Polar Column Research Articles

A series of esters of diastereomeric menthols: Comprehensive mass spectral libraries and gas chromatographic data

A comprehensive data collection of mass spectral and gas chromatographic data of a series of esters of diastereomeric menthols (menthol, isomenthol, neomenthol, and neoisomenthol), in total 150 chemical entities, obtained by GC–MS on commonly used capillary columns of different polarity (non-polar HP-5MS and polar HP-Innowax), was created. Also, MS libraries containing electron ionization MS recorded on single quadrupole as well as on quadrupole ion-trap mass detectors together with the RIs on non-polar and polar columns were compiled (available as supplementary materials). The results point out to frequent misidentification of neoisomenthyl acetate as isomenthyl acetate in the literature, and the means of how to resolve this issue was suggested. The outcomes of this study provide chemists and food technologists with a useful tool in the field of food analysis of compounds with important food aroma properties.

Separation performance of the copolymer and homopolymer of aliphatic polycarbonate diols as the stationary phases for capillary gas chromatography

This work presents the investigation of two aliphatic polycarbonate diols (CAPC and HAPC) as the stationary phases for capillary gas chromatography (GC). The CAPC and HAPC capillary columns showed moderate polarity and high column efficiency of 3704 – 4545 plates/m measured by n-octanol and naphthalene at 120 °C. It was found that despite their similar chemical compositions, CAPC and HAPC differ largely in their selectivity towards the isomers of alkanes, methylpyridines and xylenes. As demonstrated, the CAPC column exhibits advantageous comprehensive performance over the HAPC column and the commercial PEG column. Particularly, the CAPC column exhibits higher resolving performance towards the isomers indicated above and the Grob mixture than the HAPC column. Also, it shows distinct advantages over the PEG column in separating the Grob mixture, the isomers of diethylbenzenes and cymenes, and practical analysis of chemical products and the essential oil from the leaves of Rhododendron dauricum L. Additionally, the CAPC column has excellent repeatability and reproducibility on analyte retention times with the relative standard deviation (RSD) values in the range of 0.05% – 0.08% for run-to-run, 0.12% – 0.19% for day-to-day and 2.6% – 4.9% for column-to-column, respectively. Its applications to purity test of chemical products and GC–MS analysis of the essential oil demonstrate its promising future for practical GC analyses.

GC Analysis of Seven Seed Oils Containing Conjugated Fatty Acids

The fatty acid compositions, including isomer compositions, of seven seed oils containing conjugated fatty acids (CFA) were determined. Seed oils were extracted using a modified Folch extraction, converted to fatty acid methyl esters, and analyzed by gas chromatography (GC) with mass spectrometry and flame ionization detection. The MS detector was operated in positive-ion chemical ionization mode using methane reagent gas. GC was performed using two columns providing different retention characteristics: a poly(ethylene glycol) column and a more polar biscyanopropyl column. The complimentary information provided by the two columns was crucial to peak identification in several cases. The major CFA species in the samples are well known but all contained lesser amounts of additional CFA that have not been widely reported. All samples contained multiple species of conjugated linolenic acid, and two samples also contained small amounts of conjugated linoleic acid. The seed oils of Jacaranda mimosifolia and Calendula officinalis were found to contain 8c,10t,12t-18:3, the natural occurrence of which has only been recently reported in some other samples. The seed oil of Impatiens balsamina has been reported to contain four conjugated 18:4 species, and we present evidence for a fifth conjugated 18:4 isomer.

