Tetrahydro-beta-carboline alkaloids occur in fruits and fruit juices. Activity as antioxidants and radical scavengers

Tetrahydro-beta-carbolines are biologically active alkaloids that occur and accumulate in mammalian tissues, fluids, and brain, but their ultimate origin or biological role is still uncertain. Four tetrahydro-beta-carboline alkaloids: 1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid, 1-methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid, 1-methyl-1,2,3,4-tetrahydro-beta-carboline, and 6-hydroxy-1-methyl-1,2,3,4-tetrahydro-beta-carboline, are found as naturally occurring substances in some fruit and fruit juices. These compounds occur in the microg/g level in those products, and a characteristic and distinct profile appears to exist depending on the type of fruit and juice involved. Thus, 1-methyl-1,2,3,4-tetrahydro-beta-carboline may appear in tomato, tomato juice, and kiwi; 6-hydroxy-1-methyl-1,2,3,4-tetrahydro-beta-carboline in bananas, pineapple, tomato, and their corresponding juices; and 1-methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid in oranges and grapefruits, although it also occurred in most juices. Fruit-occurring tetrahydro-beta-carboline alkaloids acted as antioxidants and free radical scavengers in the ABTS assay when compared with ascorbic acid and Trolox. This suggests that tetrahydro-beta-carboline alkaloids might act as antioxidants when absorbed and accumulated in the body, contributing to the antioxidant effect of fruit products containing these compounds.     

Tetrahydro-beta-carbolines, potential neuroactive alkaloids, in chocolate and cocoa

Tetrahydro-beta-carbolines (THbetaCs), potential neuroactive alkaloids, were found in chocolate and cocoa. 6-Hydroxy-1-methyl-1,2, 3,4-tetrahydro-beta-carboline (6OHMTHbetaC), 1,2,3, 4-tetrahydro-beta-carboline-3-carboxylic acid (THCA), 1-methyl-1,2,3, 4-tetrahydro-beta-carboline-3-carboxylic acid (MTCA) in both diastereoisomers (1S,3S and 1R,3S), and 1-methyl-1,2,3, 4-tetrahydro-beta-carboline (MTHbetaC), besides serotonin and tryptamine biogenic amines, were identified and quantified in dark chocolate, milk chocolate, cocoa, and chocolate-containing cereals by RP-HPLC-fluorescence and HPLC-MS. For each THbetaC, the concentration ranges were determined: 6OHMTHbetaC (0.16-3.92 microg/g), THCA (0.01-0.85 microg/g), 1S,3S-MTCA (0.35-2 microg/g), 1R,3S-MTCA (0.14-0.88 microg/g), and MTHbetaC (nd-0.21 microg/g). The highest content was generally found in chocolates and cocoas, but cereals containing chocolate also showed an appreciable amount of THbetaCs. The possible biological implications of this novel group of alkaloids in chocolate are discussed.     

1-methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid and 1,2, 3,4-tetrahydro-beta-carboline-3-carboxylic acid in fruits

1,2,3,4-Tetrahydro-beta-carboline-3-carboxylic acid (THCA) and 1-methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (MTCA), as two diastereoisomers (1S,3S and 1R,3S), occurred in commercial fruits. Citrus fruits exhibited the highest content; other fruits contained very low levels or none at all. The content of MTCA was as follows: orange, 0.35-2.47 microg/g; lemon, 0.15-2.05 microg/g; grapefruit, 1.12-8.37 microg/g; mandarin, 0.57-2.5 microg/g; banana, nd-0.74 microg/g; pear, nd-0.017 microg/g; grape, 0.01-0.22 microg/g, tomato, 0.05-0.25 microg/g; and apple, nd-0.012 microg/g). THCA, if present, usually occurred at <0.05 microg/g. Fruit ripening and softening during storage were accompanied with a significant increase of MTCA, in both pears and bananas. Those and previous results confirm that foods are an important source of tetrahydro-beta-carbolines in humans.     

