A laser desorption ionization time-of-flight mass spectrometry investigation into triacylglycerols oxidation during thermal stressing of edible oils

Laser desorption ionization time-of-flight mass spectrometry (LDI–TOF MS) was used to characterize olive and sunflower oils before and after thermally assisted oxidation in order to develop a rapid fingerprinting method for oil that contains unchanged and oxidized components. No matrix was used to assist laser desorption, and simplified mass spectra were obtained in the mass range of interest (m/z 500–1000), where triacyl- and diacylglycerol ions were observed. Sample preparation was reduced to dissolving oil in chloroform saturated with NaCl. Sodiated triacylglycerols (TAGs), their epoxy/hydroxy and hydroperoxy derivatives, as well as TAGs with shortened chain fatty acids (β-scission products) were clearly observed in the spectra. LDI–TOF MS rapidly provides semiquantitative information about the oxidation level of edible oil, and thus represents a very useful quality control tool. Dedicated to Professor Pier Giorgio Zambonin on the occasion of his 72nd birthday.     

Direct detection of chlorpropham on potato skin using desorption electrospray ionization

Most pesticides, herbicides and other plant treatment agents are applied to the crop surface. Direct mass spectrometric methods, such as desorption electrospray ionization (DESI), offer new ways to analyze plant samples directly and rapidly. A strategy for the development and optimization of a DESI method for the direct determination of chemicals on complex surfaces is described. Chlorpropham (CP) was applied to potato surfaces as an example for a crop protection agent and analyzed using a self‐made DESI source. Aspects such as instrument selectivity, sensitivity and reproducibility were investigated. The MS⁴ fragmentation pattern of CP was analyzed to achieve the necessary detection selectivity, and is discussed in detail. Similar fragmentation was found in the ESI and DESI mass spectra, indicating that the mechanisms of ESI and DESI are closely related. A DESI method for semi‐quantification of CP on potatoes was developed. Detection limits of 6.5 µg/kg were found using MS/MS. The reproducibility, in the range of 12% (signal variation), appears to be sufficient for semi‐quantitative measurements. Copyright © 2013 John Wiley & Sons, Ltd.     

Identification of 1‐palmitoyl‐2‐linoleoyl‐phosphatidylethanolamine modifications under oxidative stress conditions by LC‐MS/MS

Phosphatidylethanolamines are a major class of phospholipids found in cellular membranes. Identification of the alterations in these phospholipids, induced by free radicals, could provide new tools for in vivo diagnosis of oxidative stress. In this study, 1‐palmitoyl‐2‐linoleoyl‐phosphatidylethanolamine oxidation products, induced by the hydroxyl radical, were studied using LC‐MS and LC‐MS/MS. Data obtained allowed the identification and separation of isomeric oxidative products with modifications in the sn‐2 acyl chain, attributed to long‐ and short‐chain products. Among long‐chain products keto, keto‐hydroxy, hydroxy, poly‐hydroxy, peroxy and hydroxy–peroxy derivatives were identified. Product ions formed by loss of two H₂O molecules vs loss of HOOH, allowed the identification of, respectively, di‐ (or poli‐) hydroxy vs peroxy derivatives. Location of functional groups was determined by the product ions formed by cleavage of C–C bonds, in the vicinity of the oxidation positions, allowing the identification of C9, C12 and C13 as the predominant substituted positions. Short‐chain products identified comprised aldehydes, hydroxy‐aldehydes and carboxylic derivatives, with modified sn‐2 acyl lengths of C7–C9 and C11, C12. Among the short‐chain products identified, C9 products showed higher relative abundance. Copyright © 2009 John Wiley & Sons, Ltd.     

Analysis of lipids with desorption atmospheric pressure photoionization‐mass spectrometry (DAPPI‐MS) and desorption electrospray ionization‐mass spectrometry (DESI‐MS)

