Capillary supercritical fluid chromatography-atmospheric pressure chemical ionization mass spectrometry of triacylglycerols in berry oils

A capillary supercritical fluid chromatograph (SFC), combined with a triple-quadrupole mass spectrometer (MS) via a liquid chromatography-atmospheric pressure chemical ionization (LC-APCI) interface, was utilized in the analysis of berry oil triacylglycerols. No modification of the commercially available interface was required. Vapor of different solvents, such as methanol, isopropanol, water, or ammonium hydroxide in methanol, was introduced in the sheath gas flow in the APCI source to achieve adequate ionization of triacylglycerols. The separation of triacylglycerols according to acyl carbon number and degree of unsaturation was accomplished on a 20 m × 50 μm i.d. SB-Cyanopropyl-25 column. The resolution of triacylglycerols in the reconstructed ion chromatogram and the sensitivity of the SFC-(APCI)MS system was comparable to or slightly better than that obtained with a flame ionization detector. No baseline drifting was observed during the SFC density programming. Triacylglycerols formed diagnostic [M + H]⁺ and [M - RCOO]⁺ ions with all tested reactant ion solvents except with ammonium hydroxide in methanol, which formed abundant [M + 18]⁺ ions instead of [M + H]⁺ ions. The abundance of the [M + H]⁺ ion increased with increasing degree of unsaturation of a triacylglycerol, whereas the abundance of the [M - RCOO]⁺ ion depended on the regiospecific distribution of the fatty acid moiety between the sn-1/3 positions and the sn-2 position and on the number of double bonds.     

Soybean oil triacylglycerol analysis by reversed-phase high-performance liquid chromatography coupled with atmospheric pressure chemical ionization mass spectrometry

Soybean oil triacylglycerols from genetically modified soybean lines were conclusively identified by reversed-phase high-performance liquid chromatography coupled with mass spectrometry with atmospheric pressure chemical ionization. Atmospheric pressure chemical ionization is a soft ionization technique which gives simple spectra for triacylglycerols. Spectral identification of the triacylglycerols was based on the molecular [M+1]⁺ ion and the 1(2)-, 2(3)- and 1(3)-diacylglycerol fragments. Triacylglycerols identified in high-stearic and high-palmitic soybean varieties were quantitated by reversed-phase high-performance liquid chromatography with flame-ionization detection. There was excellent agreement between the fatty acid composition calculated from the triacylglycerol composition and the fatty acid composition obtained by gas chromatography of the transmethylated oils. The oils of the modified soybean varieties, compared to typical soybean oil, contained increased content of triacylglycerols known to be more oxidatively stable, such as linoleoyloleoylstearoyl, linoleoylpalmitoylstearoyl, and linoleoyldipalmitoyl glycerols, and less triacylglycerols like trilinoleoylglycerol, known to decrease oxidative stability. This study showed that the atmospheric pressure chemical ionization technique is suitable for mass spectral identification of neutral molecules, such as triacylglycerols, which do not contain a chargeable functional group.     

Critical Ratios for structural analysis of triacylglycerols using mass spectrometry

Recent developments have finally allowed fragment behaviors using APCI‐MS to be elucidated after twenty years of literature reports. Critical Ratios have been defined that correspond to various aspects of triacylglycerol (TAG) analysis, from overall degree of unsaturation to localization of fatty acids on the glycerol backbone (regioisomers), to grouping of unsaturated fatty acids. The same ratios also constitute a compact library of TAG mass spectra.     

Separation of Enantiomeric Triacylglycerols by Chiral‐Phase HPLC

Liquid chromatography–atmospheric pressure chemical ionization mass spectrometry (LC–MS/APCI) with a chiral phase was used for separation of triacylglycerols (TAG) obtained either by organic synthesis or isolated from different algal species. We present chromatographic characteristics and tandem mass spectra of enantiomers and positional isomers (regioisomers) of C16, C18 and C20 polyunsaturated fatty acids. The retention time was found to depend on the structure of the given TAG, increasing with increasing number of double bonds and decreasing with increasing number of the carbons in TAG, with the exception of dieicosapentaenoyl‐palmitoyl‐glycerols.     

