Composition and nutritional values of fatty acids in marine organisms by one-step microwave-assisted extraction/derivatization and comprehensive two-dimensional gas chromatography -flame ionization detector.

This work reports the characterization of the lipidic fraction of seven species of marine organisms gathered along the shoreline of the Po Delta Park of Emilia-Romagna Region (Italy) and of the north Adriatic Sea. Two species of oysters (Crassostrea gigas and Ostrea edulis), two species of clams (Chamelea gallina and Ruditapes philippinarum), one species of mussel (Mytilus galloprovincialis), one species of macroalgae (Ulva rigida), and one species of spiny dogfish (Squalus acanthias) were analyzed to characterize their fatty acids profile and related nutritional value. The lipid fraction was simultaneously extracted and transesterified into fatty acid methyl esters (FAMEs) by using a recently developed one-step microwave-assisted extraction/derivatization (MAED) method. The obtained FAMEs extract was analyzed by a rapid comprehensive multidimensional gas chromatography (GC × GC) method (30 min). The system was equipped with a reverse set of columns (polar × non-polar) connected through a reversed fill/flush flow modulator. The GC × GC system was coupled with a flame-ionization detector (FID) for both qualitative and quantitative purposes. The MAED- GC × GC-FID methodology was suitable in the context of samples containing high percentages of omega-3 PUFA. A total of 82 FAMEs were tentatively identified using standards, literature data, and the two-dimensional plot location. FAME profiles obtained with the proposed approach were comparable with reference methods (AOCS Ce 2b-11), showing no significant differences. Moreover, to determine the food nutritional value of the samples investigated, the most common nutritional indices (index of atherogenicity, index thrombogenicity, hypocholesterolemic/hypercholesterolemic ratio, health-promoting index, unsaturation index, and the fish lipid quality index) were calculated from FAME profiles. Among the samples investigated, Squalus acanthias presented the best nutritional score, while Ruditapes philippinarum had the worst score in 3 out of 6 indices.

Current Developments of Analytical Methodologies for Aflatoxins' Determination in Food during the Last Decade (2013-2022), with a Particular Focus on Nuts and Nut Products.

This review aims to provide a clear overview of the most important analytical development in aflatoxins analysis during the last decade (2013-2022) with a particular focus on nuts and nuts-related products. Aflatoxins (AFs), a group of mycotoxins produced mainly by certain strains of the genus Aspergillus fungi, are known to impose a serious threat to human health. Indeed, AFs are considered carcinogenic to humans, group 1, by the International Agency for Research on Cancer (IARC). Since these toxins can be found in different food commodities, food control organizations worldwide impose maximum levels of AFs for commodities affected by this threat. Thus, they represent a cumbersome issue in terms of quality control, analytical result reliability, and economical losses. It is, therefore, mandatory for food industries to perform analysis on potentially contaminated commodities before the trade. A full perspective of the whole analytical workflow, considering each crucial step during AFs investigation, namely sampling, sample preparation, separation, and detection, will be presented to the reader, focusing on the main challenges related to the topic. A discussion will be primarily held regarding sample preparation methodologies such as partitioning, solid phase extraction (SPE), and immunoaffinity (IA) related methods. This will be followed by an overview of the leading analytical techniques for the detection of aflatoxins, in particular liquid chromatography (LC) coupled to a fluorescence detector (FLD) and/or mass spectrometry (MS). Moreover, the focus on the analytical procedure will not be specific only to traditional methodologies, such as LC, but also to new direct approaches based on imaging and the ability to detect AFs, reducing the need for sample preparation and separative techniques.

Analysis of mixed plastic pyrolysis oil by comprehensive two-dimensional gas chromatography coupled with low- and high-resolution time-of-flight mass spectrometry with the support of soft ionization.

According to the annual production of plastics worldwide, in 2020 about 370 million tons of plastic were produced in the world. Chemical recycling, particularly pyrolysis of plastic wastes, could be a valuable solution to resolve these problems and provide an alternative pathway to produce "recycled" chemical products for the petrochemical industry. Nevertheless, the pyrolysis oils need a detailed characterization before the upgrading test to re-use them to generate new recycled products. Multidimensional gas chromatography coupled with both low- and high-resolution time-of-flight mass spectrometers was employed for a detailed investigation among and within different chemical classes present in bio-plastic oil. The presence of several isomeric species as well as homologs series did not allow a reliable molecular identification, except for a few compounds that showed both MS similarity >800/1000 and retention index within ±20. Indeed, the identification of several isomeric species was assessed by high-resolution mass spectrometry equipped with photoionization interface. This soft ionization mode was an additional filter in the identification step allowing unambiguous identification of analytes not identified by the standard electron ionization mode at 70 eV. The injection method was also optimized using a central composite design to successfully introduce a wide range of carbon number compounds without discrimination of low/high boiling points.