The Role of Ionic Liquid Interaction in the Separation of Fatty Acid Methyl Esters—Polyunsaturated Geometric Isomers in GC–MS

Knowledge of the type and level of saturated and unsaturated fatty acids in food and clinical matrices is of practical importance, but the wide variety of fatty acids makes analyses very complex. The discrimination of the geometric isomers of fatty acid needs proper and effective separation conditions. The efficiency of three different stationary phases was evaluated by GC–MS methods in the separation of fatty acids in their methyl ester forms. Significant differences were observed in the efficiencies of polysiloxane-based (non-polar HP-5MS and medium/high polarity DB-225MS) and ionic liquid-based (SLB-IL111) columns. Baseline separation of the geometric isomers of linoleic acid methyl ester was obtained by the extremely polar SLB-IL111 column, showing a preference over the other two columns. The optimization of the experimental conditions (response linearity, limit of detection, limit of quantification, system suitability, intraday and interday repeatability and accuracy) showed the separation power of the ionic liquid interaction in the analyses by using short (25–30 m long) columns. By deducting the general principles of the interaction, predictions can be made for the separation of other isomers. The results facilitate the precise identification of various types of fatty acids in real samples for nutritional information.

Unraveling the Complex Olefin Isomer Mixture Using Two-Dimensional Gas Chromatography-Photoionization-Time of Flight Mass Spectrometry

Commercial dodecenes are a complex chemical mixture with a majority of C12 olefins and minority of C8-18 olefins. Structurally, dodecene products may consist of straight-chain alkenes, branched alkenes, as well as cyclic hydrocarbons. Due to the difference of feeds and catalysts used in the oligomerization reaction, the composition of the dodecenes is complex and their properties are very different. Knowing the complex composition of dodecenes can help tune the production process and select the appropriate products according to their end use. To reveal the complex profile of dodecenes, an analytical method using two-dimensional gas chromatography (GC×GC) coupled photoionization (PI) - time of flight mass spectrometry (TOFMS) was developed in this study. A conventional (nonpolar × polar) column combination (non-polar column as 1st dimension and mid-polar column as 2nd dimension) was selected. The analytical condition of GC was optimized using fractional factorial experimental design (DoE). Olefin congener grouping by carbon chain length and double bond equivalent (DBE) was achieved based on the detection of molecular ions by PI-TOFMS. Grouping of dodecenes by linear, mono-branched, di- and tri-branched subgroups was achieved based on branching index (BI) under the assumption of no retention time (RT) overlap among subgroups. Certain dodecene isomers were identified by retention index (RI) and further confirmed by PI mass spectra. The information altogether provided a multimodal characterization possibility to be used with statistical tools. Principal component analysis (PCA) and hierarchical clustering analysis (HCA) of seventeen dodecene samples explained the composition variance between catalysts solid phosphoric acid and zeolite, as well as between feeds with C4 and without C4.

Establishment of LC-MS/MS method for determination of aloperine in rat plasma and its application in preclinical pharmacokinetics

Aloperine, a novel natural active alkaloid derived from Sophora alopecuroides L., has attracted much attention for its anti-inflammatory, antiviral, anti-tumor, anti-allergy and other pharmacological activities. In this study, we first established and validated an efficient and sensitive high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of aloperine in rat plasma. Cytisine was used as the internal standard (IS). The separation of aloperine and IS was conducted on a Phenomenex Luna Omega Polar C18 column (2.1 × 50 mm, 1.6 μm) with 0.3% (v/v) formic acid aqueous (containing 5 mM ammonium acetate) and 0.3% (v/v) formic acid acetonitrile using isocratic elution condition at a flow rate of 0.20 mL/min. Aloperine and IS were determined under the transitions of m/z 233.2 → 98.1 and m/z 191.2 → 148.2 (positive ionization mode), respectively. The calibration curve of aloperine was established in the range of 5 (LLOQ) to 2000 ng/mL (r2 = 0.994). The well validated method was full compliance with the bioanalytical method validation of FDA, and was applied to the pharmacokinetic study of aloperine in Sprague-Dawley rats after 50 mg/kg oral administration and 5 mg/kg intravenous injection. This study provides valuable references for the further study of Sophora alopecuroides L., especially for the drug development and clinical application of aloperine.

Determination of Cytotoxic Activity of Sanguinaria canadensis Extracts against Human Melanoma Cells and Comparison of Their Cytotoxicity with Cytotoxicity of Some Anticancer Drugs.

