Quantitative Analysis Of Fatty Acids Research Articles

Omega-3 Long-Chain Polyunsaturated Fatty Acids in Nonseafood and Estimated Intake in the USA: Quantitative Analysis by Covalent Adduct Chemical Ionization Mass Spectrometry.

Omega-3 long-chain polyunsaturated fatty acids (LCPUFA) play critical roles in human development and health. Their intake is often effectively estimated solely based on seafood consumption, though the high intake of terrestrial animal-based foods with minor amounts of LCPUFA may be significant. Covalent adduct chemical ionization (CACI) tandem mass spectrometry is one approach for de novo structural and quantitative analysis of minor unsaturated fatty acids (FA), for which standards are unavailable. Here, CACI-MS and MS/MS are used to identify and quantify minor omega-3 LCPUFA of terrestrial animal foods based on the application of measured response factors (RFs) to various FA. American mean intakes of pork, beef, chicken, and eggs contribute 20, 27, 45, and 71 mg/day of docosahexaenoic acid (DHA), respectively. The estimated intake of omega-3 DHA, eicosapentaenoic acid, and docosapentaenoic acid from nonseafood sources is significant, at 164, 103, and 330 mg/day, greater than most existing estimates of omega-3 LCPUFA intake.

Impact of FADS genotype on polyunsaturated fatty acid content in human milk extracellular vesicles: A genetic association study.

Extracellular vesicles in human milk are critical in supporting newborn growth and development. Bioavailability of dietary extracellular vesicles may depend on the composition of membrane lipids. Single-nucleotide polymorphisms (SNPs) in the fatty acid desaturase gene cluster impact the content of long-chain polyunsaturated fatty acids in human milk phospholipids. This study investigated the relation between variation in FADS1 and FADS2 with the content of polyunsaturated fatty acids in extracellular vesicles from human milk. Milk was obtained from a cohort of mothers (N = 70) at 2-4 weeks of lactation. SNPs in the FADS gene locus were determined using pyrosequencing for rs174546 in FADS1 and rs174575 in FADS2. Quantitative lipidomic analysis of polyunsaturated fatty acids in human milk and extracellular vesicles from human milk was completed by gas chromatography-mass spectrometry. The rs174546 and rs174575 genotypes were independent predictors of the arachidonic acid content in extracellular vesicles. The rs174546 genotype also predicted eicosapentaenoic acid and docosahexaenoic acid in extracellular vesicles. The reduced content of long-chain polyunsaturated fatty acids in extracellular vesicles in human milk may be due to lower fatty acid desaturase activity in mothers who are carriers of the A allele in rs174546 or the G allele in rs174575. The polyunsaturated fatty acid composition of milk extracellular vesicles is predicted by the FADS genotype. These findings yield novel insights regarding extracellular vesicle content and composition that can inform the design of future research to explore how lipid metabolites impact the bioavailability of human milk extracellular vesicles.

Development and validation of a gas chromatography method for quantitative analysis of fatty acids in vegetable oils

Development and validation of a gas chromatography method for quantitative analysis of fatty acids in vegetable oils

Evaluation of Lipids for the Study of Photosynthetic Membranes.

The biological role of lipids goes far beyond the formation of a structural membrane bilayer platform for membrane proteins and controlling fluxes across the membranes. For example, in photosynthetic thylakoid membranes, lipids occupy well-defined binding niches within protein complexes and determine the structural organization of membrane proteins and their function by controlling generic physicochemical membrane properties. In this chapter, two-dimensional thin-layer chromatography (2D TLC) and gas chromatography (GC) techniques are presented for quantitative analysis of lipid classes and fatty acids in thylakoid membranes. In addition, lipid extraction methods from isolated thylakoid membranes and leaves are described together with a procedure for the derivatization of fatty acids to fatty acid methyl esters (FAME) that is required for GC analysis.

Chemical markers in marine food web: A simple workflow based on methyl tert-butyl ether extraction for fatty acids and stable isotopes assessment in plankton samples

Fatty acids (FAs) are used, often in combination with stable isotopes (SIs), as chemical biomarkers to assess the contribution of different prey to the diet of consumers and define food web structure and dynamics. Extraction of lipids is traditionally carried out using methanol (MeOH) combined with chloroform or dichloromethane, these latter being well-known environmental pollutant and potential carcinogenic agents. Recently, extraction protocols based on methyl tert-butyl ether (MTBE) and MeOH have been proposed as an alternative to halogenated solvents in lipidomic studies. However, no specific investigation has been performed to assess MTBE suitability in marine ecological studies including FA analysis together with SI measurements. We used an analytical workflow for qualitative and quantitative analysis of FAs and SIs in field samples of phytoplankton, zooplankton and the scyphomedusa Pelagia noctiluca, applying MTBE in comparison with chloroform- and dichloromethane-based protocols for total lipid extraction. Our analysis suggested that MTBE is a reliable substitute for lipid extraction in trophic ecology studies in marine planktonic organisms.