Analysis of triacylglycerols by silver-ion high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry

Triacylglycerols of the seed oils rich in α- and/or γ-linolenic acid moieties were separated by silver-ion high-performance liquid chromatography (HPLC) followed by on-line atmospheric pressure chemical ionization-mass spectrometric (APCI-MS) detection. Mass spectra of most triacylglycerols exhibited abundant [M + H]⁺ and [M − RCO₂]⁺ ions, which defined the molecular weight and the molecular association of fatty acyl residues of a triacylglycerol, respectively. Silver ions formed weaker complexes with triacylglycerols containing γ-linolenic acid than with those containing α-linolenic acid, i.e., the elution order of molecules wasXYT _γ>XYT _α’,XT _γ T _α>XT _α T _α>, andT _γ T _γ T _γ>T _γ T _γ T _α>T _γ T _α T _α>T _α T _α T _α, whereT _α=α-linolenic acid,T _γ=γ-linolenic acid, andX, Y=fatty acids different from linolenic acid. Furthermore, silver-ion HPLC resulted in partial separation within equally unsaturated triacylglycerols according to differences in the combined number of acyl carbons. Regioisomeric forms of triacylglycerols were not determined from the seed oil samples, although differences were measured with reference compounds in the relative abundances of [M − RCO₂]⁺ ions formed by a loss of a fatty acyl residue from thesn-2 position and thesn-1/3 positions. Silverion HPLC/APCI-MS provided valuable information for structure elucidation of seed oil triacylglycerols: 43 molecular species were identified from cloudberry seed oil, 39 from evening primrose oil, 79 from borage oil, 44 from alpine currant, and 56 from black currant seed oils. The quantitation requires to be studied further, especially in those cases where several molecular weight species of triacylglycerols eluted in a single chromatographic peak.     

Analysis of triglycerides using atmospheric pressure chemical ionization mass spectrometry

Atmospheric pressure chemical ionization (APCI) mass spectrometry was investigated as a new method for analysis of a mixture of triglycerides separated by reverse-phase high-performance liquid chromatography. A mixture of homogeneous (monoacid) triglyceride standards containing fatty acids with zero to three double bonds was analyzed to demonstrate the quality of mass spectra obtained by using the APCI interface. The mass spectra showed that minimal fragmentation occurs, resulting primarily in diglyceride [M−RCOO]⁺ ions and [M+1]⁺ protonated molecular ions. The degree of unsaturation within the acyl chains had a marked effect on the proportion of diglyceride ions vs. the [M+1]⁺ ions formed in the APCI source. The mass spectra of triglycerides containing fatty acids with two or three double bonds showed predominantly protonated triglyceride ions, with diglyceride peaks representing 13 to 25% of the base peak. The triglycerides containing singly unsaturated fatty acids gave diglyceride ions as the base peak, and [M+1]⁺ ions with an intensity 20 to 28% that of the base peak. Only diglyceride ions were observable in the spectra of triglycerides containing saturated fatty acids.     

Comparative analysis of triacylglycerols from Olea europaea L. fruits using HPLC and MALDI‐TOFMS

MALDI‐TOFMS and HPLC are two analytical methods that were used to characterize triacylglycerols (TAG) of the Meski, Sayali, and Picholine Tunisian olive varieties. The HPLC chromatograms of the oils showed the presence of 15 TAG species, among which triolein (OOO) was the most abundant (21–48%). In the Sayali cultivar, OOO was the predominant TAG species followed by POO and LOO. However, the minor TAG molecules were represented by LnLO and LnLP. MALDI mass spectra produced sodiated ([M + Na]⁺) and potassiated ([M + K]⁺) TAG molecules; only the major TAG were potassiated [OOO + K] ([OOO + K]⁺, [POO + K]⁺, and [LOO + K]⁺). In contrast to the HPLC chromatograms, the MALDI mass spectra showed 13 peaks of TAG. The major peak was detected at m/z 907, which corresponds to OOO with an Na⁺ adduct. The results from both HPLC and MALDI techniques predict the fatty acid composition and their percentages for each olive variety. Practical applications: TAG are the main components in vegetable oils. These biomolecules determine the physical, chemical, and nutritional properties of the oils. The nutritional benefits of TAG are related to DAG (moderate plasma lipid level) and esterified FA, which are intermediate biosynthetic molecules of TAG. TAG analysis is necessary to discriminate between oils of different origin, since some oils have similar FA profiles. Olive products, oils, and table olives, are the main diet sources of TAG in the Mediterranean countries. In this work, chromatographic and spectrometric methods were used for TAG analysis and characterization of Tunisian olive varieties.     