Melanoma is an enormous global health burden, and should be effectively addressed with better therapeutic strategies. Therefore, new therapeutic agents are needed for the management of this disease. The aim of this study was the investigation of cytotoxic activity of some isoquinoline alkaloid standards and extracts obtained from Sanguinaria canadensis—collected before, during, and after flowering—against three different human melanoma cells (A375, G361, SK-MEL-3). The cytotoxicity of these extracts was not previously tested on these melanoma cell lines. Determination of alkaloid contents was performed by HPLC-DAD using Polar RP column and mobile phase containing acetonitrile, water, and 1-butyl-3-methylimidazolium tetrafluoroborate. The cytotoxicity of alkaloid standards was investigated by determination of cell viability and calculation of IC50 values. Significant differences were observed in the alkaloids content and cytotoxic activity of the extracts, depending on the season of collection of the plant material. In the Sanguinaria canadensis extracts high contents of sanguinarine (from 4.8543 to 9.5899 mg/g of dry plant material) and chelerythrine (from 42.7224 to 6.8722 mg/g of dry plant material) were found. For both of these alkaloids, very high cytotoxic activity against the tested cell lines were observed. The IC50 values were in the range of 0.11–0.54 µg/mL for sanguinarine and 0.14 to 0.46 µg/mL for chelerythrine. IC50 values obtained for Sanguinaria canadensis extracts against all tested cell lines were also very low (from 0.88 to 10.96 µg/mL). Cytotoxic activity of alkaloid standards and Sanguinaria canadensis extracts were compared with the cytotoxicity of anticancer drugs—etoposide, cisplatin, and hydroxyurea. In all cases except the one obtained for cisplatin against A375, which was similar to that obtained for Sanguinaria canadensis after flowering against the same cell line, IC50 values obtained for anticancer drugs were higher than the IC50 values obtained for sanguinarine, chelerythrine, and Sanguinaria canadensis extracts. Our results showed that Sanguinaria canadensis extracts and isoquinoline alkaloids, especially sanguinarine and chelerythrine, could be recommended for further in vivo experiments in order to confirm the possibility of their application in the treatment of human melanomas.

Polar and Helical Isomorphous Crystals of Proline Derivatives: Influence of a Fluorine Atom on the Electric Susceptibility

Two novel nonlinear optical isomorphous crystals of proline derivatives with alkyne functionality have been obtained (Boc-L-ProNH(CH2)2CCH and Boc-cis-4-fluoro-L-ProNH(CH2)2CCH). Both derivatives, which differ only by the substitution of a H atom to a F atom, adopt the same polar and columnar right-handed helix arrangement in the crystalline state. In addition, adjacent polar helical columns all point in the same direction, thus generating a macrodipole and a crystalline system conducive for second harmonic generation (SHG) properties. This isomorphous crystal system constitutes an interesting tool to study the effect of the fluorine atom on the dipole moment and on the first hyperpolarizability. Starting from the PBC optimized geometries of the crystals, the macroscopic second-order nonlinearity, χ(2), of the newly synthesized crystals has been estimated by quantum chemical calculations. These χ(2) responses are of the same order of magnitude as those of inorganic proline derivatives while smaller than those observed in crystals of push–pull π-conjugated molecules.Graphic

Congener-specific partition properties of chlorinated paraffins evaluated with COSMOtherm and gas chromatographic retention indices

Chlorinated Paraffins (CPs) are high volume production chemicals and have been found in various organisms including humans and in environmental samples from remote regions. It is thus of great importance to understand the physical–chemical properties of CPs. In this study, gas chromatographic (GC) retention indexes (RIs) of 25 CP congeners were measured on various polar and nonpolar columns to investigate the relationships between the molecular structure and the partition properties. Retention measurements show that analytical standards of individual CPs often contain several stereoisomers. RI values show that chlorination pattern have a large influence on the polarity of CPs. Single Cl substitutions (–CHCl–, –CH2Cl) generally increase polarity of CPs. However, many consecutive –CHCl– units (e.g., 1,2,3,4,5,6-C11Cl6) increase polarity less than expected from the total number of –CHCl– units. Polyparameter linear free energy relationship descriptors show that polarity difference between CP congeners can be explained by the H-bond donating properties of CPs. RI values of CP congeners were predicted using the quantum chemically based prediction tool COSMOthermX. Predicted RI values correlate well with the experimental data (R2, 0.975–0.995), indicating that COSMOthermX can be used to accurately predict the retention of CP congeners on GC columns.

