Quantitative Analysis Of Fatty Acids Research Articles

Investigating the amount of TFAs, CLAs and ω6/ω3 of various brands of corn oil heated at different time-temperature treatments.

A qualitative and quantitative analysis of trans fatty acids (TFAs), conjugated linoleic acids (CLAs) and linoleic acid/α-linolenic acid (ω6/ω3) in eleven different brands of fresh and heated corn oil was investigated. Corn oil was subjected to thermal treatment at 180, 210 and 230°C for 4, 8 and 12h. The kinds of fatty acids were almost the same in all eleven brands of corn oil, but there were differences in the quantities of TFAs, CLAs and ω6/ω3 among the brands. The formation of TFAs in different brands of corn oil increased slowly at 180°C and rapidly at 230°C with the maximum increase of 28 and 374%, respectively. The formation of CLAs also increased during the heating with the amount of 0.00-0.32g/100g in fresh corn oil and 0.71-1.12g/100g in corn oil heated at 230°C for 12h. The value of ω6/ω3 was high in corn oil and increased with heating temperature and time. The largest rise of ω6/ω3 can reach upwards of 239:1. The results showed that temperature control was essential for maintaining the quality and nutritional value of corn oil.

High-Throughput Quantitative Lipidomics Analysis of Nonesterified Fatty Acids in Plasma by LC-MS.

Nonesterified fatty acids are important biological molecules which have multiple functions such as energy storage, gene regulation, or cell signaling. Comprehensive profiling of nonesterified fatty acids in biofluids can facilitate studying and understanding their roles in biological systems. For these reasons, we have developed and validated a high-throughput, nontargeted lipidomics method coupling liquid chromatography to high-resolution mass spectrometry for quantitative analysis of nonesterified fatty acids. Sufficient chromatographic separation is achieved to separate positional isomers such as polyunsaturated and branched-chain species and quantify a wide range of nonesterified fatty acids in human plasma samples. However, this method is not limited only to these fatty acid species and offers the possibility to perform untargeted screening of additional nonesterified fatty acid species.

Fast Fatty Acid Analysis by Core-Shell Reversed-Phase Liquid Chromatography Coupled to Evaporative Light-Scattering Detector

A high-performance liquid chromatography analysis method with an evaporative light-scattering detector has been developed for the separation and quantitative analysis of free fatty acids in biological matrices. Core-shell reversed-phase high-performance liquid chromatography separation of 10 free fatty acids is achieved within 10.5 min using a methanol/water (0.05% trifluoroacetic acid) eluent gradient. After optimization, the drift tube and nebulization temperature of the evaporative light-scattering detector was set at 35°C, nitrogen flow-rate at 1.1 standard liter per minute and column temperature at 25°C. All calibration curves showed good regression (r2 > 0.9975). A validation procedure following the International Conference on Harmonisation guidelines was implemented to certify the method. Relative standard deviations did not exceed 1.5% and 4.25% for repeatability and reproducibility respectively.

Near Infrared Spectrometry of Clinically Significant Fatty Acids Using Multicomponent Regression

We have developed methods for determining the content of clinically important fatty acids (FAs), primarily saturated palmitic acid, monounsaturated oleic acid, and the sum of polyenoic fatty acids (eicosapentaenoic + docosahexaenoic), in oily media (food products and supplements, fish oils) using different types of near infrared (NIR) spectrometers: Fourier-transform, linear photodiode array, and Raman. Based on a calibration method (regression) by means of projections to latent structures, using standard samples of oil and fat mixtures, we have confirmed the feasibility of reliable and selective quantitative analysis of the above-indicated fatty acids. As a result of comparing the calibration models for Fourier-transform spectrometers in different parts of the NIR range (based on different overtones and combinations of fatty acid absorption), we have provided a basis for selection of the spectral range for a portable linear InGaAs-photodiode array spectrometer. In testing the calibrations of a linear InGaAs-photodiode array spectrometer which is a prototype for a portable instrument, for palmitic and oleic acids and also the sum of the polyenoic fatty acids we have achieved a multiple correlation coefficient of 0.89, 0.85, and 0.96 and a standard error of 0.53%, 1.43%, and 0.39% respectively. We have confirmed the feasibility of using Raman spectra to determine the content of the above-indicated fatty acids in media where water is present.

