Boron Trifluoride-methanol Research Articles

Evaluation of five methods for derivatization and GC determination of a mixture of very long chain fatty acids (C 24:0–C 36:0)

D003 is a new active ingredient consisting of a mixture of very long chain saturated fatty acids (C 24:0–C 36:0) in a definite proportion, which shows antioxidant, antiosteoporotic, antiplatelet and cholesterol-lowering effects in experimental models. Five derivatization methods for determining these fatty acids by gas chromatography (GC), using diazomethane, sulphuric acid–methanol, hydrochloric acid–methanol, boron trifluoride–methanol and N-methyl- N-trimethylsilyltrifluoroacetamide were evaluated. GC analysis was carried out using a BPX-5 wide-bore column and 1-nonadecanoic acid (C 19:0) as internal standard. Methods were similar on account of the fatty acid content determined (84.2–86.6%). However, whereas the hydrochloric acid–methanol method needed 90 min to complete the derivatization, the other methods only required 10 min. Considering costs, speed, safety and GC response, the method using sulphuric acid–methanol was found the most appropriate for determining these fatty acids. The validation of this method: linearity over a range 40–160%, accuracy assessed through a recovery study, precision within day and inter-day, and specificity, even for samples subject to stress conditions, proved it is suitable for quality control and stability studies of the very long chain fatty acids composing this active ingredient.

Accelerated Silane Water‐Crosslinking Kinetics of Ethylene–Propylene Copolymer by Boron Trifluoride Complexes

AbstractThe hydrolysis and overall water‐crosslinking kinetics of vinyltrimethoxysilane‐grafted ethylene–propylene copolymer (EPR‐g‐VTMS) containing boron trifluoride monoethylamine complex (BF3–MEA), boron trifluoride methanol complex (BF3–MeOH), or di‐n‐butyltin(II) dilaurate (DBTL) as a silane‐crosslinking catalyst was followed by ATR‐FT‐IR and measuring the gel fraction, respectively. Kinetics results by ATR‐FT‐IR technique showed that using BF3–MEA provided much lower activation energy for the hydrolysis reactions than that of DBTL; however, BF3–MeOH had little catalytic effect on both the hydrolysis and overall water‐crosslink reaction of EPR‐g‐VTMS. We propose that the actual catalytic effect of ethylamine molecule dissociated from BF3–MEA results in the displacement of SiOCH3 groups via an SN2‐Si mechanism.magnified image

Evidence for in situ ethanolamine phospholipid adducts with hydroxy-alkenals

Hydroxy-alkenals, such as 4-hydroxy-2(E)-nonenal (4-HNE; from n-6 fatty acids), are degradation products of fatty acid hydroperoxides, including those generated by free radical attack of membrane polyunsaturated fatty acyl moieties. The cytotoxic effects of hydroxy-alkenals are well known and are mainly attributable to their interaction with different molecules to form covalent adducts. Indeed, ethanolamine phospholipids (PEs) can be covalently modified in a cellular system by hydroxy-alkenals, such as 4-HNE, 4-hydroxy-2(E)-hexenal (4-HHE; from n-3 fatty acids), and 4-hydroxy-dodecadienal (4-HDDE; from the 12-lipoxygenase product of arachidonic acid), to form mainly Michael adducts. In this study, we describe the formation of PE Michael adducts in human blood platelets in response to oxidative stress and in retinas of streptozotocin-induced diabetic rats. We have successfully characterized and evaluated, for the first time, PEs coupled with 4-HHE, 4-HNE, and 4-HDDE by gas chromatography-mass spectrometry measurement of their ethanolamine moieties. We also report that aggregation of isolated human blood platelets enriched with PE-4-hydroxy-alkenal Michael adducts was altered. These data suggest that these adducts could be used as specific markers of membrane disorders occurring in pathophysiological states with associated oxidative stress and might affect cell function.

