39 LIPID FINGERPRINTING OF INDIVIDUAL BOVINE BLASTOCYSTS BY DESORPTION IONIZATION ELECTROSPRAY MASS SPECTROMETRY

2012 ◽  
Vol 24 (1) ◽  
pp. 132 ◽  
Author(s):  
C. R. Ferreira ◽  
L. S. Eberlin ◽  
J. E. Hallett ◽  
R. G. Cooks
Keyword(s):  
Mass Spectrometry ◽  
Fatty Acids ◽  
Fatty Acid ◽  
Free Fatty Acids ◽  
Mass Spectrometer ◽  
Lipid Composition ◽  
Mass Spectra ◽  
Ms Analysis ◽  
Desi Ms ◽  
Complex Lipids

Mass spectrometry (MS) allows the detection and structural characterisation of intact molecules such as fatty acids and complex lipids. Desorption electrospray ionization (DESI) is an ambient ionization technique used for MS analysis and profiling and imaging of drugs, metabolites and lipids directly from biological samples with no sample preparation. With the recent introduction of morphologically friendly DESI-MS solvent systems, it is also possible to acquire DESI-MS data non-destructively. Due to the extractive nature of these solvent combinations, enough ion intensity can be generated to chemically profile samples of microscopic dimensions. The objective of this work was to perform chemical profiling on intact bovine blastocysts by DESI-MS, focusing on lipid distributions. Blastocysts produced in vitro were washed 3 times in PBS + 0.1% polyvinyl alcohol to remove lipids present in the culture medium, were placed in PBS/methanol 50% and stored under –20°C for 1 week. For DESI-MS analysis, the embryos were simply placed in glass slides and allowed to dry at room temperature. Mass spectra were acquired in the negative ion mode at the mass/charge range from m/z 150 to 1000, using as solvents a combination of 1:1 (vol/vol) ethanol:dimethylformamide (DMF) or acetonitrile:DMF. The mass spectrometer used was a LTQ linear ion trap mass spectrometer controlled by Xcalibur 2.0 software (Thermo Fisher Scientific, San Jose, CA, USA). The lipid species detected included deprotonated free fatty acids such as palmitic acid (m/z 255.2), stearic acid (m/z 283.2), arachidonic acid (m/z 311.2) and docosanoic acid (m/z 339.3). Free fatty acid dimers appear in the region from m/z 500 to 650 and complex lipids represented mainly by glycerophospholipid classes appear in the region from m/z 700 to 1000 and include phosphatidylinositols (PI 38:1; m/z 788.7), phosphatidylserines (PS 36:1, m/z 885.7) and also the chlorinated phosphatidylcholines (PC 36:1; m/z 794.7). After recording the mass spectra, embryos could still be observed in the glass slide with evident dehydration due to the action of the organic solvent. Since lipid composition of bovine embryos is closely related to cryosensitivity and due to the limited amount of analytes (each embryo is estimated to have a mass of 15 pg of total lipids) lipid analysis usually involves the pooling of individuals to have a large enough amount of analytes. Traditionally, gas chromatography is used for fatty acid residue analysis in oocytes and embryos pooled are submitted to lipid extraction and derivatization. Mass spectrometry by DESI, however, allows direct analysis of intact and single embryos and the profiling of not only free fatty acids but also complex lipids, represented mainly by 3 glycerophospholipid classes (PC, PI and PS). We envisage that DESI-MS will likely become a routine tool for the analysis of lipid composition in mammalian embryos and will contribute significantly to the development of culture systems that produce embryos with higher cryoresistance. Support from the Purdue University Center for Cancer Research Small Grants Program is gratefully acknowledged.

scholarly journals Characterisation ofStaphylococcus aureuslipids by nanoelectrospray ionisation tandem mass spectrometry (nESI-MS/MS)

2019 ◽  
Author(s):  
Simon A. Young ◽  
Andrew P. Desbois ◽  
Peter J. Coote ◽  
Terry K. Smith
Keyword(s):  
Mass Spectrometry ◽  
Fatty Acids ◽  
Fatty Acid ◽  
Tandem Mass Spectrometry ◽  
Lipid Composition ◽  
Opportunistic Pathogen ◽  
Lipid Profiles ◽  
Gas Chromatography Mass Spectrometry ◽  
Tandem Mass ◽  
Low Sensitivity