High-Throughput Quantitative Lipidomics Analysis of Nonesterified Fatty Acids in Human Plasma.

We present a high-throughput, nontargeted lipidomics approach using liquid chromatography coupled to high-resolution mass spectrometry for quantitative analysis of nonesterified fatty acids. We applied this method to screen a wide range of fatty acids from medium-chain to very long-chain (8 to 24 carbon atoms) in human plasma samples. The method enables us to chromatographically separate branched-chain species from their straight-chain isomers as well as separate biologically important ω-3 and ω-6 polyunsaturated fatty acids. We used 51 fatty acid species to demonstrate the quantitative capability of this method with quantification limits in the nanomolar range; however, this method is not limited only to these fatty acid species. High-throughput sample preparation was developed and carried out on a robotic platform that allows extraction of 96 samples simultaneously within 3 h. This high-throughput platform was used to assess the influence of different types of human plasma collection and preparation on the nonesterified fatty acid profile of healthy donors. Use of the anticoagulants EDTA and heparin has been compared with simple clotting, and only limited changes have been detected in most nonesterified fatty acid concentrations.

Simple Methodology for the Quantitative Analysis of Fatty Acids in Human Red Blood Cells

In the last years, there has been an increasing interest in evaluating possible relations between fatty acid (FA) patterns and the risk for chronic diseases. Due to the long life span (120 days) of red blood cells (RBCs), their FA profile reflects a longer term dietary intake and was recently suggested to be used as an appropriate biomarker to investigate correlations between FA metabolism and diseases. Therefore, the aim of this work was to develop and validate a simple and fast methodology for the quantification of a broad range of FAs in RBCs using gas chromatography with flame ionization detector, as a more common and affordable equipment suitable for biomedical and nutritional studies including a large number of samples. For this purpose, different sample preparation protocols were tested and compared, including a classic two-step method (Folch method) with modifications and different one-step methods, in which lipid extraction and derivatization were performed simultaneously. For the one-step methods, different methylation periods and the inclusion of a saponification reaction were evaluated. Differences in absolute FA concentrations were observed among the tested methods, in particular for some metabolically relevant FAs such as trans elaidic acid and eicosapentaenoic acid. The one-step method with saponification and 60 min of methylation time was selected since it allowed the identification of a higher number of FAs, and was further submitted to in-house validation. The proposed methodology provides a simple, fast and accurate tool to quantitatively analyse FAs in human RBCs, useful for clinical and nutritional studies.

Contenido de ácidos grasos en carne de bovinos cebados en diferentes sistemas de producción en el trópico colombiano

Background: Meat is a highly nutritious food and a major component of the basic food basket. Consumers select foods not only for taste and satisfaction, but also for their effects on human nutrition and health. In addition there is very little knowledge about the quality of meat produced in the Colombian systems and their fatty acid content which undoubtedly affect human health. Objetive: To determine the content and composition of fatty acids in meat from steers managed in different Colombian systems. Methods: Meat (Longissimus dorsi) samples were obtained from Zebu cattle from four production systems inthe Colombian tropics: Two silvopastoral arrangements (Cotove Research Center, National University of Colombia and Montenegro, Quindio), improved pastures (Montenegro, Quindio) and a traditional grazing system (Monteria, Cordoba). Fat was extracted from 64 meat samples in the laboratory of Analytical Chemistry of the National University of Colombia, and a quantitative analysis of fatty acids was performed by gas chromatography in the laboratory of Instrumental Analysis of the National University of Colombia in Medellin. The results were evaluated following a complete randomized design and means were separated by the Duncan test. Results: There were significant differences in the content of myristic and palmitic acids between meat from the traditional and improved pasture system, with both acids being higher in the traditional system. There were also differences in the content of linoleic and linolenic acid, being lower in the Cotove silvopastoral system. There were significant differences in the content of unsaturated acids, with the meat from silvopastoral system Quindio and the improved pastures showing the highest fatty acid saturated: polyunsaturated ratio (0.19). In the linoleic: α-linolenic ratio there were no differences. Conclusions: Although the composition and the fatty acid content in beef varied between systems evaluated, the main components of such systems fat were palmitic acid and oleic acid (30.73 and 35.62 g/100 g fatty acid).

