Reduced graphene-decorated covalent organic framework as a novel coating for solid-phase microextraction of phthalate esters coupled to gas chromatography-mass spectrometry

Two analytical methods were developed using electrochemical and spectrometric techniques for the simultaneous determination of endocrine disruptors triclosan and methylparaben in the monitoring of personal care products. For the electroanalytical analyses, a sensitive electrode based on graphene quantum dots supported in chitosan was employed. Under optimized conditions and a working potential of typically + 0.60 V for triclosan and + 0.81 V (vs. Ag/AgCl) for methylparaben, the calibration plots obtained by differential pulse voltammetry were linear in the range 0.10 to 10.0 μmol L−1. The detection limits were 0.03 and 0.04 μmol L−1 for triclosan and methylparaben, respectively. For the spectrometric method, UV/VIS spectrometry was used with a mathematical processing of non-linear deconvolution. This processing was used to solve the problem of overlapping absorption bands of triclosan (282 nm) and methylparaben (257 nm), which enabled simultaneous determination. The calibration plots by UV/VIS spectrometry were linear in the range 1.0 to 14.0 μmol L−1 with detection limits of 0.42 and 0.37 μmol L−1, respectively, for triclosan and methylparaben. Similar results obtained from the calibration plots of individual analytes suggest that the methods can be applied for individual or simultaneous determination of these species. Both methods were employed in the analysis of five samples of personal care products: toothpaste, antiseptic soap, antiseptic deodorant, shampoo, and a bath kit (soap and shampoo). The statistical tests indicated that there were no significant differences regarding the accuracy and precision of the data provided by the two methods described herein.     

Liquid chromatography-mass spectrometry approach for the characterization and purification of crude synthetic peptide hormones

In this study, conditions for the optimal separation by LC-UV and characterisation by LC-ES-MS of crude mixtures generated during SPPS of several peptide hormones are compiled. The linear solvation energy relationship (LSER) methodology has been used to predict the retention of the target peptides and their side products and then to develop a separation LC methodology with applicability on both the analytical and preparative scale. Identification of these side products by LC-ES-MS analysis has been made on the basis of their calculated molecular masses. This method may be regarded as a key tool for the optimisation of the synthetic procedures and for complying with regulatory agencies' requirements before commercialisation of a safe and effective peptide-based pharmaceutical drug.Awarded a prize as outstanding poster on the occasion of Jornadas de Análisis Instrumental (JAI), November 26 to 29, 2002, Barcelona, Spain     

Liquid chromatography-mass spectrometry in occupational toxicology: a novel approach to the study of biotransformation of industrial chemicals

Biological monitoring and biomarkers are used in occupational toxicology for a more accurate risk assessment of occupationally exposed people. Appropriate and validated biomarkers of internal dose, like urinary metabolites, besides to be positively correlated with external exposure, have a predictive value to the risk of adverse effects. The application of liquid chromatography-mass spectrometry (LC-MS) in occupational and environmental toxicology, although relatively recent, has been demonstrated valid in the determination of traditional biomarkers of exposure, as well as in metabolism studies aimed at investigating minor metabolic routes and new more specific biomarkers. This review presents selected applications of LC-MS to the study of the metabolism of industrial chemicals, like n-hexane, benzene and other aromatic hydrocarbons, styrene and other monomers employed in plastic industry, as well as to other chemicals used in working environments, like pesticides used by farmers, and antineoplastic agents prepared by hospital personnel. Analytical and pre-analytical factors, which affect quantitative determination of urinary metabolites, i.e. sample preparation, matrix effect, ion suppression, use of internal standards, and calibration, are emphasized.     

Liquid chromatography-mass spectrometry in the study of the metabolism of drugs and other xenobiotics

The application of liquid chromatography-mass spectrometry (LC/MS) to the study of metabolism of drugs and other xenobiotics is reviewed. Original research papers covering the period from 1998 to early 2000 and concerning the use of LC/MS in the study of xenobiotic metabolism in humans and other mammalian species are reviewed. LC/MS interfaces, sample preparation steps, column types, mobile phases and additives, and the type of metabolites detected are summarized and discussed in an attempt to identify the current and future trends in the use of LC/MS for metabolism studies. Applications are listed according to the parent xenobiotic type and include substances used in therapeutics, drug candidates, compounds being evaluated in clinical trials, environmental pollutants, adulterants and naturally occurring substances.     