Conceptual Structural Design and Comparative Power System Analysis of Ozone Dynamics Investigation Nano-satellite (ODIN)

The Ozone Dynamics Investigation Nano-Satellite (ODIN) is a CubeSat design proposed by Chungnam National University as contribution to the CubeSat Competition 2019 sponsored by the Korean Aerospace Research Institute (KARI). The main objectives of ODIN are (1) to observe the polar ozone column density (latitude range of 60° to 80° in both hemispheres) and (2) to investigate the chemical dynamics between stratospheric ozone and ozone depleting substances (ODSs) through spectroscopy of the terrestrial atmosphere. For the operation of ODIN, a highly efficient power system designed for the specific orbit is required. We present the conceptual structural design of ODIN and an analysis of power generation in a sun synchronous orbit (SSO) using two different configurations of 3U solar panels (a deployed model and a non-deployed model). The deployed solar panel model generates 189.7 W through one day which consists of 14 orbit cycles, while the non-deployed solar panel model generates 152.6 W. Both models generate enough power for ODIN and the calculation suggests that the deployed solar panel model can generate slightly more power than the non-deployed solar panel model in a single orbit cycle. We eventually selected the non-deployed solar panel model for our design because of its robustness against vibration during the launch sequence and the capability of stable power generation through a whole day cycle.

Blue Tansy Essential Oil: Chemical Composition, Repellent Activity Against Aedes aegypti and Attractant Activity for Ceratitis capitata

Blue tansy essential oil (BTEO) ( Tanacetum annuum L.) was analyzed by GC-MS and GC-FID using two different capillary column stationary phases. Sabinene (14.0%), camphor (13.6%), myrcene (8.0%), β-pinene (7.7%), and chamazulene (6.9%) were the main components using an SE52 column (non-polar). On a polar CW20M phase column, sabinene (15.1%), camphor (14.4%), α-phellandrene (7.9%), β-pinene (7.7%), and myrcene (6.9%) were the most abundant compounds. To assess the oil for potential applications in integrated pest management strategies, behavioral bioassays were conducted to test for repellency against yellow fever mosquito Aedes aegypti, and for attractant activity for Mediterranean fruit fly Ceratitis capitata. Results showed that BTEO was not effective in repelling Ae. aegypti (minimum effective dosage [MED]: 0.625 ± 0.109 mg/cm2 compared with the standard insect repellent DEET (N,N-diethyl-3-methylbenzamide). In assays with male C. capitata, BTEO displayed mild attraction compared with two positive controls (essential oils from tea tree Melaleuca alternifolia and African ginger bush Tetradenia riparia). Additional studies are needed to identify the specific attractant chemicals in BTEO and to determine if they confer a synergistic effect when combined with other known attractants for C. capitata. To the best of our knowledge, this study represents the first investigation of BTEO for repellency against the mosquito vector Ae. aegypti and for attractancy to C. capitata, a major agricultural pest worldwide.

Feasibility of using gas chromatography-ion mobility spectrometry to identify characteristic volatile compounds related to brandy aging

Herein, gas chromatography-ion mobility spectrometry (GC-IMS) employing FS-SE-54-CB (non-polar) and WAX (polar) columns is used to determine the volatile compound profile of brandy samples aged up to nine years and characteristic markers related to aging are identified. Non-polar GC column showed better separation of the aromatic markers. The marker compounds of brandy included esters, higher alcohols, aldehydes, and fatty acids. Isopentyl acetate, ethyl butyrate, ethyl-2-methylpropanoate, ethyl propanoate, and furfurol were identified as the characteristic volatile compounds in the brandy samples aged for six years. Gas chromatography-mass spectrometry (GC–MS) was also used to analyse the aromatic compound profile of aged brandy samples and verify the GC-IMS results. Based on the identification of ten common volatiles detected using GC-IMS and GC–MS, it was concluded that GC-IMS could be used to determine the characteristic markers related to brandy aging; However, low responses were observed for terpenes and volatile phenols.

