Matrix matching in liquid chromatography–mass spectrometry with stable isotope labelled internal standards-Is it necessary?

Matrix matching is used in analysis to compensate for matrix effects that influence analytical response. It has been a widely discussed topic in electro-spray mass spectrometry where the ionization suppression is a major problem in accurate quantitative analysis. However, the unique strength of mass spectrometry to detect and quantify accurately a co-eluting stable isotope labelled internal standard offers an easy solution to the ionization suppression problem. Given the fact that it is impossible to match the matrix of the calibration standards with all samples, mass spectrometry allows accurate quantitation without the need for matrix matching, as long as the internal standard co-elutes with the analyte of interest. If the analyte and internal standard co-elute, the slope of the calibration curve analyte response/internal standard vs. analyte concentration is independent of the matrix composition, eliminating the need for matrix matching.     

Multidimensional Mass Spectrometry of Synthetic Polymers and Advanced Materials

Multidimensional mass spectrometry interfaces a suitable ionization technique and mass analysis (MS) with fragmentation by tandem mass spectrometry (MS²) and an orthogonal online separation method. Separation choices include liquid chromatography (LC) and ion‐mobility spectrometry (IMS), in which separation takes place pre‐ionization in the solution state or post‐ionization in the gas phase, respectively. The MS step provides elemental composition information, while MS² exploits differences in the bond stabilities of a polymer, yielding connectivity and sequence information. LC conditions can be tuned to separate by polarity, end‐group functionality, or hydrodynamic volume, whereas IMS adds selectivity by macromolecular shape and architecture. This Minireview discusses how selected combinations of the MS, MS², LC, and IMS dimensions can be applied, together with the appropriate ionization method, to determine the constituents, structures, end groups, sequences, and architectures of a wide variety of homo‐ and copolymeric materials, including multicomponent blends, supramolecular assemblies, novel hybrid materials, and large cross‐linked or nonionizable polymers.     

基质中加入少量盐类改善MALDI-TOF MS的谱图信噪比

自20世纪80年代发明基质辅助激光解吸电离(Matrix assisted laser desorption ionization,MALDI)质谱以来,该技术已在生物分子分析方面得到了广泛应用．作为一种离子化方法,MALDI具有灵敏度高,对样品要求低,能耐高浓度盐和缓冲剂等优点．测定过程中使用合适的基质不仅能提高测试灵敏度和分辨率,还能扩增测试样品的种类。     

Group IVB metallocene poly(ether ester) polymers containing alpha-cyano-4-hydroxycinnamic acid that act as self-matrix materials in MALDI MS

Polymers derived from reacting Group IVB metallocene dihalides and the matrix alpha-cyano-4-hydroxycinnamic acid act as their own matrix agent when performing MALDI MS. Ion fragments containing two repeat units and greater are formed. The results are similar to those obtained employing graphite as the matrix material. The advantage of employing graphite as a comparative standard is that non-interfering ion fragment clusters are not produced by graphite. This is the second report of the inclusion of alpha-cyano-4-hydroxycinnamic acid into a condensation polymer and the use of the polymer itself as a matrix material indicating that this approach can be successfully applied to other systems. The addition of graphite as a matrix material allows the mass range to increase for the reflective mode.     

Applications of Tandem Mass Spectrometry (MS/MS) in Protein Analysis for Biomedical Research

Mass Spectrometry (MS) allows the analysis of proteins and peptides through a variety of methods, such as Electrospray Ionization-Mass Spectrometry (ESI-MS) or Matrix-Assisted Laser Desorption Ionization-Mass Spectrometry (MALDI-MS). These methods allow identification of the mass of a protein or a peptide as intact molecules or the identification of a protein through peptide-mass fingerprinting generated upon enzymatic digestion. Tandem mass spectrometry (MS/MS) allows the fragmentation of proteins and peptides to determine the amino acid sequence of proteins (top-down and middle-down proteomics) and peptides (bottom-up proteomics). Furthermore, tandem mass spectrometry also allows the identification of post-translational modifications (PTMs) of proteins and peptides. Here, we discuss the application of MS/MS in biomedical research, indicating specific examples for the identification of proteins or peptides and their PTMs as relevant biomarkers for diagnostic and therapy.     