Molecular Characterization of Compost at Increasing Stages of Maturity. 1. Chemical Fractionation and Infrared Spectroscopy

Composted organic biomasses at 60, 90, and 150 days of maturity were studied for changes in molecular composition. Compost samples were subjected to a mild sequential fractionation based on (1) organic solvent extraction, (2) transesterification with boron trifluoride in methanol (BF3-CH3OH), and (3) methanolic alkaline hydrolysis (KOH-CH3OH). The general chemical variations in compost residues following fractionation were monitored by DRIFT spectroscopy, whereas the molecular components separated along the fractionation steps were identified by GC-MS. DRIFT spectra suggested a progressive decrease of biolabile compounds such as alkyls, carbohydrates, and proteinaceous materials with compost maturity. Extraction of unbound components in an organic solvent indicated a considerable reduction of linear and branched alkanoic acids, both saturated and unsaturated, n-alkanes, and n-alkanols with enhancing compost maturity. Extracts of weakly bound molecules by transesterification revealed a decrease, with compost maturity, of components from more recalcitrant plant polyesters, such as omega-, di-, and trihydroxy acids, dioic acids, and n-alkanols. Extracts of strongly bound molecules by alkaline hydrolysis indicated a lower decrease of the same components, suggesting their reduced availability when in stable hydrophobic domains of progressively mature compost. The largest decrease in molecular components occurred when compost was stabilized from 60 to 90 days, whereas its composition did not significantly vary after stabilization at 150 days. The molecular structures of a number of steroids and terpenes appeared to be less susceptible to transformation with composting maturity, thereby resulting as useful biomarkers to trace the fate of composted organic matter in the environment. This work showed that a detailed molecular characterization of compost by a stepwise chemical fractionation enables the evaluation of compost maturity and origin of composted biomasses, as well as the identification of environmental tracers.

Quantitative analysis of bacterial medium-chain-length poly([R]-3-hydroxyalkanoates) by gas chromatography

Standard chromatographic methods for the quantification of bacterial poly(3-hydroxyalkanoate) (PHA) proved to be inappropriate for the analysis of medium-chain-length PHA (mcl-PHA). Transesterification catalyzed by protic acids is not quantitative for mcl-PHA under common conditions due to slow reaction kinetics and formation of side-products in case of functionalized side-chains. To circumvent these limitations, an improved method for the quantification of mcl-PHA by GC-FID was developed. Boron trifluoride in methanol was successfully applied to quantitatively methanolyse different mcl-PHA (recovery >94%). This novel method is well-suited for the analysis of purified mcl-PHA as well as for mcl-PHA in biomass.

Separation of molecular constituents from a humic acid by solid-phase extraction following a transesterification reaction

A selective removal of humic constituents involved in ester groups was conducted by a boron trifluoride–methanol transesterification reaction after removal of structurally unbound lipids. An analytical separation of subfractions containing specific classes of compound liberated from the humic matrix simplifies their identification by gas chromatography–mass spectrometry. We compared the traditional liquid–liquid separation into phenolic and aliphatic fractions with the modern and versatile fractionation technique using solid-phase extraction (SPE) on aminopropyl-bonded phases. Our results showed that both methods ensured separation of the same compounds, such as di- and tri-hydroxyalkanoic acids, α-, β- and ω-hydroxy fatty acids, alkanoic acids, α,ω-alkanedioic acids, n-alkanols, phenolic acids and sterols. Moreover, the SPE method not only provided a larger recovery of compounds, but involved smaller sample and solvent requirements, and larger ease and rapidity of sample handling than the traditional liquid–liquid separation. The SPE method should be thus recommended in structural studies of natural organic matter.

Statistical evaluation of an analytical GC/MS method for the determination of long chain fatty acids

In-depth evaluation of an analytical method to detect and quantify long chain fatty acids (C 8–C 16) at trace level concentrations (25–1000 μg/l) is presented. The method requires derivatization of the acids with methanolic boron trifluoride, separation, and detection by gas chromatography–mass spectrometry. The calibration experiments passed all the tested performance criteria such as linearity, homoscedasticity, and ruggedness. The detection limits and related quantities were computed by applying the method detection limit, and the calibration line approximation. The values obtained by applying the latter approach were more reliable and consistent with the actual statistical theory of detection decisions and yielded the following concentrations: C 8, 87.6 μg/l; C 10, 45.2 μg/l; C 11, 39.9 μg/l; C 12, 37.7 μg/l; C 14, 41.4 μg/l and C 16, 40.6 μg/l. Two different gas–liquid chromatographic columns were tested and similar results achieved, which shows the ruggedness of the method.

