Recent Applications of Solid Phase Microextraction Coupled to Liquid Chromatography

Solid phase microextraction (SPME) is one of the most popular sample preparation methods which can be applied to organic compounds allowing the simultaneous extraction and pre-concentration of analytes from the sample matrix. It is based on the partitioning of the analyte between the extracting phase, generally immobilized on a fiber substrate, and the matrix (water, air, etc.), and has numerous advantages such as rapidity, simplicity, low cost, ease of use and automation, and absence of toxic solvents. Fiber SPME has been widely used in combination with various analytical instrumentation even if most of the work has been done coupling the extraction technique with gas and liquid chromatography (GC and LC). This manuscript presents an overview of the recent works (from 2010 to date) of solid phase microextraction coupled to liquid chromatography (SPME-LC) relevant to analytical applications performed using commercially available fibers or lab-made fibers already developed in previous papers, and to improved instrumental systems and approaches

Measurement of squalene in olive oil by fractional crystallization or headspace solid phase microextraction coupled with gas chromatography

Squalene is the most abundant component in the unsaponifiable fraction of olive oil with strong antioxidant properties. Its concentration in olive oils varies between 0.2 and 16.2 g/kg depending on the cultivar(s) used. The propose of this work was to determine the effectiveness of two different extraction methods for squalene determination by gas chromatography (GC) coupled to a flame ionization detector (FID) or to mass spectrometry (MS). In a first approach, oil samples were dissolved in methanol/acetone mixture 7:3 (v/v) and triglycerides separated by fractional crystallization at −20°C. The organic layer was removed, reduced to dryness and the residue reconstituted in n-heptane (containing squalane as external standard) and analyzed by GC-FID. A headspace (HS) solid phase microextraction (SPME) GC-MS method has been also developed in order to have an environmentally friendly (i.e. solventless) extraction procedure. The linear range investigated with both methods was 1.0-10 g/kg. Within-day and between days precision values, expressed as RSD%, were 4 and 7% (GC-FID), and 3 and 6% (GC-MS), respectively. The limit of detection (LOD) at a signal-to-noise (S/N) ratio of 3 were 0.019 (GC-FID) and 0.003 (GC-MS) g/kg; the limit of quantification (LOQ) calculated at S/N = 10 were 0.063 (GC-FID) and 0.008 (GC-MS) g/kg, well below the typical squalene concentration levels found in olive oils. The obtained percentage recoveries were 70 ± 2 (GC-FID) and 98 ± 3 (GC-MS), and were not concentration dependent. The potential of the method has been demonstrated by the analysis of several different olive oil samples produced from different cultivars and different locations

Analytical investigations on lindane bioremediation capability of the demosponge Hymeniacidon perlevis

Lindane is an organochlorine pesticide that has been widely used to treat agricultural pests. It is of particular concern because of its toxicity, persistence and tendency to bioaccumulate in terrestrial and aquatic ecosystems. In this context, we investigated the ability of the demosponge Hymeniacidon perlevis to bioremediate lindane polluted seawater during in vitro experimentation. Lindane was extracted by solid-phase micro-extraction (SPME) and determined by gas chromatography–mass spectrometry (GC–MS). Furthermore, we assessed the role exerted in lindane degradation by bacteria isolated from the sponge. Sponges showed low mortality in experimental conditions (lindane concentration 1 μg/L) and were able to remove about 50% of the lindane content from seawater in 48 h. Bacteria isolated from sponges showed a remarkable remediating capacity (up to 97% of lindane removed after 8-days). A lindane metabolite was identified, 1,3,4,5,6-pentachloro-cyclohexene. The results obtained are a prelude to the development of future strategies for the in situ bioremediation of this pollutan

Development, Optimization, and Comparison of Different Sample Pre-Treatments for Simultaneous Determination of Vitamin E and Vitamin K in Vegetables