Deeper investigation of oxygen-containing compounds in oleaginous feedstock (animal fat) by preparative column chromatography and comprehensive two-dimensional gas chromatography coupled with high-resolution time-of-flight mass spectrometry.

The production of renewable fuels as biodiesel and bio-jet fuel is usually originated by the transformation and processing of oleaginous feedstocks, mainly composed of triacylglycerols. Currently, a significant part of the triacylglycerol production relies on grassy oil crops or other woody oil plants, representing more than 120 million metric tons every year. Considering that the worldwide triacylglycerol demand is expected to rise in the future, alternative routes are necessary to ensure a sustainable biodiesel industry and limit diesel price volatility. In this context, the use of animal fats could be an interesting alternative for biodiesel production as the production of animal byproducts represents nearly 17 million tons per year in the European Union only (2020). Animal fats, however, contain large amounts of no-esterified fatty acids and other oxygen compounds, reducing the yield of biodiesel. Therefore, a specific pretreatment is needed before the trans-esterification process. The setup of such appropriate pretreatments requires detailed upstream characterization of the minor components present in the feedstock. For this purpose, the minor component profile of animal fat was investigated by comprehensive two-dimensional gas chromatography coupled with high-resolution time-of-flight mass spectrometry. This was preceded by an innovative sample fractionation and focalization of these minor components by a preparative liquid chromatographic column method. The overall method permitted to extract different levels of information from the two-dimensional chromatograms, leading to a tentative identification of more than 150 compounds, mainly oxygenated, belonging to different chemical classes.

Comprehensive Chemical Characterization of Chia (Salvia hispanica L.) Seed Oil with a Focus on Minor Lipid Components.

A comprehensive chemical characterization of different lipid components, namely fatty acid composition after derivatization in fatty acid methyl esters (FAMEs), triacylglycerols (TAGs), phospholipids (PLs), free fatty acids (FFAs), sterols, carotenoids, tocopherols, and polyphenols in Chia seed oil, obtained by Soxhlet extraction, was reported. Reversed phase liquid chromatography (RP-LC) coupled to UV and mass spectrometry (MS) detectors was employed for carotenoids, polyphenols, and TAGs determination; normal phase-LC in combination with fluorescence detector (FLD) was used for tocopherols analysis; PL and FFA fractions were investigated after a rapid solid phase extraction followed by RP-LC-MS and NanoLC coupled to electron ionization (EI) MS, respectively. Furthermore, gas chromatography (GC)-flame ionization (FID) and MS detectors were used for FAMEs and sterols analysis. Results demonstrated a significant content of bioactive compounds, such as the antioxidant tocopherols (22.88 µg mL-1), and a very high content of essential fatty acids (81.39%), namely α-linolenic (62.16%) and linoleic (19.23%) acids. In addition, for the best of authors knowledge, FFA profile, as well as some carotenoid classes has been elucidated for the first time. The importance of free fatty acids in vegetable matrices is related to the fact that they can be readily involved in metabolic processes or biosynthetic pathways of the plant itself. For a fast and reliable determination of this chemical class, a very innovative and sensitive NanoLC-EI-MS analytical determination was applied.

Exploring 20 eV electron impact ionization in gas chromatography-tandem mass spectrometry for the determination of estrogenic compounds.