Characterization of α- and γ-linolenic acid oils by reversed-phase high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry

Triacylglycerols of oils rich in α- and/or γ-linolenic acids were analyzed by reversed-phase high-performance liquid chromatography (HPLC) coupled with atmospheric pressure chemical ionization mass spectrometry [(APCI)MS]. The (APCI)MS spectra of most triacylglycerols exhibited [M + H]⁺ and [M - RCOO]⁺ ions, which defined the molecular weight and the molecular association of fatty acyl residues, respectively. Reversed-phase HPLC resulted in, at least, partial separation of triacylglycerols containing α- and/or γ-linolenic acid moieties. Molecules containing α-linolenic acid residues exhibited a relatively weaker retention by the stationary phase than the corresponding molecules containing γ-linolenic acid residues. Good separation of triacylglycerols of cloudberry seed oil, evening primrose oil, borage oil, and black-currant seed oil was achieved. The molecular species identification of separated components was based on the (APCI)MS data as well as on the elution properties of triacylglycerols on reversed-phase HPLC. This study demonstrated the efficiency of HPLC-(APCI)MS in determining the molecular species compositions of triacylglycerols in complex natural mixtures. Good quality mass spectra could be extracted even from minor chromatographic peaks representing 0.2% or less of the total triacylglycerols.     

Analysis of Indigo-type compounds in natural dyes by negative ion atmospheric pressure photoionization mass spectrometry

Atmospheric Pressure Photoionization (APPI) in Mass Spectrometry (MS) has been utilized for a number of indigo-related compounds and was found to exhibit an excellent response. All structures were ionized in negative ion mode yielding almost exclusively deprotonated molecules. Their product ion mass spectra were also recorded and showed characteristic losses mainly of small neutrals such as CO, HBr and CONH₂. APPI-MS was applied further to the analysis of indigo dyestuffs of historical importance. HPLC with single ion monitoring (SIM) was employed for the separation and detection of the compounds. A simple HPLC gradient that separated the components in less than 10 min was developed. MS/MS spectra of the colouring components were also recorded and compared to that of the reference substances. The composition of Tyrian purple originating from Murex trunculus (Hexaplex trunculus), was by far the most complex, whereas some of the structures were also detected in Purpura haemastoma (Stramonita haemastoma) and Plicopurpura pansa (Plicopurpura patula subs. pansa). Further, a number of synthetic indigo dyes, produced at different times of the 19thcentury by different manufacturers, were analyzed; similar spectra were obtained suggesting that these were highly pure. Overall, the developed analytical procedure was very efficient offering high sensitivity and selectivity. APPI was proved suitable for ionizing the species under investigation producing clear mass spectra with characteristic fragment ions and may be used successfully in the future for the characterization of similar structures in historical art objects.     