Sudden Stratospheric Warmings

AbstractSudden stratospheric warmings (SSWs) are impressive fluid dynamical events in which large and rapid temperature increases in the winter polar stratosphere (∼10–50 km) are associated with a complete reversal of the climatological wintertime westerly winds. SSWs are caused by the breaking of planetary‐scale waves that propagate upwards from the troposphere. During an SSW, the polar vortex breaks down, accompanied by rapid descent and warming of air in polar latitudes, mirrored by ascent and cooling above the warming. The rapid warming and descent of the polar air column affect tropospheric weather, shifting jet streams, storm tracks, and the Northern Annular Mode, making cold air outbreaks over North America and Eurasia more likely. SSWs affect the atmosphere above the stratosphere, producing widespread effects on atmospheric chemistry, temperatures, winds, neutral (nonionized) particles and electron densities, and electric fields. These effects span both hemispheres. Given their crucial role in the whole atmosphere, SSWs are also seen as a key process to analyze in climate change studies and subseasonal to seasonal prediction. This work reviews the current knowledge on the most important aspects of SSWs, from the historical background to dynamical processes, modeling, chemistry, and impact on other atmospheric layers.

Application of HPLC-DAD for In Vitro Investigation of Acetylcholinesterase Inhibition Activity of Selected Isoquinoline Alkaloids from Sanguinaria canadensis Extracts.

Isoquinoline alkaloids may have a wide range of pharmacological activities. Some of them have acetylcholinesterase activity inhibition. Nowadays, neurodegenerative disorders such as Alzheimer’s disease have become a serious public health problem. Searching for new effective compounds with inhibited acetylcholinesterase activity is one of the most significant challenges of modern scientific research. The aim of this study was the in vitro investigation of acetylcholinesterase activity inhibition of extracts obtained from Sanguinaria canadensis collected before, during and after flowering. The acetylcholinesterase activity inhibition of these extracts has not been previously tested. The aim was also to quantify selected alkaloids in the investigated extracts by high performance liquid chromatography (HPLC). The analyses of alkaloid content were performed using HPLC in reversed phase (RP) mode using Polar RP column and mobile phase containing acetonitrile, water and ionic liquid (IL). The acetylcholinesterase activity inhibition of the tested plant extracts and respective alkaloid standards were examined using high performance liquid chromatography with diode-array detector (HPLC-DAD) for the quantification of 5-thio-2-nitro-benzoic acid, which is the product of the reaction between the thiocholine (product of the hydrolysis of acetylthiocholine reaction) with Ellman reagent. The application of the HPLC method allowed for elimination of absorption of interfering components, for example, alkaloids such as sanguinarine and berberine. It is revealed that the HPLC method can be successfully used for the evaluation of the acetylcholinesterase inhibitory activity in samples such as plant extracts, especially those containing colored components adsorbing at wavelength in the range 405–412 nm. The acetylcholinesterase inhibition activity synergy of pairs of alkaloid standards and mixture of all investigated alkaloids was also determined. Most investigated alkaloids and all Sanguinaria canadensis extracts exhibited very high acetylcholinesterase activity inhibition. IC50 values obtained for alkaloid standards were from 0.36 for berberine to 23.13 µg/mL for protopine and from 61.24 to 89.14 µg/mL for Sanguinaria canadensis extracts. Our investigations demonstrated that these plant extracts can be recommended for further in vivo experiments to confirm their acetylcholinesterase activity inhibition.