In electron ionization mass spectrometry (MS), the generation of characteristic fragmentation patterns allows reliable and sensitive identification of compounds. However, loss or a less intense signal of the molecular ion (or more diagnostic ions) can often be observed, which can be detrimental for identification and/or sensitivity, even when MS/MS approaches are applied for quantification. The benefits of applying lower ionization energy (i.e., 20 eV compared to 70 eV) using a gas chromatography (GC) - tandem MS (MS/MS) instrument were investigated in the detection of three estrogenic compounds, namely estrone (E1), 17ß-estradiol (E2), and 17α-ethynylestradiol (EE2), emerging aquatic pollutants included in the European Commission Watch List. As expected, the relative intensity of molecular ions (M+.) or high-mass fragments closely related (M+.-CH3) increased significantly at 20 eV compared to 70 eV (from 4.6 % to 35.0 % for EE2, from 22.5 % to 87.3 % for E2, and from 76 % to 100 % for E1). This change in the spectrum profile led to an overall increase in the sensitivity of the compounds when detected using the multiple reaction monitoring mode. These results were compared with the instrumental limit of quantification obtained in liquid chromatography - MS/MS showing a limit of quantification of about 100-folds lower for GC-MS/MS and a completely neglectable matrix effect, thus posing the base for the development of a miniaturized sample preparation method (with an overall lower concentration factor) to achieve the challenging low limits of detection required by the EU regulation for estrogenic compounds.

Advanced mono- and multi-dimensional gas chromatography-mass spectrometry techniques for oxygen-containing compound characterization in biomass and biofuel samples.

A wide variety of biomass, from triglycerides to lignocellulosic-based feedstock, are among promising candidates to possibly fulfill requirements as a substitute for crude oils as primary sources of chemical energy feedstock. During the feedstock processing carried out to increase the H:C ratio of the products, heteroatom-containing compounds can promote corrosion, thus limiting and/or deactivating catalytic processes needed to transform the biomass into fuel. The use of advanced gas chromatography techniques, in particular multi-dimensional gas chromatography, both heart-cutting and comprehensive coupled to mass spectrometry, has been widely exploited in the field of petroleomics over the past 30 years and has also been successfully applied to the characterization of volatile and semi-volatile compounds during the processing of biomass feedstock. This review intends to describe advanced gas chromatography-mass spectrometry-based techniques, mainly focusing in the period 2011-early 2020. Particular emphasis has been devoted to the multi-dimensional gas chromatography-mass spectrometry techniques, for the isolation and characterization of the oxygen-containing compounds in biomass feedstock. Within this context, the most recent advances to sample preparation, derivatization, as well as gas chromatography instrumentation, mass spectrometry ionization, identification, and data handling in the biomass industry, are described.

Exhaled human breath analysis in active pulmonary tuberculosis diagnostics by comprehensive gas chromatography-mass spectrometry and chemometric techniques.

Tuberculosis (TB) is the deadliest infectious disease, and yet accurate diagnostics for the disease are unavailable for many subpopulations. In this study, we investigate the possibility of using human breath for the diagnosis of active TB among TB suspect patients, considering also several risk factors for TB for smokers and those with human immunodeficiency virus (HIV). The analysis of exhaled breath, as an alternative to sputum-dependent tests, has the potential to provide a simple, fast, non-invasive, and readily available diagnostic service that could positively change TB detection. A total of 50 individuals from a clinic in South Africa were included in this pilot study. Human breath has been investigated in the setting of active TB using the thermal desorption-comprehensive two-dimensional gas chromatography-time of flight mass spectrometry methodology and chemometric techniques. From the entire spectrum of volatile metabolites in breath, three machine learning algorithms (support vector machines, partial least squares discriminant analysis, and random forest) to select discriminatory volatile molecules that could potentially be useful for active TB diagnosis were employed. Random forest showed the best overall performance, with sensitivities of 0.82 and 1.00 and specificities of 0.92 and 0.60 in the training and test data respectively. Unsupervised analysis of the compounds implicated by these algorithms suggests that they provide important information to cluster active TB from other patients. These results suggest that developing a non-invasive diagnostic for active TB using patient breath is a potentially rich avenue of research, including among patients with HIV comorbidities.

SPME-GC×GC-TOF MS fingerprint of virally-infected cell culture: Sample preparation optimization and data processing evaluation.

Untargeted metabolomics study of volatile organic compounds produced by different cell cultures is a field that has gained increasing attention over the years. Solid-phase microextraction has been the sampling technique of choice for most of the applications mainly due to its simplicity to implement. However, a careful optimization of the analytical conditions is necessary to obtain the best performances, which are highly matrix-dependent. In this work, five different solid-phase microextraction fibers were compared for the analysis of the volatiles produced by cell culture infected with the human respiratory syncytial virus. A central composite design was applied to determine the best time-temperature combination to maximize the extraction efficiency and the salting-out effect was evaluated as well. The linearity of the optimized method, along with limits of detection and quantification and repeatability was assessed. Finally, the effect of i) different normalization techniques (i.e. z-score and probabilistic quotient normalization), ii) data transformation (i.e. in logarithmic scale), and iii) different feature selection algorithms (i.e. Fisher ratio and random forest) on the capability of discriminating between infected and not-infected cell culture was evaluated.

