Analytica Chimica Acta (v.657, #1)

Post-column photochemical reaction systems have developed into a common approach for enhancing conventional methods of detection in HPLC. Photochemical reactions as a means of ‘derivatization’ have a significant number of advantages over chemical reaction-based methods, and a significant effort has been demonstrated to develop an efficient photochemical reactor. When coupled to electrochemical (EC) detection, the technique allows for the sensitive and selective determination of a variety of compounds (e.g., organic nitro explosives, beta-lactam antibiotics, sulfur-containing antibiotics, pesticides and insecticides). This review will focus on developments and methods using post-column photochemical reaction systems followed by EC detection in liquid chromatography. Papers are presented in chronological order to emphasize the evolution of the approach and continued importance of the application.
Keywords: Post-column; Photochemical reaction systems; Photolysis; Electrochemical detection; Photo-assisted electrochemical detection; Liquid chromatography;

Recent analytical approaches in quality control of traditional Chinese medicines—A review by Yong Jiang; Bruno David; Pengfei Tu; Yves Barbin (9-18).
Traditional Chinese medicines (TCMs) are gaining more and more attention all over the world, due to their specific theory and long historical clinical practice. But the uncontrollable quality is a bottleneck for its modernization and globalization. This paper reviewed the recent analytical methods in the quality control of TCMs, including screening strategies of bioactive markers from TCMs through biochromatographic methods, the traditional chromatographic methods, DNA methods, as well as the spectroscopic methods, including FT-IR, NIR and NMR. The comprehensive methods, such as fingerprint and multi-component quantification are emphasized; hyphenated techniques, like HPLC-MS, GC-MS, CE-MS, LC-NMR, chemometric methods, and combination of chemical and biological methods, such as biofingerprint, metabolic fingerprint are now more and more widely used in TCMs. In a few word, the analysis and quality control of TCMs are moving towards an integrative and comprehensive direction, in order to better address the inherent holistic nature of TCMs.
Keywords: Traditional Chinese medicines; Quality control; Chromatographic method; Spectroscopic method; Bioactive marker; Hyphenated technology; Fingerprint;

Multivariate curve resolution alternating least squares (MCR-ALS) is shown to be a powerful chemometric method for the analysis of environmental monitoring data sets. It allows for the investigation, resolution, identification, and description of pollution patterns distributed over a particular geographical area, time and environmental compartment. An integrated interpretation of the main features characterizing pollution patterns of organic contaminants affecting the Ebro River basin (Catalonia, NE Spain) is attempted using the results obtained by MCR-ALS analysis of surface water, groundwater, sediment and soil data sets obtained in a 3-year extensive monitoring study. Agricultural practices were identified as the main source of surface and groundwater diffuse pollution, while sediments and soils appeared mostly polluted by a contamination pattern mainly loaded by polycyclic aromatic hydrocarbons (PAHs) of possible pyrolitic origin. Additionally, a third pollution pattern related to past and ongoing industrial activities was detected to be principally stored in the sediment compartment. Geographical and temporal distributions of these pollution sources are given.
Keywords: Chemometrics; Multivariate curve resolution alternating least squares; Organic pollution; Water; Sediment; Soil;

A simple analytical procedure for the simultaneous determination of eight endogenous steroids (testosterone, androstenedione, 17β-estradiol, estrone, pregnenolone, progesterone, dihydroandrostenedione, and dihydrotestosterone) in aquatic molluscs by solid-phase extraction (SPE) and gas chromatography/mass spectrometry (GC–MS) has been developed. After a microwave-assisted extraction, samples were further extracted and purified using two successive SPE (EnviChrom-P and NH2) cartridges. Steroids were derivatized with a mixture of N-methyl-N(trimethylsilyl)trifluoroacetamide (MSTFA)/mercaptoethanol/ammonium iodide (NH4I) and determined by GC–MS in selective ion monitoring mode. Recoveries were in the range 85–114%, although slightly lower for dihydrotestosterone, and the repeatability of the procedure, expressed as the coefficient of variation, was lower than 16%. The limits of detection determined in digestive glands of mussels were in the range 0.1–0.4 ng g−1 wet weight for all the steroids. The developed procedure was then applied to the monitoring of steroid profiles in the digestive glands of mussels from the Arcachon Bay (France) during two reproductive cycles. In parallel, two physiological parameters (lipid content and the condition index of mussels) were also monitored, as well as the seawater temperature and salinity. Only progesterone and pregnenolone were detected in the digestive glands of mussels, and the seasonal variations of progesterone levels seemed to be related to the spawning periods of Mytilus sp. in the Bay. The current challenge for the determination of natural steroids in aquatic invertebrates is also briefly discussed.
Keywords: Steroids; Digestive gland; Mussels; Arcachon Bay; Solid-phase extraction; Gas chromatography/mass spectrometry;

