Separations, Volume 7, Issue 1 (March 2020) – 19 articles

An ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method, in combination with a mixed matrix membrane microextraction method for the quantification of nonsteroidal anti-inflammatory drugs (NSAIDs) in environmental water samples, is reported. The extraction device was prepared by casting well-dispersed polymeric bonded octadecyl (C₁₈) particles in a cellulose triacetate matrix solution onto commercially available 200 μL micropipette tips. The membrane formed contains 25% of the adsorbent loading amount and was firmly attached to the inner wall of the membrane tip. The dynamic extraction was performed by withdrawing and dispensing the sample solution through the tip device for effective analyte adsorption, followed by the analyte desorption process into 40 μL of methanol and acetonitrile (1:1) prior to UPLC–MS/MS analysis. NSAIDs—namely diclofenac, ibuprofen, indoprofen, naproxen and sulindac—were chosen as targeted analytes. Several extraction parameters were comprehensively optimized, including sample pH value, ionic strength, dynamic extraction cycle, desorption solvent and desorption time. The optimized conditions demonstrated a linear range from 0.25 to 500 ng L⁻¹, with correlation coefficients (r²) from 0.9988 to 0.9992 and detection limits ranging from 0.08 to 0.40 ng L⁻¹. The recoveries of the spiked water samples were between 92% and 99% and exhibited excellent precision relative to standard deviations (RSDs ≤ 4.9%), and enrichment factors (EFs) were at 201–249 for the developed approach. Full article

In this study, a metabolomic approach was used to investigate the effect of seasonality on the chemical composition and yield of anti-inflammatory active principle, α-humulene, in the essential oil of three genotypes of Varronia curassavica Jacq. (Syn. Cordia verbenaceae). The essential oils were extracted by hydrodistillation and analyzed by comprehensive two-dimensional gas chromatography coupled to mass spectrometry (GC×GC-MS). The GC×GC approach a three-fold improvement in qualitative analysis (48 compounds were identified by GC-MS versus 135 by GC×GC-MS). The improved resolving power of GC×GC resolved important coelutions and enabled the detection of unusual substances in V. curassavica essential oil. The chromatographic data was analyzed by using peak table-based chemometrics, namely, principal component analysis (PCA) and hierarchical cluster analysis (HCA). The metabolic study showed that seasonality has a significant effect on the chemical composition. The α-humulene content was affected by genotype and season. Spring and summer were the best harvest seasons for the yield of the active ingredient, found in higher concentrations in the VC2 genotype. The proposed metabolomic workflow was successfully applied to terpene analysis found in V. curassavica essential oil, and such results have broadened our understanding of the influence of seasonal factors on the specialized metabolism of the species. Full article

In this research paper, we report a hyphenated technique based on ultra-high performance liquid chromatography–tandem mass spectrometry for the determination of twelve Perfluorinated Alkylated Substances in surface and groundwater samples from Lombardia Region during the monitoring activities in 2018 as new emerging and toxic pollutants. A green analytic method, developed by using an online Solid Phase Extraction coupled with UHPLC-MS/MS and previously validated, was applied for 4992 determinations conducted on 416 samples from 109 different sampling stations. Among the results, PFOS, PFOA, PFBA, PFBS, PFPeA and PFHxA were identified as the most abundant analytes detected. PFASs concentrations, in most cases, were below the limits of quantification and, in the cases where the limits of quantification have been exceeded, the values found were lower than Italy directive. PFOS is an exception and in fact this compound was detected in 76% of analyzed samples (surface and ground waters). Solid phase extraction with high performance liquid chromatography–tandem Mass Spectrometry has proved to be a very good Hyphenated techniques able to detect low concentrations of pollutants in surface and groundwater samples. Full article

In the present study, we report the results of our investigation of the oxidative forced degradation of glutathione in its nutraceutical formulations by two validated analytical methods. The first is based on the reaction of glutathione with o-phthalaldehyde through an automated zone fluidics flow platform and fluorimetric detection (λ_ex/λ_em = 340/425 nm). The second is based on the separation of glutathione and its oxidation product by a green reversed-phase HPLC method coupled to direct UV detection, at 210 nm. A solution of 3% w/v H₂O₂ provided fast oxidation of more than 95% of glutathione to yield oxidized glutathione in a time period of 180 min. The mechanism of the oxidation was proved to follow pseudo-first order kinetics. The k, t₉₀ and t_1/2 values were calculated. Full article