Development and validation of a GC–FID method for quantitative analysis of oleic acid and related fatty acids

Oleic acid is a common pharmaceutical excipient that has been widely used in various dosage forms. Gas chromatography (GC) has often been used as the quantitation method for fatty acids normally requiring a derivatization step. The aim of this study was to develop a simple, robust, and derivatization-free GC method that is suitable for routine analysis of all the major components in oleic acid USP-NF (United States Pharmacopeia-National Formulary) material. A gas chromatography–flame ionization detection (GC–FID) method was developed for direct quantitative analysis of oleic acid and related fatty acids in oleic acid USP-NF material. Fifteen fatty acids were separated using a DB-FFAP (nitroterephthalic acid modified polyethylene glycol) capillary GC column (30m×0.32mm i.d.) with a total run time of 20min. The method was validated in terms of specificity, linearity, precision, accuracy, sensitivity, and robustness. The method can be routinely used for the purpose of oleic acid USP-NF material analysis.

Trophic relationships in an estuarine environment: A quantitative fatty acid analysis signature approach

In order to better understand the functioning of aquatic environments, it is necessary to obtain accurate diet estimations in food webs. Their description should incorporate information about energy flow and the relative importance of trophic pathways. Fatty acids have been extensively used in qualitative studies on trophic relationships in food webs. Recently a new method to estimate quantitatively single predator diet has been developed. In this study, a model of aquatic food web through quantitative fatty acid signature analysis was generated to identify the trophic interactions among the species in the Rocha Lagoon. The biological sampling over two consecutive annual periods was comprehensive enough to identify all functional groups in the aquatic food web (except birds and mammals). Heleobia australis seemed to play a central role in this estuarine ecosystem. As both, a grazer and a prey to several other species, probably H. australis is transferring a great amount of energy to upper trophic levels. Most of the species at Rocha Lagoon have a wide range of prey items in their diet reflecting a complex food web, which is characteristic of extremely dynamic environment as estuarine ecosystems. QFASA is a model in tracing and quantitative estimate trophic pathways among species in an estuarine food web. The results obtained in the present work are a valuable contribution in the understanding of trophic relationships in Rocha Lagoon.

Quantitative Analysis of Fatty Acids from Safflower by Selective and Sensitive Pre-Column Derivatization Method Using HPLC-FLD and Online APCI/MS

Quantitative Analysis of Fatty Acids from Safflower by Selective and Sensitive Pre-Column Derivatization Method Using HPLC-FLD and Online APCI/MS

Application of solid phase microextraction for quantitation of polyunsaturated fatty acids in biological fluids.

Development of a straightforward strategy for simultaneous quantitative analysis of nonesterified fatty acids (NEFA) species in biofluids is a challenging task because of the extreme complexity of fatty acid distribution in biological matrices. In this study, we present a direct immersion solid phase microextraction method coupled to a liquid chromatography-mass spectrometry platform (DI-SPME- HPLC-ESI -MS) for determination of unconjugated fatty acids (FA) in fish and human plasma. The proposed method was fully validated according to bioanalytical method validation guidelines. The LOD and LOQ were in the range of 0.5-2 and 5-12 ng/mL, respectively, with a linear dynamic range of 100 fold for each compound. Absolute and relative matrix effects were comprehensively evaluated and found to be in the acceptable range of 91-116%. The affinity constant (Ka) of individual FAs to protein albumin was determined to be 9.2 × 10(4) to 4.3 × 10(5) M(-1). The plasma protein binding (PPB%) was calculated and found to be in the range of 98.0-99.7% for different polyunsaturated fatty acids (PUFAs). The PUFAs under study were found at a high concentration range in fish plasma, whereas only a few were within quantification range in control human plasma. The method was successfully applied for monitoring PUFA changes during the operation in plasma samples obtained from patients undergoing cardiac surgery with the use of cardiopulmonary bypass (CPB). The most significant contribution induced by surgery was noticed in the concentration level of α-linolenic acid (18:3, ALA), arachidonic acid (20:4, AA), and docosahexanoic acid (22:6, DHA) soon after administration of CPB in all cases.