Gas chromatography and gas chromatography/mass spectrometry for the characterization of complex mixtures of large oligosaccharides

High temperature gas chromatography and gas chromatography/mass spectrometry are used in the characterization of complex natural mixtures of permethylated oligosaccharides released from proteins or lipids. The high resolution allows separation of isomeric compounds and the mass range extends to oligosac-charides around molecular mass 2000 daltons or 10 sugar residues for isomalto-oligosaccharides. The mass spectra of permethylated oligosaccharide alditols from mucin glycopeptides are very informative and the approach allows a simple and rapid characterization of these complex components.     

Tandem Mass Spectrometric Characterization of Fetuin Sialylated Glycopeptides Enriched by TiO2 Microcolumn

Sialylation of glycoproteins is vital for the function or physicochemical properties of a protein. It becomes more and more important to develop approaches that can be used to efficiently isolate and identify sialylated glycopeptides or glycoproteins for monitoring changes in glycoproteome. In the present study, we analyze intact structures of the enriched sialylated glycopeptides of bovine fetuin by matrix‐assisted laser desorption/ionization‐tandem mass spectrometry (MALDI‐MS/MS), without any chemical derivation. The experimental data show that the optimal loading buffer for TiO₂ as matrix is 80% acetonitrile/2% TFA (trifluoroacetic acid)/100 mg/mL DHB (2,5‐dihydroxybenzoic acid) which is also compatible with MALDI‐mass spectrometric analysis. This study indicates that the improved enrichment approach combined with MALDI‐MS/MS may be a powerful tool to analyze intact structures and components of the sialylated glycopeptides from complex peptide mixture.     

Tandem mass spectrometry of synthetic polymers

The detailed characterization of macromolecules plays an important role for synthetic chemists to define and specify the structure and properties of the successfully synthesized polymers. The search for new characterization techniques for polymers is essential for the continuation of the development of improved synthesis methods. The application of tandem mass spectrometry for the detailed characterization of synthetic polymers using the soft ionization techniques matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐MS) and electrospray ionization mass spectrometry (ESI‐MS), which became the basic tools in proteomics, has greatly been increased in recent years and is summarized in this perspective. Examples of a variety of homopolymers, such as poly(methyl methacrylate), poly(ethylene glycol), as well as copolymers, e.g. copolyesters, are given. The advanced mass spectrometric techniques described in this review will presumably become one of the basic tools in polymer chemistry in the near future. Copyright © 2009 John Wiley & Sons, Ltd.     

Sensitive inductively coupled plasma mass spectrometry assay for the determination of platinum originating from cisplatin, carboplatin, and oxaliplatin in human plasma ultrafiltrate

We present a highly sensitive, rapid method for the determination of platinum originating from the anticancer agents cisplatin, carboplatin, and oxaliplatin in human plasma ultrafiltrate. The method is based on the quantification of platinum by inductively coupled plasma mass spectrometry and allows quantification of 7.50 ng l-1 platinum in only 150 microl of matrix. Sample pretreatment involves dilution of samples with 1% HNO3. Validation fulfilled the most recent FDA guidelines for bioanalytical method validation. Validated ranges of quantification were 7.50 ng l-1 to 1.00x10(5) ng l-1 in plasma ultrafiltrate for all three platinum compounds. The assay is now successfully used to support pharmacokinetic studies in cancer patients treated with cisplatin, carboplatin, or oxaliplatin.     

Direct tissue analysis using matrix-assisted laser desorption/ionization mass spectrometry: practical aspects of sample preparation

Practical guidelines for the preparation of tissue sections for direct analysis by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry are presented. Techniques for proper sample handling including tissue storage, sectioning and mounting are described. Emphasis is placed on optimizing matrix parameters such as the type of matrix molecule used, matrix concentration, and solvent composition. Several different techniques for matrix application are illustrated. Optimal instrument parameters and the necessity for advanced data analysis approaches with regards to direct tissue analysis are also discussed.     