Development and validation of a HPLC-MS/MS method for the analysis of fatty acids - in the form of FAME ammonium adducts - in human whole blood and erythrocytes to determine omega-3 index

Recent scientific studies in the field of health and nutrition have unanimously affirmed the importance of consuming the omega-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), because of their cardioprotective properties. Fatty acid profiling in erythrocyte membranes allows the omega-3 index, which is a recognized indicator of the risk of developing cardiovascular disease, to be calculated. One consequence of the upward trend in healthy lifestyles and longevity is an increase in the number of studies into the omega-3 index, which requires a reliable method for the quantitative analysis of fatty acids. This article describes the development and validation of a sensitive and reproducible liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method for the quantitative analysis of 23 fatty acids (in the form of fatty acid methyl esters, FAMEs) in 40 µl of whole blood and erythrocytes. The list of acids includes saturated, omega-9 unsaturated, omega-6 unsaturated and omega-3 unsaturated fatty acids as well as their trans-isomers. The limit of quantitation was 250 ng ml−1 for C12:0, C16:0 and C18:0; and 62.5 ng ml−1 for other FAMEs, including EPA, DHA and trans-isomers of FAME C16:1, C18:1 and C18:2 n-6. Sample preparation for fatty acid (FA) esterification/methylation with boron trifluoride-methanol (BF3) has been optimized. Chromatographic separation has been carried out on a C8 column in gradient mode using a mixture of acetonitrile, isopropanol and water with the addition of 0.1% formic acid and 5 mM ammonium formate. As a result, the problem of separating the cis- and trans-isomers of FAME C16:1, C18:1 and C18:2 n-6 has been solved. The electrospray ionization mass spectrometry (ESI-MS) detection of FAMEs, in the form of ammonium adducts, has been optimized for the first time, which has made the method more sensitive that when the protonated species are used. This method has been applied to 12 samples from healthy subjects that consumed omega-3 supplements and has proven to be a reliable tool for determining the omega-3 index.

Bone marrow fatty acids affect osteoblastic differentiation through miR-92b-3p in the early stages of postmenopausal osteoporosis

Osteoporosis is partially caused by dysfunctions in the commitment, differentiation or survival of osteoblasts. Bone marrow fatty acids affect bone resorption and formation. In this study, we aimed to explore the role of fatty acids in the early stages of postmenopausal osteoporosis and determine whether they influence osteogenic differentiation through microRNAs. A quantitative analysis of bone marrow fatty acids early after ovariectomy or sham surgery in a rat osteoporotic model was performed using gas chromatography/mass spectrometry. The results showed that palmitoleate was significantly decreased on postoperative day 3 while both pentadecanoate and palmitoleate were significantly decreased on postoperative day 5 in rats in the ovariectomized group compared with those in the sham group. Palmitoleate promotes osteogenic differentiation, whereas pentadecanoate inhibits this process. Palmitoleate levels were higher than those of pentadecanoate; therefore, the early overall effect of significant bone marrow fatty acid changes was a decrease in osteogenic differentiation. We also found that miR-92b-3p inhibited osteoblastogenesis via the miR-92b-3p/phosphatase and tensin homolog regulatory axis. Palmitoleate, pentadecanoate, and palmitate influenced the osteoblastogenesis of MC3T3-E1 cells through miR-92b-3p. Taken together, we propose that miR-92b-3p mediates the effect of bone marrow fatty acids on osteoblast differentiation in the early stages of osteoporosis. These findings may provide molecular insights for the treatment of osteoporosis.

