GC/EI-MS method for the determination of phytosterols in vegetable oils.

Sterols are a highly complex group of lipophilic compounds present in the unsaponifiable matter of virtually all living organisms. In this study, we developed a novel gas chromatography with mass spectrometry selected ion monitoring (GC/MS-SIM) method for the comprehensive analysis of sterols after saponification and silylation. A new referencing system was introduced by means of a series of saturated fatty acid pyrrolidides (FAPs) as internal standards. Linked with retention time locking (RTL), the resulting FAP retention indices (RIFAP) of the sterols could be determined with high precision. The GC/MS-SIM method was based on the parallel measurement of 17 SIM ions in four time windows. This set included eight molecular ions and seven diagnostic fragment ions of silylated sterols as well as two abundant ions of FAPs. Altogether, twenty molecular ions of C27- to C31-sterols with 0-3 double bonds were included in the final method. Screening of four common vegetable oils (sunflower oil, hemp oil, rapeseed oil, and corn oil) enabled the detection of 30 different sterols and triterpenes most of which could be identified.

Assessment of hepatic fatty acids during non-alcoholic steatohepatitis progression using magnetic resonance spectroscopy.

INTRODUCTION AND OBJECTIVES: Non-alcoholic fatty liver disease (NAFLD) encompasses a spectrum of liver abnormalities including steatosis, steatohepatitis, fibrosis, and cirrhosis. Liver biopsy remains the gold standard method to determine the disease stage in NAFLD but is an invasive and risky procedure. Studies have previously reported that changes in intrahepatic fatty acids (FA) composition are related to the progression of NAFLD, mainly in its early stages. The aim of this study was to characterize the liver FA composition in mice fed a Choline-deficient L-amino-defined (CDAA) diet at different stages of NAFLD using magnetic resonance spectroscopy (MRS). METHODS: We used in-vivo MRS to perform a longitudinal characterization of hepatic FA changes in NAFLD mice for 10 weeks. We validated our findings with ex-vivo MRS, gas chromatography-mass spectrometry and histology. RESULTS: In-vivo and ex-vivo results showed that livers from CDAA-fed mice exhibit a significant increase in liver FA content as well as a change in FA composition compared with control mice. After 4 weeks of CDAA diet, a decrease in polyunsaturated and an increase in monounsaturated FA were observed. These changes were associated with the appearance of early stages of steatohepatitis, confirmed by histology (NAFLD Activity Score (NAS) = 4.5). After 10 weeks of CDAA-diet, the liver FA composition remained stable while the NAS increased further to 6 showing a combination of early and late stages of steatohepatitis. CONCLUSION: Our results suggest that monitoring lipid composition in addition to total water/fat with MRS may yield additional insights that can be translated for non-invasive stratification of high-risk NAFLD patients.

Toward higher extraction and enrichment factors via a double-reservoirs microfluidic device as a micro-extractive platform.

In this study, firstly, a double-reservoir and switchable prototype of a micro-chip along with the respective holders were fabricated. A cyclic desorption process using microliter volume of organic solvent was adopted to prevent any outdoor contamination. As extractive phases, two identical sheets of electrospun polyamide/polypyrrole/titania nanofibers were synthesized using core-shell electro-spinning technique and utilized for determination of memantine in plasma samples. Field emission scanning electron microscopy images showed a high degree of porosity and homogeneity throughout the sheet structure. Also, energy dispersive X-ray analysis confirmed the presence of titania, while the recorded Fourier transform infrared spectra proved the chemical structures of the polymeric mats. The incorporation of titania as well as polypyrrole in the composition of polyamide nanofibers improved both mechanical stability and extraction capacity of the extractive phase and therefore facilitated the extraction/desorption process. The limits of detection and quantification were 0.01 and 0.04 ng/mL, respectively. In addition, the interday and intraday precisions were lower than 6.7% (n = 3). The linearity was in the range of 0.14-75.00 ng/mL, while recoveries were between 94.1 and 98.4% with the regression coefficient of 0.9987.

Applications of solid-phase micro-extraction with mass spectrometry in pesticide analysis.