Stearoyl-CoA Desaturase Is Involved in the Control of Proliferation,Anchorage-independent Growth, and Survival in Human TransformedCells

Saturated and monounsaturated fatty acids are the most abundant fatty acid species in mammalian organisms, and their distribution is regulated by stearoyl-CoA desaturase, the enzyme that converts saturated into monounsaturated fatty acids. A positive correlation between high monounsaturated fatty acid levels and neoplastic transformation has been reported, but little is still known about the regulation of stearoyl-CoA desaturase in cell proliferation and apoptosis, as well as in cancer development. Here we report that simian virus 40-transformed human lung fibroblasts bearing a knockdown of human stearoyl-CoA desaturase by stable antisense cDNA transfection (hSCDas cells) showed a considerable reduction in monounsaturated fatty acids, cholesterol, and phospholipid synthesis, compared with empty vector transfected-simian virus 40 cell line (control cells). hSCDas cells also exhibited high cellular levels of saturated free fatty acids and triacylglycerol. Interestingly, stearoyl-CoA desaturase-depleted cells exhibited a dramatic decrease in proliferation rate and abolition of anchorage-independent growth. Prolonged exposure to exogenous oleic acid did not reverse either the slower proliferation or loss of anchorage-independent growth of hSCDas cells, suggesting that endogenous synthesis of monounsaturated fatty acids is essential for rapid cell replication and invasiveness, two hallmarks of neoplastic transformation. Moreover, apoptosis was increased in hSCDas cells in a ceramide-independent manner. Finally, stearoyl-CoA desaturase-deficient cells were more sensitive to palmitic acid-induced apoptosis compared with control cells. Our data suggest that, by globally regulating lipid metabolism, stearoyl-CoA desaturase activity modulates cell proliferation and survival and emphasize the important role of endogenously synthesized monounsaturated fatty acids in sustaining the neoplastic phenotype of transformed cells.

Endothelial lipase releases saturated and unsaturated fatty acids of high density lipoprotein phosphatidylcholine

We assessed the ability of endothelial lipase (EL) to hydrolyze the sn-1 and sn-2 fatty acids (FAs) from HDL phosphatidylcholine. For this purpose, reconstituted discoidal HDLs (rHDLs) that contained free cholesterol, apolipoprotein A-I, and either 1-palmitoyl-2-oleoylphosphatidylcholine, 1-palmitoyl-2-linoleoylphosphatidylcholine, or 1-palmitoyl-2-arachidonylphosphatidylcholine were incubated with EL- and control (LacZ)-conditioned media. Gas chromatography analysis of the reaction mixtures revealed that both the sn-1 (16:0) and sn-2 (18:1, 18:2, and 20:4) FAs were liberated by EL. The higher rate of sn-1 FA cleavage compared with sn-2 FA release generated corresponding sn-2 acyl lyso-species as determined by MS analysis. EL failed to release sn-2 FA from rHDLs containing 1-O-1'-hexadecenyl-2-arachidonoylphosphatidylcholine, whose sn-1 position contained a nonhydrolyzable alkyl ether linkage. The lack of phospholipase A(2) activity of EL and its ability to liberate [(14)C]FA from [(14)C]lysophosphatidylcholine (lyso-PC) led us to conclude that EL-mediated deacylation of phosphatidylcholine (PC) is initiated at the sn-1 position, followed by the release of the remaining FA from the lyso-PC intermediate. Thin-layer chromatography analysis of cellular lipids obtained from EL-overexpressing cells revealed a pronounced accumulation of [(14)C]phospholipid and [(14)C]triglyceride upon incubation with 1-palmitoyl-2-[1-(14)C]linoleoyl-PC-labeled HDL(3), indicating the ability of EL to supply cells with unsaturated FAs.