In this article, the effect of spray solvent on the analysis of selected lipids including fatty acids, fat‐soluble vitamins, triacylglycerols, steroids, phospholipids, and sphingolipids has been studied by two different ambient mass spectrometry (MS) methods, desorption electrospray ionization‐MS (DESI‐MS) and desorption atmospheric pressure photoionization‐MS (DAPPI‐MS). The ionization of the lipids with DESI and DAPPI was strongly dependent on the spray solvent. In most cases, the lipids were detected as protonated or deprotonated molecules; however, other ions were also formed, such as adduct ions (in DESI), [M‐H]⁺ ions (in DESI and DAPPI), radical ions (in DAPPI), and abundant oxidation products (in DESI and DAPPI). DAPPI provided efficient desorption and ionization for neutral and less polar as well as for ionic lipids but caused extensive fragmentation for larger and more labile compounds because of a thermal desorption process. DESI was more suitable for the analysis of the large and labile lipids, but the ionization efficiency for less polar lipids was poor. Both methods were successfully applied to the direct analysis of lipids from pharmaceutical and food products. Although DESI and DAPPI provide efficient analysis of lipids, the multiple and largely unpredictable ionization reactions may set challenges for routine lipid analysis with these methods. Copyright © 2012 John Wiley & Sons, Ltd.     

Desorption electrospray ionization ‐ orbitrap mass spectrometry of synthetic polymers and copolymers

Desorption ElectroSpray Ionization (DESI) ‐ Orbitrap Mass Spectrometry (MS) was evaluated as a new tool for the characterization of various industrial synthetic polymers (poly(ethylene glycol), poly(propylene glycol), poly(methylmethacrylate), poly(dimethylsiloxane)) and copolymers, with masses ranging from 500 g.mol⁻¹ up to more than 20 000 g.mol⁻¹. Satisfying results in terms of signal stability and sensitivity were obtained from hydrophobic surfaces (HTC Prosolia) with a mixture water/methanol (10/90) as spray solvent in the presence of sodium salt. Taking into account the formation of multiplied charged species by DESI‐MS, a strategy based on the use of a deconvolution software followed by the automatic assignment of the ions was described allowing the rapid determination of M_n, M_w and PDI values. DESI‐Orbitrap MS results were compared to those obtained from matrix‐assisted laser desorption/ionization‐ time‐of‐flight MS and gel permeation chromatography. An application of DESI‐Orbitrap MS for the detection and identification of polymers directly from cosmetics was described. Copyright © 2012 John Wiley & Sons, Ltd.     

Modified MALDI MS fatty acid profiling for bacterial identification

Bacterial fatty acid profiling is a well‐established technique for bacterial identification. Ten bacteria were analyzed using both positive‐ and negative‐ion modes with a modified matrix‐assisted laser desorption ionization mass spectrometry (MALDI MS) approach using CaO as a matrix replacement (metal oxide laser ionization MS (MOLI MS)). The results show that reproducible lipid cleavage similar to thermal in situ tetramethyl ammonium hydroxide saponification/derivatization had occurred. Principal component analysis showed that replicates from each organism grouped in a unique space. Cross validation (CV) of spectra from both ionization modes resulted in greater than 94% validation of the data. When CV results were compared for the two ionization modes, negative‐ion data produced a superior outcome. MOLI MS provides clinicians a rapid, reproducible and cost‐effective bacterial diagnostic tool. Copyright © 2013 John Wiley & Sons, Ltd.     

Detection of the Short‐Lived Radical Cation Intermediate in the Electrooxidation of N,N‐Dimethylaniline by Mass Spectrometry

The N,N‐dimethylaniline (DMA) radical cation DMA^.+, a long‐sought transient intermediate, was detected by mass spectrometry (MS) during the electrochemical oxidation of DMA. This was accomplished by coupling desorption electrospray ionization (DESI) MS with a waterwheel working electrode setup to sample the surface of the working electrode during electrochemical analysis. This study clearly shows that DESI‐based electrochemical MS is capable of capturing electrochemically generated intermediates with half‐lives on the order of microseconds, which is 4–5 orders of magnitude faster than previously reported electrochemical mass spectrometry techniques.     

Dual parallel electrospray ionization and atmospheric pressure chemical ionization mass spectrometry (MS), MS/MS and MS/MS/MS for the analysis of triacylglycerols and triacylglycerol oxidation products