Pesticides, widely applied in agriculture, can produce a variety of transformation products and their continuous use causes deleterious effects to ecosystem. Efficient and sensitive analytical techniques for enrichment and analysis of pesticides samples are highly required. Compared with other extraction methods, solid-phase micro extraction is a solvent free, cost effective, robust, versatile, and high throughput sample preparation technique, especially for the analysis of pesticides from complicated matrices. Coupling of solid-phase micro extraction with gas chromatography and mass spectrometry and liquid chromatography-mass spectrometry has been extensively applied in pesticide analysis. On the other hand, in recent years, combination of fast separation using solid-phase micro extraction and rapid detection using ambient mass spectrometry is providing highly efficient pesticide screening. This article summarizes the applications of solid-phase micro extraction coupled to mass spectrometry for pesticides analysis.

Chemical Composition of Two Different Lavender Essential Oils and Their Effect on Facial Skin Microbiota.

Lavender oil is one of the most valuable aromatherapy oils, its anti-bacterial and anti-fungal activities can be explained by main components such as linalool, linalyl acetate, lavandulol, geraniol, or eucalyptol. The aim of the study was to assess the anti-microbial effects of two different lavender oils on a mixed microbiota from facial skin. The commercial lavender oil and essential lavender oil from the Crimean Peninsula, whose chemical composition and activity are yet to be published, were used. Both oils were analysed by gas chromatography coupled to mass spectrometry. The composition and properties of studied oils were significantly different. The commercial ETJA lavender oil contained 10% more linalool and linalyl acetate than the Crimean lavender oil. Both oils also had different effects on the mixed facial skin microbiota. The Gram-positive bacilli were more sensitive to ETJA lavender oil, and Gram-negative bacilli were more sensitive to Crimean lavender oil. However, neither of the tested oils inhibited the growth of Gram-positive cocci. The tested lavender oils decreased the cell number of the mixed microbiota from facial skin, but ETJA oil showed higher efficiency, probably because it contains higher concentrations of monoterpenoids and monoterpenes than Crimean lavender oil does.

Determination of sugars and cyclitols isolated from various morphological parts of Medicago sativa L.

Plant research interest has increased all over the world, and a large body of evidence has been collected to show the huge potential of medicinal plants in various disease treatments. Medicago sativa L., known as alfalfa, is a rich source of biologically active components and secondary metabolites and was frequently used from the ancient times both as fodder crop and as a traditional medicine in the treatment of various diseases. Cyclitols, naturally occurring in this plant, have a particular interest for us due to their significant anti-diabetic, antioxidant, anti-inflammatory, and anti-cancer properties. In the present study we revealed the isolation, the identification, and the quantification of some cyclitols and sugars extracted from different morphological parts of alfalfa plant. Soxhlet extraction and solid phase extraction were used as extraction and purification methods, while for the analyses derivatization followed by gas chromatography with mass spectrometry was involved. The obtained results showed significant differences in the quantities of cyclitols and sugars found in the investigated morphological parts, ranging between 0.02 and 13.86 mg/g of plant in case of cyclitols, and in the range of 0.09 and 40.09 mg/g of plant for sugars. However, roots have the richest part of cyclitols and sugars in contrast to the leaves.

Determination of ultraviolet filter compounds in environmental water samples using membrane-protected micro-solid-phase extraction.

A method based on membrane-protected micro-solid-phase extraction coupled with gas chromatography and mass spectrometry was developed for the determination of six ultraviolet filter compounds in various aqueous media. Multiwalled carbon nanotubes as the sorbent were encapsulated in a sealed polypropylene membrane packet and immersed in the sample to extract the analytes, and then dichloromethane was used for desorption purpose. The method was sensitive enough for quantitative analysis of the target analytes, with limits of quantification between 0.01 and 0.06 µg/L, and produced a linear response (R2  > 0.991) over the calibration range (0.05-6 µg/L). The obtained reproducibility was practically suitable with relative standard deviation values of less than 14% in pure water (spiked at 0.20/µg L) and less than 15% in real samples. The optimized method was applied for the analysis of real water samples with varying matrix complexity: tap, river, and dam water; geothermal spa; and sewage treatment plant effluent. Various levels and patterns of contamination were observed in the examined samples, while the sample from spa was the most contaminated, regarding the target analytes. Matrix spikes and matrix spike replicates were also analyzed to validate the technique for analysis of real aqueous samples, and satisfactory results were achieved.