Estimation of Lipid and Fatty Acids Composition over the Ground Part of Nepeta olgae Regel (L.) Plants Growing in Namangan Region

The article is devoted to the study of lipids and fatty acid composition of the above-ground part of the Nepeta olgae Regel (L.) plant of the Laminaceae family. It was found that the content of neutral lipids (NL) is 5.54%, PL - 6.12%, and total lipids (NL, PL) - 11.66%. Of the neutral lipids, the unsaponifiable substances (HB) had a bright yellow color, which is explained by a small amount of carotenoids (88.87 mg%). Glycolipids dominate in PL. Among the unsaponifiable substances were found biologically active components such as hydrocarbons, carotenoids, aliphatic alcohols, sterols and triterpenols. Phytosterols were the main component of unsaponifiable NS.
 Quantitative and qualitative analysis of fatty acids from the plant Nepeta olgae Regel (L.) was carried out by gas chromatography (GC). 28 acids were identified, of which 11 compounds are saturated, and 7 compounds are unsaturated fatty acids. Of the fatty acids, the main ones are linolenic 18:3 (35.48), palmitic 16:0 (33.38%), as well as ω-3 polyunsaturated fatty acids, including eicosanoic 20:1, cis-11,14-eicosadienoic 20:2, 8,11,14-eicosatriene 20:3 + arachidonic 20:4.
 Extracts of Nepeta olgae Regel (L.) were distinguished by a high content of polyunsaturated acids, which determines their potential biological activity.

Quantitative analysis of fatty acids and vitamin E and total lipid profiling of dietary supplements from the German market

Certain polyunsaturated fatty acids with n-3 double bonds are essential nutrients for the human body and are part of the bilayer of cell membranes or precursors of tissue hormones. The most abundant dietary n-3 fatty acids in human nutrition are α-linolenic, eicosapentaenoic, and docosahexaenoic acid and can be taken up through dietary sources such as vegetable oils or fish or, alternatively, dietary supplements with high levels of n-3 fatty acids. In previous studies, considerable variation of lipid patterns and quantities of n-3 fatty acids were observed. In this study, 33 dietary supplements from the German market, based on fish-, krill-, microalgae, and plant oil, have been analyzed. Lipid profiling (LC–MS) revealed triacylglycerols as the dominant lipid species in most samples. However, krill oil was rich in phospholipids and samples containing fatty acid concentrates featured abundant fatty acid ethyl esters and diacylglycerols. Furthermore, total lipid profiles showed considerable variance depending on the lipid sources (e.g., fish or plant oil), which was also apparent in fatty acid analysis. The contents of n-3 fatty acids ranged between 150 and 570 mg/g capsule content (GC–MS) and vitamin E (α-tocopherol and tocopheryl acetate) were found in quantities ranging from 1.2 to 86.1 mg/g capsule content (HPLC–UV/Vis). While our analyses indicated a good agreement between labeled and present quantities of total n-3 fatty acids and vitamin E for the majority of samples, significant differences in agreement between individual fatty acids were observed, as well as frequent mismatches between declared and present vitamin E derivatives.

Quantitative Trait Loci (QTL) Mapping and Marker Analysis of Fatty Acids in Peanut

Quantitative Trait Loci (QTL) Mapping and Marker Analysis of Fatty Acids in Peanut

Electroactive biofilm-based sensor for volatile fatty acids monitoring: A review

• Directly convert chemical energy into electric signal form the basis of biosensors. • Electroactive biofilm-based biosensor is feasible and attractive for VFA monitoring. • VFA biosensors’ configuration and operation greatly impact their performance. • Dynamic changes tested off or on-line by biosensor can reflect bio-process status. • Microbial analysis and introducing machine learning are critical in optimization. Quantitative analysis of volatile fatty acids (VFAs) is critical for the monitoring, control and optimization of biological processes. However, commonly used detection techniques such as titration and chromatography are too complex, time-consuming or costly, making them not optimal for industrial applications. Biosensors, which can directly convert chemical energy stored in organics into electrical signals, have emerged as an attractive alternative for VFA measurements. They are simple, sensitive and low-cost, and have shown great potential for online monitoring. Among them, the electroactive biofilm (EAB)-based sensor is the most adopted. Based on a summary of the frontiers in EAB sensor development, direct/indirect contact of biofilm with VFA in tests can largely determine their performance. Various applications can illustrate the feasibility of such biosensors for off- or on-line VFA monitoring in different biological processes. These remarkable progresses have been made in recent years, but they are still facing several challenges that need to be addressed for field applications. Microbial analysis of EAB, introduction of machine learning, and optimization of biosensor design and operation will be interesting for future studies. This review can provide guidelines and references for the research, development and application of EAB sensors for online VFA monitoring.