AbstractStaphylococcus aureusis a major opportunistic pathogen that is exposed to antimicrobial innate immune effectors and antibiotics that can disrupt its cell membrane. An understanding ofS. aureuslipid composition and its role in defending the cell against membrane-disrupting agents is of fundamental importance. Common methods for characterising lipid profiles suffer shortcomings such as low sensitivity of detection and inferior resolution of the positional assignments of fatty acid chains in lipids. This present study developed a rapid and sensitive nano-electrospray ionisation tandem mass spectrometry (nESI-MS/MS) method to characterise the lipid composition of three commonly studiedS. aureusisolates: Newman, Mu50 and BB270. Confirming previous studies, nESI-MS/MS revealed that phosphatidylglycerols were most abundant inS. aureusmembranes, while diglucosyldiacylglycerols and lysyl-phosphatidylglycerols were also detected. Positional assignments for individual fatty acid chains within these lipids were also determined. Concomitantly, gas chromatography mass spectrometry of the fatty acids validated the molecular characterization and showed the principal species present in each strain were predominately anteiso- and iso-branched chain fatty acids. Though the fatty acid and lipid profiles were similar between theS. aureusstrains, this method was sufficiently sensitive to distinguish minor differences in lipid composition. In conclusion, this nESI-MS/MS methodology can characterise the role of lipids in antimicrobial resistance, and may even be applied to the rapid diagnosis of drug-resistant strains in the clinic.

scholarly journals The Role of Lipid Composition in the Sensory Attributes and Acceptability of the Salted and Dried Mullet Roes (Bottarga): A Study in Human and Animal Models

Nutrients ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 3454
Author(s):  
Antonella Rosa ◽  
Raffaella Isola ◽  
Mariella Nieddu ◽  
Carla Masala
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Free Fatty Acid ◽  
Free Fatty Acids ◽  
Lipid Composition ◽  
Dietary Lipids ◽  
Taste Perception ◽  
Human Model ◽  
Potential Contribution ◽  
Fatty Food

A taste component is implicated in the oro-sensory detection of dietary lipids and free fatty acids seem to be involved in fatty food recognition. Bottarga, the salted and semi-dried ovary product of mullet (Mugil spp.), is a rich-fat food. A comparative sensory assessment of different commercial bottarga samples was performed in insect and human models in relation to their lipid composition. The bottarga attractant effect to Ceratitis capitata was assessed by behavioral tests. The subjective odor and taste perception of bottarga samples was investigated in human determining the rate of pleasantness, familiarity, and intensity dimensions using the 7-points Likert-type scale. Bottarga samples showed similar lipid profiles, but differences emerged in total and free fatty acid levels. Significant differences were observed in the attractant effect/acceptability of samples to medflies, negatively correlated to their total and free fatty acids. Insect female exhibited the ability to select among bottarga samples based on their visual and olfactory properties. In the human model, a potential contribution of free fatty acid amount in the pleasantness and familiarity dimensions of taste of bottarga samples was evidenced. Women exhibited a greater ability than men to select bottarga samples based on their better olfactory perception. Our results increase the knowledge about this outstanding product with nutritional and nutraceutical properties.

scholarly journals Composition Characterization of Fatty Acid Zinc Salts by Chromatographic and NMR Spectroscopic Analyses on Their Fatty Acid Methyl Esters

2019 ◽  
Vol 2019 ◽  
pp. 1-11
Author(s):  
Kwang Seo Park ◽  
Yun Ju Kim ◽  
Eun Kyung Choe
Keyword(s):  
Mass Spectrometry ◽  
Fatty Acids ◽  
Gas Chromatography ◽  
Fatty Acid ◽  
Free Fatty Acids ◽  
Fatty Acid Methyl Esters ◽  
Methyl Esters ◽  
Gas Chromatography Mass Spectrometry ◽  
Acid Methyl ◽  
Zinc Salts

