Exclusive quantification of methyl-branched fatty acids and minor 18:1-isomers in foodstuff by GC/MS in the SIM mode using 10,11-dichloroundecanoic acid and fatty acid ethyl esters as internal standards

Minor fatty acids (iso- and anteiso-fatty acids, vaccenic acid, elaidic acid) in foodstuff (seafood, milk, and dairy products) were analyzed by gas chromatography with electron ionisation mass spectrometry in the selected ion monitoring mode (GC/EI-MS-SIM). For this purpose, lipids were obtained by accelerated solvent extraction (ASE) and the fatty acid constituents were converted into methyl esters. Instead of the determination of the relative contribution of the minor fatty acids to the sum of all fatty acids detected (the so-called 100% method), we exclusively quantified the minor fatty acids, which was possible by using two types of internal standards (IS-1 and IS-2). For recovery checks during the extraction and/or the transesterification step we added the novel 10,11-dichloroundecanoic acid (DC-11:0) as IS-1. DC-11:0, which has never been detected in foodstuff, was synthesized by electrophilic addition of chlorine to 10-undecenoic acid (11:1n-10). The novel IS eluted in the range of 23:0 from the polar GC column used and showed the same properties as fatty acids in foodstuff during sample preparation. Recovery rate of DC-11:0 was generally >96% in the various samples analyzed. Ethyl esters (FAEE) of a12:0, a14:0, a15:0, a16:0, a17:0, and a18:0 (IS-2) were added to both the external standard (a quantitative mixture of methyl esters of methyl-branched fatty acids and 18:1n-9trans) in order to determine their response factors relative to FAEE and to the food samples. With this technique, (only) methyl-branched fatty acids (MBFAs) as well as vaccenic acid (18:1n-7) and elaidic acid (18:1n-9trans) were quantified in a range of dairy products (including twelve cheeses) and seafood. All samples were analyzed in triplicates, and good standard deviations (concentrations 0.002–5 g/100 g; standard deviations 0.00–0.03) were obtained in all cases. MBFAs were detected in all samples analyzed. The highest content of MBFAs (3.0 g/100 g) was determined in red-smear of romadur cheese. In all except two cheeses, i17:0 was the most abundant MBFA. The highest amount of 18:1n-9trans was found in feta (2.84 g/100 g) whereas 0.03 g/100 g in big eye snapper (Pricanthus tayenus) marked the lowest record of this minor fatty acid. Seal oil contained the highest amount of 18:1n-7 with 5.00 g/100 g, whereas emmental cheese was the sample with the lowest content of this monoenoic fatty acid. The combination of suitable IS and a sensitive GC/EI-MS-SIM method proved to be well suited for the quantification of minor fatty acids in foodstuff. When only a set of fatty acids is going to be analyzed, this method is less time consuming compared to “100% methods” and less prone to false results due to the higher selectivity of GC/MS compared to GC in combination with flame ionisation detection (GC/FID).     

Iron ore reduction in a continuously operated multistage lab-scale fluidized bed reactor—Mathematical modeling and experimental results

Industrial-scale fluidized bed processes for iron ore reduction (e.g., FIOR and FINMET) are operated by continuous feeding of ore, while laboratory tests are mostly performed under batchwise operation. The reduction behavior under continuous operation is influenced by both the residence time of the iron ore particles and the reduction kinetics, which is obtained by batch tests. In a mathematical model for such a process, the effect of both phenomena has to be considered. The residence time distribution of iron ore particles in a laboratory fluidized bed reactor was obtained by measuring the response of a step input and described by mathematical models similar to a continuously stirred tank reactor. In the same reactor, reduction tests with continuous feeding of iron ore were performed. Based on batch tests in a fluidized bed reactor, a mathematical model was developed to describe the kinetics of iron ore reduction under fluidized bed conditions. This kinetic model was combined with the fluidized bed reactor model to describe continuous iron ore reduction. In this detailed model, the change of gas composition while rising in the fluidized bed was considered. The degree of reduction and the gas conversion for reactors in series were calculated. The results obtained by the mathematical model were compared with experimental data from the laboratory-scale reactor.     

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.     

Enantioselective determination of anteiso fatty acids in food samples

Anteiso fatty acids (aFAs)-long-chain carboxylic acids with a methyl branch on the (n - 2)-carbon-are among the most simple fatty acids that are chiral. The most frequently occurring aFAs in food are 12-methyltetradecanoic acid (a15:0) and 14-methylhexadecanoic acid (a17:0), structures where the asymmetric carbon is more than 10 carbons separated from the polar head group. Previously, only enantioseparation of 4-methyl-substituted carboxylic fatty acids has been reported by gas chromatography. Here we present the first direct partial enantioresolution of synthesized racemic a15:0-a17:0 on a capillary column coated with 50% heptakis(6-O-tert-butyldimethylsilyl-2,3-di-O-methyl)-beta-cyclodextrin diluted in OV1701. Synthesized (S)-(+)-enantiomers were used to demonstrate that the elution order was (R)- prior to (S)-enantiomers. Using this system, food samples (butter, goat's milk fat, suet, human milk, seal oil, cod liver oil) known to contain aFAs were analyzed. Prior to the enantioselective gas chromatography, unsaturated fatty acids were preseparated by urea complexation, silver ion high performance liquid chromatography (Ag+-HPLC), or both from food samples. The fractions of the food samples enriched with methyl-branched fatty acids were then analyzed by GC/MS in the SIM mode. The measurements confirmed that the (S)-enantiomer of a15:0 (ee >96%), a16:0, and a17:0 (ee >90%, respectively) dominated in all samples. While the (R)-enantiomers could not be identified in samples from ruminants and human milk, their presence could be established in cod liver and seal oil (ee <86%).