Quantitative and carbon isotope ratio analysis of fatty acids isolated from human brain hemispheres.

Our knowledge surrounding the overall fatty acid profile of the adult human brain has been largely limited to extrapolations from brain regions in which the distribution of fatty acids varies. This is especially problematic when modeling brain fatty acid metabolism, therefore, an updated estimate of whole-brain fatty acid concentration is necessitated. Here, we sought to conduct a comprehensive quantitative analysis of fatty acids from entire well-characterized human brain hemispheres (n= 6) provided by the Douglas-Bell Canada Brain Bank. Additionally, exploratory natural abundance carbon isotope ratio (CIR; δ13 C, 13 C/12 C) analysis was performed to assess the origin of brain fatty acids. Brain fatty acid methyl esters (FAMEs) were quantified by gas chromatography (GC)-flame ionization detection and minor n-6 and n-3 polyunsaturated fatty acid pentafluorobenzyl esters by GC-mass spectrometry. Carbon isotope ratio values of identifiable FAMEs were measured by GC-combustion-isotope ratio mass spectrometry. Overall, the most abundant fatty acid in the human brain was oleic acid, followed by stearic acid (STA), palmitic acid (PAM), docosahexaenoic acid (DHA), and arachidonic acid (ARA). Interestingly, cholesterol as well as saturates including PAM and STA were most enriched in 13 C, while PUFAs including DHA and ARA were most depleted in 13 C. These findings suggest a contribution of endogenous synthesis utilizing dietary sugar substrates rich in 13 C, and a combination of marine, animal, and terrestrial PUFA sources more depleted in 13 C, respectively. These results provide novel insights on cerebral fatty acid origin and concentration, the latter serving as a valuable resource for future modeling of fatty acid metabolism in the human brain.

Quantitative Analysis and Structural Elucidation of Fatty Acids by Isobaric Multiplex Labeling Reagents for Carbonyl-Containing Compound (SUGAR) Tags and m-CPBA Epoxidation.

In this study, a novel analytical method was developed to investigate fatty acids (FAs) for relative quantification, carbon-carbon double-bond localization, and cis-/trans-geometry differentiation by isobaric multiplex labeling reagents for carbonyl-containing compound (SUGAR) tag conjugation and meta-chloroperoxybenzoic acid (m-CPBA) epoxidation. FAs are essential components of cells and have diverse functions in energy storage and as complex lipid constituents. It has been reported that FAs play different roles in various biological processes such as the functional development of the brain. The comprehensive characterization and quantification of FAs are crucial to further elucidate their biological roles. However, it is challenging to perform relative quantification and structural elucidation of FAs using integrated mass spectrometry (MS)-based methods. Recently, our group developed isobaric multiplex SUGAR tags for quantitative glycomics. Besides aldehyde/ketone groups on glycans, hydrazide groups also possess reactivity toward carboxylic acids on FAs. In this study, we extended SUGAR tag labeling with FAs for the quantitative analysis by liquid chromatography (LC)-MS/MS in the positive ion mode and applied this strategy for the comparative analysis of FAs hydrolyzed from oil samples. In addition, to comprehensively elucidate the structures of unsaturated FAs, epoxidation by m-CPBA was performed before SUGAR tag labeling to enable carbon-carbon double-bond localization. Moreover, the cis- and trans-geometries of carbon-carbon double bonds in multiple pairs of monounsaturated FAs could also be differentiated in higher-energy collisional dissociation (HCD)-MS/MS. This study developed a high-throughput comprehensive FA analysis platform, which could be widely applied and utilized in biological and clinical studies.

Treatment effects of Radix ginseng-Schisandra chinensis herb pair on Alzheimer’s disease: An investigation of MS-based metabolomics investigation

Traditional Chinese medicine (TCM) plays a synergistic and comprehensive pharmacodynamic role of multi-channel and multi-target through its multi-components, showing unique therapeutic advantages in chronic and multi-gene complex diseases. Herb pair is a unique combination of two relatively fixed herbs, which embodies the integrity of TCM theory. In this study, untargeted fecal metabolomics based on MS was used to investigate the action mechanism of Radix ginseng and Schisandra chinensis (GS) herb pair on the complex disease of Alzheimer's disease (AD), and further analyze the therapeutic effects of small molecular components and saccharides of GS on AD. Quantitative analysis of bile acids (BAs) and short-chain fatty acids (SCFAs) further verified the conclusion of untargeted metabolomics. The results of the pharmacodynamics evaluation showed that the AD model was successfully constructed, and each TCM group had a different degree of improvement compared with the AD group. PCA analysis based on untargeted fecal metabolomics showed that the metabolic disorders in AD rats changed significantly over time, and there were different degrees of callback in each TCM group. The result indicated that the GS herb pair can regulate metabolic disorders of AD. Further analysis of therapeutic biomarkers showed that GS mainly regulated the metabolism of bile acid biosynthesis, sphingolipid metabolism, porphyrin and chlorophyll metabolism, etc. to treat AD. This study will help to further understand the pathogenesis of AD from metabolomics, and provide beneficial support for the further study of GS and the clinical treatment of complex diseases with TCM.