A simple HPLC-MS/MS method for the analysis of multi-mycotoxins in betel nut

ABSTRACT Mycotoxins are naturally occurring toxins produced by certain molds and can be found in nuts, cereals, fruits, and oils. Furthermore, current food processing systems cannot wholly remove mycotoxins. The adverse health effects of chewing betel nut containing mycotoxins range from long-term consequences such as immune deficiency. In this study, a rapid solid-phase extraction (SPE) method to LC-MS/MS system was developed to determine nine mycotoxins in commonly used five types of betel nut products. The SPE eluate was injected into a Phenomenex UHPLC Kinetex Polar C18 Column (2.1 × 100 mm, 2.6 mm) and detected by an Agilent 6470 triple-quadrupole mass spectrometer. All mycotoxins were separated under 6.8 min with electrospray ionization in multiple reaction monitoring modes. The calibration curves ranged from 0.5 to 600 ng/mL (r2>0.99), LOQ values were between 0.25 to10 ng/mL, and LOD values were between 5 to 20 ng/mL. All the QCs have shown precision, and the accuracy range for intra-day and inter-day did not exceed 14.3% (RSD) and 91.2 to 117.6% (relative error). The matrix effect and extraction recovery for five types of betel nuts were in the range of 81.5 to 99.2% and 70.1 to 113.9%, respectively (except deoxynivalenol).

Through-pore polymerization in polar high-performance liquid chromatography columns allowing scanning electron microscopy based imaging of the packing order

To allow an enhanced understanding of the order in packed HPLC columns, in this work a methodology for immobilizing native polar silica particles is developed based on the polymerization of a methyl methacrylate (MMA) and ethylene glycol dimethacrylate (EGDMA) as a cross-linker in the interstitial pores of HPLC columns. Subsequent mechanical cutting then allows scanning electron microscopy (SEM) based imagery of cross-sections of the packed bed. In this way, the packing efficiency of home-made and commercial HPLC columns with 4.6 mm inner diameter and 150 mm length comprising the same packing material of 5 µm silica particles are compared. The methodology is developed for native silica used in e.g. hydrophilic interaction liquid chromatography (HILIC) and in normal phase LC. In order to confirm the feasibility of the developed methodology, the conventional methods for the evaluation of column, efficiency and porosity, are also employed. The obtained porosity information is compared and showed the same trend with the external porosity measurements obtained via inverse size exclusion approach, illustrating its potential application to study the micro-heterogeneity of packed HPLC columns and to guide the optimization of the packing process of HPLC columns.

A Novel Chemical Profile of a Selective In Vitro Cholinergic Essential Oil from Clinopodium taxifolium (Kunth) Govaerts (Lamiaceae), a Native Andean Species of Ecuador.

A novel chemical profile essential oil, distilled from the aerial parts of Clinopodium taxifolium (Kunth) Govaerts (Lamiaceae), was analysed by Gas Chromatography-Mass Spectrometry (GC-MS, qualitative analysis) and Gas Chromatography with Flame Ionization Detector (GC-FID, quantitative analysis), with both polar and non-polar stationary phase columns. The chemical composition mostly consisted of sesquiterpenes and sesquiterpenoids (>70%), the main ones being (E)-β-caryophyllene (17.8%), α-copaene (10.5%), β-bourbonene (9.9%), δ-cadinene (6.6%), cis-cadina-1(6),4-diene (6.4%) and germacrene D (4.9%), with the non-polar column. The essential oil was then submitted to enantioselective GC analysis, with a diethyl-tert-butyldimethylsilyl-β-cyclodextrin diluted in PS-086 chiral selector, resulting in the following enantiomeric excesses for the chiral components: (1R,5S)-(−)-α-thujene (67.8%), (1R,5R)-(+)-α-pinene (85.5%), (1S,5S)-(−)-β-pinene (90.0%), (1S,5S)-(−)-sabinene (12.3%), (S)-(−)-limonene (88.1%), (S)-(+)-linalool (32.7%), (R)-(−)-terpinen-4-ol (9.3%), (S)-(−)-α-terpineol (71.2%) and (S)-(−)-germacrene D (89.0%). The inhibition activity against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) of C. taxifolium essential oil was then tested, resulting in selective activity against BChE with an IC50 value of 31.3 ± 3.0 μg/mL (positive control: donepezil, IC50 = 3.6 μg/mL).