Preliminary investigation of human exhaled breath for tuberculosis diagnosis by multidimensional gas chromatography - Time of flight mass spectrometry and machine learning.

Tuberculosis (TB) remains a global public health malady that claims almost 1.8 million lives annually. Diagnosis of TB represents perhaps one of the most challenging aspects of tuberculosis control. Gold standards for diagnosis of active TB (culture and nucleic acid amplification) are sputum-dependent, however, in up to a third of TB cases, an adequate biological sputum sample is not readily available. The analysis of exhaled breath, as an alternative to sputum-dependent tests, has the potential to provide a simple, fast, and non-invasive, and ready-available diagnostic service that could positively change TB detection. Human breath has been evaluated in the setting of active tuberculosis using thermal desorption-comprehensive two-dimensional gas chromatography-time of flight mass spectrometry methodology. From the entire spectrum of volatile metabolites in breath, three random forest machine learning models were applied leading to the generation of a panel of 46 breath features. The twenty-two common features within each random forest model used were selected as a set that could distinguish subjects with confirmed pulmonary M. tuberculosis infection and people with other pathologies than TB.

Analysis of lipid profile in lipid storage myopathy.

Lipid dysmetabolism disease is a condition in which lipids are stored abnormally in organs and tissues throughout the body, causing muscle weakness (myopathy). Usually, the diagnosis of this disease and its characterization goes through dosage of Acyl CoA in plasma accompanied with evidence of droplets of intra-fibrils lipids in the patient muscle biopsy. However, to understand the pathophysiological mechanisms of lipid storage diseases, it is useful to identify the nature of lipids deposited in muscle fiber. In this work fatty acids and triglycerides profile of lipid accumulated in the muscle of people suffering from myopathies syndromes was characterized. In particular, the analyses were carried out on the muscle biopsy of people afflicted by lipid storage myopathy, such as multiple acyl-coenzyme A dehydrogenase deficiency, and neutral lipid storage disease with myopathy, and by the intramitochondrial lipid storage dysfunctions, such as deficiencies of carnitine palmitoyltransferase II enzyme. A single step extraction and derivatization procedure was applied to analyze fatty acids from muscle tissues by gas chromatography with a flame ionization detector and with an electronic impact mass spectrometer. Triglycerides, extracted by using n-hexane, were analyzed by high performance liquid chromatography coupled to mass spectrometer equipped with an atmospheric pressure chemical ionization interface. The most representative fatty acids in all samples were: C16:0 in the 13-24% range, C18:1n9 in the 20-52% range, and C18:2n6 in the 10-25% range. These fatty acids were part of the most representative triglycerides in all samples. The data obtained was statistically elaborated performing a principal component analysis. A satisfactory discrimination was obtained among the different diseases. Using component 1 vs component 3 a 43.3% of total variance was explained. Such results suggest the important role that lipid profile characterization can have in supporting a correct diagnosis.

Characterisation of minor components in vegetable oil by comprehensive gas chromatography with dual detection.

The profile of minor compounds, such as alcohols, sterols, free and alkyl fatty acids, waxes, etc., was investigated in different vegetable oils by a comprehensive gas chromatographic system, coupled with a simultaneous dual detection (flame ionisation detector and mass spectrometer) for quantitative and qualitative purposes. Such a system generated a unique two-dimensional chromatogram to be used as a chemical fingerprint. Multi-level information, due not only to a more "comprehensive" preparation technique, but also thanks to the exploitation of a more powerful and sensitive analytical determination allowed the extrapolation of diagnostic information from the minor components profile of different vegetable oils, along with their characteristic profile. Furthermore, an admixture of an extra virgin olive oil with a low amount of sunflower and palm oils was evaluated, attesting to the powerful diagnostic information provided by the proposed approach.

Chemical characterisation of old cabbage (Brassica oleracea L. var. acephala) seed oil by liquid chromatography and different spectroscopic detection systems.

