Flavor-protein binding: disulfide interchange reactions between ovalbumin and volatile disulfides

The irreversible binding of selected sulfur-containing flavor compounds to proteins was investigated in aqueous solutions containing ovalbumin and a mixture of disulfides (diethyl, dipropyl, dibutyl, diallyl, and 2-furfuryl methyl) using solid-phase micro-extraction (SPME). In systems which had not been heated, the recovery of disulfides from the headspace above the protein at the native pH (6.7) was similar to that from an aqueous blank. However, significant losses were observed when the pH of the solution was increased to 8.0. When the protein was denatured by heating, much greater losses were observed and some free thiols were produced. In similar heat-denatured systems at pH 2.0, no losses of disulfides were observed. Disulfides containing allyl or furfuryl groups were more reactive than saturated alkyl disulfides. Interchange reactions between protein sulfhydryl groups and the disulfides are believed to be responsible for the loss of the disulfides.     

Volatile compounds thermally generated from S-propylcysteine and S-propylcysteine sulfoxide-aroma precursors of Allium vegetables.

Two nonvolatile flavor precursors occurring in Allium vegetables, S-propyl-L-cysteine and its sulfoxide, were heated in closed model systems at different temperatures (from 80 to 200 degrees C) in the presence of variable amounts of water (0-98%) for 1-60 min. It seems to be indisputable that thermally generated breakdown products of both S-propyl-L-cysteine and particularly S-propyl-L-cysteine sulfoxide can significantly participate in the aroma formation of culinary processed Alliumvegetables. Dipropyl disulfide, dipropyl trisulfide, propylthiol, and dipropyl thiosulfonate were identified as the predominant volatile compounds generated by thermal degradation of S-propylcysteine sulfoxide. Dipropyl disulfide and 2-(propylthio)ethylamine were the major breakdown products formed from S-propylcysteine. Substantial amounts of various alkyl- and alkylthio-substituted pyridines were also generated from both S-propylcysteine and its sulfoxide.     

Structure and function relationship study of allium organosulfur compounds on upregulating the pi class of glutathione S-transferase expression

Allium organosulfides are potential chemopreventive compounds due to their effectiveness on the induction of phase II detoxification enzyme expression. In this study, we examined the structure and function relationship among various alk(en)yl sulfides on the expression of the pi class of glutathione S-transferase (GSTP) in rat Clone 9 cells, and what mechanism is involved. Cells were treated with 300 μM dipropyl sulfide (DPS), dipropyl disulfide (DPDS), propyl methyl sulfide (PMS), and propyl methyl disulfide (PMDS) for 48 h. DPDS and PMDS displayed more potency on GSTP protein and mRNA induction than that of DPS and PMS. Next, we compared the effectiveness of DPDS, PMDS, and diallyl disulfide (DADS), which have the same number of sulfur atoms but differ in the side alk(en)yl groups. The maximum increases on protein expression, mRNA level, and enzyme activity were noted in cells treated with DADS, followed by DPDS and PMDS. A reporter assay showed that three disulfides increased GSTP enhancer I (GPE I) activity (P < 0.05) in the order DADS > DPDS ≥ PMDS. Electromobility gel shift assays showed that the DNA binding of GPE I to nuclear proteins reached a maximum at 1 to 3 h after alk(en)yl disulfide treatment. Supershift assay revealed that c-jun bound to GPE I. Silencing of extracellular signal-regulated kinase (ERK) 2 expression inhibited c-jun activation and GSTP induction. Results suggest that both the type of alk(en)yl groups and number of sulfur atoms are determining factors of allium organosulfides on inducing GSTP expression, and it is likely related to the ERK-c-Jun-GPE I pathway.     

Investigation of the reaction between 4-hydroxy-5-methyl-3(2H)-furanone and cysteine or hydrogen sulfide at pH 4.5

Reaction of 4-hydroxy-5-methyl-3(2H)-furanone (HMF) with cysteine or hydrogen sulfide at pH 4.5 for 60 min at 140 degrees C produced complex mixtures of volatile compounds, the majority of which contained sulfur. Sixty-nine compounds were identified, some tentatively, by GC/MS. These included disulfides (26), thiols (7), dithiolanones (6), thiophenones (4), dithianones (3), and thienothiophenes (6). The main non-sulfur compounds were 2, 3-pentanedione, 2,4-pentanedione, and 3,4-hexanedione. Both systems produced approximately the same total quantity of volatile compounds, but the reaction containing cysteine gave the larger number of individual compounds, with thiols quantitatively the dominant components. By comparison, the major products formed in the reaction with hydrogen sulfide were the dithiolanones. Reaction pathways are presented for the major products and, where applicable, possible reasons for the differences in composition of the two systems are discussed. The contribution of these reactions, and their products, to the flavor of roasted foods is considered.     