Determination of siloxanes in water samples employing graphene oxide/Fe3 O4 nanocomposite as sorbent for magnetic solid-phase extraction prior to GC-MS.

A new, fast, simple, and environmentally friendly analytical method has been developed to determine six siloxanes in water samples: octamethyltrisiloxane, octamethylcyclotetrasiloxane, decamethyltetrasiloxane, decamethylcyclopentasiloxane, dodecamethylpentasiloxane and dodecamethylcyclohexasiloxane. The analytical method consists of magnetic solid-phase extraction employing graphene oxide/Fe3 O4 as sorbent for the separation and preconcentration of siloxanes prior to GC-MS determination. The extraction procedure was optimized by means of a Plackett-Burman design. Under the optimized extraction conditions (graphene oxide/Fe3 O4 , 20 mg; extraction time, 10 min; eluent volume, 0.5 mL ACN; elution time, 2.5 min; sample volume, 20 mL), the method rendered repeatability levels with a relative standard deviation between 9 and 20% (n = 6, 10 µg/L). Methodological limits of detection ranged from 0.003 to 0.1 µg/L. The linearity of the method was studied between the methodological limit of quantification and 100 µg/L, obtaining correlation coefficient values between 0.990 and 0.999. The applicability of the method was assessed by analyzing drinking, river and wastewater samples. Relative recovery values ranged between 70 and 120% (1 and 60 µg/L spiking level) showing that the matrix had a negligible effect on extraction. Finally, the greenness of this method was confirmed by the semiquantitative Eco-Scale metrics.

Magnetic solid-phase extraction or dispersive liquid-liquid microextraction for pyrethroid determination in environmental samples.

The determination of 15 pyrethroids in soil and water samples was carried out by gas chromatography with mass spectrometry. Compounds were extracted from the soil samples (4 g) using solid-liquid extraction and then salting-out assisted liquid-liquid extraction. The acetonitrile phase obtained (0.8 mL) was used as a dispersant solvent, to which 75 µL of chloroform was added as an extractant solvent, submitting the mixture to dispersive liquid-liquid microextraction. For the analysis of water samples (40 mL), magnetic solid-phase extraction was performed using nanocomposites of magnetic nanoparticles and multiwalled carbon nanotubes as sorbent material (10 mg). The mixture was shaken for 45 min at room temperature before separation with a magnet and desorption with 3 mL of acetone using ultrasounds for 5 min. The solvent was evaporated and reconstituted with 100 µL acetonitrile before injection. Matrix-matched calibration is recommended for quantification of soil samples, while water samples can be quantified by standards calibration. The limits of detection were in the range of 0.03-0.5 ng/g (soil) and 0.09-0.24 ng/mL (water), depending on the analyte. The analyzed environmental samples did not contain the studied pyrethroids, at least above the corresponding limits of detection.

Determination of phthalate esters in soil using a quick, easy, cheap, effective, rugged, and safe method followed by GC-MS.

A quick, easy, cheap, effective, rugged, and safe procedure was designed to extract pesticide residues from fruits and vegetables with a high percentage of water. It has not been used extensively for the extraction of phthalate esters from sediments, soils, and sludges. In this work, this procedure was combined with gas chromatography with mass spectrometry to determine 16 selected phthalate esters in soil. The extraction efficiency of the samples was improved by ultrasonic extraction and dissolution of the soil samples in ultra-pure water, which promoted the dispersion of the samples. Furthermore, we have simplified the extraction step and reduced the risk of organic solvent contamination by minimizing the use of organic solvents. Different extraction solvents and clean-up adsorbents were compared to optimize the procedure. Dichloromethane/n-hexane (1:1, v/v) and n-hexane/acetone (1:1, v/v) were selected as the extractants from the six extraction solvents tested. C18/primary secondary amine (1:1, m/m) was selected as the sorbent from the five clean-up adsorbents tested. The recoveries from the spiked soils ranged from 70.00 to 117.90% with relative standard deviation values of 0.67-4.62%. The proposed approach was satisfactorily applied for the determination of phthalate esters in 12 contaminated soil samples.