Ochratoxin A in wine and grape juice sold in Canada

Ochratoxin A (OTA) was determined in 251 samples of wines and grape juice collected over 3 years in Canada. In total, 25/84 samples of red wine, 22/96 samples of white wine, 3/46 red grape juices and 1/25 white grape juices contained OTA levels above the limit of quantitation (LOQ). Canadian wines, when compared with imported products, showed both a lower OTA occurrence, noted as positive (19 versus 48% above the limit of detection (LOD) for wines), and a lower level of OTA contamination (upper bound mean of 17.5 versus 163 pg ml−1 for wines). Wines from the USA contained no quantifiable levels of ochratoxin A. OTA was found in Canadian and US grape juice samples, with 12.9% above the LOD and an upper bound mean of 13.3 pg ml−1. It was extracted from a wine or grape juice sample by passing it through an immunoaffinity column. The sample matrix was washed off the column with water. OTA was eluted from the column with methanol and quantitatively determined by liquid chromatography using a fluorescence detector. The presence of OTA was confirmed by esterification with boron trifluoride-methanol. The LOQ of OTA was estimated as 20 pg ml−1 in white wine (S/N 10:1) and 40 pg ml−1 in red wine, white grape juice and red grape juice (S/N 20:1). The LOD was estimated as 4 pg ml−1 for white wine and 8 pg ml−1 for red wine and white and red grape juices (S/N 3:1).

Phenyl biliverdin isomers obtained by chemical oxidation of iron(III) complex of 5-phenyl protoporphyrin IX

Oxidative cleavage of synthetic 5-phenyl protohemin IX in pyridine solution in the presence of ascorbic acid (coupled oxidation), followed by esterification of the products with boron trifluoride-methanol rendered mainly three isomeric biliverdins. These were identified by MS and 1D and 2D1H NMR as 15-phenyl biliverdin IXβ (1), 10-phenyl biliverdin IXγ (2) and 5-phenyl biliverdin IXδ (3) dimethyl esters. The fact that biliverdin IXα dimethyl ester derivative is not obtained indicates that oxidation fails to occur in the α-meso-carbon bearing the phenyl group.

Unified gas chromatographic–mass spectrometric method for quantitating tyrosine metabolites in urine and plasma

Tyrosine and many of its catabolites play significant roles in the in the toxicity associated with acquired and congenital forms of hypertyrosinemia. We now report a specific and sensitive GC/MS method for the simultaneous determination of tyrosine metabolites maleylacetone (MA), fumarylacetone (FA), succinylacetone (SA), fumarate and acetoacetate in urine and plasma. Tyrosine metabolites and an internal standard, 2-oxohexanoic acid (OHA), in urine or plasma samples were derivatized to their methyl esters with a 12% boron trifluoride–methanol complex (12%BF 3–MeOH). The reaction mixture was extracted with methylene chloride and analyzed by GC/MS, using a selected ion monitoring (SIM) mode. The detection limits were in the range of 0.08–0.4 ng and the quantitation limits were 0.2–2 ng. Most of the intraday and interday coefficients of variation for three concentrations (low, medium and high) of the analytes were below 10%. Sensitivity and selectivity are superior to existing HPLC or enzymatic methods and derivatization of samples is simpler than the traditional silylation of organic acids used for analysis by GC/MS or derivatization to oximes, followed by silylation in the case of the ketoacids, such as SA. Furthermore, the current procedure can be performed in aqueous solution, which results in a high percentage yield without appreciable analyte degradation or formation of side products. Thus far, the method has been successfully applied in the analysis of over 5000 urine and plasma samples from humans and rodents.

A New Method for the Synthesis of Bishydroperoxides Based on a Reaction of Ketals with Hydrogen Peroxide Catalyzed by Boron Trifluoride Complexes.

AbstractFor Abstract see ChemInform Abstract in Full Text.

A new method for the synthesis of bishydroperoxides based on a reaction of ketals with hydrogen peroxide catalyzed by boron trifluoride complexes

A reaction of cycloalkanone, alkanone and alkyl aryl ketone ketals with H 2O 2 catalyzed by boron trifluoride etherate and boron trifluoride–methanol complexes was studied. A new versatile method for the synthesis of bishydroperoxides and their derivatives, viz. 1,1′-dihydroperoxyperoxides and 1,2,4,5-tetraoxacyclohexanes, was developed based on this reaction.

Boron Trifluoride—Methanol Complex — Mild and Powerful Reagent for Deprotection of Labile Acetylated Amines.

AbstractFor Abstract see ChemInform Abstract in Full Text.