Identification of diacylglycerols and triacylglycerols in a structured lipid sample by atmospheric pressure chemical ionization liquid chromatography/mass spectrometry

Atmospheric pressure chemical ionization liquid chromatography/mass spectrometry was used in the identification of diacylglycerol and triacylglycerol (TAG) molecular species in a sample of a structured lipid. In the study of acylglycerol standards, the most distinctive differences between the diacylglycerol and TAG molecules were found to be the molecular ion and the relative intensity of monoacylglycerol fragment ions. All saturated TAG ranging from tricaproin to tristearin, and unsaturated TAG including triolein, trilinolein, and trilinolenin, had ammonium adduct molecular ions [M+NH₄]⁺. Protonated molecular ions were also produced for TAG containing unsaturated fatty acids and the intensity increased with increasing unsaturation. Diacylglycerol fragment ions were also formed for TAG. The ammonium adduct molecular ion was the base peak for TAG containing polyunsaturated fatty acids, whereas the diacylglycerol fragment ion was the base peak for TAG containing saturated and monounsaturated medium- and long-chain fatty acids; the relative intensities of the ammonium adduct molecular ions were between 14 and 58%. The most abundant ion for diacylglycerols, however, was the molecular ion [M−17]⁺, and the relative intensity of the monoacylglycerol fragment ion was also higher than that for TAG. These distinctive differences between the diacylglycerol and TAG spectra were utilized for rapid identification of the acylglycerols in the sample of a structured lipid.     

High‐performance liquid chromatography‐mass spectrometry of epoxy resins

The application of liquid chromatography coupled to mass spectrometry (LC‐MS) for the analysis of epoxy resins is shown in two examples. Electro spray (ESI) and atmospheric pressure chemical ionization (APCI) are compared with respect to the ionization of diglycidylether of bisphenol A‐based (DGEBA) epoxy resins. By‐products in a typical modified solid DGEBA‐based epoxy resin and in a new weatherable crosslinker for powder coating applications are characterized and discussed. © 1999 John Wiley & Sons, Inc. J Appl Polym Sci 72: 913–925, 1999     

Characterization of the effects of β‐carotene on the thermal oxidation of triacylglycerols using HPLC‐ESI‐MS

RP HPLC method coupled to ESI‐MS was used for the analysis and characterization of the oxidation of model triacylglycerols (TAGs) in presence of β‐carotene. β‐Carotene was added to the TAGs and oxidized in the Rancimat at 110°C. The samples were separated isocratically using a mixture of isopropanol with methanol and a Phenomenex C18 column. β‐Carotene degradation was measured using high performance TLC. We found that β‐carotene plays an important role during the thermal degradation of high oleic acid model TAGs. Half of the β‐carotene was degraded before 3 h of thermal treatment. β‐Carotene significantly increases the peroxide value of the TAGs after the third hour, suggesting a pro‐oxidant action. However, different TAGs show different activity toward thermal treatment and β‐carotene. The LLL was found to be less stable, OLL and OLO were stable till 10 and 12 h respectively, while POO, OOO, and OSO were the stable TAGs till 14 h. In TAGs, replacing linoleic acid by oleic acid, the stability of the corresponding TAG was found to increase by 2 h. A new class of oxidized TAGs was reported for the first time, together with previously reported species. The proposed mechanism of formation and identification of the newly identified species have been explained. Among the oxidized species of TAGs, mono‐hydroperoxides, bis‐hydroperoxides, epoxy‐epidioxides, and epoxides were the major compounds identified.     

A novel template-free wet chemical synthesis method for economical production of zinc oxide microrods under atmospheric pressure

This work reports a template-free wet chemical synthesis method for economical production of zinc oxide (ZnO) microrods by using an inexpensive polypropylene beaker as a reactor under atmospheric pressure. The morphology and crystal structure of the as-prepared ZnO microrods were investigated by scanning electron microscopy (SEM), X-ray powder diffraction (XRD) and Raman scattering. Fourier-transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA), Ultraviolet–visible (UV/Vis) spectroscopy and methylene blue photocatalytic degradation test were also conducted to study their physical and chemical properties. The results show that the as-prepared ZnO microrods have a hexagonal crystal structure. Under the optimal synthesis condition, they have a diameter of 0.8–2.6 μm and a length of 10–40 μm. Compared with commercial ZnO powders, the as-prepared ZnO microrods have a higher whiteness. Meanwhile, the as-prepared ZnO microrods have very low photocatalytic activity, indicating that they have good photostability and are unlikely to cause the photodegradation of polymers such as binders. As a result, they will be an excellent and economical white pigment with great potential in coating applications.     