Development of Gas-Chromatographic Method for Simultaneous Determination of Cannabinoids and Terpenes in Hemp.

An original gas chromatographic method has been developed for simultaneous determination of major terpenes and cannabinoids in plant samples and their extracts. The main issues to be addressed were the large differences in polarity and volatility between both groups of analytes, but also the need for an exhaustive decarboxylation of cannabinoid acidic forms. Sample preparation was minimised, also by avoiding any analyte derivatisation. Acetone was found to be the most appropriate extraction solvent. Successful chromatographic separation was achieved by using a medium polarity column. Limits of detection ranged from 120 to 260 ng/mL for terpenes and from 660 to 860 ng/mL for cannabinoids. Parallel testing proved the results for cannabinoids are comparable to those obtained from established HPLC methods. Despite very large differences in concentrations between both analyte groups, a linear range between 1 and 100 µg/mL for terpenes and between 10 and 1500 µg/mL for cannabinoids was determined.

Quantification of mycophenolic acid in human plasma by liquid chromatography with time-of-flight mass spectrometry for therapeutic drug monitoring.

This study presents, for the first time, the development and validation of a liquid chromatography and time-of-flight mass-spectrometry (LC-TOF-MS) based assay to quantify mycophenolic acid (MPA) in patient samples as part of a routine therapeutic drug monitoring service. MPA was extracted from 50 μl human plasma by protein precipitation, using sulindac as internal standard (IS). Separation was obtained on a Luna™ Omega polar C18 column kept at 40°C. The mobile phase consisted of a mixture of acetonitrile-deionized water (50:50, v/v) with 0.1% formic acid at a flow rate of 350 μl/min. Analyte and IS were monitored on a TOF-MS using a Jet-Stream™ (electrospray) interface running in positive mode. Assay performance was evaluated by analysing patient plasma (N = 69) and external quality assessment (N = 6) samples. The retention times were 2.66 and 2.18 min for MPA and IS, respectively. The lower limit of quantification of MPA was 0.1 μg/ml. The within- and between-assay reproducibility results ranged from 1.81 to 10.72%. Patient and external quality assessment sample results were comparable with those obtained previously by an in-house validated LC-MS/MS method. This method showed satisfactory analytical performance for the determination of MPA in plasma over the calibration range of 0.1-15.0 μg/ml.

Tutorial for the Characterization of Fatty Acid Methyl Esters by Gas Chromatography with Highly Polar Capillary Columns.

The fatty acid composition of fats and oils is commonly determined by gas chromatography after preparing fatty acid methyl esters (FAME). Capillary columns coated with polyethylene glycol emerged as the preferred separation tool for the quantification of the polyunsaturated fatty acids contained primarily in marine oils. However, their selectivity is inadequate for measuring the trans fatty acids (TFA) contained in refined vegetable oils, dairy fats, and marine oils. Highly polar 100% poly(biscyanopropyl siloxane) capillary columns provide the necessary selectivity, but small differences in the phase polarity caused by column age, conditioning, or manufacturing variations affect the reproducibility of their separations of these complex samples. In this study, a simple procedure is described to compensate for small variations in column selectivity by adjusting the elution temperature. The balance between the dipole-induced dipole interactions and dispersive interactions was determined by measuring selectivity factors [SF(i)] corresponding to the elution of an unsaturated FAME such as 18:3n-3 relative to two saturated FAME such as 20:0 and 22:0. Knowing the SF(i) provided by the installed capillary column at a given elution temperature, and the SF(i) of the target separation, we propose a simple calculation to determine the necessary elution temperature adjustment to achieve (or restore) the desired separation. After determining the SF(i) which provides the optimal separation of TFA, the novel methodology was applied to the separation of refined vegetable oils, butter fats, and marine oils.