A simplified method for the quantification of total cholesterol in lipids using gas chromatography

In this study, a quick and simple protocol for determination of total cholesterol in fats and oils using gas chromatography was established. Practically, about 0.1 g of lipid was saponified with 0.5 n NaOH in methanol, followed by methylation in a 10% boron trifluoride methanol solution (BF 3/MeOH). The methylated sample was then extracted with 2 ml ether, added with 100 μl 0.4% (w/v) 5α-cholestane as internal standard, and analyzed by gas chromatography. The whole procedure for the quantification of cholesterol in lipids by the above method would take only about 30 min. Standard addition test revealed that the recovery of cholesterol ranged from 94 to 107% with a coefficient variance not more than 7.1%. Comparisons of cholesterol content in 12 different fats and oils pre-treated with AOAC saponification and BF 3 methylation followed by GC analysis also showed no significant difference. These results suggested that, without losing accuracy, BF 3 methylation method for total cholesterol quantification not only consumes less solvent but also reduces by more than half the time required for the AOAC method.

Boron trifluoride–methanol complex—mild and powerful reagent for deprotection of labile acetylated amines

A set of amino-group possessing cyanine dyes is obtained from their N-acetyl derivatives via deprotection with boron trifluoride–methanol complex in good yields.

Development of an Analytical Scheme for Simazine and 2,4-D in Soil and Water Runoff from Ornamental Plant Nursery Plots

The transport and fate of pesticides applied to ornamental plant nursery crops are not well documented. Methodology for analysis of soil and water runoff samples concomitantly containing the herbicides simazine (1-chloro-4,6-bis(ethylamino)-s-triazine) and 2,4-D ((2,4-dichlorophenoxy)acetic acid) was developed in this research to investigate the potential for runoff and leaching from ornamental nursery plots. Solid-phase extraction was used prior to analysis by gas chromatography and liquid chromatography. Chromatographic results were compared with determination by enzyme-linked immunoassay analysis. The significant analytical contributions of this research include (1) the development of a scheme using chromatographic mode sequencing for the fractionation of simazine and 2,4-D, (2) optimization of the homogeneous derivatization of 2,4-D using the methylating agent boron trifluoride in methanol as an alternative to in situ generation of diazomethane, and (3) the practical application of these techniques to field samples.

Gas Chromatographic Analysis of Infant Formulas for Total Fatty Acids, Including trans Fatty Acids

Twelve powdered and 13 liquid infant formulas were analyzed by using an extension of AOAC Official Method 996.01 for fat analysis in cereal products. Samples were hydrolyzed with 8 N HCl and extracted with ethyl and petroleum ethers. Fatty acid methyl esters were prepared by refluxing the mixed ether extracts with methanolic sodium hydroxide in the presence of 14% boron trifluoride in methanol. The extracts were analyzed by gas chromatography. In powdered formulas, saturated fatty acid (SFA) content (mean +/- SD; n = 12) was 41.05 +/- 3.94%, monounsaturated fatty acid (MUFA) content was 36.97 +/- 3.38%, polyunsaturated fatty acid (PUFA) content was 20.07 +/- 3.08%, and total trans fatty acid content was 1.30 +/- 1.27%. In liquid formulas, SFA content (mean +/- SD; n = 13) was 42.29 +/- 2.98%, MUFA content was 36.05 +/- 2.47%, PUFA content was 20.65 +/- 2.40%, and total trans fatty acid content was 0.88 +/- 0.54%. Total fat content in powdered formulas ranged from 4.4 to 5.5 g/100 kcal and linoleic acid content ranged from 868 to 1166 mg/100 kcal. In liquid formulas, total fat content ranged from 4.1 to 5.1 g/100 kcal and linoleic acid content ranged from 820 to 1100 mg/100 kcal. There were no significant differences between powdered and liquid infant formulas in concentrations of total fat, SFA, MUFA, PUFA, or trans fatty acids.

Chemical Oxidation of Synthetic Iron(III)-Complex of 15-Phenyl Protoporphyrin IX

Oxidative cleavage of 15-phenyl protohemin IX in pyridine solution in the presence of ascorbic acid (coupled oxidation), followed by esterification of the products with boron trifluoride-methanol rendered only three isomeric biliverdins. These were identified by 1 H NMR and MS spectrometry as 10-phenyl biliverdin IXa (1), 5-phenyl biliverdin IXβ (2) and 15-phenyl biliverdin IX8 (3) dimethyl esters. The fact that biliverdin IXy dimethyl ester derivative is not obtained indicates that oxidation fails to occur in the γ-meso-carbon bearing the phenyl group.