Quantitation of selected odor-active constituents in dry fermented sausages prepared with different curing salts

The odor-active compounds of dry-fermented sausages with added nitrite or nitrate as curing agents were identified by gas chromatography-olfactometry (GC-O) applying the detection frequency (DF) method. The quantification of these compounds in the sausage was determined by multiple headspace solid-phase microextraction (multiple HS-SPME). There were no specific odor-active compounds related to the use of nitrite or nitrate although there were differences in the DF value of several compounds. The nitrite-added sausages presented higher DF values for ethanol, 1-hexanol, propanoic acid, 2-heptenal, and nonanal while the nitrate-added sausages had higher DF values for phenylacetaldehyde and 3-methyl-butanal. Eighteen compounds were quantified by multiple HS-SPME. Most of them were above their air detection thresholds, but only hexanal, heptanal, and 1-octen-3-ol were in a concentration higher than their oil threshold values. These compounds would probably be the main contributors to the aroma of fermented sausages.     

Influence of the simultaneous addition of the protease flavourzyme and the lipase novozym 677BG on dry fermented sausage compounds extracted by SDE and analyzed by GC-MS

A dry fermented sausage (chorizo de Pamplona) was elaborated with the simultaneous addition of a lipase (Novozym 677BG) and a protease (Flavourzyme) and ripened during 21 days, in contrast to the control without enzymes and ripened during 35 days. Faster and more intense lipolytic and proteolytic activities were observed in the modified sausage, despite its shorter maturation time. At the end of the ripening, a determination of the profile of compounds extracted by simultaneous distillation-extraction with dichloromethane was carried out. The total amounts of extracted compounds (expressed in milligrams of dodecane per gram of dry matter) were 2.5 in the sausage with enzymes and 1.9 in the control. The chemical groups showing increments due to the use of enzymes were esters (103.5% increment) and acids (87% increment) in both cases due to the greater presence of long-chain fatty acid products. However, development of substances originated from further degradation process of amino acids and free fatty acids did not seem to have taken place.     

辐照处理提高猪肉火腿肠保鲜效果

辐照是一种非常有效的食品杀菌保鲜技术,近年来在即食肉制品保鲜方面的应用逐渐引起了人们的关注。为了探讨γ辐照技术对猪肉火腿肠的杀菌保鲜效果,试验采用1、3、5、7、9 k Gy 5个剂量分别对火腿肠进行辐照处理,辐照结束后样品在4℃条件下冷藏,然后跟踪测定其在冷藏过程中脂肪和蛋白质氧化、颜色、p H值、挥发性盐基氮(total volatile basic nitrogen,TVB-N)以及菌落总数的变化,分析不同剂量辐照处理对火腿肠保鲜效果的影响。结果表明,辐照可以显著降低火腿肠脂肪和蛋白质的氧化稳定性,促进其在储藏过程中的氧化。1 k Gy剂量辐照对火腿肠的色泽影响不显著;3~9 k Gy剂量辐照,可以使样品的红度值(a*值)显著降低、亮度值(L*值)显著升高(P0.05),而在冷藏过程中a*值与L*值均随储藏时间的延长而降低,且高剂量辐照组要比低剂量组降低更快一些。火腿肠p H值受辐照影响不显著(P0.05),且在冷藏过程中所有处理组火腿肠的p H值变化也不显著,均在6.0~6.5范围内。5 k Gy剂量辐照即可对猪肉火腿肠中的微生物起到有效抑制作用。     

Degradation of tetrahydro-beta-carbolines in the presence of nitrite: HPLC-MS analysis of the reaction products.

Motivated by the identification of numerous novel tetrahydro-beta-carboline-carboxylic acids in food samples, we studied the reactions of tetrahydro-beta-carbolines in the presence of nitrosating agents. The anticipated formation of nitroso derivatives from unsubstituted tetrahydro-beta-carbolines, and from tetrahydro-beta-carboline-3-carboxylic acids was indicated by HPLC-MS/MS analysis and validated by the characteristic product ion spectra of the respective nitroso compounds. In addition, oxidative decarboxylation resulted in formation of the corresponding dihydro-beta-carbolines, and in the generation of the beta-carbolines harman or norharman. Subsequently, we studied the reactivity of tetrahydro-beta-carboline-1-carboxylic acids derived from the Pictet-Spengler condensation of indole amines with alpha-oxo acids. Again, in the presence of nitrosating agents the rapid disappearance of the starting material was obvious, but no nitroso derivatives could be observed. Instead, further HPLC-MS/MS studies demonstrated that dihydro-beta-carbolines were the major products of tetrahydro-beta-carboline-1-carboxylic acids. Finally, we demonstrated that freshly isolated nitroso-precursors spontaneously decomposed to yield harman alkaloids. In conclusion, we revealed that nitroso-tetrahydro-beta-carbolines can represent intermediates involved in the generation of beta-carbolines, and we established a novel pathway for the formation of harman alkaloids from nutritional tetrahydro-beta-carbolines.     