Two mass spectrometers, in parallel, were employed simultaneously for analysis of triacylglycerols in canola oil, for analysis of triolein oxidation products, and for analysis of triacylglycerol positional isomers separated using reversed-phase high-performance liquid chromatography. A triple quadrupole mass spectrometer was interfaced via an atmospheric pressure chemical ionization (APCI) interface to two reversed-phase liquid chromatographic columns in series. An ion trap mass spectrometer was coupled to the same two columns using an electrospray ionization (ESI) interface, with ammonium formate added as electrolyte. Electrospray ionization mass spectrometry (ESI-MS) under these conditions produced abundant ammonium adduct ions from triacylglycerols, which were then fragmented to produce MS/MS spectra and then fragmented further to produce MS/MS/MS spectra. ESI-MS/MS of the ammoniated adduct ions gave product ion mass spectra which were similar to mass spectra obtained by APCI-MS. ESI-MS/MS produced diacylglycerol fragment ions, and additional fragmentation (MS/MS/MS) produced [RCO](+) (acylium) ions, [RCOO+58](+) ions, and other related ions which allowed assignment of individual acyl chain identities. APCI-MS of triacylglycerol oxidation products produced spectra like those reported previously using APCI-MS. APCI-MS/MS produced ions related to individual fatty acid chains. ESI-MS of triacylglycerol oxidation products produced abundant ammonium adduct ions, even for those molecules which previously produced little or no intact molecular ions under APCI-MS conditions. Fragmentation (MS/MS) of the [M+NH(4)](+) ions produced results similar to those obtained by APCI-MS. Further fragmentation (MS/MS/MS) of the diacylglycerol fragments of oxidation products provided information on the oxidized individual fatty acyl chains. ESI-MS and APCI-MS were found to be complementary techniques, which together contributed to a better understanding of the identities of the products formed by oxidation of triacylglycerols.     

常压火焰离子化质谱技术对食用植物油快速分析的初探

建立了常压火焰离子化质谱(Ambient flame ionization mass spectrometry,AFI-MS)快速分析食用植物油(橄榄油、芝麻油、花生油和葵花籽油)的方法。AFI-MS检出食用植物油(橄榄油、芝麻油、花生油和葵花籽油)中的26种甘油三酯和11种甘油二脂。AFI-MS分析显示,不同的食用植物油(橄榄油、芝麻油、花生油和葵花籽油)得到的质谱图轮廓信息不同。通过对不同食用植物油的甘油三酯相对峰强度进行分析,可初步归纳出食用植物油的类型。AFI-MS分析食用植物油的操作简单,普通的打火机就可以作为离子源用于食用植物油的分析。这种便捷的离子化技术可以用于食用植物油的快速分析。     

Quantitative analysis of positional isomers of triacylglycerols via electrospray ionization tandem mass spectrometry of sodiated adducts

Herein we report a reversed‐phase high‐performance liquid chromatography tandem mass spectrometry (RP‐HPLC/MS/MS) method for the analysis of positional isomers of triacylglycerols (TAGs) in vegetable oils. The fragmentation behavior of [M + X]⁺ ions (X = NH₄, Li, Na or Ag) was studied on a quadrupole‐time‐of‐flight (Q‐TOF) mass spectrometer under low‐energy collision‐induced dissociation (CID) conditions. Mass spectra that were dependent on the X⁺ ion and the nature and position of the acyl substituents were observed for four pairs of 'AAB/ABA'‐type TAGs, namely PPO/POP, OOP/OPO, LLO/LOL and OOL/OLO (where P is 16:0, palmitic acid; O is 18:1, oleic acid; and L is 18:2, linoleic acid). For the majority of [M + X]⁺ adducts, the loss of the fatty acid in the outer positions (sn‐1 or sn‐3) was favored over the loss in the central position (sn‐2), which enabled the determination of the fractional abundance of the isomers. Ratios of the intensity of fragment ions at various AAB/ABA compositions produced linear calibration curves with positive slopes, comparable to those obtained traditionally by ESI‐MS/MS of [M + NH₄]⁺ adducts. The only exceptions were the [M + Ag]⁺ adducts of the PPO/POP system, which produced calibration curves with negative slopes. Sodium adducts provided the most consistent level of isomeric discrimination for the TAGs studied and also offered the most convenience in that they required no additive to the mobile phase. Therefore, calibration curve data derived from [M + Na]⁺ adducts were applied to the quantification of TAG regioisomers in sunflower and olive oils. The regiospecific analysis showed that palmitic acid was typically located at positions sn‐1 or sn‐3, whereas unsaturated fatty acids, oleic and linoleic acids were mostly found at the sn‐2 position. Copyright © 2010 Crown in the right of Canada. Published by John Wiley & Sons, Ltd.     