Influence of filter ventilation on the chemical composition of cigarette mainstream smoke by Thomas Adam; John McAughey; Christoph Mocker; Conor McGrath; Ralf Zimmermann (36-44).
Total yields of cigarette smoke constituents are greatly influenced by smoking behaviour, the tobacco blend as well as a variety of cigarette design parameters. Thereby, filter ventilation, i.e. diluting the smoke by providing a zone of microscopic holes around the circumference of the filter is one method to reduce the yield of ‘tar’ and other smoke compounds. However, little is known how these design variations influence the combustion conditions, and therefore, the overall chemical pattern of the smoke. In this paper single photon ionization-time-of-flight mass spectrometry (SPI-TOFMS) is used to characterize and compare cigarettes on a puff-by-puff basis, which differ only in filter ventilation magnitude. The research cigarettes investigated were made from Virginia tobacco and featured filter ventilations of 0% (no ventilation), 35%, and 70%. The cigarettes were smoked under two different puffing regimes, one using the puffing parameters of the conventional International Organization for Standardization (ISO) smoking regime and a more intense smoking condition. Results show that every variation entails a change of the chemical pattern, whereby, in general, cigarettes with 0% filter ventilation as well as the intense smoking regime lead to a more complete combustion compared to the ISO smoking conditions and the high ventilated cigarettes. Changes in the overall patterns can also be observed during the smoking for individual puffs. Some substances dominate the first puff, some species are more pronounced in the middle puffs, whereas others are preferably formed in the last puffs. This demonstrates the high complexity of the occurring processes. Results might help to understand the formation and decomposition reactions taking place when a cigarette is smoked and offer scope for targeted reduction strategies for specific toxicants or groups of toxicants in the smoke.
Keywords: Single photon ionization; Cigarette smoke; Filter ventilation; Smoking regime;

Simultaneous analysis of fourteen tertiary amine stimulants in human urine for doping control purposes by liquid chromatography–tandem mass spectrometry and gas chromatography–mass spectrometry by Jianghai Lu; San Wang; Ying Dong; Xiaobing Wang; Shuming Yang; Jianli Zhang; Jing Deng; Yang Qin; Youxuan Xu; Moutian Wu; Gangfeng Ouyang (45-52).
A method for the simultaneous screening and confirmation of the presence of fourteen tertiary amine stimulants in human urine by gas chromatography–mass spectrometry (GC–MS) in combination with liquid chromatography–tandem mass spectrometry (LC–MS/MS) has been developed and validated. Solid phase extraction (SPE) and liquid–liquid extraction (LLE) approaches were utilized for the pre-treatment of the urine samples. The study indicated that the capillary temperature played a significant role in the signal abundances of the protonated molecules of cropropamide and crotethamide under positive ion electrospray ionization (ESI) conditions. In addition, comparison studies of two different pre-treatment approaches as well as the two ionization modes were conducted. The LODs of the developed method for all the analytes were lower than the minimum required performance limit (MRPL) as set forth in the World Anti-Doping Agency (WADA) technical document for laboratories. The human urine sample obtained after oral administration of prolintane·HCl was successfully analyzed by the developed method, which demonstrated the applicability and reliability of the method for routine doping control analysis.
Keywords: Tertiary stimulants; Doping analysis; Gas chromatography–mass spectrometry; Liquid chromatography–tandem mass spectrometry;

Rheostatic control of tryptic digestion in a microscale fluidic system by Andrew J. Percy; David C. Schriemer (53-59).
Integrated fluidic systems that unite bottom-up and top-down proteomic approaches have the potential to deliver complete protein characterization. To circumvent fraction collection, as is conducted in current blended approaches, a technique to regulate digestion efficiency in a flow-through system is required. The present study examined the concept of regulating tryptic digestion in an immobilized enzyme reactor (IMER), incorporating mixed solvent systems for digestion acceleration. Using ovalbumin, cytochrome c, and myoglobin as protein standards, we demonstrate that tryptic digestion can be efficiently regulated between complete digestion and no digestion extremes by oscillating between 45 and 0% acetonitrile in the fluid stream. Solvent composition was tuned using programmable solvent waveforms in a closed system consisting of the IMER, a sample delivery stream, a dual gradient pumping system and a mass spectrometer. Operation in this rheostatic digestion mode provides access to novel peptide mass maps (due to substrate unfolding hysteresis) as well as the intact protein, in a reproducible and stable fashion. Although cycle times were on the order of 90 s for testing purposes, we show that regulated digestion is sufficiently rapid to be limited by solvent switching efficiency and kinetics of substrate unfolding/folding. Thus, regulated digestion should be useful in blending bottom-up and top-down proteomics in a single closed fluidic system.
Keywords: Mass spectrometry; Tryptic digestion; Peptides; Microscale fluidics; Protein identification; Proteomics;

