Lothar W. Kroh

Caramelisation in food and beverages

The decomposition of sugars leads to the formation of volatile (caramel aroma) and brown-coloured compounds (caramel colours). The reaction can be effected by heat and is catalysed by acids and bases. The colours and aromas depend on the sugar used (i.e. whether mono-, oligo- or polysaccharide) and are formed mostly through deoxyosuloses, O-heterocyclic and carbocyclic intermediates as well as low-molecular-weight sugar fragments. The typical caramel aromas are traced back to cyclopentanone (cyclotene) and hydroxymethylfuranone (furaneol) and their formation and analysis, discussed. The structures of coloured products of caramelisation are still not fully understood. The formation of brown products of caramelisation in dessert wine is discussed.

Investigation of the influence of reaction conditions on the elementary composition of melanoidins

Abstract The elementary composition of melanoidins produced in solvent-free reaction mixtures differs, markedly, depending on whether pentoses (ribose) or hexoses (glucose, fructose) are used. In the case of ribose, more than 4 mol sugar are incorporated into the melanoidin per mol amino acid, whereas, under the same reaction conditions, only about half the number of moles for hexoses are involved. The fundamental composition of model melanoidins under constant reaction conditions is only negligibly influenced by the molar ratio of the reactants. Reaction conditions have a significant influence on the composition of the polymers. Depending on the reaction temperature in a solvent-free milieu, at least 2 mol glucose per mol amino acid are incorporated into the polymer. In aqueous solution this ratio decreases to less than 1:1. In agreement with results of the elementary composition of melanoidins, a new fundamental structure of the polymer which is formed by reaction of dicarbonyl compounds with one another or with amino components is proposed. The structure is supported by IR, UV and CP-MAS NMR studies.

Composition of Phenolic Compounds and Glycoalkaloids α-Solanine and α-Chaconine during Commercial Potato Processing

The influence of a commercial production process for dehydrated potato flakes on the content of free phenolic compounds, total phenolics, and glycoalkaloids in potatoes during the subsequent processing steps was determined. Processing byproducts, such as potato peel (steam peeling), mashed potato residues, and side streams (blanching and cooking waters), have also been investigated. A high-performance liquid chromatography (HPLC) method was developed to separate and quantify caffeic acid, gallic acid, ferulic acid, p-coumaric acid, p-hydoxybenzoic acid, protocatechuic acid, vanillic acid, catechin, and three isomers of caffeoylquinic acid: chlorogenic, neochlorogenic and cryptochlorogenic acid. Determination of the glycoalkaloids alpha-solanine and alpha-chaconine was performed by using a high-performance thin-layer chromatography (HPTLC) method. The deliverables reveal that processing potatoes to potato flakes remarkably diminishes the content of the analyzed compounds, mainly due to peeling and leaching. The influence of thermal exposure is less significant. About 43% of the initial phenolic acids and 10% of the glycoalkaloids remain after processing. The results of the total phenolic content assay by Folin-Ciocalteu reagent are proportional to the content of phenolic compounds determined by HPLC. Steam peeling has a higher influence on glycoalkaloid losses compared to that on phenolics. The highest amounts of phenolic compounds and glycoalkaloids were found in peeling byproduct. During processing, the amount of chlorogenic acid decreased, whereas the concentration of neochlorogenic acid increased due to isomerization. The impact of the results on potato processing technology is discussed.

Identification of complex, naturally occurring flavonoid glycosides in kale (Brassica oleracea var. sabellica) by high‐performance liquid chromatography diode‐array detection/electrospray ionization multi‐stage mass spectrometry