In situ analysis of soybeans and nuts by probe electrospray ionization mass spectrometry

The probe electrospray ionization (PESI) is an ESI‐based ionization technique that generates electrospray from the tip of a solid metal needle. In the present work, we describe the PESI mass spectra obtained by in situ measurement of soybeans and several nuts (peanuts, walnuts, cashew nuts, macadamia nuts and almonds) using different solid needles as sampling probes. It was found that PESI‐MS is a valuable approach for in situ lipid analysis of these seeds. The phospholipid and triacylglycerol PESI spectra of different nuts and soybean were compared by principal component analysis (PCA). PCA shows significant differences among the data of each family of seeds. Methanolic extracts of nuts and soybean were exposed to air and sunlight for several days. PESI mass spectra were recorded before and after the treatment. Along the aging of the oil (rancidification), the formation of oxidated species with variable number of hydroperoxide groups could be observed in the PESI spectra. The relative intensity of oxidated triacylglycerols signals increased with days of exposition. Monitoring sensitivity of PESI‐MS was high. This method provides a fast, simple and sensitive technique for the analysis (detection and characterization) of lipids in seed tissue and degree of oxidation of the oil samples. Copyright © 2015 John Wiley & Sons, Ltd.     

Analysis of sexual assault evidence: statistical classification of condoms by ambient mass spectrometry

Desorption electrospray ionization mass spectrometry (DESI‐MS) and easy ambient sonic‐spray ionization mass spectrometry (EASI‐MS) are employed here in the forensic analysis of chemical compounds found in condoms and relative traces, and their analytical performances are compared. Statistical analysis of data obtained from mass spectra only was applied in order to obtain classification rules for distinguishing ten types of condoms. In particular, two supervised chemometric techniques [linear discriminant analysis (LDA) and soft independent modeling of class analogy (SIMCA)] were carried out on absolute and relative intensity values to test the performances of statistical models in terms of predictive capacity. The achieved classification of samples was excellent because of the high prediction percentages of the method used both for DESI and EASI mass spectrometry analyses, confirming these two as potential ambient ionization techniques for forensic analyses in case of sexual assault crimes. EASI‐MS showed 99% prediction ability for LDA using relative data and 100% prediction ability for SIMCA using both absolute and relative ones, while DESI showed 94% prediction ability for both LDA and SIMCA. The absence of any sample preparation technique gives advantages in terms of sample preservation and reduced contamination, allowing successive analyses to be performed on the same sample by other techniques. Copyright © 2015 John Wiley & Sons, Ltd.     

Identification of intact oxidation products of glycerophospholipids in vitro and in vivo using negative ion electrospray iontrap mass spectrometry

Free radical‐induced oxidation products of polyunsaturated fatty acids esterified to phospholipids have been implicated in a number of human diseases including atherosclerosis and neurodegenerative diseases. Some of these phospholipid oxidation products have potent biological activities and likely contribute to human pathophysiological conditions. Oxidation products have also been used as markers of oxidative stress in vivo. Identification and quantification of phospholipid oxidation products are often performed by analyzing the oxidized free fatty acid moieties after hydrolysis from the phospholipids head groups by gas chromatography–mass spectrometry (GC–MS) or liquid chromatography–mass spectrometry (LC–MS). We now describe the definitive identification of intact oxidized products of glycerophospholipids including glycerophosphatidylcholine (GPC), glycerophosphatidylethanolamine (GPE), and glycerophosphatidylserine (GPS) in vitro and in vivo using iontrap MS. For these analyses, the negative ions of the oxidation products of phospholipids are fragmented to MSⁿ and unequivocal structural characterization is obtained based on collision‐induced dissociation (CID) of the sn‐2 carboxylate ion. This technique overcomes the need to hydrolyze fatty acids from phospholipids in the analysis. The method has been used to identify a number of oxidation products of glycerophospholipids including hydroxyeicosatetraenoates (HETEs) and isoprostanes (IsoPs) esterified to different classes of glycerophospholipids in vitro and in vivo. These studies thus provide a new approach to identify the intact oxidation products of glycerolphospholipids. Copyright © 2009 John Wiley & Sons, Ltd.     