The early life gut microbiota and atopic disease

Background Asthma is the most prevalent of all childhood diseases and accounts for the majority of hospitalizations and school absences in children [1]. Current mouse model research has identified the early life gut microbiota as a potential therapeutic target for the prevention of asthma and atopic diseases [2-4]. We hypothesize that the early life gut microbiota could play a similar preventative role against atopic disease development in humans. Methods 1262 children enrolled in the Canadian Healthy Infant Longitudinal Development (CHILD) Study with complete skin prick test and wheeze data at one year were grouped into four clinically relevant phenotypes: atopy + wheeze, atopy only, wheeze only, and control. Bacterial 16S rDNA from 3-month and 1-year stool samples of 319 children in these four phenotypes was extracted, amplified, and subjected to high throughput Illumina sequencing. Quantitative polymerase chain reaction (qPCR) and short chain fatty acid (SCFA) analysis were also conducted on 44 children in the two extreme phenotypes (atopy + wheeze vs. control). Results 16S sequence analysis of our sample cohort (319 subjects) identified bacterial populations that differed in abundance in the atopy + wheeze group at 3-months of age but not at 1-year of age. Additionally, significant changes in the abundance of certain bacterial genera were found in the atopy + wheeze group when compared to controls by qPCR at 3-months of age only. Changes in stool short chain fatty acid production between the atopy + wheeze group and the control group were also observed at 3months of age only. Conclusions Shifts in the relative abundance of certain gut bacterial populations and differences in the levels of stool SCFAs before 3-months of age are associated with atopy and wheeze at one year of age.

Diet estimation of Paralichthys orbignyanus in a coastal lagoon via quantitative fatty acid signature analysis

Quantitative fatty acid analysis (QFASA) is a statistical model designed to quantitatively estimate predator diets using fatty acid (FA) signatures among the predator and its potential prey. QFASA estimated the diet of a migratory flatfish Paralichthys orbignyanus over its fattening stage in the Rocha lagoon (a semi-closed estuary) where all prey available to this top predator species are well known. A 20-week controlled feeding trial obtained calibration coefficients (CC) for P. orbignyanus fed two types of prey (silverside and menhaden). Several subsets of FA were tested in order to elucidate which is the most suitable for applying QFASA to this species. QFASA was applied to all CC and FA subsets to validate the model. The model predicts better the consumed diet with silverside CC than with menhaden CC. The subset which best adjusts the diet over the validation process, includes approximately 34% of total FA, containing mainly dietary FA. The diet estimation in nature for P. orbignyanus varied according to whether the model is applied with or without CC. When the diet was estimated without CC, results were similar to those based on stomach content analysis (reported in previous studies); it fed mainly on silverside (~88%), but also some minor soft-body species that are only evident using this kind of methodology (QFASA). When the diet was estimated with silverside CC, a higher presence of silverside (~97%) was observed. These results seem to indicate a tendency to overestimate the presence of the item used as prey for CC calculations.

Quantitative analysis of free fatty acids in human serum using biexciton auger recombination in cadmium telluride nanoparticles loaded on zeolite.

Nanometer-sized semiconductor particles (CdTe) were used as an inorganic matrix for the laser desorption/ionization mass spectrometry of fatty acids. The excitation power dependence of the peak intensity of stearic acid (Ste), [Ste - H](-), was observed, and it was proportional to the square of the excitation power. The peak intensity of [Ste - H](-) decreased by addition of a hole scavenger (KSCN). It was understood that the ionization of fatty acids were due to the biexciton Auger recombination and electron ejection from CdTe. CdTe were then loaded on zeolite surface. The peak intensity enhancement of the deprotonated ion of fatty acid were observed. This phenomenon was explained by measuring the carrier lifetime for Auger recombination in CdTe. In addition, reproducibility of fatty acid ions was highly improved reflecting homogeneous distribution of CdTe on zeolite surface. CdTe/HM20 was successfully applied to the quantitative analysis of Ste in human serum by isotope dilution using (13)C18-Ste. The concentration of Ste in human serum samples was estimated to be 76.62 mg/kg with the standard deviation (SD) of 2.37 mg/kg.