Aging effects and grape variety dependence on the content of sulfur volatiles in wine

Thirteen sulfur compounds (boiling points from 35 to 231 degrees C), usually considered as possible off-flavoring volatiles, were quantified by a concurrent headspace-solid phase microextraction method coupled with gas chromatography-mass spectrometry (HS-SPME/GC-MS) on 80 not off-flavoring wines of four varieties (Merlot, Marzemino, and Teroldego as red wines and Chardonnay as a white one) and of five vintages produced in the North Italian Trentino region. The results of the research, the first Italian data-bank per variety on such volatiles, allow us to make a comparison with the data of other winegrowing areas, to investigate the aging effect on the considered volatiles, and, finally, to try a variety discrimination using statistical procedures. Dimethyl sulfide, 3-(methylthio)-1-propanol, diethyl sulfide, and diethyl disulfide were found to increase with time whereas 2-mercaptoethanol and ethylmercaptan showed a decreasing trend. Furthermore, the concentration of several compounds was found to be dependent on the variety. For instance, sulfide, disulfides, benzothiazole, and thioalcohols are at higher levels in Merlot wines, whereas thiols and thioacetates are more abundant in Marzemino and Teroldego wines. Chardonnay products, well apart from the other wines, are the poorest in 3-(methylthio)-1-propanol and rather rich in dimethyl disulfide and in diethyl disulfide, mostly in the aged wines. Applying the principal component analysis to the data, it was possible to demonstrate that Chardonnay and Merlot wines are well-discriminated from the Italian native varietal wines, which on their turn are only partially distinguishable among them. A contribution of these compounds to the variety characteristics of wine is reasonable.     

Determination of odor active aroma compounds in freshly cut leek (Allium ampeloprasum Var. bulga) and in long-term stored frozen unblanched and blanched leek slices by gas chromatography olfactometry analysis

The odor active compounds in freshly cut leek slices and in blanched and unblanched leek slices stored for 12 months were investigated by a detection frequency method. Fifteen judges were evaluating the three samples randomized. The most important aroma compounds in the freshly cut leek slices were dipropyl disulfide, methyl propenyl disulfide, pentanal, decanal, and propyl propenyl disulfide in order of priority. When stored frozen and unblanched for 12 months, the aroma composition changed and the most important compounds became pentanal, decanal, 2,5-dimethyl furan, unknown compound I, and dipropyl disulfide. Blanching before freezing prevented to some degree these changes but also reduced the perceived intensity of the aroma compounds. The most important aroma compounds in the blanched sample were dipropyl disulfide, unknown compound I, pentanal, 2,5-dimethyl furan, and propyl propenyl disulfide.     

Structure-activity relationship of neuroprotective and reactive oxygen species scavenging activities for allium organosulfur compounds

The neuroprotective and antioxidative activities of five organosulfur compounds with a thioallyl structure (-S-CH2CH=CH2) were characterized in terms of structure-activity relationships. Among five organosulfur compounds, only S-allyl-L-cysteine (SAC) having the alanyl group (-CH2CH-NH2-COOH) and lacking the oxo (O=) group with in between molecular properties, was effective in protecting cell death induced by both oxygen glucose deprivation and global cerebral ischemia. Conversely, lipophillic organosulfur compounds including diallyl sulfide, diallyl disulfide, and diallyl trisulfide were devoid of in vitro and in vivo neuroprotective activities. Furthermore, a significant correlation was only found between the in vivo neuroprotective activity and the OH- scavenging activity (gamma = 0.55 and p = 0.032) among reactive oxygen species scavenging activities. These results indicate that the presence of the alanyl group and the absence of the oxo group are essential for the manifestation of neuroprotective activity against ischemic insults and scavenging of OH radical, with SAC surfacing as a potent neuroprotectant.     