Validation of quantitative fatty acid signature analysis for estimating the diet composition of free-ranging killer whales

Accurate diet estimates are necessary to assess trophic interactions and food web dynamics in ecosystems, particularly for apex predators like cetaceans, which can regulate entire food webs. Quantitative fatty acid analysis (QFASA) has been used to estimate the diets of marine predators in the last decade but has yet to be implemented on free-ranging cetaceans, from which typically only biopsy samples containing outer blubber are available, due to a lack of empirically determined calibration coefficients (CCs) that account for fatty acid (FA) metabolism. Here, we develop and validate QFASA for killer whales using full blubber from managed-care and free-ranging individuals. First, we compute full, inner, and outer blubber CCs from the FA signatures across the blubber layers of managed-care killer whales and their long-term diet items. We then run cross-validating simulations on the managed-care individuals to evaluate the accuracy of diet estimates by comparing full-depth and depth-specific estimates to true diets. Finally, we apply these approaches to subsistence-harvested killer whales from Greenland to test the utility of the method for free-ranging killer whales, particularly for the outer blubber. Accurate diet estimates for the managed-care killer whales were only achieved using killer whale-specific and blubber-layer-specific CCs. Modeled diets for the Greenlandic killer whales largely consisted of seals (75.9 ± 4.7%) and/or fish (20.4 ± 2.4%), mainly mackerel, which was consistent with stomach content data and limited literature on this population. Given the remote habitats and below surface feeding of most cetaceans, this newly developed cetacean-specific QFASA method, which can be applied to outer-layer biopsies, offers promise to provide a significant new understanding of diet dynamics of free-ranging odontocetes and perhaps other cetacean species throughout the world’s oceans.

Quantitative analysis of fatty acids and monosaccharides composition in Chamerion angustifolium L. by GC/MS method

Chamaenerion angustifolium L. widely known as fireweed, has significant natural resources in Ukraine. It has long been used in folk medicine as a sedative, analgesic, astringent, antimicrobial, anti-inflammatory, antisclerotic, wound healing, and diaphoretic agent. These properties are the result of the availability of many groups of biologically active compounds such as flavonoids, hydroxycinnamic acids, tannins, ascorbic acid, polyphenols, macro- and microelements. The aim of the study was to determine the content of fatty acids and monosaccharides of the herb. The qualitative composition and quantitative content of these biologically active compounds were determined using a GC/MS method. The results of the analysis showed that the herb of fireweed has monosaccharides, such as D-ribose, D-arabinose, D-mannose, D-glucose, D-galactose, and Myo-inositol. D-glucose presented in raw materials in the highest amount, it is content in the herb was 11.23 mg/kg. Among fatty acids, the mono-, polyunsaturated and saturated acids were determined. The content of polyunsaturated fatty acids of the total fatty acids was 53.19%, monounsaturated – 4.92%, and saturated – 41.89%. Linolenic, palmitic, and linoleic acids dominated among the determined twelve fatty acids, their content was 1.82 mg/kg, 1.74 mg/kg, and 1.10 mg/g.

Quantitative analysis of short-chain fatty acids in human plasma and serum by GC-MS.