This paper describes the effects of oral administration of non-steroidal anti-inflammatory drugs on the endogenous and synthetic anabolic androgenic steroids urinary excretion as assessed by gas-chromatography mass-spectrometry. Experiments were carried out on 5 male subjects, with pathologies and/or diseases, treated with non-steroidal anti-inflammatory drugs. To set up the individual baseline variability of testosterone and its main metabolites, urine samples were collected for 3 days, every 2 h prior to the administration of the drug(s); whereas the study of the effects of a single dose of each drug, here considered, on the endogenous androgen steroid urinary concentrations, was assessed by collecting urine samples for 2 days, every 2 h. Data obtained after drugs administration were then evaluated taking into account the individual baseline variability. The results showed that, only in the case of propyphenazone administration, the relative urinary concentrations of some testosterone metabolites were significantly altered. More specifically, the urinary levels of dehydroepiandrosterone, 11keto-etiocholanolone, 11β-hydroxyandrosterone, 11β-hydroxyetiocholanolone, androsterone, etiocholanolone and some metabolite ratios decrease significantly, generally between 2 and 10 h after administration of the drug, whereas no effects were observed on urinary calculated concentrations of testosterone, epitestosterone, 5α-androstane-3α,17β-diol, 5β-androstane-3α,17β-diol and testosterone/epitestosterone ratio. The observed effects do not depend on alterations on pharmacokinetics (excretion/metabolism), but on steroid sample preparation steps (hydrolysis and derivatization) inhibition. More specifically the significant decrease of dehydroepiandrosterone and testosterone metabolites urinary levels was due to a reduced yield of the steroid derivatization step for the presence in urine of the main metabolites of propyphenazone, namely hydroxyl-propyphenazone metabolites.
Keywords: Androgenic anabolic steroids; Derivatization; NSAID; Gas-chromatography mass-spectrometry; Anti-doping analysis;

A simple, fast, low cost and sensitive direct β-correction spectrophotometric assay of cyanide ions based on its reaction with the reagent 4-hydroxy-3-(2-oxoindolin-3-ylideneamino)-2-thioxo-2H-1,3-thiazin-6(3H)-one, abbreviated as HOTT in aqueous media of pH 7–10 is described. The electronic spectrum of the produced brown-red colored species showed well defined and sharp peak at λ max  = 466 nm. The effective molar absorptivity for the produced cyano compound was 2.5 × 104  L mol−1  cm−1. Beer's law and Ringbom's plots were obeyed in the concentration range 0.05–2.0 and 0.30–1.5 μg mL−1 cyanide ions, respectively. The proposed method offers 16.0 and 50.3 μg L−1 lower limits of detection (LOD) and quantification (LOQ) of the cyanide ion, respectively. The analytical utility of the method for the analysis of cyanide ions in tap and drinking water samples was demonstrated and the results were compared successfully with the conventional cyanide ion selective electrode. The short time response and the detection by the naked eye make the method available for the detection and quantitative determination of cyanide in a variety of samples e.g. fresh and drinking water. Moreover, the structure of the produced colored species was determined with the aid of spectroscopic measurements (UV–Vis, IR, 1H and 13C NMR) and elemental analysis.
Keywords: Cyanide ions; Industrial wastewater; Determination; β-Correction spectrophotometry; Nucleophilic addition;

Feasibility of gymnodimine and 13-desmethyl C spirolide detection by fluorescence polarization using a receptor-based assay in shellfish matrixes by Eva S. Fonfría; Natalia Vilariño; Begoña Espiña; M. Carmen Louzao; Mercedes Álvarez; Jordi Molgó; Rómulo Aráoz; Luis M. Botana (75-82).
The detection of toxins in shellfish through reliable methods is essential for human health preservation and prevention of economic losses in the aquaculture industry. Although no human intoxication has been unequivocally linked to gymnodimines or spirolides, these phycotoxins are highly toxic by intraperitoneal injection causing false positives in lipophilic toxin detection by the mouse bioassay. Based on the detection of molecular interactions by fluorescence polarization an inhibition assay was developed using fluorescent α-bungarotoxin and nicotinic acetylcholine receptor-enriched membranes of Torpedo marmorata to detect gymnodimine and 13-desmethyl C spirolide. Both toxins, classified into the cyclic imine group, inhibit the interaction of α-bungarotoxin with Torpedo nicotinic acetylcholine receptors in the nM range. In this study we analyze the matrix effect of four shellfish species on the fluorescence polarization assay. Mussels, clams, cockles and scallops were extracted with acetone and sequentially partitioned with n-hexane and chloroform. The interference of these shellfish extracts with the α-bungarotoxin fluorescence or its binding to the nicotinic acetylcholine receptor was lower than 11%. The average recovery rates of gymnodimine and 13-desmethyl C spirolide using these solvents were 90.6 ± 7.8% and 89.6 ± 3.2%, respectively with variations among species. The quantification range of this fluorescence polarization assay for gymnodimine and 13-desmethyl C spirolide in all tested species was 80–2000 μg kg−1 and 85–700 μg kg−1 of shellfish meat, respectively. This assay format can be used to detect gymnodimine and 13-desmethyl C spirolide in shellfish as a screening assay.
Keywords: Cyclic imines; Gymnodimine; Spirolide; Fluorescence polarization; Nicotinic acetylcholine receptors; Shellfish;