Photolytic covalent binding of indoleacrylic acid to DNA

E-beta-Indol-3-ylacrylic acid (IA), radiolabelled at the 2 position with 14C, and calf thymus DNA have been irradiated with UV light (lambda greater than 280 nm) in phosphate buffer (pH 7.0). Re-isolation of the DNA indicates covalent binding of IA at levels of up to 295 nmol IA/mg native DNA (0.097 IA/base). Binding is observed for both native and heat denatured DNA, but is more efficient with the latter. Quantum efficiencies of 2.60 X 10(-5) and 2.30 X 10(-4) mol IA bound to native DNA/mol photon absorbed have been measured at 308 and 266 nm, respectively. Studies with the four polyribonucleotides indicate a strong preference for binding to poly[U]. Photolysis of either untreated or enzymatically degraded labeled native DNA with 254 nm light leads to the reformation of IA, and a 2 + 2 photocycloadduct of IA and thymidine has been isolated and characterized and matched by HPLC to a DNA derived adduct. Equilibrium dialysis studies provide no evidence for preassociation of IA to DNA.     

Liquid chromatography-mass spectrometry identification of imidacloprid photolysis products

Photolysis of imidacloprid by near ultraviolet (UV) radiation was investigated using reversed phase liquid chromatography (RPLC) and liquid chromatography-electrospray ionization/mass spectrometry (LC-ESI/MS) operated in the positive ion mode. The decomposition of imidacloprid by near UV light is first order with a half life of 10.18 h. Photo-degradation products of imidacloprid identified in this study included imidacloprid urea, imidacloprid olefin, and imidacloprid desnitro. A degradation pathway is proposed for imidacloprid from an examination of the product decay curves obtained in this monitoring study which was performed in a batch test mode.     

Liquid chromatography-mass spectrometry of hydroxy and non-hydroxy fatty acids as amide derivatives.

A useful method for analyzing fatty acids by liquid chromatography-mass spectrometry with an atmospheric-pressure chemical-ionization interface system has been developed. The sensitivity of six kinds of palmitamide derivatives monitored by a single ion of [M+H]+ was, in decreasing order: N-n-propylamide greater than anilide greater than N,N-diethylamide, amide greater than N,N-diphenylamide greater than N-1-naphthylamide. Individual fatty acids were identified from a mixture of amide derivatives of authentic fatty acids from C16:0 to C30:0 on a mass chromatogram. This method was used to detect both hydroxy and non-hydroxy fatty acids. Many kinds of fatty acid, including hydroxy fatty acids of the rat brain, were detected in a single run.     

A general approach to calculating isotopic distributions for mass spectrometry

Fundamental principles for obtaining mass spectral isotopic distributions are applied to a general computer program that can be used to calculate and present in tabular and graphic form the isotopic contributions for any molecular formula. A unique feature is the retention of the isotopic distribution, exact mass, and absolute abundance for all individual peaks at each mass. Special considerations have been made for the large number of isotopic combinations that occur for many higher mass compounds. The computer program accepts the input of a molecular formula followed by interactive input of a number of parameters that affect the final presentation of the theoretical distribution profile.     

On-line solid-phase extraction coupled to hydrophilic interaction chromatography-mass spectrometry for the determination of polar drugs

The present study describes the first fully automated method based on on-line solid-phase extraction (SPE) coupled to hydrophilic interaction chromatography-electrospray-mass spectrometry (HILIC-(ESI)MS) to determine a group of polar drugs that includes illicit drugs (such as cocaine, morphine, codeine and metabolites) and pharmaceuticals in environmental water samples. The SPE was performed using a highly retentive polymeric sorbent. The HILIC separation was optimised and the initial high organic content of the chromatographic mobile phase, was also suitable for the proper on-line elution of the analytes retained in the SPE column and for enhancing the ESI ionisation efficiency. This method allows the loading of samples of up to 250ml of ultrapure water or 10ml of environmental water samples spiked at low ngl(-1) levels of the analytes. The method yields near 100% recoveries for all the analytes. The method was also validated with environmental water samples with linear ranges from 5 to 1000ngl(-1) and limits of detection ≤2ngl(-1) for most of the compounds.     