Short-chain fatty acids (SCFAs) are volatile fatty acids produced by gut microbial fermentation of dietary nondigestible carbohydrates. Acetate, propionate, and butyrate SCFA measures are important to clinical and nutritional studies for their established roles in promoting healthy immune and gut function. Additionally, circulating SCFAs may influence the metabolism and allied function of additional tissues and organs. The accurate quantification of SCFAs in plasma/serum is critical to understanding the biological role of SCFAs. The low concentrations of circulating SCFAs and their volatile nature present challenges for quantitative analysis. Herein, we report a sensitive method for SCFA quantification via extraction with methyl tert-butyl ether after plasma/serum acidification. The organic extract of SCFAs is injected directly with separation and detection using a polar GC column coupled to mass spectrometry. The solvent-to-sample ratio, plasma volume, and amount of HCl needed for SCFA protonation were optimized. Method validation shows good within-day and inter-day repeatability. The limit of detection was 0.3-0.6µg/mL for acetate and 0.03-0.12µg/mL for propionate and butyrate. Successful application of this method on clinical plasma and serum samples was demonstrated in six datasets. By simplifying the sample preparation procedure, the present method reduces the risk of contamination, lowers the cost of analysis, increases throughput, and offers the potential for automated sample preparation.

Assessing the concentration of conjugated fatty acids within pomegranate seed oil using quantitative nuclear magnetic resonance (qNMR).

Pomegranate seed is rich in oil, and seed oil of pomegranate consists of conjugated fatty acids with different percentages. The current contribution covers how to determine percentages of different isomeric conjugated fatty acids. The percentages of these isomers are analysed by quantitative nuclear magnetic resonance (qNMR) using benzoic acid as an internal reference chemical with a well-defined amount. Linear mathematical equations are developed for the quantitative analysis of fatty acids found in pomegranate seed oil. The developed approach is utilised for the pomegranate seed oils prepared in the laboratory and tested for commercial samples. Among the oils derived at the laboratory, the Yeni Hicaz pomegranate cultivar seeds yielded the highest fraction of punicic acid. Among the acids, punicic acid was the one with the highest fraction, while linolenic acid was the one with the lowest percentage. These results are important in identifying pomegranate seed oils. Among the commercial samples tested with the current approach, only one of them showed similar content analysis as in the laboratory-derived oils.

A feasibility quantitative analysis of free fatty acids in polished rice by fourier transform near-infrared spectroscopy and chemometrics.

Free fatty acids (FFAs) are an important indicator of the freshness and quality of rice. In this study, the vibration response of C-H chemical bonds (-CH3 , -CH2 , H-C=C-H) of FFAs in the near-infrared region was determined by analyzing the standard reagent. In addition, the spectral data of different physical forms of rice and chemometrics, such as partial least squares (PLS), synergy interval-PLS, and competitive adaptive reweighted sampling (CARS), were applied to develop an optimal regression model for rice FFAs determination. The performance of the FFAs model established by using the polished rice granule spectrum (PRG) combined with CARS was the best, the correlation coefficients of the calibration set and prediction set were 0.99 (root mean squared errors of the calibration=2.00mg/100g)and 0.98 (root mean squared errors of the prediction=3.21mg/100g),respectively, and the ratio of performance-to-deviation was 4.50. Compared with the rice powder spectral, the PRG spectral can better retain the information of FFAs. The result shows that NIRS can rapidly, non-destructively, and accurately detect FFAs in rice granules, which will help rice business and food regulatory authorities to establish an early warning mechanism of rice aging. PRACTICAL APPLICATION: Free fatty acids (FFAs) in rice are an important indicator for evaluating the freshness of rice, and their high responsiveness to the deterioration of rice quality. The real-time detection of FFAs in rice can timely adjust the parameters of the rice storage environment, which is very meaningful to ensure the quality of rice.

Profiling and quantitative analysis of underivatized fatty acids in Chlorella vulgaris microalgae by liquid chromatography-high resolution mass spectrometry.

Chlorella vulgaris is a popular microalga used for biofuel production; nevertheless, it possesses a strong cell wall that hinders the extraction of molecules, especially lipids within the cell wall. For tackling this issue, we developed an efficient and cost‐effective method for optimal lipid extraction. Microlaga cell disruption by acid hydrolysis was investigated comparing different temperatures and reaction times; after hydrolysis, lipids were extracted with n‐hexane. The best recoveries were obtained at 140°C for 90 min. The microalgae were then analyzed by an untargeted approach based on liquid chromatography with high‐resolution mass spectrometry, providing the tentative identification of 28 fatty acids. First, a relative quantification on the untargeted data was performed using peak area as a surrogate of analyte abundance. Then, a targeted quantitative method was validated for the tentatively identified fatty acids, in terms of recovery (78‐100%), intra‐ and interday relative standard deviations (<10 and <9%, respectively) and linearity (R 2 > 0.98). The most abundant fatty acids were palmitic, palmitoleic, oleic, linoleic, linolenic, and stearic acids.