Identification and occurrence of the novel alkaloid pentahydroxypentyl-tetrahydro-beta-carboline-3-carboxylic acid as a tryptophan glycoconjugate in fruit juices and jams

The novel carbohydrate-derived beta-carboline, 1-pentahydroxypentyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid, was identified in fruit- and vegetable-derived products such as juices, jams, and tomato sauces. This compound occurred as two diastereoisomers, a cis isomer (the major compound) and a trans isomer, ranging from undetectable amounts to 6.5 microg/g. Grape, tomato, pineapple, and tropical juices exhibited the highest amount of this alkaloid (up to 3.8 mg/L), whereas apple, banana, and peach juices showed very low or nondetectable levels. This tetrahydro-beta-carboline was also found in jams (up to 0.45 microg/g), and a relative high amount was present in tomato concentrate (6.5 microg/g) and sauce (up to 1.8 microg/g). This beta-carboline occurred in fruit-derived products as a glycoconjugate from a chemical condensation of D-glucose and L-tryptophan that is highly favored at low pH values and high temperature. Production, processing treatments, and storage of fruit juices and jams can then release this beta-carboline. Fruit-derived products and other foods containing this compound might be an exogenous dietary source of this glucose-derived tetrahydro-beta-carboline.     

Smoke-derived taint in wine: effect of postharvest smoke exposure of grapes on the chemical composition and sensory characteristics of wine

Although smoke exposure has been associated with the development of smoke taint in grapes and subsequently in wine, to date there have been no studies that have demonstrated a direct link. In this study, postharvest smoke exposure of grapes was utilized to demonstrate that smoke significantly influences the chemical composition and sensory characteristics of wine and causes an apparent 'smoke taint'. Verdelho grapes were exposed to straw-derived smoke for 1 h and then fermented according to two different winemaking treatments. Control wines were made by fermenting unsmoked grapes. Sensory studies established a perceivable difference between smoked and unsmoked wines; smoked wines were described as exhibiting 'smoky', 'dirty', 'earthy', 'burnt' and 'smoked meat' characteristics. Quantitative analysis, by means of gas chromatography-mass spectrometry, identified guaiacol, 4-methylguaiacol, 4-ethylguaiacol, 4-ethylphenol, eugenol, and furfural in each of the wines made from smoked grapes. However, these compounds were not detected in the unsmoked wines, and their origin is therefore attributed to the application of smoke. Increased ethanol concentrations and browning were also observed in wines made from grapes exposed to smoke.     

Liquid chromatography-tandem mass spectrometry analysis of 2-amino-1-methyl-6-(4-hydroxyphenyl)imidazo[4,5-b]pyridine in cooked meats

Several cooked meats such as beef (fried, coated-fried), pork (fried, coated-fried), and chicken (fried, griddled, coated-fried, roasted) were analyzed for the heterocyclic amine 2-amino-1-methyl-6-(4-hydroxyphenyl)imidazo[4,5- b]pyridine (4'-OH-PhIP) not commonly determined in food and 2-amino-1-methyl-6-phenylimidazo[4,5- b]pyridine (PhIP). The highest content of 4'-OH-PhIP was found in fried and griddled chicken breast, the concentration being 43.7 and 13.4 ng/g, respectively, whereas the corresponding PhIP concentrations were 19.2 and 5.8 ng/g. The estimated concentration of both pyridines in fried pork loin, in fried pork sausages, and in coated-fried chicken was below 2.5 ng/g. In the rest of the samples, 4'-OH-PhIP was not detected. The analyses were performed by solid-phase extraction and LC-MS/MS. The fragmentation of 4'-OH-PhIP in an ion trap mass analyzer was studied in order to provide information for the identification of 4'-OH-PhIP. Additionally, the effect of red wine marinades on the formation of 4'-OH-PhIP in fried chicken was examined, finding a notable reduction (69%) in the amine's occurrence.     