Rapid analysis of ultraviolet filters using dispersive liquid-liquid microextraction coupled to headspace gas chromatography and mass spectrometry.

An ionic-liquid-based in situ dispersive liquid-liquid microextraction method coupled to headspace gas chromatography and mass spectrometry was developed for the rapid analysis of ultraviolet filters. The chemical structures of five ionic liquids were specifically designed to incorporate various functional groups for the favorable extraction of the target analytes. Extraction parameters including ionic liquid mass, molar ratio of ionic liquid to metathesis reagent, vortex time, ionic strength, pH, and total sample volume were studied and optimized. The effect of the headspace temperature and volume during the headspace sampling step was also evaluated to increase the sensitivity of the method. The optimized procedure is fast as it only required ∼7-10 min per extraction and allowed for multiple extractions to be performed simultaneously. In addition, the method exhibited high precision, good linearity, and low limits of detection for six ultraviolet filters in aqueous samples. The developed method was applied to both pool and lake water samples attaining acceptable relative recovery values.

Microwave-assisted-demulsification dispersive liquid-liquid microextraction for the determination of triazole fungicides in water by gas chromatography with mass spectrometry.

A novel microwave-assisted-demulsification dispersive liquid-liquid microextraction technique was established for determination of three triazole fungicides in environmental water samples by gas chromatography with mass spectrometry. Importantly, microwave irradiation has been applied in demulsification to achieve the phase separation and enrichment of triazole fungicides in water samples successfully with low-density toluene as extractant. The experimental variables, including microwave power, microwave time, ultrasonic time, type and volume of extraction solvent, and effect of salting out were investigated. Under the optimized conditions, the method showed good linearity for myclobutanil, tebuconazole, and difenoconazole in the range of 1-100 µg/L. The limits of detection and the limits of quantification were within the range of 0.14-0.27 and 0.47-0.90 µg/L, respectively. The suitable enrichment factors for three triazole pesticides were in the range of 425-636. The recoveries were between 89.3 and 108.7%, and the relative standard deviations were from 5.4 to 8.6%. Finally, environmental water samples were used to verify the applicability of the proposed method for analysis of triazole fungicides targets. It can be concluded that the developed microwave-assisted-demulsification dispersive liquid-liquid microextraction gas chromatography with mass spectrometry method was a rapid, efficient, reliable, and environmental friendly way for analysis of triazole fungicides in water.

Rapid identification and global characterization of multiple constituents from the essential oil of Cortex Dictamni based on GC-MS.

The root of Dictamnus dasycarpus Turcz., also known as Cortex Dictamni, is a Chinese herbal medicine that has been commonly used in the treatment of inflammation, microbial infection, cancer, and other diseases in China for thousands of years. Recently, the essential oil of Cortex Dictamni has been widely studied, and a large number of volatile constituents have been discovered. However, the research of the essential oil of Cortex Dictamni in vivo remains unknown, especially the constituents absorbed into blood after oral administration. Hence, a sensitive and rapid method using gas chromatography with mass spectrometry combined with MassHunter software and the National Institute of Standards and Technology 2014 database was used to investigate the absorbed components in rat serum after oral administration of the essential oil of Cortex Dictamni. With the established method, a total of 36 compounds were screened and identified in the essential oil of Cortex Dictamni based on the mass spectrometry data and compound database. Among them, eight compounds, elemol, thymol methyl ether, ß-eudesmol, ß-cyclocostunolid, guaiazulene, trans-4-hydroxystilbene, ethyl oleate, and monoelaidin, were tentatively characterized in rat serum. This work demonstrated that the established method proved to be a powerful technique for rapid, simple, reliable, and automated identification of bioactive components of herbal medicine.

Nanostructured gemini-based supramolecular solvent coupled with ultrasound-assisted back extraction as a preconcentration step before GC-MS.