Foraging diversity in lactating New Zealand sea lions: insights from qualitative and quantitative fatty acid analysis

Lactating New Zealand (NZ) sea lions (Phocarctos hookeri) exhibit different foraging patterns during their foraging trips, with benthic divers spending more energy at sea than mesopelagic conspecifics. We compared blubber fatty acids (FAs) of 14 benthic and 12 mesopelagic females captured at the Auckland Islands, NZ subantarctic, in late January 2000 using an analysis of similarities (ANOSIM). FA profiles between foraging types were significantly different (global R = 0.30, p = 0.001), suggesting a different use in prey resources. We then compared the diet predictions by quantitative FA signature analysis (QFASA) by using a prey FA library available in the region. Overall, diet predictions were significantly distinct between benthic and mesopelagic females (global R = 0.17, p = 0.022), although the diets consisted of the same prey but in different contributions. The results suggest benthic females do not compensate their higher foraging costs by feeding on prey with higher energy densities. Foraging areas of benthic females are not exploited by the trawling fishery; therefore, the benthic tactic might be a trade-off between a higher foraging cost and less resource competition.

A Rapid Multidimensional GC–Flame-Ionization Detector Method for Determination of Fatty Acid Methyl Esters

Fatty acids play important roles in the pathogenesis of many diseases. In order to investigate a wide spectrum of fatty acids in biological matrix, we developed an approach for quantitative analysis of fatty acids based on a heart-cutting 2D gas chromatographic-flame ionization detector system. This system uses a Dean Switch interface between the primary polar and secondary nonpolar capillary column. Most of the standards were separated by the primary column. Unresolved components are selectively transferred to the secondary column, where they are completely separated. This method was further evaluated using plasma obtained from 11 healthy and 11 chronic coronary artery disease patients. Our results prove that this approach is sensitive, precise and specific, capable of measuring 37 specific fatty acids. It indicates that this approach offers a reliable and sensitive technical platform for comprehensive quantification of fatty acids.

High-throughput and high-sensitivity quantitative analysis of serum unsaturated fatty acids by chip-based nanoelectrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry: Early stage diagnostic biomarkers of pancreatic cancer

In this study, Fourier transform ion cyclotron resonance mass spectrometry (FTICR MS) coupled with chip-based direct-infusion nanoelectrospray ionization source (CBDInanoESI) in a negative ion mode is first employed to evaluate the effect of serum and its corresponding supernatant matrixes on the recoveries of serum free fatty acids (FFAs) based on spike-and-recovery experimental strategy by adding analytes along with analog internal standard (IS). The recoveries between serum (69.8-115.6%) and the supernatant (73.6-99.0%) matrixes are almost identical. Multiple point internal standard calibration curves between the concentration ratios of individual fatty acids to ISs, (C(17:1) as IS of C(16:1), C(18:3), C(18:2), or C(18:1) or C(21:0) as IS of C(20:4) or C(22:6)) versus their corresponding intensity ratios were constructed for C(16:1), C(18:3), C(18:2), C(18:1), C(20:4) and C(22:6), respectively, with correlation coefficients of greater than 0.99, lower limits of detection between 0.3 and 1.8 nM, and intra- and inter-day precision (relative standard deviations <18%), along with the linear dynamic range of three orders of magnitude. Sequentially, this advanced analytical platform was applied to perform simultaneous quantitative and qualitative analysis of multiple targets, e.g., serum supernatant unsaturated FFAs from 361 participants including 95 patients with pancreatic cancer (PC), 61 patients with pancreatitis and 205 healthy controls. Experimental results indicate that the levels of C(18:1), C(18:2), C(18:3), C(20:4) and C(22:6), as well as the level ratios of C(18:2)/C(18:1) and C(18:3)/C(18:1) of the PC patients were significantly decreased compared with those of healthy controls and the patients with pancreatitis (p < 0.01). It is worth noting that the ratio of C(18:2)/C(18:1), polyunsaturated fatty acids (PUFAs) (C(18:2), C(18:3), C(20:4), and C(22:6)), panel a (C(16:1), C(18:3), C(18:2), C(20:4) and C(22:6)) and panel b (C(18:2)/C(18:1) and C(18:3)/C(18:1)) performed excellent diagnostic ability, with an area under the receiver operating characteristic curve of ≥0.869, sensitivity of ≥85.7%, and specificity of ≥86.7% for differentiating the early stage PC from non-cancer subjects, which are greatly higher than those of clinically used serum biomarker CA 19-9. More importantly, this platform can also provide a fast and easy way to quantify the levels of FFAs in less than 30 s per sample.