Antiplatelet activity of soy sauce as functional seasoning.

In seeking the functionality of foodstuffs applicable to medicine, soy sauce was found to show antiplatelet activity. Therefore, the active components in soy sauce were purified, structurally identified, and studied for their inhibitory effects on the aggregation of human platelets. Aqueous 2-fold diluents of soy sauce inhibited platelet aggregation induced by collagen and epinephrine depending on the dilution factor. Since a basic extract with diethyl ether completely inhibited collagen-induced aggregation, it was subjected to serial extractions and multistep HPLC fractionations for purifying antiplatelet components. The finally obtained isolates were identified as 1-methyl-1,2,3,4-tetrahydro-beta-carboline and 1-methyl-beta-carboline on the basis of EI-MS, (1)H NMR, diode array, and fluorescence spectra. Their spectral data and chromatographic behaviors were the same as those of synthetic ones. 1-Methyl-1,2,3, 4-tetrahydro-beta-carboline showed mean concentrations (n = 5-6) of 4.6, 4.2, 28.6, 11.6, and 65.8 microgram/mL to produce 50% inhibition of the maximal aggregation response induced by epinephrine, platelet-activating factor, collagen, adenosine 5'-diphosphate, and thrombin, respectively. Its inhibitory effect was much greater than that of 1-methyl-beta-carboline on platelet aggregation by all the tested inducers. The quantitative HPLC analysis revealed that the significant amounts of both antiplatelet compounds were uniformly contained in commercially available soy sauce. From these results, soy sauce may be referred to as functional seasoning containing alkaloidal components with the potent preventive effect on thrombus formation.     

基于超声成像技术的火腿肠质构分析与等级判别

为了研究超声成像技术在火腿肠质构分析与等级判别方面应用的可行性。通过对火腿肠蛋白质、淀粉等理化指标的测定将其分为特级、优级、普通级,并采集2个品牌3个等级的火腿肠共240份超声图像信息,在Matlab 7.0环境下提取图像角二阶矩、平均值等纹理特征值,最后利用线性判别式分析(linear discriminant analysis,LDA)和支持向量机(support vector machine,SVM)建立火腿肠的等级判别模型。结果表明:同品牌不同等级火腿肠超声图像、纹理特征值均具有较大差异,而同等级不同品牌火腿肠差异较小。建立的识别模型中:SVM优于LDA模型,当主成分为3时,SVM模型对应的校正集、预测集识别率均为100%,模型效果最佳。因此,超声成像技术可实现火腿肠内部质构的分析和等级的快速准确识别,研究结果可为超声成像技术在火腿肠内部质构分析和等级判别方面的应用提供参考。     

Sulfuric acid/hydrogen peroxide digestion and colorimetric a collaborative study

Seven laboratories participated in a collaborative study of a method for determination of phosphorus in meat and meat products. Samples are digested in sulfuric acid and hydrogen peroxide; digestion is complete in approximately 10 min. Phosphorus is determined by colorimetric analysis of a dilute aliquot of the sample digest. The collaborators analyzed 3 sets of blind duplicate samples from each of 6 classes of meat (U.S. Department of Agriculture classifications): smoked ham, water-added ham, canned ham, pork sausage, cooked sausage, and hamburger. The calibration curve was linear over the range of standard solutions prepared (phosphorus levels from 0.05 to 1.00%); levels in the collaborative study samples ranged from 0.10 to 0.30%. Standard deviations for repeatability (sr) and reproducibility (SR) ranged from 0.004 to 0.012 and 0.007 to 0.014, respectively. Corresponding relative standard deviations (RSDr and RSDR, respectively) ranged from 1.70 to 7.28% and 3.50 to 9.87%. Six laboratories analyzed samples by both the proposed method and AOAC method 24.016 (14th Ed.). One laboratory reported results by the proposed method only. Statistical evaluations indicated no significant difference between the 2 methods. The method has been adopted official first action by AOAC.     