Liquid-phase microextraction based on gemini-based supramolecular solvent was successfully applied as a preconcentration step before gas chromatography with mass spectrometry. To eliminate the interferences of gemini surfactant, the analytes were back-extracted into an immiscible organic solvent in the presence of ultrasonic sound waves. Three phthalate esters (di-n-butyl-, butylbenzyl-, bis(2-ethylhexyl)-, and di-n-octyl phthalatic esters) were used as target analytes. The effective parameters on extraction efficiency of the target analytes (i.e., the amount of surfactant and volume of propanol as major components making up the supramolecular solvent, ionic strength, hexane volume, and ultrasound time) were investigated and optimized by a one-variable-at-a-time method. Under the optimum conditions, the preconcentration factors of the analytes were in the range of 95-182. The linear dynamic range of 0.05-200.00 µg/L with a correlation of determination of (R2 ) ≥ 0.9935 was obtained. The proposed method had an excellent limit of detection (S/N = 3) of 0.01 for di-n-octyl and 0.02 µg/L for butylbenzyl- and di-n-butyl-phthalatic ester. Good relative recoveries in the range of 85.7-105.2% guaranteed the accuracy of the amount of phthalates distinguished in the nonspiked samples.

Gas chromatography with mass spectrometry for the determination of phthalates preconcentrated by microextraction based on an ionic liquid.

A new procedure is proposed for the analysis of migration test solutions obtained from plastic bottles used in the packaging of edible oils. Ultrasound-assisted emulsification microextraction with ionic liquids was applied for the preconcentration of six phthalate esters: dimethylphthalate, diethylphthalate, di-n-butylphthalate, n-butylbenzylphthalate, di-2-ethylhexylphthalate, and di-n-octylphthalate. The enriched ionic liquid was directly analyzed by gas chromatography and mass spectrometry using direct insert microvial thermal desorption. The different factors affecting the microextraction efficiency, such as volume of the extracting phase (30 µL of the ionic liquid) and ultrasound application time (25 s), and the thermal desorption step, such as desorption temperature and time, and gas flow rate, were studied. Under the selected conditions, detection limits for the analytes were in the 0.012-0.18 µg/L range, while recovery assays provided values ranging from 80 to 112%. The use of butyl benzoate as internal standard increased the reproducibility of the analytical procedure. When the release of the six phthalate esters from the tested plastic bottles to liquid simulants was monitored using the optimized procedure, analyte concentrations of between 1.0 and 273 µg/L were detected.

Development and validation of a subcritical 1,1,1,2-tetrafluoroethane extraction technique: Gas chromatography-mass spectrometry method for the simultaneous determination of polycyclic aromatic hydrocarbons, polychlorinated biphenyls, and polybrominated biphenyl ethers in aquatic products.

A simple, rapid, and green method was developed for the simultaneous analysis of polycyclic aromatic hydrocarbons, polychlorinated biphenyls, and polybrominated biphenyl ethers in aquatic products using subcritical 1,1,1,2-tetrafluoroethane extraction coupled with gas chromatography and mass spectrometry. Effects of the extraction temperature, pressure, and cosolvent volume on the extraction efficiency were investigated by extracting spiked oyster samples. The results show that the maximum extraction efficiency was obtained at 40°C, 12 MPa, and a cosolvent (dichloromethane) volume of 5.0 mL. Under these conditions, the calibration curves had good linearity with square of the correlation larger than 0.998 in the concentration range of 5-800 ng/mL; limit of detection and limit of quantitation were 0.16-2.83 and 0.55-9.43 ng/g, respectively. At spiked levels of 10, 30, and 50 ng/g, the average recoveries were 70.4-80.4% for polycyclic aromatic hydrocarbons, 74.0-83.6% for polychlorinated biphenyls, and 66.9-78.0% for polybrominated biphenyl ethers, with average relative standard deviations of less than 16.3%. The established method has no significant differences in recovery compared to traditional methods and is suitable for the analysis of real samples.

Electrospun magnetic polybutylene terephthalate nanofibers for thin film microextraction.

A thin film microextraction method using elecrospun magnetic polybutylene terephthalate nanofibers is developed and implemented to isolate some selected triazines. Due to the high mechanical stability of these nanofibers, they are repeatedly used under harsh magnetic stirring and ultrasonic conditions without any damage and structure degradation. The presence of magnetic nanoparticles within the nanofiber structure increases the extraction efficiency while the fibers could be collected by an external magnet. The synthesized nanocomposite showed strong affinity toward the selected analytes. Apart from the concentration of magnetic nanoparticles within the nanocomposite network, the effect of different parameters on the extraction and desorption processes including the sample pH, extraction time, sample volume, type of desorption solvent, solvent volume, and desorption time were optimized. Eventually, the detection limits were in the range of 0.02-0.05 ng/mL, while the limits of quantification were between 0.1 and 0.2 ng/mL. The linear dynamic range was 0.1-100 ng/mL, and the relative standard deviations were 4-9% (n = 3). The developed method was extended to the real water samples, and the relative recoveries were in the range of 86-103%, indicating that the prepared sorbent is suitable for extraction of triazines from environmental samples.