Sensitivity enhancement in tandem liquid chromatographic mass spectrometric assays by summation of two transition ion pairs – perspectives

The introduction of tandem liquid chromatographic mass spectrometric assays has boosted the sensitivity of the bioanalytical determination for the scores of analytes as compared to other conventional modes of detection [1–6]. Additionally, many intelligent variants such as improved ionization potential, suitable adduct formation, and/or identification of a precursor ion, switching of modality of detection (+ve to –ve and vice versa), and derivatization steps (to aid fragmentation and/or impart ionization property) within tandem mass spectrometric assays have further aided in improving the sensitivity. Some researchers have also expressed thoughts of using matrix effects in a beneficial manner so that there may be ion enhancement for improving the sensitivity of the assay.     

Characterization of polyethylenimine by electrospray ionization and matrix-assisted laser desorption/ionization

Branched polyethylenimines (PEIs) with lower average molecular weights (600, 1200 and 1800 Da) have been studied by Electrospray Ionization (ESI) and Matrix‐Assisted Laser Desorption/Ionization (MALDI) mass spectrometry. In both, ESI and MALDI mass spectra, the main distribution arises from protonated PEI oligomers with NH₂ end groups, [PEI + H]⁺, which are observed at m/z 43n + 18. A trace of sodium contamination in the PEI samples results in the presence of a series that appears at m/z 43n + 40 [PEI + Na]⁺. However, only the MALDI mass spectra show a [PEI + K]⁺ series at m/z 43n + 56, because of matrix contamination with potassium, and a series generated by condensation of the matrix with PEI at m/z 43n + 30. Collisionally activated dissociation tandem mass spectrometry (CAD (MS/MS)) of protonated PEI oligomers is shown to yield three fragment ion series b_n, and K_n. The experiments have demonstrated the capabilities of these mass spectrometry techniques, along with CAD MS/MS to detect and characterize such polar synthetic polymers. Copyright © 2011 John Wiley & Sons, Ltd.     

Recent Advances in MALDI Mass Spectrometry of Polymers

Matrix-Assisted Laser Desorption/Ionization (MALDI) allows the identification of repeat units and end groups, the structural analysis of linear and cyclic oligomers, and the estimate of composition and sequence for copolymers. MALDI has also been applied to the measurement of molar mass distributions in polymers and to the study of thermal and oxidative processes in polymers. This paper illustrates the detection of self-association in macromolecules made by coupling MALDI and Size Exclusion Chromatography (SEC), the investigation of polymer oxidation phenomena, and the characterization of copolymers formed in the processing of reactive polymer blends.     

Structure-Property Relationships in Rubber-Modified Styrenic Polymers

Styrenic polymers and copolymers are often impact modified with rubber particles. The efficiency of rubber toughening depends mainly on the size of the rubber particles and the degree of cross-linking. The deformation rate, the temperature, the orientation of the polymer molecules and the efficiency of rubber grafting also influence rubber toughening. It is thought that on impact, cavitation inside the rubber particles occurs which reduces the detrimental dilatational stress in the bulk polymer without forming cracks in the brittle matrix or at the rubber-matrix interface. Crazing and shearing are facilitated if the rubber particles can easily cavitate. This can be achieved by either avoiding too much cross-linking or by adding oil (silicone oil in the case of ABS) into the rubber particles, which acts as nuclei for void formation. An electron spectroscopic imaging method is described which allows visualizing the location of the oil. Already after cooling silicone oil modified ABS samples down to liquid nitrogen temperature rubber cavitation is observed. This cavitation is caused by the thermal stress developing due to the differences in thermal expansion coefficient between the rubber phase and the SAN-matrix and is facilitated by silicone oil. Voiding also leads to an increase of light scattering, which can be detected by an optical microscope using dark field illumination.     