Analysis of triacylglycerols in refined edible oils by isocratic HPLC‐ESI‐MS

A simple, fast and reproducible reversed‐phase high performance liquid chromatography (HPLC) method coupled to electrospray ionization mass spectrometry (ESI‐MS) for the analysis of triacylglycerols (TAGs) species in the commercial edible oils has been developed. The TAGs species were separated using isocratic 18% isopropanol in methanol and a Phenomenex C18 column. The ESI‐MS conditions were optimized using flow injection analysis of standard TAG. Fifteen, fourteen, and sixteen TAGs were separated and identified in corn oil, rapeseed oil, and sunflower oil, respectively. The presence of intense protonated molecular (M + H⁺), ammonium (M + ${\rm NH}_{4}^{ + } $ ), and sodium (M + Na⁺) adducts ions and their respective diacylglycerols ions in the ESI‐MS spectra showed correct identification of TAGs. Some minor potassium adducts (M + K⁺) were also found. In addition, the identity of the fatty acid, position of each fatty acid, and the location of the double bond in the fatty acid moiety were explained. It was found that this isocratic method is useful for fast screening and identification of triacylglycerols in lipids.     

A gas chromatography‐mass spectrometry method for the detection of adipocere in grave soils

Adipocere refers to a decomposition product, which forms due to the post‐mortem conversion of adipose tissue into a lipid mixture. During the conversion process, triglycerides present in the adipose tissue convert to free fatty acids via hydrolysis. Under the right conditions, hydrogenation of the free fatty acids will yield adipocere, which is chiefly comprised of saturated fatty acids. In a burial environment, adipocere may form macroscopically on the decomposing remains, but may also leach into the surrounding soil. This paper describes the development of a gas chromatography‐mass spectrometry method which allows for the quantitative analysis of trace levels of adipocere in grave soils.     

B‐stage characterization of o‐cresol novolac epoxy resin system using raman spectroscopy and matrix‐assisted laser desorption/ionization mass spectrometry

The B‐stage of the o‐cresol novolac epoxy resin–phenol novolac hardener–triphenylphosphine (TPP) catalyst system was characterized using Raman spectroscopy and matrix‐assisted laser desorption/ionization (MALDI) mass spectrometry. The consistent decreasing intensities of characteristic epoxy resin peaks in MALDI mass and Raman spectra according to the melt mixing time were observed, which is due to the formation of the epoxy–phenol–TPP complex and the propagation reaction between them and with another epoxy resin. Our microscopic analysis method will provide a useful tool to control the optimum condition of the melt mixing process in the B‐stage. © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 76: 1940–1946, 2000     

Quantification of linolenic acid isomers by gas chromatography‐mass spectrometry and deconvolution of overlapping chromatographic peaks

The use of a technique for deconvolution of overlapping chromatographic peaks, Gentle, has been evaluated for quantification of alpha linolenic acid isomers analysed by gas chromatography‐mass spectrometry. Mixtures containing varying amounts of linolenic acid methyl ester isomers with two or three trans double bonds were analysed by two different temperature programs. Overlapping chromatographic peaks were resolved by Gentle, and the areas of the resolved peaks were compared with reference values calculated by use of internal standards. The results show that the small differences that exist between the mass spectra of the analysed isomers are sufficient to achieve deconvolution of severely overlapping peaks. The errors were larger than seen for quantification of chromatographically resolved peaks. Especially for small peaks in a peak cluster, the errors relative to the peak size were large.     

Influence of liquid viscosity and bed porosity on two‐phase pressure drop in concurrent gas‐liquid upflow through packed beds

It was observed in the experimental investigations that the concurrent upflow of air‐Monoethanol amine system through the packed bed gave higher pressure drop in bubble flow regime than the air‐water system. But when the flow regime changed to spray flow, air‐water system showed higher pressure drop than the other. This phenomenon was observed for the two column packing used in this study. An attempt is made to explain this phenomenon.