We report an extensive chemical characterisation of fatty acids, triacylglycerols, tocopherols, carotenoids and polyphenols contained in the oil extracted from old cabbage (Brassica oleracea L. var. acephala) by cold-pressing of the seeds. Analyses were performed by GC-FID combined with mass spectrometry, HPLC with photodiode array, fluorescence and mass spectrometry detection. The 94% of the total fatty acids were unsaturated, rappresented by erucic acid (more than 50%) followed by linoleic, linolenic and oleic acids accounting for approximately 10% each. The most abundant triacylglycerols (>13%) were represented by erucic-gadolenic-linoleic, erucic-eruci-linoleic and erucic-erucic-oleic. Among tocopherols, γ-tocopherol accounted for over 70% of the total content. Thirteen carotenoids and 11 polyphenols were identified and measured. In particular, the total content in carotenoids was 10.9 ppm and all-E-lutein was the main component (7.7 ppm); among polyphenols, six hydroxycinnamic acids and five flavonoids, were identified by combining information from retention times, PDA and MS data.

Improvement of mineral oil saturated and aromatic hydrocarbons determination in edible oil by liquid-liquid-gas chromatography with dual detection.

Mineral oils, which are mainly composed of saturated hydrocarbons and aromatic hydrocarbons, are widespread food contaminants. Liquid chromatography coupled to gas chromatography with flame ionization detection represents the method of choice to determine these two families. However, despite the high selectivity of this technique, the presence of olefins (particularly squalene and its isomers) in some samples as in olive oils, does not allow the correct quantification of the mineral oil aromatic hydrocarbons fraction, requiring additional off-line tools to eliminate them. In the present research, a novel on-line liquid chromatography coupled to gas chromatography method is described for the determination of hydrocarbon contamination in edible oils. Two different liquid chromatography columns, namely a silica one (to retain the bulk of the matrix) and a silver-ion one (which better retains the olefins), were coupled in series to obtain the mineral oil aromatic hydrocarbons hump free of interfering peaks. Furthermore, the use of a simultaneous dual detection, flame ionization detector and triple quadrupole mass spectrometer allowed us not only to quantify the mineral oil contamination, but also to evaluate the presence of specific markers (i.e. hopanes) to confirm the petrogenic origin of the contamination.

Reliability of the ΔECN42 limit and global method for extra virgin olive oil purity assessment using different analytical approaches.

Two data elaboration approaches for evaluating olive oils authenticity were compared: (I) determination of the difference between the theoretical and actual amounts of triacylglycerols with partition number 42 (ΔECN42 ⩽ |0.2|); and (II) the global method, which considers also partition numbers 44 and 46 (returning a "correct"/"not correct" result). Analysis of 31 genuine extra virgin olive oil samples was performed using different analytical methods, namely liquid chromatography (LC) coupled with a refractive index detector (RID) and LC coupled with a mass spectrometry (MS), and the results compared. Several false positives were highlighted using the ΔECN42 limit with both instrumental approaches. The global method algorithm returned "correct" results for all the samples analysed (except two that gave no results) with LC-MS; on the other hand, 10 false positives were obtained elaborating data deriving from NARP-LC-RID analysis.

Non-polar lipids characterization of Quinoa (Chenopodium quinoa) seed by comprehensive two-dimensional gas chromatography with flame ionization/mass spectrometry detection and non-aqueous reversed-phase liquid chromatography with atmospheric pressure chemical ionization mass spectrometry detection.

A chemical characterization of major lipid components, namely, triacylglycerols, fatty acids and the unsaponifiable fraction, in a Quinoa seed lipids sample is reported. To tackle such a task, non-aqueous reversed-phase high-performance liquid chromatography with mass spectrometry detection was employed. The latter was interfaced with atmospheric pressure chemical ionization for the analysis of triacylglycerols. The main triacylglycerols (>10%) were represented by OLP, OOL and OLL (P = palmitoyl, O = oleoyl, L = linoleoyl); the latter was present in the oil sample at the highest percentage (18.1%). Furthermore, fatty acid methyl esters were evaluated by gas chromatography with flame ionization detection. 89% of the total fatty acids was represented by unsaturated fatty acid methyl esters with the greatest percentage represented by linoleic and oleic acids accounting for approximately 48 and 28%, respectively. An extensive characterization of the unsaponifiable fraction of Quinoa seed lipids was performed for the first time, by using comprehensive two-dimensional gas chromatography with dual mass spectrometry/flame ionization detection. Overall, 66 compounds of the unsaponifiable fraction were tentatively identified, many constituents of which (particularly sterols) were confirmed by using gas chromatography with high-resolution time-of-flight mass spectrometry.