Liquid chromatography-mass spectrometry method for the determination of venlafaxine in human plasma and application to a pharmacokinetic study

A sensitive and selective liquid chromatographic-mass spectrometric (LC-MS) method for the determination of venlafaxine in human plasma has been developed. Samples were prepared using liquid-liquid extraction and analyzed on a C(18) column interfaced with a triple quadrupole mass spectrometer. Positive electrospray ionization was employed as the ionization source. The mobile phase was methanol-water containing 10 mmol/L ammonium acetate, pH 7.9 adjusted with aqueous ammonia (80:20, v/v) at the flow rate of 1.0 mL/min. The analyte and internal standard clozapine were both detected by use of selected ion monitoring mode. The method was linear in the concentration range of 1.0-200.0 ng/mL. The lower limit of quantification (LLOQ) was 1.0 ng/mL. The intra- and inter-day relative standard deviation across three validation runs over the entire concentration range was less than 10.1%. The accuracy determined at three concentrations (5.0, 50.0 and 150.0 ng/mL for venlafaxine) was within +/-10.0% in terms of relative error (RE). The method was successfully applied for the evaluation of pharmacokinetic profiles of venlafaxine capsule in 20 healthy volunteers. The results show AUC, T(max), C(max) and T(1/2) between the testing formulation and reference formulation have no significant difference (p > 0.05). Relative bioavailability was 103.4 +/- 14.1%.     

Liquid chromatography-mass spectrometry and strategies for trace-level analysis of polar organic pollutants

Liquid chromatography–mass spectrometry using atmospheric pressure ionization (LC–API-MS) has drastically changed the analytical methods used to detect polar pollutants in water. The present status of application of this technique to organic water constituents is reviewed. The selection of the appropriate LC conditions, whether reversed-phase liquid chromatography, ion-pair chromatography, capillary electrophoresis or ion chromatography, and of the most sensitive ionization mode, electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI), depends upon the polarity and acidity of the analytes. Strongly acidic compounds such as aromatic sulfonates, sulfonated dyes, haloacetic acids, linear alkylbenzene sulfonates, aliphatic sulfonates and sulfates and complexing agents, weakly acidic compounds such as carboxylates and phenols, neutral compound classes, namely alkylphenol ethoxylates, alcohol ethoxylates and polycyclic aromatic hydrocarbons and the basic toxins, quaternary ammonium compounds and organometallic compounds are considered. The selection of the mass spectrometer depends upon the analytical task: triple-quadrupole mass spectrometers are highly suited for sensitive quantitation and for qualitative analyses, ion traps are especially suited for structure elucidation, whereas time-of-flight mass spectrometers and quadrupole time-of-flight mass spectrometers with their higher mass resolution are ideal for the determination of molecular formulas of unknown compounds and for screening purposes. While large steps have already been made, future efforts with respect to water analysis may be directed at fine-tuning the methodical arsenal for increased sensitivity and selectivity and to extend LC–MS application to transformation products.     

Liquid chromatography-mass spectrometry method for quantification of caspofungin in clinical plasma samples