Pesticides and veterinary pharmaceuticals are used for effective crop production and prevention of livestock diseases; these chemicals are released into the environment via various pathways. Although the chemicals are typically present in trace amounts post-release, they could disturb aquatic ecosystems and public health through resistance development toward drugs or diseases, e.g., reproductive disorders. Thus, the residues of pesticides and veterinary pharmaceuticals in the environment must be managed and monitored. To that end, we developed a simultaneous analysis method for 41 target chemicals in environmental water samples using ultra-high-performance liquid chromatography (UHPLC)–quadrupole-orbitrap high-resolution mass spectrometry (HRMS) coupled with an on-line solid-phase extraction system. Calibration curves for determining linearity were constructed for 10–750 ng∙L⁻¹, and the coefficient of determination for each chemical exceeded 0.99. The method’s detection and quantitation limits were 0.32–1.72 ng∙L⁻¹ and 1.02–5.47 ng∙L⁻¹, respectively. The on-line solid-phase extraction system exhibited excellent method reproducibility and reduced experimental error. As the proposed method is applicable to the monitoring of pesticides and veterinary pharmaceuticals in surface water and groundwater samples acquired near agricultural areas, it allows for the management of chemicals released into the environment. Full article

Historically, one-dimensional gas chromatography combined with mass spectrometry (GC/MS) has been employed in the analysis of petrochemical samples such as diesel, crude oil and bitumen. With increasingly complex samples, obtaining detailed information can be difficult with this method due to the large number of coelutions. By implementing comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GC×GC-TOFMS), the limitations of GC/MS can be overcome, due to the ability of this method to separate mixtures using two different separation mechanisms and obtain full mass spectra. Furthermore, this enables an investigation of biomarkers, compounds which aid in the identification of geological and environmental processes, potentially differentiating crude oil samples. Cryogenic-based thermal modulators are typically used for this application due to their superior focusing effect; however, some platforms require expensive cryogenic consumables. The solid-state modulator (SSM), a cryogen-free thermal platform, was employed for the first time for the group and biomarker analysis of Alberta oil sands bitumen. Evaluation of the SSM performance was based on published literature data on bitumen analysis. Extracted ion chromatograms (EIC) and molecular ion peaks were used for the confirmation of the groups’ and individual’s analytes. Identification of the characteristic biomarkers responsible for determining thermal maturity, source rock or oil origin was achieved. These results indicate the successful analysis of bitumen by consumable-free, solid-state modulation-based GC×GC-TOFMS. Full article

Thirty years since the invention and public disclosure of solid phase microextraction (SPME), the technology continues evolving and inspiring several other green extraction technologies amenable for the collection of small molecules present in complex matrices. In this manuscript, we review the fundamental and operational aspects of a novel SPME geometry that can be used to “hunt” target molecules in complex matrices: the SPME Arrow. In addition, a series of applications in environmental, food, cannabis and forensic analysis are succinctly covered. Finally, special emphasis is placed on novel interfaces to analytical instrumentation, as well as recent developments in coating materials for the SPME Arrow. Full article

Presently, results from a study carried out in this area using the essential oil from the Calyptranthes concinna species, a representative from the Myrtaceae family, are reported. The essential oil was obtained by hydrodistillation and gas chromatography coupled to mass spectrometry was used to identify its chemical constituents. Antibacterial activity was determined using the broth microdilution method, thus obtaining the Minimal Inhibitory Concentration (MIC) value, from which the subinhibitory concentration (MIC/8) was derived. The C. concinna essential oil presented antibacterial activity against both standard and multiresistant bacteria. In addition, the oil demonstrated an antibiotic activity potentiation against Staphylococcus aureus and Escherichia coli when in combination with the antibiotic gentamicin, reducing the MIC from 141.38 μg/mL and 208.63 μg/mL to 64 μg/mL and 128 μg/mL, respectively. Conclusions: Findings from the present study suggest this oil is promising in terms of its antimicrobial activity. Full article

