The mass spectrometry of iso and anteiso monoenoic fatty acids

The normal, iso, and anteiso Δ⁸- and Δ⁹-17:1 fatty acid methyl esters were synthesized and their electron impact-induced fragmentation was studied by mass spectrometry. The mass spectra of the preterminal branched monoenoic fatty acid methyl esters present characteristic fragment ions, now understood to be indicative of the position of the methyl group. These fragment ions are in the iso compound m/e 227 [M-55]⁺, m/e 195 [M-87]⁺, and m/e 177 [M-105]⁺, while in the anteiso compound these fragments are shifted by 14 mass units to m/e 213, m/e 181, and m/e 163. The 15-D-iso Δ⁸- and Δ⁹-17:1 methyl esters were synthesized because the characteristic fragment ions in the methyl branched compounds indicated a key role of the tertiary hydrogen atom in the rearrangement process. A fragmentation mechanism consisting of a double bond migration triggered by the tertiary hydrogen and an allylic cleavage assuming a displacement mechanism is proposed.     

New natural 2-acetoxy fatty acids using chemical ionization and electron impact mass spectrometry

The phospholipids of the spongePolymastia gleneni contain saturated long chain (C_22–30)-acetoxy fatty acids. Their structures were assigned based on chromatographic and spectrometric data as well as comparison with a synthetic sample. The use of capillary gas chromatography combined with chemical ionization and electron impact mass spectrometry was instrumental in the eludication of structures, since only a very small amount of crude lipids was available. Part 10 of “Phospholipids in Marine Organisms.” For Part 9 in this series, see reference 12.     

Analysis of α-branched fatty acids by gas chromatography and mass spectrometry

Gas liquid chromatography and mass spectrometry were utilized in combination to identify isomeric α-branched chain fatty acid methyl esters. In a given isomeric series equivalent chain length, values decreased with increase in the number and size of α-alkyl substituents. Mass spectra of the α-monoalkyl derivatives are characterized by prominent McLafferty rearrangement ion peaks, whereas those of the α,α-dialkyl isomers contain the former ions plus an α-cleavage ion. Presented at the AOCS Meeting, Chicago, September 1973.     

Photoelectron resonance capture ionization mass spectrometry of fatty acids in olive oil

Photoelectron resonance capture ionization (PERCI) mass spectrometry has been developed for the direct online analysis of organics, including lipids. Analysis is conducted without the need for sample preparation or chemical derivatization such as methylation, foregoing the use of harmful or toxic chemicals. PERCI is currently being adapted towards the analysis of edible oils. Herein, as a proof of principle of the simplicity and potential utility of this method towards the analysis of edible oils, we present the analysis of the prevalent fatty acids (FA) in Tuscan extra‐virgin olive oil along with triolein and linolenic acid (LNA) standards. Ionization of olive oil results in little fragmentation of the prevalent FA, which are measured as their molecular ions, [FA–H]^?. The relative concentrations of these FA determined by PERCI were in good agreement with established values. Further utility of PERCI was demonstrated by interrogation of ozonized LNA and olive oil, with 13 of the 18 anticipated products of the ozonolysis of LNA measured as their molecular anions, [M–H]^?. Similarly, the PERCI mass spectrum of ozonized olive oil showed all the anticipated ions of the predominant FA, oleic acid, as well as many molecular ions arising from less abundant unsaturated FA.     

Derivatization of keto fatty acids: I. Synthesis and mass spectrometry of thiazolidinones

The synthesis of alkyl chain-substituted thiazolidinones from oxo acids is described. Reactions of mercaptoacetic acid with three oxoesters, methyl 10-oxoundecanoate, methyl 12-oxooctadecanoate and methyl 9,10-dioxooctadecanoate gives excellent yields of the corresponding thazolidinones. Mass spectral fragmentation patterns of these long-chain thiazolidinone derivatives are discussed. Presented at the ISF/AOCS World Congress, 1980, New York.     

Analysis of fatty acids by mass spectrometry in the selected ion monitoring mode

GC/EI‐MS‐SIM (gas chromatography / electron ionisation – mass spectrometry with selected ion monitoring) can be easily applied to the determination of fatty acids as methyl esters (FAME). SIM masses should have a high relative abundance and for this reason we suggest m/z 74 and m/z 87 for saturated and monoenoic fatty acids as well as m/z 79 and m/z 81 for FAME with more than one double bond. The relevance and ratio of these fragment ions along with GC retention times enables a good identification of a FAME represented by a peak detected in the chromatograms. Compared to GC with a flame ionisation detector, this simple method provides a higher selectivity and better sensitivity. It also offers the possibilities of using a wide range of internal standards which enables the individual quantification of FAME instead of the determination of their contributions to the fatty acid pattern.     

Analysis of insect triacylglycerols using liquid chromatography‐atmospheric pressure chemical ionization‐mass spectrometry

An HPLC non‐aqueous reversed‐phase separation system was adapted for analyzing insect triacylglycerols (TAG). The method uses two conventional Nova‐Pak C18 columns connected in series, for a total length of 45 cm. The mobile phase gradient is mixed from acetonitrile and 2‐propanol, and the flow rate is programmed from 1.0 to 0.7 mL/min. TAG are detected by atmospheric pressure chemical ionization‐mass spectrometry. The method ensures efficient separation of isomers and analysis of high‐molecular‐weight TAG with equivalent chain lengths up to 72. The method performance is demonstrated on analysis of TAG isolated from the fat body of the bumblebee Bombus lucorum.     