Caspofungin [(CASPO) MK-0991] is the first broad-spectrum anti-fungal agent of the echinocandin class approved for clinical use. Measurement of CASPO levels in blood might help monitor therapy in patients who are critically ill, in particular, if high-dose regimens or combinations of CASPO with other anti-fungals are used. The objective of this study was to develop a fast method for the measurement of CASPO levels in clinical blood samples using liquid chromatography coupled to a triple-quadrupole mass spectrometer. Stock solutions were prepared in plasma to avoid CASPO adsorption to glass and plastic surfaces during processing. CASPO and the internal standard (IS) were extracted from 100 microl of plasma using acetonitrile protein precipitation. The supernatant was diluted and directly injected into an analytical column (C8; 2.1 x 30 mm). The total run time was 15 min. CASPO was ionized by electrospray in the positive mode. CASPO and IS [M + 2H]2+ parent ions (m/z 547.3 and 547.8, respectively) and specific product ions (m/z 137.1 and 62.2, respectively) were used for the ion transitions. No carry over or cross-talk was observed on the column. The mean method recovery was 90 +/- 3%. Neither blood from different individuals (n = 6) nor the presence of concomitant drugs (n = 33) in plasma samples interfered with CASPO quantification. Quantification over time of the CASPO levels in plasma and whole blood was investigated at different pre-analysis storage conditions. The calibration curve included the clinically relevant CASPO concentration range from 0.04 to 20 microg/ml. Mean intra- and inter-day accuracy was 96.1 +/- 2.2% and 102.5 +/- 2.4%, respectively. Mean intra- and inter-day precision was 7.9 +/- 3.2% and 6.3 +/- 1.8%, respectively. This simple and robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method may easily be implemented for monitoring CASPO therapy.     

Liquid chromatography-mass spectrometry for simultaneous analyses of iminodipeptides containing an N-terminal or a C-terminal proline

Simultaneous analyses of synthetic iminodipeptides containing an N-terminal proline or a C-terminal proline have been demonstrated using liquid chromatography-mass spectrometry with an atmospheric pressure ionization interface system. The separation of iminodipeptides was carried out on a reversed-phase high-performance liquid chromatographic column using 0.1% aqueous trifluoroacetic acid-methanol (75:25, v/v, pH 2.0) as mobile phase. Very intense protonated molecular ions [M + H]+ of various synthetic iminodipeptides, Pro-Gly, Gly-Pro, Pro-Ala, Ala-Pro, Pro-Val, Val-Pro, Pro-Leu and Leu-Pro, were observed. Pro-Gly (Pro-X) and Gly-Pro (X-Pro) have the same protonated molecular ion (m/z 173), but the peaks of these compounds on the mass chromatograms were clearly distinguished by the differences of the retention times and mass spectra. The synthetic iminodipeptides containing an N-terminal proline added to urine samples from a patient with prolidase deficiency were also distinguished from iminodipeptides containing a C-terminal proline in urine samples from a patient with prolidase deficiency by scanning the [M + H]+ ion of each iminodipeptide. We established the method to measure simultaneously the various iminodipeptides containing an N-terminal or a C-terminal proline in biological samples.     

Liquid chromatography-mass spectrometry of trace compounds with a moving-belt interface and multi-dimensional chromatography

A high-performance liquid chromatographic method with on-line mass spectrometric detection is described for the structural analysis of a number of synthetic impurities, present at trace levels in almitrine. To obtain mass spectra with various ionization methods and high-resolution mass measurements, a moving-belt liquid chromatograph-mass spectrometer interface is used. A two-column switching system allows the injection of large amounts of almitrine, from which the trace compounds are trapped on a second column, while discarding the major component. This permits the introduction of the impurities into the mass spectrometer by elution of the second column, without the risk of introducing too large an amount of the major compound into the mass spectrometer. The mass spectra thus obtained are of sufficient quality to permit a correct structural assignment of the impurities.     

Photosensitized reduction and DNA covalent binding of aziridinylquinones

Photolysis of anaerobic aqueous mixtures (at wavelength maxima above 600 nm and at pH 7.4) containing either aluminum phthalocyanine tetrasulfonate (AlPcS₄), chlorin e6 (CHLORIN), pheophorbide-a (PHEO) or a novel tetracationic phthalocyanine derivative (TETCHLORIN) in the presence of the quinones diaziquone (AZQ), carboquone (CARBOQ) or 2,5-dicloro-diaziridinyl-1,4-benzoquinone (AZDClQ) produces the corresponding semiquinones. Photolysis of these mixtures under the conditions stated above, but in the presence of DNA and at pH 5.5 produces quinone–DNA covalent adducts. Absorption bands seen in irradiated solutions suggest binding of these quinones to DNA through the open aziridine ring. In general, the quinone CARBOQ yielded the largest amounts of adducts photosensitized by the dyes studied here. No quinone–DNA adducts were detected if samples were irradiated at pH 7.4.Thus, both photoreduction of these quinones and an acidic environment are needed for these quinones to bind DNA. These results suggest a potential mode of therapy with special applications to hypoxic regions in solid tumors which are characterized by an acidic environment.     