Identification of characteristic aroma components of Thai fried chili paste

Three forms of Thai fried chili pastes (CP) were prepared, consisting of an unheated CP (UH-CP), a CP heated at 100 degrees C for 25 min (H25-CP, typical product), and a CP excessively heated for 50 min (H50-CP). The potent odorants in the CPs were investigated by two gas chromatography-olfactometry methods: dynamic headspace dilution analysis (DHDA) and aroma extract dilution analysis (AEDA). DHDA revealed that the predominant odorants in heated CPs were mainly sulfur-containing compounds, followed by lipid-derived compounds, Strecker aldehydes, and Maillard reaction products. Dimethyl sulfide, allyl mercaptan, 2- (or 3-) methylbutanal, ally methyl sulfide, 2,3-butanedione, 3,3'-thiobis(1-propene), and methyl propyl disulfide were among the most potent headspace odorants detected by DHDA. By AEDA, 2-vinyl-4 H-1,3-dithiin and diallyl trisulfide had the highest FD factors in H25-CP. On the basis of their high FD factors by both GCO methods, the predominant odorants in H25-CP were 3-vinyl-4 H-1,2-dithiin, allyl methyl disulfide, and allyl methyl trisulfide. Furthermore, dimethyl trisulfide and diallyl disulfide had the highest odor activity values in H25-CP, suggesting that these were also potent odorants in CP. In addition, methional, 3-methylbutanoic acid, 4-hydroxy-2,5-dimethyl-3-(2 H)-furanone, and 3-hydroxy-4,5-dimethyl-2( 5H)-furanone (sotolon) were indicated as potent thermally derived odorants of H25-CP.     

Effect of Number of Carbons in Added Disulfide on Breadmaking Quality

Breadmaking properties such as bread height and specific volume were improved in bread from wheat flour with added disulfides such as dimethy disulfide (M₂S₂), diethyl disulfide (E₂S₂), dipropyl disulfide (Pro₂S₂), and dibutyl disulfide (B₂S₂). However, the amount of disulfides for maximum breadmaking properties was various and strongly related to the number of carbons (C) in the disulfide. Brabender farinographs of disulfide‐wheat flour showed an interesting profile, and the properties of the modification of the width of the tail. Size‐exclusion high performance liquid chromatography (SE‐HPLC) of the wheat proteins in the control and the disulfide‐added flours showed profiles of low, medium, and high molecular weight peaks. The area of the high molecular weight peak was larger in the disulfide‐added flours than in the control, indicating that this protein was important for the improvement of breadmaking properties.     

Sulfur compounds reduce potato toxins during extrusion cooking.

Free sulfhydryl groups in sulfur compounds have been reported to act directly on natural toxins to reduce toxicity. The objective of this study was to reduce protease inhibitors and glycoalkaloids in simulated snack foods by the addition of sulfur-containing compounds prior to extrusion. Thiamine, methionine, and benzyl disulfide were added to potato flakes at levels of 0.5% or 1.0% prior to twin-screw extrusion. Total and free thiols and protease inhibitors were monitored before and after extrusion by colorimetric assays. Potato glycoalkaloids were analyzed by HPLC and by immunoassay. Extrusion reduced potato flake disulfide bonds; disulfide bonds were higher in samples containing added sulfur compounds. Trypsin inhibitor activity was reduced by as much as 79% by extrusion plus methionine. Extrusion significantly reduced carboxypeptidase inhibitor, but only when benzyl disulfide and 0.5% methionine were not added. One percent methionine and thiamine resulted in 60% reductions in glycoalkaloids.     

两种叶菜类蔬菜对毒死蜱的吸收转移规律

采用水培方法,研究了毒死蜱对两种叶菜类蔬菜菠菜和生菜生长的影响、在不同培养液中的降解速度以及在蔬菜中的吸收和转移规律。结果表明,低浓度毒死蜱（1.0和10.0mg·L^-1）对两种供试蔬菜的生长没有明显影响,但高浓度毒死蜱（100.0mg·L^-1）对两种蔬菜的生长均有一定的影响,而且生菜对毒死蜱较菠菜更为敏感。两种蔬菜均能明显促进毒死蜱在溶液中的降解,在不同溶液中的降解速度如下：菠菜-培养液〉生菜-培养液〉塘水〉培养液。两种供试蔬菜对毒死蜱均有很强的吸收能力,而且具有相似的吸收规律。毒死蜱在菠菜根中达到最大吸收值所需的时间比生菜根所需的时间短,但在茎和叶中所需的时间两种蔬菜相同。     

Temperature-independent pectin gel method for aerobic plate count in dairy and nondairy food products: collaborative study

