Dieter H. Welti

Composition of carrageenan blends inferred from 13C-NMR and infrared spectroscopic analysis

Abstract Carrageenan blends mainly composed of kappa, iota and lambda carrageenan were investigated by 13C-NMR and infrared (IR) spectroscopy. 13C-NMR was shown to be a powerful tool for quantifying the kappa-iota ratio in a carrageenan blend. The technique, however, was not helpful for identifying lambda carrageenan. Consequently, IR spectroscopy was used to achieve a semi-quantitative determination of lambda carrageenan. By combining both techniques, the carrageenan composition of several commercial samples was established.

Synthesis of Deuterated Volatile Lipid Degradation Products To Be Used as Internal Standards in Isotope Dilution Assays. 2. Vinyl Ketones

The isotopically labeled compounds [5,6-(2)H(2)]-(Z)-1, 5-octadien-3-one (d-I) and [1-(2)H(1;2),2-(2)H(1;1)]-1-octen-3-one (d-II), as well as the unlabeled reference compound (Z)-1, 5-octadien-3-one (I) were prepared by improved synthesis procedures. Labeling position, chemical purity, and isotopic distribution of the compounds were characterized by various MS and NMR techniques. These molecules are used as internal standards in quantification experiments based on isotope dilution assay. The newly prepared compound d-II was synthesized in a simple two-step procedure, and formation of the main isotopomers was studied in model systems.

Oxidation of caffeine and related methylxanthines in ascorbate and polyphenol-driven Fenton-type oxidations.

Caffeine and related methylxanthines were subjected to free radical mediated oxidation by incubation with Fe(3+)-EDTA/ascorbate and Fe(3+)-EDTA/polyphenolics. The reaction mixtures were analysed by reverse-phase HPLC, revealing the corresponding C-8 hydroxylated analogues as the major products of hydroxyl radical mediated attack. Further oxidation products of caffeine, analysed by liquid chromatography-mass spectrometry (LC-MS), were the N1-, N3- and N7-demethylated methylxanthine analogues theobromine, paraxanthine and theophylline, respectively. Isolable amounts of the imidazole ring operated 6-amino-5-(N-formylmethyl-amino)-1,3-dimethyl-uracil (1,3,7-DAU) derivative were also detected, which was characterised by 1H NMR and mass spectroscopy. The identified products indicate that the pertinent chemical reactions, i.e. C-8 hydroxylation, demethylations, and C8-N9 bond scission, are comparable to the primary metabolic pathways of caffeine in humans. The influence of pH, transition metals, hydrogen peroxide, free radical scavengers and metal chelators on caffeine oxidation was studied. This report illustrates that natural food-borne reactants can aid in identifying specific chemical markers of free radical induced damage. Furthermore, potentially anti-and pro-oxidative reactions can be elucidated which may be important in assessing the impact of nutrient additives and supplements on the shelf life and stability of foods and beverages.

Synthesis of trans-4,5-epoxy-(E)-2-decenal and its deuterated analog used for the development of a sensitive and selective quantification method based on isotope dilution assay with negative chemical ionization.

The volatile compound trans-4,5-epoxy-(E)-2-decenal (1) was synthesized in two steps with good overall yields. The newly developed method is based on trans-epoxidation of (F)-2-octenal with alkaline hydrogen peroxide followed by a Wittig-type chain elongation with the ylide formylmethylene triphenylphosphorane. For the synthesis of [4,5-2H2]-trans-4,5-epoxy-(E)-2-decenal (d-1), [2,3-2H2]-(E)-2-octenal was prepared by reduction of 2-octyn-1-ol with lithium aluminum deuteride and subsequent oxidation of [2,3-2H2]-(E)-2-octen-1-ol with manganese oxide. Compound d-1 was used as internal standard for the quantification of 1 by isotope dilution assay. Among various mass spectrometry (MS) ionization techniques tested, negative chemical ionization with ammonia as reagent gas gave best results with respect to both sensitivity and selectivity. The detection limit was found to be at about 1 pg of the analyte introduced into the gas chromatography-MS system.

Identification of potent odorants formed by autoxidation of arachidonic acid: structure elucidation and synthesis of (E,Z,Z)-2,4,7-tridecatrienal.