Separation of cytokinin isomers with a partial filling-micellar electrokinetic chromatography-mass spectrometry approach

A new method based on partial filling-MEKC (PF-MEKC) directly coupled to ESI-MS was developed for the simultaneous separation and determination of 13 structurally similar cytokinins, including various geometric and positional isomers of cytokinins. On the basis of the resolution of the neighboring isomer peaks, different parameters (i.e., pH and concentration of buffer, surfactant concentrations, length of the injected micellar plug, organic modifier, and applied separation voltage) were optimized to achieve a satisfactory PF-MEKC separation. Under optimum conditions, the separation of 13 cytokinin standards was accomplished within 25 min. MS/MS with multiple reaction monitoring detection was carried out to obtain sufficient selectivity. PF-MEKC-MS/MS allowed for the direct identification and confirmation of the cytokinins present in banana (Musa spp.) pulp sample after extraction and purification. Finally, trans-zeatin riboside (ZR) and trans-zeatin (Z) were unambiguously identified in banana pulp. It is anticipated that the current PF-MEKC-MS method can be applied to analyze cytokinins in a wide range of biological samples.     

Liquid chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry determination of N-methylpyrrolidinone in riverine biofilms

A rugged procedure utilizing reversed-phase liquid chromatography with positive-ion electrospray ionization mass spectrometry (LC-MS) along with tandem MS is described for the quantification and confirmation of N-methylpyrrolidinone (NMP) in methanolic extracts of riverine biofilm. The LC-MS method provided a 100-fold improvement in detection limits (2 ng g(-1) with a repeatability of 80-95% based on triplicate analyses) compared to a conventional LC-UV detection procedure and was applicable to quantitative analysis of biofilm samples with little or no clean up. Under low-energy collision induced dissociation (CID) conditions (17 V, laboratory frame of reference, with argon as the collision gas), two product-ions of the [M+H]+ ion were formed at m/z 69 [MH-CH3NH2]+ and m/z 58 [MH-CH3NCH]+ with relative abundances of 30% and 5%, respectively. These CID transitions were used to demonstrate that biofilm uptake of a photocatalytically-generated mixture of NMP was rapid once acclimation was achieved.     

Determination of acidic drugs in sewage water by gas chromatography-mass spectrometry as tert.-butyldimethylsilyl derivatives

A procedure is described for the determination of five acidic non-steroidal anti-inflammatory pharmaceuticals (ibuprofen, naproxen, ketoprofen, tolfenamic acid and diclofenac) in sewage water. The analytical method involves the concentration of water samples using a solid-phase extraction polymeric sorbent, functionalized with N-vinylpyrrolidone. Analytes were eluted with ethyl acetate. derivatized using N-methyl-N-(tert.-butyldimethylsilyl)trifluoroacetamide (MTBSTFA) and analyzed by GC-MS. Influence of time, temperature and volume of MTBSTFA in the yield of the derivatization step were studied in detail using a factorial central composite design. Quantification limits of the analytical procedure for 500 ml of sewage water ranged from 20 to 50 ng/l. Recoveries from 90 to 115% were found for sewage water samples spiked with the studied compounds at the low ng/ml level. Results obtained for real samples show the presence of ibuprofen and naproxen in both influent and effluent of a sewage water treatment plant.     

Liquid chromatography-mass spectrometry and related techniques for purity assessment in early drug discovery

The combination of HPLC and MS has become the most valuable analytical tool to determine the identity and purity of a drug substance in the drug discovery arena over the past decade. This article describes different LC/MS configurations and their broad applicability to meet the fundamental analytical requirements involved in discovering new drugs. In addition, the value of chemiluminescence nitrogen detection for absolute purity determination and the convenience of CE as an orthogonal separation technique to HPLC are also discussed. In summary, LC/MS-based methodologies that implicate automation, various levels of throughput and open access systems have proved to be an integral part of the drug discovery process. As a result, the paradigm of high-quality/-quantity information is fulfilled in a timely fashion.