Crude tall oil (CTO) is the third largest by-product at kraft pulp and paper mills. Due the large presence of value-added fatty and resin acids, CTO has a huge valorization potential as a biobased, readily available, non-food, and low-cost biorefinery feedstock. The objective of this work was to present a method for the isolation of high-value linoleic acid (LA), an omega (ω)-6 essential fatty acid, from CTO using a combination of pretreatment, fractionation, and purification techniques. Following the distillation of CTO to separate the tall oil fatty acids (TOFAs) from CTO, LA was isolated and purified from TOFAs by urea complexation (UC) and low-temperature crystallization (LTC) in the temperature range between −7 and −15 °C. The crystallization yield of LA from CTO in that range was 7.8 w/w at 95.2% purity, with 3.8% w/w of ω-6 γ-linolenic acid (GLA) and 1.0% w/w of ω-3 α-linolenic (ALA) present as contaminants. This is the first report on the isolation of LA from CTO. The approach presented here can be applied to recover other valuable fatty acids. Furthermore, once the targeted fatty acid(s) are isolated, the rest of the TOFAs can be utilized for the production of biodiesel, biobased surfactants, or other valuable bioproducts. Full article

The present study was aimed to validate the phenolic, flavonoids, alkaloids and tannins profile of Aspleniuma ethiopicum (Burm. f.) Becherer methanolic extracts using HPTLC (High-performance thin-layer chromatography). The chromatographic separation was performed using the standard method. The separation methodology was optimized and phytoconstituents of A. aethiopicum methanolic extracts were separated. The methanolic extract of A. aethiopicum showed several bands identified as known phenolic compounds. The obtained flavonoids profiles were identified. HPTLC alkaloids profile of A. aethiopicum revealed 11 types of alkaloids. The tannin profile of methanolic extracts of A. aethiopicum demonstrated 11 different types of tannins. The observed profiles will be used as phytomarker to identify the chemical constituents of A. aethiopicum methanolic extracts. These profiles will act as a fingerprint of A. aethiopicum and help us to distinguish from its adulterants. The observed profile will help us to identify the crude drugs and improve the therapeutic potentials of A. aethiopicum. Full article

In vivo solid-phase microextraction (SPME) has been recently proposed for the extraction, clean-up and preconcentration of analytes of biological and clinical concern. Bioanalysis can be performed by sampling exo- or endogenous compounds directly in living organisms with minimum invasiveness. In this context, innovative and miniaturized devices characterized by both commercial and lab-made coatings for in vivo SPME tissue sampling have been proposed, thus assessing the feasibility of this technique for biomarker discovery, metabolomics studies or for evaluating the environmental conditions to which organisms can be exposed. Finally, the possibility of directly interfacing SPME to mass spectrometers represents a valuable tool for the rapid quali- and quantitative analysis of complex matrices. This review article provides a survey of in vivo SPME applications focusing on the extraction of tissues, cells and simple organisms. This survey will attempt to cover the state-of- the-art from 2014 up to 2019. Full article

Globally, the rapid decline in wildlife species has many causes. The illegal trafficking of fauna and flora is a major contributor to species decline and continues to grow at an alarming rate. To enable the prosecution of those involved in the trafficking of illegal wildlife, accurate and reliable identification is paramount. Traditionally, morphology and DNA amplification are used. This paper investigates a novel application of volatilome profiling using comprehensive two-dimensional gas chromatography coupled with time of flight mass spectrometry for wildlife sample detection. Known samples of elephant-derived ivory, other dentine samples, and bone (a common ivory substitute) were used as reference samples for volatilome profiling. Subsequently, specimens that were suspected ivory from border control seizures were obtained and analysed. Confirmatory DNA analyses were conducted on seized samples to establish the reliability parameters of volatilome profiling. The volatilome method correctly identified six of the eight seized samples as elephant ivory, which was confirmed through DNA analysis. There was also clear distinction of African elephant ivory parts from the bone and dentine samples from other species, as shown through PCA and discriminant analyses. These preliminary results establish volatilome profiling through GC×GC-TOFMS as a novel screening method used for the identification of unknown wildlife contraband. Full article