Simultaneous determination of dextrose, sucrose, maltose, and lactose in sausage products by liquid chromatography

A liquid chromatographic (LC) method for the simultaneous determination of dextrose, sucrose, maltose, and lactose in sausage products has been developed. Dextrose, sucrose, maltose, and lactose are extracted from comminuted meat products with 52% ethanol. After filtration, the extracts are purified by passing them through a C18 Sep-Pak cartridge and 2 ion exchange resin Econo-columns in series. After concentration and filtration, extracts are analyzed by LC using a normal phase amino column and a differential refractometer detector. Homogeneously ground samples of cooked and fresh sausages are fortified with dextrose, sucrose, maltose, and lactose at 4 different concentrations. Average recovery for dextrose, sucrose, maltose, and lactose at all 4 levels of fortification was greater than 80% with a coefficient of variation less than 10%.     

Comparison of odor-active volatile compounds of fresh and smoked salmon

The odorant volatile compounds of raw salmon and smoked salmon have been investigated by two gas chromatography-olfactometry methods (frequency detection and odorant intensity) and gas chromatography-mass spectrometry. After simultaneous steam distillation-solvent extraction with diethyl ether and the recovery of the aromatic extract in ethanol, qualitative olfactometric characterization and identification followed by a quantitative assessment of the odorant volatile compounds were carried out. The origin of many odorant compounds of smoked salmon can be attributed to wood smoke. Another part of smoked salmon aroma is due either to the odorant compounds of the raw fish flesh or to an evolution of fish flesh aroma thanks to the smoking process conditions. Forty-nine odorant compounds have been identified in fresh salmon and 74 in smoked salmon. Carbonyl compounds, such as heptanal or (E,Z)-2,6-nonadienal, show a high detection frequency and odorant intensity in unsmoked fish, giving the flesh its typical fishy odor. For smoked salmon, phenolic compounds, such as cresol or guaiacol, and furanic compounds seem to be responsible for the smoked odor.     

辐照对发酵香肠品质特性的影响

本文以0、2、4、6kGy剂量60Co γ射线辐照发酵香肠,以水分活度、水分含量、pH值、含盐量、氨基酸总量、非蛋白氮、总氮、硫代巴比妥酸值、挥发性风味物质、肌浆蛋白与肌原纤维蛋白降解情况为指标,研究辐照对香肠品质的影响。结果表明：发酵香肠经不同剂量辐照后水分含量差异显著(P<0.05),氯化钠含量差异并不显著,而0、2kGy辐照后发酵香肠的pH值显著高于4、6kGy辐照,4kGy剂量辐照的发酵香肠中氨基酸总量显著高于2、6kGy处理的样品(P<0.05),4kGy剂量辐照香肠烯烃、酸和酮类物质含量最多,6kGy辐照组中醇类物质含量最多。辐照处理的发酵香肠中肌浆蛋白和肌原蛋白的降解情况低于对照。     

Flavor characterization of ripened cod roe by gas chromatography, sensory analysis, and electronic nose

Characterization of the flavors of ripened roe products is of importance to establish a basis for a standardized product. Flavor profiles of commercially processed ripened roe from Iceland and Norway were studied by sensory analysis, gas chromatography-olfactometry (GC-O), gas chromatography-mass spectrometry (GC-MS), and an electronic nose to characterize the headspace of ripened roe. Sensory analysis showed that ripened roe odor and flavor in combination with caviar flavor and whey/caramel-like odor give the overall positive effect of the complex characteristic roe flavor. Analysis of volatiles by GC-MS and electronic nose confirmed the presence of aroma compounds contributing to the typical ripening and spoilage flavors detected by the sensory analysis. Methional, 1-octen-3-ol, and 2,6-nonadienal were the most important compounds contributing to ripened roe odor. Spoilage flavors were partly contributed by 3-methyl-1-butanol and 3-methylbutanal, which can be measured by the electronic nose and are suggested as quality indicators for objectively assessing the ripening of roe. Principal component analysis of the overall data showed that GC-O correlated well with sensory evaluation and the electronic nose measurements.     