A core-shell titanium dioxide polyaniline nanocomposite for the needle-trap extraction of volatile organic compounds in urine samples.

We synthesized a titanium dioxide-polyaniline core-shell nanocomposite and implemented it as an efficient sorbent for the needle-trap extraction of some volatile organic compounds from urine samples. Polyaniline was synthesized, in the form of the emeraldine base, dissolved in dimethyl acetamide followed by diluting with water at pH 2.8, using the interfacial polymerization method. The TiO2 nanoparticles were encapsulated inside the conducting polymer shell, by adapting the in situ dispersing approach. The surface characteristics of the nanocomposite were investigated by Fourier transform infrared spectrometry, scanning electron microscopy, and transmission electron microscopy. After obtaining acceptable preliminary results, some selected volatile compounds, including chloroform, benzene, toluene, ethylbenzene, xylene, and chlorobenzenes were used as model analytes to validate the enrichment properties of the prepared sorbent in conjunction with gas chromatography mass spectrometric detection. Important parameters influencing the extraction process such as extraction temperature, ionic strength, sampling flow rate, extraction time, desorption temperature, and time were optimized. The limits of detection and limits of quantification values were in the range of 0.5-3  and 2-5 ng/L, respectively, using time-scheduled selected ion monitoring mode. The relative standard deviation percent with three replicates was in the range of 5-10%. The applicability of the developed needle-trap method was examined by analyzing urine samples and the relative recovery percentages for the spiked samples were in the range of 81-105%.

Application of solid-phase microextraction with gas chromatography and mass spectrometry for the early detection of active moulds on historical woollen objects.

The goal of this work was to determine the microbial volatile organic compounds emitted by moulds growing on wool in search of particular volatiles mentioned in the literature as indicators of active mould growth. The keratinolytically active fungi were inoculated on two types of media: (1) samples of wool placed on broths, and (2) on broths containing amino acids that are elements of the structure of keratin. All samples were prepared inside 20 mL vials (closed system). In the first case (1) the broths did not contain any sources of organic carbon, nitrogen, or sulfur, i.e. wool was the only nutrient for the moulds. A third type of sample was historical wool prepared in a Petri dish without a broth and inoculated with a keratinolytically active mould (open system). The microbial volatiles emitted by moulds were sampled with the headspace solid-phase microextraction method. Volatiles extracted on solid-phase microextraction fibers were analyzed in a gas chromatography with mass spectrometry system. Qualitative and quantitative analyses of chromatograms were carried out in search of indicators of metabolic activity. The results showed that there are three groups of volatiles that can be used for the detection of active forms of moulds on woollen objects.

Fatty acid profiling of blood cell membranes by gas chromatography with mass spectrometry.

Fatty acids, which are well-known for their influence on human metabolism and signal transduction, are also a substantial component of cellular membranes and regulate the basic properties and functions of membranes. Owing to their multiple functions, fatty acid profiles of cell membranes are of great interest to those who are studying the relationship between membrane biochemical compositions and functions. A HCl-catalyzed derivation method and a gas chromatography with mass spectrometry analysis method were developed to accurately profile the fatty acids in cell membranes of erythrocytes, leukocytes, and platelets. The detection limits of all 35 fatty acids ranged from 0.58 to 22 ng/mL and the limits of quantitation were between 2.1 and 72 ng/mL. Finally, the established method was used to profile the membrane fatty acids of 44 healthy volunteers from the north and south of China. Results revealed significant differences in the fatty acid profiles from the two regions, particularly those of the erythrocytes. This technique may be applied to cell membrane studies to generate new biological hypotheses concerning fatty acid composition and membrane functions as well as to construct related disease profiles.