DESI analysis of mammalian cell cultures – sample preparation and method optimisation

Desorption electrospray ion source (DESI) is widely used as an MS imaging technique. It is a rapid and convenient method of surface analysis, but to date, there are methodological obstacles to its application to the analysis of cell culture. This study reported optimised conditions for the analysis of cell culture samples. Parameters such as the surface, medium removal and sample desiccation techniques were assessed as a function of output data quality. Supercharging agents, surfactants and optimal parameters for the DESI ion source were evaluated for use in cell culture analyses. Data indicated that plastic dishes or sodium glass coated with poly‐l ‐lysine and washing cell cultures with 150 mM ammonium acetate followed by drying with inert gas were superior for DESI analyses. The addition of 1 μM surfactin to the DESI spray solvent significantly improved the results for negative and positive ion modes. Copyright © 2014 John Wiley & Sons, Ltd.     

Quantification of triacylglycerol regioisomers by ultra‐high‐performance liquid chromatography and ammonia negative ion atmospheric pressure chemical ionization tandem mass spectrometry

The regioisomer composition of triacylglycerols (TAGs) in various vegetable oils was determined with a new liquid chromatography/tandem mass spectrometry (LC/MS/MS method). A direct inlet ammonia negative ion chemical ionization (NICI) MS/MS method was improved by adapting it to LC negative ion (NI) atmospheric pressure chemical ionization (APCI) MS/MS system using ammonia as nebulizer gas. The method is based on the preferential formation of [M–H–RCOOH–100]^? ions during collision‐induced dissociation by loss of sn‐1/3 fatty acids from [M–H]^? ions. Calibration curves were created from nine reference TAGs: Ala/L/L, Gla/L/L, L/L/O, L/O/O, P/O/O, P/P/O, Po/Po/V, Po/Po/O, and C/O/O. The calibration curves were used to quantify the regioisomer compositions of selected TAGs in rapeseed oil, sunflower seed oil, palm oil, black currant seed oil, and sea buckthorn pulp oil. The method discriminates the different regioisomers and the results obtained by this method were in good agreement with previous results. This proves that this new method can be used for the determination of regiospecific distribution of fatty acids in TAGs. Copyright © 2009 John Wiley & Sons, Ltd.     

Desorption electrospray ionization mass spectrometry: direct toxicological screening and analysis of illicit Ecstasy tablets

Desorption electrospray ionization mass spectrometry (DESI-MS) was used as a simple and rapid way to analyze drug tablets and powders without sample preparation. Experiments were performed with a home-made DESI source coupled to a triple-quadrupole linear-ion trap (QqQ(LIT)) mass spectrometer. Twenty-one commercial drugs as well as some illicit Ecstasy tablets and powders were analyzed. MS spectra almost exclusively showed the protonated or deprotonated ion of the drug after directing the pneumatically assisted electrospray onto the tablet's surface. With some tablets, inhomogeneity of the surface resulted in different spectra depending on the spot analyzed, thus showing that DESI could be used for imaging. Directly triggered MS/MS spectra were used for confirmatory analysis, with analysis times often below 10 s per tablet. For illicit Ecstasy tablets, DESI-MS, GC/MS and LC/MS analyses provided similar qualitative results for the main analytes. With MS/MS spectra library comparison or exact mass measurements, this technique could become very powerful for the rapid analysis of unknown tablets and shows the great potential of desorption techniques as an alternative to solution-based analysis.     

A new enone‐containing triglyceride derivative as precursor of thermosets from renewable resources

A novel triglyceride containing α,β‐unsaturated ketone was prepared through photoperoxidation from high oleic sunflower oil by two steps one pot environmentally friendly procedure. This new enone‐containing triglyceride was crosslinked with diaminodiphenylmethane (DDM) via aza‐Michael addition. A kinetic study of the reaction of p‐toluidine with either enone‐containing methyl oleate or epoxidized methyl oleate, as model compounds, allowed us to establish the higher reactivity of the former, thus confirming this curing system as an alternative to amine‐cured epoxidized vegetable oils. The thermal properties of thermosets from enone‐ and epoxy‐containing triglycerides with DDM have been evaluated. © 2008 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: 6843–6850, 2008     

Rapid and sensitive determination of fatty acids in edible oil by liquid chromatography-electrospray ionization tandem mass spectrometry