To implement EU REACH- (Registration, Evaluation, and Authorization of Chemicals-) like chemical legislations in various countries of which the purpose is human and environment safety, the first step is substance identification followed by the hazard and risk assessments. Although both structural and composition identifications are required, the latter can more importantly result in the essential data to fill out the required substance information such as purity and concentrations of constituents, as well as impurities. With fatty acid zinc salts (FAZSs) as an exemplary industrial chemical of which chromatographic and nuclear magnetic resonance (NMR) analyses were impossible due to their insolubility in water and any organic solvents, the composition characterization was tried by preparing their fatty acid methyl esters (FAMEs) using the conc. HCl/methanol/toluene method. This acid-catalyzed methyl esterification was optimized with zinc stearate as a surrogate substance. Gas chromatography-mass spectrometry (GC-MS) and NMR analyses on methyl-esterified products revealed that the optimum conditions were at 90°C for 10 min or 45°C for 30 min with two equivalent HCl as well as at 45°C for 10 min with five equivalent HCl. Almost all zinc stearates were converted into the corresponding fatty acids with 97–99% conversion rates. Free fatty acids (FFAs) were detected in extracted ion chromatograms of pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS) in the methyl-esterified products with incomplete conversions of 73∼79%. The optimized conc. HCl/methanol/toluene method of direct one-step reaction from FAZSs was compared with the two-step NaOH saponification/BF3-methanol method after acidic hydrolysis of FAZSs. The mechanism of fatty acid zinc salts into free fatty acids and fatty acid methyl esters was suggested with the evidence of the formation of Zn(OH)2.

scholarly journals Investigation of the Membrane Fluidity Regulation of Fatty Acid Intracellular Distribution by Fluorescence Lifetime Imaging of Novel Polarity Sensitive Fluorescent Derivatives

2021 ◽  
Vol 22 (6) ◽  
pp. 3106
Author(s):  
Giada Bianchetti ◽  
Salome Azoulay-Ginsburg ◽  
Nimrod Yosef Keshet-Levy ◽  
Aviv Malka ◽  
Sofia Zilber ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Free Fatty Acids ◽  
Membrane Fluidity ◽  
Fluorescence Lifetime ◽  
Intracellular Distribution ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Fatty Acid Derivatives ◽  
Polarity Sensitive

Free fatty acids are essential structural components of the cell, and their intracellular distribution and effects on membrane organelles have crucial roles in regulating the metabolism, development, and cell cycle of most cell types. Here we engineered novel fluorescent, polarity-sensitive fatty acid derivatives, with the fatty acid aliphatic chain of increasing length (from 12 to 18 carbons). As in the laurdan probe, the lipophilic acyl tail is connected to the environmentally sensitive dimethylaminonaphthalene moiety. The fluorescence lifetime imaging analysis allowed us to monitor the intracellular distribution of the free fatty acids within the cell, and to simultaneously examine how the fluidity and the microviscosity of the membrane environment influence their localization. Each of these probes can thus be used to investigate the membrane fluidity regulation of the correspondent fatty acid intracellular distribution. We observed that, in PC-12 cells, fluorescent sensitive fatty acid derivatives with increased chain length compartmentalize more preferentially in the fluid regions, characterized by a low microviscosity. Moreover, fatty acid derivatives with the longest chain compartmentalize in lipid droplets and lysosomes with characteristic lifetimes, thus making these probes a promising tool for monitoring lipophagy and related events.

scholarly journals The regulation of glyceride synthesis in isolated white-fat cells. The effects of palmitate and lipolytic agents

1972 ◽  
Vol 128 (5) ◽  
pp. 1057-1067 ◽  
Author(s):  
E. D Saggerson
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Free Fatty Acids ◽  
Fatty Acid Synthesis ◽  
Acid Synthesis ◽  
Fat Cells ◽  
Glyceride Synthesis ◽  
Glucose Incorporation ◽  
High Concentrations ◽  
Stimulation Of