Synthesis of alkyl‐branched fatty acids

Alkyl‐branched fatty compounds are of interest for industrial products in the cosmetics and lubricant areas. In this review, clay‐ and zeolite‐catalyzed isomerizations of unsaturated fatty compounds, especially of oleic acid, are discussed. While clay‐catalyzed reactions give most complex mixtures of dimeric fatty acids and of monomeric so‐called “isostearic acid”, the zeolite‐catalyzed process yields preferentially an isomeric mixture of isostearic acids having the methyl branch on the 8–14 positions of the alkyl chain. Synthetically useful additions of alkyl radicals can only be performed on ω‐unsaturated fatty compounds, whereas perfluoroalkyl iodides were added to fatty compounds with terminal as well as internal double bonds using electron transfer‐initiated radical addition reactions. Electrophilic additions of alkyl carbenium ions generated by decomposition of alkyl chloroformates by ethylaluminum sesquichloride give well‐defined alkyl‐branched oleochemicals with good yields.     

Autoxidative rates of nonmethylene-interrupted polyenoic fatty acids

Autoxidative rates of free fatty acids and methyl esters increased in the following order: oleic (c9–18:1), c5, c9–18:2, linoleic (c9,c12–18:2), c5,c9,c12–18:3, t5,c9,c12–18:3, and linolenic (c9,c12,c15–18:3). The increase of the rates due to the Δ5-olefinic bond was much lower than that due to an olefinic bond extension in a methylene-interrupted sequence. Agreement of the oxidative rates obtained from conjugated diene content with those obtained from POV showed that oxygen attack did not occur at the isolated olefinic bond, but at the methylene-interrupted diene in the oxidation of 5,9,12–18:3. The Δ5-olefinic bond may promote the oxidation of the methylene-interrupted diene intramolecularly.     

Derivatization of keto fatty acids: VI. Synthesis,oxidation and mass spectrometry of dithiolanes

The synthesis of alkyl chain-substituted dithiolanes, bisdithiolane and disulfolanes from oxo acids and their decomposition to original oxo acids are described. Reactions of ethanedithiol with 2 oxo esters, methyl 10-oxoundecanoate and methyl 12-oxooctadecanoate, give excellent yield of the corresponding dithiolanes, which are oxidized to the respective disulfolanes by m-chloroperbenzoic acid (m-CPBA). A similar reaction of ethanedithiol with methyl 9,10-dioxooctadecanoate affords bisdithiolane. Retroreactions of the dithiolanes and bisdithiolanes under acidic conditions and of disulfolanes under alkaline conditions yield the parent oxo acids. The structures of the individual reaction products have been established from analytical and spectral data and confirmed by a study of their mass spectra.     

Measurement of the metabolic interconversion of deuterium-labeled fatty acids by gas chromatography/mass spectrometry

An analytical method that was developed to analyze deuterium-labeled fatty acids in human blood has been extended to identify labeled fatty acids from C₁₄ to C₂₄ chain length which are formed by metabolic processes such as desaturation, elongation, or shortening of the labeled fatty acids fed. A new computer and a hard-wave adder have been utilized to assure reliable data acquisition. Relative standard derivations for the analysis of labeled fatty acids were measured at 0.02, 0.03, and 0.04 at the 5%, 1%, and 0.2% levels of the labeled fatty acid methyl esters, respectively. The method makes extensive use of standards and computer processing for accuracy and high productivity. Data from a chylomicron triacylglycerol fraction are included to demonstrate the sensitivity of detection of metabolites formed by desaturation and elongation. The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

Detection of hydroxy fatty acids in biological samples using capillary gas chromatography in combination with positive and negative chemical ionization mass spectrometry

The most common method for use in the structural analysis of hydroxy fatty acids in biological samples is the gas chromatography-mass spectrometry (GC-MS) analysis of trimethylsilyl ethers of the methyl esters using electron impact ionization. A comparison of electron impact (EI) with chemical ionization mass spectrometry (CI-MS) shows that CI-MS is the superior technique. All ions necessary for structural analysis are observed at sufficiently high levels of intensity when methane or isobutane are used as reactant gases. The molecular weight can be determined from the ion group M+H, M-15 and M+H−90. The ionic series M+H−n×90 enables one to determine the number of hydroxyl groups. The position of the hydroxyl groups can be derived from the fragments of the α-cleavage of the fatty acid chain. The application of heptafluorobutyrates as derivatives for hydroxy fatty acid methyl esters shows advantages in the trace analysis of these compounds. Heptafluorobutyrates exhibit useful mass fragmentation patterns in the positive as well as in the negative CI mode. With methane as the reactant gas, M+H usually is base peak in positive mass spectra. The ionic series M+H−n×214 leads to the number of hydroxy groups in the molecule. In the negative mass spectra, M and M-20 are indicative for the molecular weight. The ion group m/z 213, 194 and 178 at high levels of intensity is typical for heptafluorobutyrates. The advantage of the application of heptafluorobutyrates is the high sensitivity which can be obtained in trace analysis using negative MS. Heptafluorobutyrates of hydroxy fatty acids gave a 20-fold higher response in the negative scan mode compared to that of the positive. The detection limit for heptafluorobutyrates in negative CI-MS was on the order of 1 fg (10⁻¹⁵ g).