The demand for the recovery of valuable metals and the need to understand the impact of heavy metals in the environment on human and aquatic life has led to the development of new methods for the extraction, recovery, and analysis of metal ions. With special emphasis on environmentally friendly approaches, efforts have been made to consider strategies that minimize the use of organic solvents, apply micromethodology, limit waste, reduce costs, are safe, and utilize benign or reusable materials. This review discusses recent developments in liquid- and solid-phase extraction techniques. Liquid-based methods include advances in the application of aqueous two- and three-phase systems, liquid membranes, and cloud point extraction. Recent progress in exploiting new sorbent materials for solid-phase extraction (SPE), solid-phase microextraction (SPME), and bulk extractions will also be discussed. Full article

An analytical method based on high-resolution quadrupole–time-of-flight (QToF) mass spectrometry has been developed as an alternative to the classical method, using a low-resolution ion trap (IT) analyzer to reduce interferences in N-nitrosamines determination. Extraction of the targeted compounds was performed by solid-phase extraction (SPE) following the United States Environmental Protection Agency (USEPA) -521 method. First, both electron impact (EI) and positive chemical ionization (PCI) using methane as ionization gas were compared, along with IT and QToF detection. Then, parameters such as limits of detection (LOD) and quantification (LOQ), linearity, and repeatability were assessed. The results showed that the QToF mass analyzer combined with PCI was the best system for the determination of the N-nitrosamines, with instrumental LOD and LOQ in the ranges of 0.2–4 and 0.6–11 ng mL⁻¹, respectively, which translated into method LOD and LOQ in the ranges of 0.2–1.3 and 0.6–3.9 ng L⁻¹, respectively. The analysis of real samples showed the presence of 6 of the N-nitrosamines in influent, effluent, and tap water. N-nitrosodimethylamine (NDMA) was quantified in all the analyzed samples at concentrations between 1 and 27 ng L⁻¹. Moreover, four additional nitrosamines were found in tap and wastewater samples. Full article

Green analytical chemistry principles aim to minimize the negative impact of analytical procedures in the environment, which can be considered both at close (to ensure the safety of the analysts) and global (to conserve our natural resources) levels. These principles suggest, among other guidelines, the reduction/minimization of the sample treatment and the use of renewable sources when possible. The first aspect is largely fulfilled by microextraction, which is considered to be among the greenest sample treatment techniques. The second consideration is attainable if natural products are used as raw materials for the preparation of new extraction phases. This strategy is in line with the change in our production system, which is being gradually moved from a linear model (take–make–dispose) to a circular one (including reusing and recycling as key terms). This article reviews the potential of natural products as sorbents in extraction and microextraction techniques from the synergic perspectives of two research groups working on the topic. The article covers the use of unmodified natural materials and the modified ones (although the latter has a less green character) to draw a general picture of the usefulness of the materials. Full article

The present study represents the determination of Ti and Cr in dry animal feeds using wet acid digestion and inductively coupled plasma–atomic emission spectrometry (ICP-AES), in order to use these metals as digestibility markers. A radiofrequency power of 1350 W and a nebulizer argon flow of 0.8 L/min was selected. The limits of detection were between 11.4 and 16.1 μg/g for titanium and between 10.7 and 38.2 μg/g for chromium. The recovery values for the aqueous solutions were 89.5–103.9% (titanium) and 85.3–104.2% (chromium), with relative standard deviations (RSD%) under 2.1% and standard errors under 2.32%, demonstrating that the method offered good accuracy and repeatability. Six different samples of commercially available feedstuffs (two cat foods, two dog foods, and two poultry foods) were analyzed and the levels of investigated metals were found to be in the ranges of 0.10 g/kg and <LOD for chromium and titanium, respectively (dog foods); 0.10–0.18 g/kg, 0.70 g/kg for chromium and titanium, respectively (cat foods); and 0.07 g/kg, 0.82–1.35 g/kg for chromium and titanium, respectively (poultry foods). Full article