The aroma composition of autoxidized arachidonic acid was characterized by aroma extract dilution analysis. The most potent odorant was trans-4,5-epoxy-(E)-2-decenal followed by 1-octen-3-one, (E,Z)-2,4-decadienal, (E,Z,Z)-2,4,7-tridecatrienal, (E,E)-2,4-decadienal, and hexanal. (E,Z,Z)-2,4,7-Tridecatrienal was unequivocally identified by mass spectrometry and nuclear magnetic resonance (NMR) data. The stereochemistry of its extended double-bond system was elucidated on the basis of NMR measurements. The target compound was synthesized in four steps starting with bromination of 2-octyn-1-ol, followed by copper-catalyzed coupling of the bromide with ethylmagnesium bromide and (E)-2-penten-4-yn-1-ol. Partial hydrogenation of the resulting C(13)-compound with triple bonds in the positions C-4 and C-7 gave rise to (E,Z,Z)-2,4,7-tridecatrien-1-ol, which was finally oxidized to the target compound. It exhibits a typical egg-white-like, marine-like odor at low concentrations, and an intense orange-citrus, animal-like odor at higher concentrations. Its odor threshold was estimated by gas chromatography-olfactometry to be 0.07 ng/L air, which is of the same order of magnitude as that reported for 1-octen-3-one and (E,E)-2,4-decadienal.

Quantification of key odorants formed by autoxidation of arachidonic acid using isotope dilution assay.

Six odor-active compounds generated by autoxidation of arachidonic acid (AA) were quantified by isotope dilution assay (IDA), i.e., hexanal (1), 1-octen-3-one (2), (E,Z)-2,4-decadienal (3), (E,E)-2,4-decadienal (4), trans-4,5-epoxy-(E)-2-decenal (5), and (E,Z,Z)-2,4,7-tridecatrienal (6). Compound 1 was the most abundant odorant with about 700 mg/100 g autoxidized AA, which corresponds to 2.2 mol% yield. Based on the odor activity values (ratio of concentration to odor threshold), odorants 3 (fatty) and 5 (metallic) showed the highest sensory contribution followed by 1 (green), 2 (mushroom-like), 6 (egg white-like), and 4 (fatty). For the first time, reliable quantitative results are reported for odorants 1–6 in autoxidized AA, in particular odorant 6, which is a characteristic compound found in autoxidized AA. Synthesis of deuterated 6, required for IDA, is described in detail. The formation of odorants 1–6 by autoxidation of AA is discussed with respect to the quantitative data.

Chemo-enzymatic synthesis of α-terpineol thioacetate and thiol derivatives and their use as flavouring compounds

Reaction of (R,S)‐α‐terpineol with thioacetic acid in food‐grade n‐hexane resulted into two α‐terpineol thioacetate derivatives with the same molecular weight. After 5 h of reaction time, (R,S)‐α‐terpineol was completely transformed and the mixture analysed by different chromatographic techniques. The aroma character of the α‐terpineol thioacetates was described as exotic, sweet, blackcurrant, roasted and sulphury. Of eight lipases and two esterases assayed, only non‐immobilized pig liver esterase (PLE) hydrolysed α‐terpineol thioacetates into the corresponding α‐terpineol thiols. When reactions were performed in 0.2 m phosphate buffer at pH 8.0 and 30 °C with non‐immobilized PLE, α‐terpineol thiols were produced in an optimal yield of 88% after 24 h of reaction time. The aroma character of α‐terpineol thiols was described as green, exotic and fresh grapefruit. Flavouring powders were prepared by freeze‐drying the α‐terpineol thioacetates and α‐terpineol thiols in the presence of maltodextrine. Preliminary applications showed that these flavouring preparations could be used to improve the flavour quality of lighter cooked notes and tropical fruit aromas. Copyright

Metabolism of the food mutagen 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline in human hepatocytes.

2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) is a heterocyclic aromatic amine formed during cooking of fish and meat (1). It is mutagenic and carcinogenic in different animal species and is suspected to play a role in the etiology of human cancer (2). Studies performed in animals and with purified human enzymes demonstrated that MeIQx is metabolized by cytochrome P450 (CYP) 1A2 in the liver and CYP1A1 and 1BI in extrahepatic tissues (3). However the metabolic pathways of MeIQx remain incompletely elucidated in humans.