The absence of vitamin E from the diet can lead to cardiovascular disease, cancer, cataracts, and premature aging. Vitamin K deficiency can lead to bleeding disorders. These fat-soluble vitamins are important nutritional factors that can be determined in different methods in vegetables. In this work, the simultaneous determination of α-tocopherol, α-tocopheryl acetate, phylloquinone, and menaquinone-4 by gas chromatography–mass spectrometry (GC–MS) has been optimized using both direct injection and solid phase microextraction (SPME). Three different sample pre-treatment approaches based on: (A) solid–liquid–liquid–liquid extraction (SLE–LLE), (B) SLE, and (C) SPME were then applied to extract the target analytes from vegetables samples using menaquinone as internal standard. All the procedures allowed the determination of the target analytes in onion, carrot, celery, and curly kale samples. Similar results were obtained with the three different approaches, even if the one based on SPME offers the best performance, together with a reduced use of solvent, time consumption, and experimental complexity, which makes it the preferable option for industrial applications

Determination of Commercial Animal and Vegetable Milks’ Lipid Profile and Its Correlation with Cell Viability and Antioxidant Activity on Human Intestinal Caco-2 Cells

none9openantonella aresta; Stefania De Santis; Alessia Carocci; Alexia Barbarossa; Andrea Ragusa; Nicoletta De Vietro; Maria Lisa Clodoveo; Filomena Corbo; Carlo ZAMBONINAresta, Antonella; De Santis, Stefania; Carocci, Alessia; Barbarossa, Alexia; Ragusa, Andrea; De Vietro, Nicoletta; Lisa Clodoveo, Maria; Corbo, Filomena; Zambonin, Carl

Bovine and soybean milk bioactive compounds: Effects on inflammatory response of human intestinal Caco-2 cells

In this study the effects of commercial bovine and soybean milks and their bioactive compounds, namely genistein, daidzein and equol, on the inflammatory responses induced by lipopolysaccharide (LPS) treatment of human intestinal Caco-2 cells were examined, in terms of nitric oxide (NO) release and inducible nitric oxide synthetase (iNOS) expression. Both milks and their bioactive compounds significantly inhibited, dose-dependently, the expression of iNOS mRNA and protein, resulting in a decreased NO production. The NF-κB activation in LPS-stimulated intestinal cells was also examined. In all cases we observed that cell pre-treatment before LPS activation inhibited the IkB phosphorylation. Accordingly, quantification of bioactive compounds by solid phase microextraction coupled with liquid chromatography has shown that they were absorbed, metabolized and released by Caco-2 cells in culture media. In conclusion, we demonstrated that milks and compounds tested are able to reduce LPS-induced inflammatory responses from intestinal cells, interfering with NF-kB dependent molecular mechanisms

Mussel Inspired Polydopamine as Silica Fibers Coating for Solid-Phase Microextraction

Commercial solid-phase microextraction fibers are available in a limited number of expensive coatings, which often contain environmentally harmful substances. Consequently, several different approaches have been used in the attempt to develop new sorbents that should possess intrinsic characteristics such as duration, selectivity, stability, and eco-friendliness. Herein we reported a straightforward, green, and easy coating method of silica fibers for solid-phase microextraction with polydopamine (PDA), an adhesive, biocompatible organic polymer that is easily produced by oxidative polymerization of dopamine in mild basic aqueous conditions. After FT-ATR and SEM characterization, the PDA fibers were tested via chromatographic analyses performed on UHPLC system using biphenyl and benzo(a)pyrene as model compounds, and their performances were compared with those of some commercial fibers. The new PDA fiber was finally used for the determination of selected PAHs in soot samples and the results compared with those obtained using the commercial PA fiber. Good reproducibility, extraction stability, and linearity were obtained using the PDA coating, which proved to be a very promising new material for SPME

Volatile Organic Compounds, Indole, and Biogenic Amines Assessment in Two Mediterranean Irciniidae (Porifera, Demospongiae)

