v/v) and then converted to their benzylamine derivates at 80 °C for 1 h. The analytes are separated on a Gemini 5 μm 100 Å column, using a gradient elution program with a solvent consisting of 0.77 mM ammonium hydroxide (pH 10.0) and acetonitrile. An API 4000 tandem mass spectrometer operated in positive ion mode and equipped with an electrospray ionization source is used as detector. This method allows for a lower limit of quantification of 0.030 ng mL⁻¹. The validation results show accuracy (%RE < 11.7) and precision (%RSD < 8.6) at a broad linear dynamic range (0.030–100 ng mL⁻¹). The simple and quantitative derivatization coupled with tandem mass spectrometric analysis yields a sensitive and robust method for the quantification of (5R)-hydroxytriptolide in Phase I pharmacokinetic studies.     

Styrene oxide DNA adducts: quantitative determination using <Superscript>31</Superscript>P monitoring

The reaction of styrene oxide, a potential carcinogen in humans, with DNA constituents has been used to develop an improved method for quantification of DNA adducts. To enable monitoring of DNA adducts caused by xenobiotics at physiological relevant levels, a robust, reliable and powerful method based on monitoring of phosphorus in nucleotides is described. An efficient enzymatic digestion step and a sample-preconcentration procedure are essential, and enable separation of alkylated nucleotides from the large excess of native nucleotides. The adducts are detected by means of the phosphorus signal measured at mass m/z=31 with an inductively-coupled-plasma mass spectrometer. Bis(4-nitrophenyl)phosphate (BNPP) serves as internal standard for quantification of the adducts. The absolute limit of detection, 45 fmol, corresponds to detection of three modified nucleotides among 10⁷ native nucleotides (the calculation is based on use of 50 g calf thymus DNA). An adduct formation ratio at the DNA of 3.6 adducts per 1000 nucleotides was measured, which is 75% lower than for reaction with monomeric 2-deoxy-nucleotides. In addition, a substantial amount of phosphate adducts were detected, but in DNA the rate of phosphate formation was lower than with monomeric nucleotides. Most probably these adducts escaped unnoticed when ³¹P-post-labelling was employed.     

Environmental applications of membrane introduction mass spectrometry

The purpose of this review is to highlight the versatility of membrane introduction mass spectrometry (MIMS) in environmental applications, summarize the measurements of environmental volatile organic compounds (VOCs) accomplished using MIMS, present developments in the detection of semi-volatile organic compounds (SVOCs) and forecast possible future directions of MIMS in environmental applications.