Kale is a member of the Brassicaceae family and has a complex profile of flavonoid glycosides. Therefore, kale is a suitable matrix to discuss in a comprehensive study the different fragmentation patterns of flavonoid glycosides. The wide variety of glycosylation and acylation patterns determines the health-promoting effects of these glycosides. The aim of this study is to investigate the naturally occurring flavonoids in kale. A total of 71 flavonoid glycosides of quercetin, kaempferol and isorhamnetin were identified using a high-performance liquid chromatography diode-array detection/electrospray ionization multi-stage mass spectrometry (HPLC-DAD/ESI-MS(n)) method. Of these 71 flavonol glycosides, 27 were non-acylated, 30 were monoacylated and 14 were diacylated. Non-acylated flavonol glycosides were present as mono-, di-, tri- and tetraglycosides. This is the first time that the occurrence of four different fragmentation patterns of non-acylated flavonol triglycosides has been reported in one matrix simultaneously. In addition, 44 flavonol glycosides were acylated with p-coumaric, caffeic, ferulic, hydroxyferulic or sinapic acid. While monoacylated glycosides existed as di-, tri- and tetraglycosides, diacylated glycosides occurred as tetra- and pentaglycosides. To the best of our knowledge, 28 compounds in kale are reported here for the first time. These include three acylated isorhamnetin glycosides (isorhamnetin-3-O-sinapoyl-sophoroside-7-O-D-glucoside, isorhamnetin-3-O-feruloyl-sophoroside-7-O-diglucoside and isorhamnetin-3-O-disinapoyl-triglucoside-7-O-diglucoside) and seven non-acylated isorhamnetin glycosides.

Characterization of pollen carotenoids with in situ and high-performance thin-layer chromatography supported resonant Raman spectroscopy

Raman signatures of the carotenoid component are studied in individual pollen grains from different species of trees. The information is obtained as differences in the strong pre-resonant Raman spectra measured before and after photodepletion of the carotenoid molecules. The results provide the first in situ evidence of interspecies differences in pollen carotenoid content, structure, and/or assembly between plant species without prior purification. The analysis of carotenoids in situ is confirmed by high-performance thin-layer chromatography (HPTLC)-supported resonance Raman data measured directly on the HPTLC plates after separation of carotenoids in pollen extracts. Utilization of the in situ, extraction-free procedure in carotenoid analysis will improve sensitivity and structural selectivity and provides insight into carotenoid structure and composition in single pollen grains.

Evaluation of Spent Coffee Obtained from the Most Common Coffeemakers as a Source of Hydrophilic Bioactive Compounds

The main hydrophilic antioxidant compounds (3-, 4-, and 5-monocaffeoylquinic and 3,4-, 3,5-, and 4,5-dicaffeoylquinic acids, caffeine, and browned compounds, including melanoidins) and the antioxidant capacity (Folin-Ciocalteu, ABTS, DPPH, Fremys salt, and TEMPO) were evaluated in Arabica and Robusta spent coffee obtained from the preparation of coffee brews with the most common coffeemakers (filter, espresso, plunger, and mocha). All spent coffee grounds, with the exception of those from the mocha coffeemaker, had relevant amounts of total caffeoylquinic acids (6.22-13.24 mg/g of spent coffee), mainly dicaffeoylquinic acids (3.31-5.79 mg/g of spent coffee), which were 4-7-fold higher than in their respective coffee brews. Caffeine ranged from 3.59 to 8.09 mg/g of spent coffee. The antioxidant capacities of the aqueous spent coffee extracts were 46.0-102.3% (filter), 59.2-85.6% (espresso), and <42% (plunger) in comparison to their respective coffee brews. This study obtained spent coffee extracts with antioxidant properties that can be used as a good source of hydrophilic bioactive compounds.

Influence of Brewing Method and Acidity Regulators on the Antioxidant Capacity of Coffee Brews

The antioxidant capacity of coffee brews prepared with different coffeemakers (filter, plunger, mocha, and espresso) was measured by colorimetric (total phenolic compounds and ABTS) and electron spin resonance (ESR) spectroscopy techniques (Fremys salt and TEMPO). The mocha coffeemaker had the highest yield in coffee antioxidant extraction per gram of ground roasted coffee, but espresso coffee was richest in terms of antioxidant intake (per milliliter of coffee brew) followed by mocha, plunger, and filter. Both Folin-Ciocalteu (total phenolic compounds) and ABTS assays reacted with standard solutions of chlorogenic acids (CGA) and melanoidins (MO-Ala and MO-Gly). However, Fremys salt was mainly scavenged by chlorogenic acids, whereas the stabilized radical TEMPO was effectively scavenged by melanoidins, but not by chlorogenic acids. Thus, ESR spectroscopy allows distinguishing between phenolic and nonphenolic antioxidants. Moreover, the addition of pH-regulator agents to coffee, such as sodium carbonate (75 ppm) and bicarbonate (75 ppm), to extend its shelf life, slightly increases the pH, modifying the antioxidant capacity in those coffee brews with the highest capacity (mocha and espresso).