LC/ESI-MS/MS detection of FAs by charge reversal derivatization with more than four orders of magnitude improvement in sensitivity

Quantitative analysis of fatty acids (FAs) is an important area of analytical biochemistry. Ultra high sensitivity FA analysis usually is done with gas chromatography of pentafluorobenzyl esters coupled to an electron-capture detector. With the popularity of electrospray ionization (ESI) mass spectrometers coupled to liquid chromatography, it would be convenient to develop a method for ultra high sensitivity FA detection using this equipment. Although FAs can be analyzed by ESI in negative ion mode, this method is not very sensitive. In this study, we demonstrate a new method of FA analysis based on conversion of the carboxylic acid to an amide bearing a permanent positive charge, N-(4-aminomethylphenyl)pyridinium (AMPP) combined with analysis on a reverse-phase liquid chromatography column coupled to an ESI mass spectrometer operating in positive ion mode. This leads to an ∼60,000-fold increase in sensitivity compared with the same method carried out with underivatized FAs. The new method is about 10-fold more sensitive than the existing method of gas chromatography/electron-capture mass spectrometry of FA pentafluorobenzyl esters. Furthermore, significant fragmentation of the precursor ions in the nontag portion improves analytical specificity. We show that a large number of FA molecular species can be analyzed with this method in complex biological samples such as mouse serum.

Sebaceous Lipid Profiling of Bat Integumentary Tissues: Quantitative Analysis of Free Fatty Acids, Monoacylglycerides, Squalene, and Sterols

White-nose syndrome (WNS) is a fungal disease caused by Pseudogymnoascus destructans and is devastating North American bat populations. Sebaceous lipids secreted from host integumentary tissues are implicated in the initial attachment and recognition of host tissues by pathogenic fungi. We are interested in determining if ratios of lipid classes in sebum can be used as biomarkers to diagnose severity of fungal infection in bats. To first establish lipid compositions in bats, we isolated secreted and integral lipid fractions from the hair and wing tissues of three species: big brown bats (Eptesicus fuscus), Eastern red bats (Lasiurus borealis), and evening bats (Nycticeius humeralis). Sterols, FFAs, MAGs, and squalene were derivatized as trimethylsilyl esters, separated by gas chromatography, and identified by mass spectrometry. Ratios of sterol to squalene in different tissues were determined, and cholesterol as a disease biomarker was assessed. Free sterol was the dominant lipid class of bat integument. Squalene/sterol ratio is highest in wing sebum. Secreted wing lipid contained higher proportions of saturated FFAs and MAGs than integral wing or secreted hair lipid. These compounds are targets for investigating responses of P. destructans to specific host lipid compounds and as biomarkers to diagnose WNS.

Determination of volatile fatty acids in wastewater by solvent extraction and gas chromatography

The purpose of this study was to develop a liquid–liquid extraction method for the analysis of volatile fatty acids collected at the elutriation units of Unit 3, 4 and 5 at Johannesburg Water-Northern Works Wastewater Treatment Plant. Liquid–liquid extraction (LLE) method employing dichloromethane (DCM) and methyl-tert-butyl-ether (MTBE) as extracting solvents was used during the quantitative analysis of volatile fatty acids namely acetic, propionic, butyric, isobutyric, valeric, isovaleric and heptanoic acid. The detection of the extracts was by gas chromatography coupled to a mass spectrometer operating under electron ionization mode (GC–EI–MS). The results showed that MTBE was a better extraction solvent than DCM as it gave much higher recoveries (>5 folds). On the other hand, the overall reactor performance for all the three units in the period when the samples were collected, which was measured by the ratio of propionic to acetic acid was good since the ratio o did not exceed 1.4 with the exception of the samples collected on the 3rd of October where the ratio exceeded 1.4 significantly. The concentration of acetic acid, another indicator for the reactor performance in all three units was way below 800mg/L thus the digester balance was on par.