Determination of the triacylglycerol fraction in fish oil by comprehensive liquid chromatography techniques with the support of gas chromatography and mass spectrometry data.

Fish oil made from menhaden (Brevoortia tyrannus) can be used as a dietary supplement for the presence of high levels of the long-chained omega-3 fatty acids, viz. epentaenoic and docosahexanoic. In this work, for the first time, two different multidimensional approaches were developed and compared, in terms of peak capacity, for triacylglycerol characterization. In particular, silver ion chromatography with a silver-ion column and non-aqueous reverse-phase liquid chromatography with a C18 column were tested in both comprehensive (stop-flow) and off-line modes. The use of mass spectra attained by atmospheric pressure chemical ionization for both LC approaches, and the fatty acids methyl esters profile of menhaden oil obtained by gas chromatography analysis, greatly supported the elucidation of the triacylglycerol content in menhaden oil. The off-line approach afforded a better separation and, thus, higher peak capacity to allow identifying and semiquantifying more than 250 triacylglycerols. Such a huge number has never been reported for a menhaden oil sample.The main disadvantage of such an approach over the stop-flow one was the longer analysis time, mainly attributable to solvent exchange between the two dimensions.

Fingerprinting of vegetable oil minor components by multidimensional comprehensive gas chromatography with dual detection.

The potentiality of a multidimensional comprehensive gas chromatographic (GC × GC) method, employing a simultaneous dual detection (FID and mass spectrometer), to generate peculiar two-dimensional chromatograms to be used as a chemical fingerprint, was investigated to characterize minor compounds in edible oil, particularly olive oil. The best column combination for this application was investigated comparing two column sets (orthogonal or reverse-type), equivalent in terms of theoretical plate number, but differing in stationary phase combination. The apolar × mid-polar set gave a superior separation power, thus was used for further characterization. Different levels of information were extrapolated from the two-dimensional chromatogram. Using the FID, reliable quantification of the alkyl esters fatty acids and waxes was obtained, comparable to the results obtained using the official method, as required by the European legislation. However, thanks to a slight modification of the sample preparation method, the increased separation power obtained using the GC × GC method, and the support of the mass spectrometer detector, further diagnostic information was extrapolated considering the free sterol and tocopherol fractions. In particular, the profiles of extra virgin olive oil samples were compared with a hazelnut oil sample, highlighting that the latter was characterized by a larger number of compounds, completely absent in the extra virgin olive oil samples, which can be used to detect illegal admixtures.

Analysis of human plasma lipids by using comprehensive two-dimensional gas chromatography with dual detection and with the support of high-resolution time-of-flight mass spectrometry for structural elucidation.

The main focus of the present research is the analysis of the unsaponifiable lipid fraction of human plasma by using data derived from comprehensive two-dimensional gas chromatography with dual quadrupole mass spectrometry and flame ionization detection. This approach enabled us to attain both mass spectral information and analyte percentage data. Furthermore, gas chromatography coupled with high-resolution time-of-flight mass spectrometry was used to increase the reliability of identification of several unsaponifiable lipid constituents. The synergism between both the high-resolution gas chromatography and mass spectrometry processes enabled us to attain a more in-depth knowledge of the unsaponifiable fraction of human plasma. Additionally, information was attained on the fatty acid and triacylglycerol composition of the plasma samples, subjected to investigation by using comprehensive two-dimensional gas chromatography with dual quadrupole mass spectrometry and flame ionization detection and high-performance liquid chromatography with atmospheric pressure chemical ionization quadrupole mass spectrometry, respectively.

Mass spectrometric elucidation of triacylglycerol content of Brevoortia tyrannus (menhaden) oil using non-aqueous reversed-phase liquid chromatography under ultra high pressure conditions.

A non-aqueous reversed phase high performance liquid chromatography method was developed, and optimized for triacylglycerol analysis in a Brevoortia tyrannus (menhaden) oil sample. Four columns were serially coupled to tackle such a task, for a total length of 60 cm of shell-packed stationary phase, and operated under ultra high pressure conditions. As detection, positive-ion atmospheric pressure chemical ionization mass spectrometry was used to attain identification of the analyzed sample components. A number of 137 triacylglycerols containing up to 19 fatty acids, with 14-22 carbon atom alkyl chain length and 0-6 double bonds, were positively identified in the complex lipidic sample. This is the first work that reports an extensive characterization of the triacylglycerol fraction of menhaden oil.