LC–MS/MS analysis of coccidiostats in meat supply chain safety

The presence of coccidiostats in meat products represents an important topic because of the animal administration of these substances, authorized as feed additives for targeted species, in order to prevent and inhibit coccidiosis. Coccidiostats include both ionophores and synthetic molecules characterized by different chemical–physical properties such as polarity. Meat is a matrix characterized by many interfering compound groups, such as proteins, phospholipids, and fats. High-performance liquid chromatography (HPLC) coupled to mass spectrometry (MS) analysis allows the required selectivity and sensitivity for discriminating analytes and matrix interferences. For these reasons, an LC–MS/MS method for the analysis of coccidiostats in meat products was developed without SPE purification steps. The correct analyte quantification is allowed by matrix-matched calibration. The method validation was performed by the replicated analysis of spiked meat samples at two different concentration levels (limit of quantification—LOQ—and a 10 times LOQ) in order to evaluate method recovery and repeatability, plus spiked samples at higher concentrations up to 10,000 μg/kg. Moreover, the metrological approach was used for the calculation of method uncertainty. The application of the developed method to real samples evidenced the presence of some non-ionophores coccidiostats in the meat and liver of chicken and rabbit species. Although, the determined concentration was below the established MRLs, the monitoring of coccidiostats in the meat supply chain is confirmed as a good strategy in order to safeguard consumer health.     

Measurement of ceftazidime concentration in human plasma by ultra‐performance liquid chromatography–tandem mass spectrometry. Application to critically ill patients and patients with osteoarticular infections

Ceftazidime is an antibiotic belonging to the third generation of the cephalosporin family. It is indicated in the treatment of serious, simple or mixed bacterial infections, and its administration in continuous or intermittent infusion allows optimization of the concentration of antibiotic to keep it above the minimum inhibitory concentration. We developed and validated a chromatographic method by ultra‐performance liquid chromatography–tandem mass spectrometry to measure ceftazidime concentration in human plasma. Following extraction with acetonitrile and 1,2‐dichloroethane, the chromatographic separation was achieved using an Acquity ® UPLC ® BEH^TM (2.1 × 100 mm i.d., 1.7 µm) reverse‐phase C₁₈ column, with a water–acetonitrile linear gradient containing 0.1% formic acid at a 0.4 mL/min flow rate. Ceftazidime and its internal standard (cefotaxime) were detected by electrospray ionization mass spectrometry in positive ion multiple reaction monitoring mode using mass‐to‐charge transitions of 547.0 → 467.9/396.1 and 456.0 → 395.8/324.1, respectively. The limit of quantification was 0.58 mg/L and linearity was observed in the range 0.58–160 mg/L. Coefficients of variation and absolute relative biases were <9.8 and 8.4%. The mean recovery for ceftazidime was 74.4 ± 8.1%. Evaluation of the matrix effect showed ion enhancement, and no carry‐over was observed. The validated method could be applied to daily clinical laboratory practice to measure the concentration of ceftazidime in plasma. Copyright © 2015 John Wiley & Sons, Ltd.     

A validated method for the determination of traces of UV filters in fish using LC–MS/MS

An analytical method for the determination of UV filter substances in fish tissue has been developed and validated using benzophenone-3, 3-(4-methylbenzylidene)-camphor, 2-ethylhexyl-2-cyano-3,3-diphenyl-2-propenoate and 2-ethylhexyl 3-(methoxyphenyl)-2-propenoate as target analytes. The fish fillets were homogenised and extracted by Soxhlet extraction. The extracts were run through a clean-up process including gel permeation chromatography followed by solid-phase extraction. Quantification of the compounds was performed using liquid chromatography with tandem mass spectrometric detection. Blank fish as well as spiked blank fish were analysed to validate the analytical method. The analytical method developed has the multiple advantages of enabling separation, simultaneous identification and quantification of each of the four selected compounds in a single run. Contamination of blank samples and abnormally high concentrations in spiked samples were avoided by taking extensive precautions during the fish preparation procedure. The method was validated in accordance with internationally accepted criteria, such as specificity, accuracy and repeatability. The combination of LC with tandem mass spectrometry ensures a high level of specificity. The accuracy of the method was reported as the mean recovery rate for the analytes in the sample matrix. Mean recoveries were in the range 86–108%. The precision is expressed as the relative standard deviation, and in all but one of the cases was 20% or below. The accuracy of the method allows residue analyses to be performed on biological matrices at ng/g levels. The determined limit of quantification for each analyte was 8 ng/g fish. For all spiking levels ≥8 ng/g, relative standard deviations were ≤ 20%.     