Difference in the volatile composition of pine-mushrooms (Tricholoma matsutake Sing.) according to their grades

The differences in volatile components of pine-mushrooms (Tricholoma matsutake Sing.) according to their grades were observed by applying multivariate statistical methods to GC-MS data sets. A total of 35 and 37 volatile components were identified in raw and cooked pine-mushrooms, respectively. The volatile components in pine-mushrooms were primarily composed of C8 species, such as 3-octanol, 1-octen-3-ol, 1-octanol, (E)-2-octen-1-ol, 3-octanone, 1-octen-3-one, (E)-2-octenal, and octanoic acid. The levels of ethyl octanoate, junipene, and 3-methyl-3-buten-2-one were much higher in raw pine-mushroom of higher grades, whereas the reverse was true for C8 components. On the other hand, furfuryl alcohol, benzyl alcohol, phenylethyl alcohol, dihydro-5-methyl-2(3H)-furanone, 2(5H)-furanone, (E)-2-methyl-2-butenal, furfural, phenylacetaldehyde, benzoic acid methyl ester, camphene, and beta-pinene were the major components of cooked mushrooms. These volatile components formed by various thermal reactions could be mainly responsible for the difference in volatile components of cooked pine-mushrooms according to their grades.     

SPME-GC-MS versus Selected Ion Flow Tube Mass Spectrometry (SIFT-MS) analyses for the study of volatile compound generation and oxidation status during dry fermented sausage processing

The use of selected ion flow tube mass spectrometry (SIFT-MS) and gas chromatography-mass spectrometry together with solid phase microextraction (GC-MS-SPME) has been compared in the analysis of volatile compounds during dry fermented sausage processing. Thus, the headspace (HS) of samples of dry fermented sausages with different fat contents was analyzed during their manufacture using both techniques, and significant and positive correlations were found between SIFT-MS and SPME-GC-MS measurements for the compounds pentanal, hexanal, 2-heptenal, octanal, 2-nonenal, 2-butanone, 2-pentanone, ethanol, acetic acid, and hexanoic acid. The oxidative status of fermented sausages during processing was also evaluated, and a significant correlation was obtained between the HS concentration of lipid autoxidation volatile compounds measured by SIFT-MS and SPME-GC-MS and the level of thiobarbituric acid reactive substances (TBARS) in the sausage. The hexanal measured by SIFT-MS resulted in a higher correlation coefficient (r = 0.936) than that obtained using SPME-GC-MS (r = 0.927). SIFT-MS is shown to be a fast, real time analytical technique for monitoring changes in the profile of volatile compounds in dry fermented sausages during processing and a useful tool to evaluate the oxidative status of meat products.     

L-tryptophan reacts with naturally occurring and food-occurring phenolic aldehydes to give phenolic tetrahydro-beta-carboline alkaloids: activity as antioxidants and free radical scavengers

The reaction between the essential amino acid l-tryptophan and flavoring or naturally occurring phenyl and phenolic aldehydes was studied, and the alkaloidal reaction products were characterized by NMR and HPLC-MS. Benzaldehyde, vanillin, syringaldehyde, salicylaldehyde, and anisaldehyde condensed with l-tryptophan in aqueous-acidic media affording the corresponding phenolic tetrahydro-beta-carboline-3-carboxylic acid as two diastereoisomers, 1S,3S-cis and 1R,3S-trans. With the exception of benzaldehyde, the rest of the aldehydes needed heating conditions (70 degrees C) to significantly form tetrahydro-beta-carbolines over time with the cyclization highly favored at low pH. This suggests a likely formation of these compounds under conditions that may occur in foods, food processing, or cooking. The new phenolic tetrahydro-beta-carboline alkaloids were assayed, for the first time, for their activity as free radical scavengers and antioxidants and showed good antioxidant properties with Trolox equivalent antioxidant capacity (TEAC) values much higher than those of ascorbic acid and the water soluble vitamin E analogue, Trolox, in the 2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) assay.