A sensitive and robust on-line LC/MS method was developed for quantitative determination of linoleic acid,docosahexaenoic acid and docosanoic acid from edible oil samples.The oil samples were dissolved in chloroform-isopropyl alcohol(20:80,v:v)solution and the three fatty acids were separated by HPLC with a C4 column using 10 mmol/L ammonium acetate-isopropyl alcohol-acetonitrile(20:40:40,v:v:v)mobile phase in isocratic elution.Electrospray ionization mass spectrometry with the selected ion recording monitoring was used to detect and quantify the fatty acid.The calibration curves were linear in the range of 10.00–5000 pg/mL for linoleic acid and docosanoic acid,and 1.000–500.0 pg/mL for docosahexaenoic acid.The limit of detection was 2.0 pg/mL for linoleic acid,3.0 pg/mL for docosanoic acid,and 0.20 pg/mL for docosahexaenoic acid.The results showed that the method described in this paper could be utilized for rapid determination of three fatty acids at picogram levels in edible oils.     

Protein analysis by desorption electrospray ionization mass spectrometry and related methods

Desorption electrospray ionization mass spectrometry (DESI‐MS) requires little to no sample preparation and has been successfully applied to the study of biologically significant macromolecules such as proteins. However, DESI‐MS and other ambient methods that use spray desorption to process samples during ionization appear limited to smaller proteins with molecular masses of 25 kDa or less, and a decreasing instrumental response with increasing protein size has often been reported. It has been proposed that this limit results from the inability of some proteins to easily desorb from the surface during DESI sampling. The present study investigates the apparent mass dependence of the instrumental response observed during the DESI‐MS analysis of proteins using spray desorption collection and reflective electrospray ionization. Proteins, as large as 66 kDa, are shown to be quantitatively removed from surfaces by using spray desorption collection. However, incomplete dissolution and the formation of protein–protein and protein–contaminant clusters appear to be responsible for the mass‐dependent loss in sensitivity for protein analysis. Alternative ambient mass spectrometry approaches that address some of the problems encountered by spray desorption techniques for protein analysis are also discussed. Copyright © 2013 John Wiley & Sons, Ltd.     

Acylglycerols (=Glycerides) from the Glandular Trichome Exudate on the Leaves of Paulownia tomentosa

Chemical investigations of the glandular trichome exudates on the leaves of Paulownia tomentosa (Scrophulariaceae) led to the identification of the thirty acylglycerols (=glycerides) 1 – 30 , including five known ones ( 2, 3, 6, 9 , and 15 ) (Fig. 1). Spectroscopic analysis combined with GC/MS studies of the glycerides and the liberated fatty acids, in the form of trimethylsilyl ether derivatives and trimethylsilylated methyl esters, respectively, established that the constituents belonged to 1,3‐di‐O‐acetyl‐2‐O‐(fatty acyl)glycerols, 1‐O‐acetyl‐2‐O‐(fatty acyl)‐sn‐glycerols, and 2‐O‐(fatty acyl)glycerols, wherein the fatty acyl moiety was either an eicosanoyl or an octadecanoyl group bearing OH and/or AcO groups at the 3‐, 3,6‐, 3,7‐, 3,8‐, or 3,9‐positions. The 1‐O‐acetyl‐2‐O‐[(3R,6S)‐3‐(acetyloxy)‐6‐hydroxyeicosanoyl]‐sn‐glycerol ( 12 ; 20% of the total glycerides), 2‐O‐[(3R,8R)‐3,8‐bis(acetyloxy)eicosanoyl]glycerol ( 17 ; 14%), 2‐O‐[(3R,9R)‐3,9‐bis(acetyloxy)eicosanoyl]glycerol ( 18 ; 12%), and 2‐O‐[(3R)‐3‐(acetyloxy)eicosanoyl]glycerol ( 10 ; 12%) were relatively abundant constituents. The configurations of the stereogenic centers of the fatty acyl moieties were determined by ¹H‐NMR analysis of the monoesters obtained from (R)‐ and (S)‐2‐(naphthalen‐2‐yl)‐2‐methoxyacetic acid ((R)‐ and (S)‐2NMA? OH and the hydroxy‐substituted fatty acid methyl esters (Fig. 2). The configuration at C(2) of the glycerol moiety of the 1‐O‐acetyl‐2‐O‐(fatty acyl)glycerols was determined to be (2S) by chemical conversion of, e.g., G‐2 (= 2 / 3 1 : 10) to (+)‐3‐O‐[tert‐butyl)diphenylsilyl]‐sn glycerol of known absolute configuration.