Free mycophenolic acid determination in human plasma ultrafiltrate by a validated liquid chromatography–tandem mass spectrometry method

The aim of this study was to develop and validate fully the liquid chromatography–tandem mass spectrometry method for free mycophenolic acid (MPA) concentration measurements in plasma ultrafiltrate that will be reliable and simple in preparation with deuterated MPA (MPA‐d3) chosen as an internal standard. The chromatographic separation was made with Zorbax Eclipse XDB‐C18 column (4.6 × 150 mm) using a gradient of two solutions as a mobile phase: (A) water and (B) methanol, each containing 0.1% formic acid and 2.5 mm ammonium acetate. Satisfactory repeatability of retention times was achieved with average values of 7.54 ± 0.20 min and 7.50 ± 0.19 min for MPA and MPA‐d3, respectively. The method was selective, with no carry‐over or matrix effect observed. The analytical range was proven for MPA ultrafiltrate concentrations of 1–500 ng/mL. The accuracy and precision fell within the acceptance criteria for intraday (accuracy: 100.63–110.46%, imprecision: 6.23–7.76%), as well as interday assay (accuracy: 98.81–110.63%; imprecision: 5.36–10.22%). The method was used for free MPA determination in plasma samples from patients treated with mycophenolate mofetil. To the best of our knowledge this is the first liquid chromatography–tandem mass spectrometry method for free MPA monitoring using MPA‐d3 that allows to measure plasma ultrafiltrate concentrations as low as 1 ng/mL.     

Hybrid Materials Based on Conducting Polymers for Nitrite Sensing: A Mini Review

Well-known as a hazardous compound, nitrite constitute a real threat to the public health. So, there is a pressing need to detect and quantify them in different matrix. Even though conventional analytical methods can be used to address this issue, electrochemistry allows a fast, sensitive, and efficient analysis. Conducting polymers continue to raise great interest among scientific communities due to their properties. Moreover, their combination with carbon nanomaterials, or metallic nanoparticles improves their properties, and provides great results. In this paper, we will focus on some revealing works devoted to the electrochemical detection of nitrite using this kind of materials.     

Development,validation and comparison of four methods for quantifying endogenous 25OH‐D3 in human plasma

To meet the increasing clinical needs for 25‐hydroxyvitamin D3 (25OH‐D3) detection, the development of an efficient and accurate high‐performance liquid chromatography–mass spectrometry (HPLC–MS) method for plasma 25OH‐D3 quantitation is important. Since 25OH‐D3 is an endogenous compound, the lack of a plasma blank increases the difficulty of accurately quantifying 25OH‐D3. Selection of a method suitable for clinical monitoring among various methods for endogenous compound quantification is necessary. Methyl tert butyl ether was chosen for the sample treatment in a liquid–liquid extraction protocol. Water as a blank matrix, 5% human serum albumin in water as a blank matrix, surrogate analyte and background subtraction were designed to address the problem of a deficiency of a plasma blank. Four liquid chromatography–tandem mass spectrometry methods were fully validated to verify the advantages and limitations owing to regulatory deficiencies for endogenous compound validation. All four methods met the criteria and could be used to monitor clinical samples. Overall 30 human plasma samples were quantified in parallel using the four methods. The difference between any two methods was <12.6% and the total relative standard deviation was <5.2%. Background subtraction and 5% human serum albumin in water as a blank matrix may be better choices considering data quality, matrix similarity, cost and practicality.