Genotypic and climatic influence on the antioxidant activity of flavonoids in kale (Brassica oleracea var. sabellica).

The influence of genotype and climatic factors, e.g. mean temperature and mean global radiation level, on the antioxidant activity of kale was investigated. Therefore, eight kale cultivars, hybrid and traditional, old cultivars, were grown in a field experiment and harvested at four different times. In addition to the investigation of the total phenolic content, the overall antioxidant activity was determined by TEAC assay and electron spin resonance spectrometry. A special aim was to characterize the contribution of single flavonoids to the overall antioxidant activity using an HPLC-online TEAC approach. The antioxidant activity and the total phenolic content were influenced by the genotype and the eco-physiological factors. The HPLC-online TEAC results showed that not all flavonol glycosides contribute to the overall antioxidant activity in the same manner. Taking the results of the structural analysis obtained by HPLC-ESI-MS(n) into account, distinct structure-antioxidant relationships have been observed.

Structurally different flavonol glycosides and hydroxycinnamic acid derivatives respond differently to moderate UV-B radiation exposure

The aim of this study was to investigate the modifying influence of moderate ultraviolet-B (UV-B) radiation exposure on structurally different flavonol glycosides and hydroxycinnamic acid derivatives during pre-harvest using kale, a leafy Brassica species with a wide spectrum of different non-acylated and acylated flavonol glycosides. Juvenile kale plants were treated with short-term (1 day), moderate UV-B radiation [0.22-0.88 kJ m⁻² day⁻¹ biologically effective UV-B (UV-B(BE))]. Twenty compounds were quantified, revealing a structure-specific response of flavonol glycosides and hydroxycinnamic acid derivatives to UV-B radiation. A dose- and structure-dependent response of the investigated phenolic compounds to additional UV-B radiation was found. The investigated quercetin glycosides decreased under UV-B; for kaempferol glycosides, however, the amount of sugar moieties and the flavonol glycoside hydoxycinnamic acid residue influenced the response to UV-B. Monoacylated kaempferol tetraglucosides decreased in the investigated UV-B range, whereas the monoacylated kaempferol diglucosides increased strongly with doses of 0.88 kJ m⁻² day⁻¹ UV-B(BE) . The UV-B-induced increase in monoacylated kaempferol triglucosides was dependent on the acylation pattern. Furthermore, the hydroxycinnamic acid glycosides disinapoyl-gentiobiose and sinapoyl-feruloyl-gentiobiose were enhanced in a dose-dependent manner under UV-B. While UV-B radiation treatments often focus on flavonol aglycones or total flavonols, our investigations were extended to structurally different non-acylated and acylated glycosides of quercetin and kaempferol.

Electron spin resonance (ESR) studies on the formation of roasting-induced antioxidative structures in coffee brews at different degrees of roast.

The antioxidative properties of coffee brew fractions were studied using electron spin resonance spectroscopy using 2,2,6,6-tetramethyl-1-piperidin-1-oxyl (TEMPO) and Fremys salt (nitrosodisulfonate) as stabilized radicals. TEMPO was scavenged by antioxidants formed during roasting and not by chlorogenic acid, whereas Fremys salt was scavenged by all antioxidants tested including chlorogenic acid. The stabilized radical TEMPO allowed the exclusive measurement of roasting-induced antioxidants. The roasting-induced antioxidant activity of coffee brews increased with increasing degree of roast, and most of these antioxidants were formed during the initial roasting stage. The majority of these roasting-induced antioxidants were present in the high molecular weight fractions, indicating that the formation of these antioxidants preferably occurs at specific high molecular weight structures, likely being arabinogalactan and/or protein moieties which might be part of the melanoidin complex. It was found that chlorogenic acids most probably do not lose their antioxidant activity and phenolic characteristics upon incorporation in coffee melanoidins. The parameter fast reacting antioxidants (FRA) was introduced as an alternative for the antioxidative potential. FRA levels showed that coffee fractions rich in roasting-induced antioxidants exposed their antioxidant activity relatively slowly, which must be a consequence of its complex structure. Finally, the melanoidin content and the roasting-induced antioxidant activity showed a positive and linear correlation for the coffee brew fractions, showing that roasting-induced antioxidants are present within melanoidins. This is the first time that the formation of roasting-induced antioxidants could be directly correlated with the extent of Maillard reaction and melanoidin formation in a complex product such as coffee.