Solid phase microextraction (SPME) coupled to gas chromatography-mass spectrometry (GC-MS) was employed for the headspace determination of the volatile organic fraction emitted by two of the most common Mediterranean demosponges, Ircinia variabilis and Sarcotragus spinosulus, and of indole and some biogenic amines released by sponges in an aqueous medium. A total of 50/30 µm divinylbenzene/carboxen/polydimethylsiloxane and 75 µm carboxen/polydimethylsiloxane fibers were used for the headspace extraction of low molecular weight sulfur compounds from a hermetically sealed vial containing sponge fragments, while the direct immersion determination of indole and biogenic amines was performed. The biogenic amines were extracted after in-solution derivatization with isobutyl chloroformate. All analytical parameters (linearity, limits of detection, and quantification, precision, and recovery) were evaluated for indole and biogenic amines. SPME-GC-MS proved to be a reliable means of highlighting the differences between molecules released by different sponges, principally responsible for their smell. The combined approaches allowed the identification of several volatile compounds in the headspace and other molecules released by the sponges in an aqueous medium, including indole and the BAs cadaverine, histamine, isobutylamine, isopentylamine, propylamine, 2-phenylethylamine, putrescine and tryptamine. The results obtained represent a further contribution to the picture of odoriferous molecules secreted by sponges

Determination of α-Tocopherol in Olive Oil by Solid-Phase Microextraction and Gas Chromatography – Mass Spectrometry

Olive oil has great human health benefits and is an important component of the Mediterranean diet. Its quality, sensory attributes and oxidative stability are linked to the presence of minor compounds. Vitamin E (α-tocopherol) is a key component in these properties. In this work, solidphase microextraction coupled to gas chromatography - mass spectrometry was employed for the determination of -tocopherol in olive oil. The analytical performance of the method have been assessed in fortified olive oil with negligible vitamin E concentrations. The calibration curve was linear from 0.020 to 0.500 mg/g. The limits of detection and quantification were 0.006 and 0.021 mg/g, respectively. Intra-day and inter-day relative standard deviations were 3.2 and 10.0, respectively, and were concentration independent. The method was employed for the determination of α-tocopherol in virgin and extra virgin olive oil, reporting average concentrations of 0.044 ± 0.03 and 0.200 ± 0.05 mg/g, respectively. Overall, the method is simple, sensitive, rapid, solvent-free, and provided high recoveries of 97.7 ± 3.1%. In addition, vitamin E stability in extra-virgin olive oil was characterized by a shelf-life study

Simultaneous determination of salicylic, 3-methyl salicylic, 4-methyl salicylic, acetylsalicylic and benzoic acids in fruit, vegetables and derived beverages by SPME-LC-UV/DAD

Salicylic and benzoic acid are phenolic acids occurring in plant cells, thus they can be present in fruit and vegetables at various levels. They possess anti-inflammatory and antimicrobial properties, however they may induce symptoms and health problems in a small percentage of the population. Therefore, a low phenolic acid diet may be of clinical benefit to such individuals. In order to achieve this goal, the concentration of these substances in different food and beverages should be assessed. The present work describes for the first time a new method, based on solid phase microextraction (polydimethylsiloxane-divinylbenzene fiber) coupled to liquid chromatography with UV diode array detection, for the simultaneous determination of salicylic acid, 3-methyl salicylic acid, 4-methyl salicylic acid, acetylsalicylic acid and benzoic acid in selected fruit, vegetables and beverages.All the aspects influencing fiber adsorption (time, temperature, pH, salt addition) and desorption (desorption and injection time, desorption solvent mixture composition) of the analytes have been investigated. An isocratic separation was performed using an acetonitrile-phosphate buffer (pH 2.8; 2. mM) mixture (70:30, v/v) as the mobile phase. The estimated LOD and LOQ values (μg/mL) were in the range 0.002-0.028 and 0.007-0.095. The within-day and day-to-day precision values (RSD%) were between 4.7-6.1 and 6.6-9.4, respectively.The method has been successfully applied to the analysis of fava beans, blueberries, kiwi, tangerines, lemons, oranges and fruit juice (lemon and blueberry) samples. The major advantage of the method is that it only requires simple homogenization and/or centrifugation and dilution steps prior to SPME and injection in the LC system