1. 0.5mm-Palmitate stimulated incorporation of [U-14C]glucose into glyceride glycerol and fatty acids in normal fat cells in a manner dependent upon the glucose concentration. 2. In the presence of insulin the incorporation of 5mm-glucose into glyceride fatty acids was increased by concentrations of palmitate, adrenaline and 6-N-2′-O-dibutyryladenosine 3′:5′-cyclic monophosphate up to 0.5mm, 0.5μm and 0.5mm respectively. Higher concentrations of these agents produced progressive decreases in the rate of glucose incorporation into fatty acids. 3. The effects of palmitate and lipolytic agents upon the measured parameters of glucose utilization were similar, suggesting that the effects of lipolytic agents are mediated through increased concentrations of free fatty acids. 4. In fat cells from 24h-starved rats, maximal stimulation of glucose incorporation into fatty acids was achieved with 0.25mm-palmitate. Higher concentrations of palmitate were inhibitory. In fat cells from 72h-starved rats, palmitate only stimulated glucose incorporation into fatty acids at high concentrations of palmitate (1mm and above). 5. The ability of fat cells to incorporate glucose into glyceride glycerol in the presence of palmitate decreased with increasing periods of starvation. 6. It is suggested that low concentrations of free fatty acids stimulate fatty acid synthesis from glucose by increasing the utilization of ATP and cytoplasmic NADH for esterification of these free fatty acids. When esterification of free fatty acids does not keep pace with their provision, inhibition of fatty acid synthesis occurs. Provision of free fatty acids far in excess of the esterification capacity of the cells leads to uncoupling of oxidative phosphorylation and a secondary stimulation of fatty acid synthesis from glucose.

Effect of insulin on free fatty acid uptake by hepatocytes in the duck

1984 ◽  
Vol 102 (3) ◽  
pp. 381-386 ◽  
Author(s):  
R. Gross ◽  
P. Mialhe
Keyword(s):  
Fatty Acids ◽  
Growth Hormone ◽  
Fatty Acid ◽  
Free Fatty Acid ◽  
Glucose Metabolism ◽  
Free Fatty Acids ◽  
Fatty Acid Uptake ◽  
Acid Uptake ◽  
Low Doses ◽  
Higher Doses

ABSTRACT To elucidate the hypolipacidaemic effect of insulin in ducks, its action on the uptake of free fatty acids (FFA) by duck hepatocytes was determined. At low doses (10 mu./l) insulin stimulated FFA uptake. This effect was not observed with higher doses of insulin (20, 30 and 50 mu./l). Growth hormone at physiological concentrations and corticosterone (14·4 nmol/l) decreased basal activity, probably by reducing glucose metabolism and consequently α-glycerophosphate (α-GP) supply. Insulin was able to reverse the inhibition induced by GH and corticosterone on both FFA uptake and α-GP production. These results therefore suggest that the hypolipacidaemic effect of insulin may be partly mediated by its action on hepatic FFA uptake. J. Endocr. (1984) 102, 381–386

scholarly journals Fatty Acids Produced by Neofusicoccum vitifusiforme and N. parvum, Fungi Associated with Grapevine Botryosphaeria Dieback

Agriculture ◽  
2018 ◽  
Vol 8 (12) ◽  
pp. 189 ◽  
Author(s):  
Maria Salvatore ◽  
Selene Giambra ◽  
Daniele Naviglio ◽  
Marina DellaGreca ◽  
Francesco Salvatore ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Fatty Acids ◽  
Gas Chromatography ◽  
Fatty Acid ◽  
Linoleic Acid ◽  
Gas Chromatography Mass Spectrometry ◽  
Fungal Pathogenicity ◽  
Botryosphaeria Dieback ◽  
Plant Pathogen Interaction ◽  
Abundant Fatty Acid

There is evidence that secondary metabolites are involved in the fungal pathogenicity and virulence of Neofusicoccum spp. Fatty acids may also influence the plant–pathogen interaction but, so far, no information is available on their production by species of Neofusicoccum associated with Botryosphaeria dieback, which is a well-known syndrome of several plants with a complex etiology. In the present paper, the production of fatty acids in liquid medium, by strains of N. vitifusiforme and N. parvum associated with declining Sicilian vine plants, was evaluated. Data, acquired via gas chromatography–mass spectrometry (GC/MS), show the presence of linoleic acid as the most abundant fatty acid produced by both examined strains. In addition, the pathogenicity of N. vitifusiforme was tested on 2-year-old grapevine plants of cv. Inzolia.

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