Ten laboratories participated in a collaborative study to compare the pectin-based plate count (PC) Redigel method with the aerobic plate count and standard plate count agar-based standard methods for the estimation of total bacterial counts in 9 different nondairy food and dairy food products. The foods were cream, homogenized milk, raw milk, cheese, raw chicken, raw oysters, frozen broccoli, flour, and spices. Each laboratory analyzed 6 samples (3 sample pairs) of each food group. Counts obtained by the pectin-based plate count and agar-based plate count methods differed significantly (P less than 0.05) only for homogenized milk, where the pectin gel method resulted in higher counts. The actual counts were higher in the pectin gel method in 8 of the 9 food groups. The log means for pectin gel and agar-based media, respectively, for the 9 food groups were: cream 8.106 and 7.844; homogenized milk 8.642 and 8.231; raw milk 8.711 and 8.423; chicken 7.654 and 7.645; oysters 7.201 and 7.180; broccoli 7.102 and 6.798; cheese 8.045 and 8.055; flour 4.112 and 3.988; spice 5.379 and 5.314. The repeatability standard deviations favored the pectin gel method in 6 of the 9 foods tested. The reproducibility standard deviations favored the pectin gel method in 7 of the 9 foods tested. These results strongly support the suitability of the pectin gel method as an alternative to agar-based plate count and other methods for total bacterial counts in nondairy and dairy food products. The pectin gel method has been adopted official first action.     

Deodorization with ku-ding-cha containing a large amount of caffeoyl quinic acid derivatives

Caffeoyl quinic acid (CQA) derivatives in ku-ding-cha, mate, coffee, and related plants were determined by HPLC. One ku-ding-cha contained a large amount of 3,5-dicaffeoylquinic acid (3,5-diCQA, 10.6% in dry weight) as well as 3-CQA (1.7%), 4-CQA (1.1%), 5-CQA (6.3%), 3,4-diCQA (1.8%), and 4,5-diCQA (4.3%). In this ku-ding-cha, the total caffeic acid moiety was 90.3 mmol/100 g of dry weight. The leaves of Ilex latifolia, which is one original species of ku-ding-cha, and another plant of the same genus, I. rotunda, also contained 3,5-diCQA (9.5 and 14.6%), 3-CQA (4.3 and 1.9%), and 5-CQA (4.8 and 3.8%), respectively, whereas raw coffee bean contained 5.5% 5-CQA and other low CQA derivatives. 3,5-DiCQA and 5-CQA with an apple acetone powder (AP) containing polyphenol oxidase showed high capturing activities toward thiols, and two addition compounds between 3,5-diCQA and methane thiol were also identified. Ku-ding-cha indicated extremely strong capturing activities toward methanethiol, propanethiol, and 2-propenethiol in the presence of apple AP. Furthermore, drinking ku-ding-cha reduced the amount of allyl methyl sulfide gas, well-known to persist as malodorous breath long after the ingestion of garlic.     

Synthesis and stable isotope dilution assay of ethanethiol and diethyl disulfide in wine using solid phase microextraction. Effect of aging on their levels in wine

Ethanethiol and diethyl disulfide (DEDS) most often occurred at levels above their olfactive threshold in wines with nauseous sulfur-linked smells. As ethanethiol is very oxidizable and chemically reactive, a stable isotopic dilution analysis of both ethanethiol and its disulfide in wines using solid phase microextraction and GC-MS was developed. The latter involved the determination of the proportion of DEDS formed by oxidation of the thiol during the analysis conditions, which was obtained by the use of two differently labeled disulfide standards. An original synthesis of labeled ethanethiol standards in conditions minimizing oxidation was developed, and the corresponding labeled diethyl disulfides were obtained from these thiols. This analytical method was used to follow the levels of these sulfur compounds during aging in a young red wine spiked with ethanethiol and added with enological tannins, with or without oxygen addition. The total levels of these two sulfur compounds were shown to decrease steadily after 60 days of aging, up to 83%. The effect of oxygen sped this decrease, but the effect of enological tannins was very slight. Residual ethanethiol was detected in its disulfide form from approximately 36% in the nonoxygenated wines to 69% in the oxygenated samples.     

硝酸稀土对菠菜中毒死蜱残留的降解作用研究

菠菜是我国蔬菜出口的重要品种之一,其毒死蜱残留量直接关系到我国农产品的出口和消费者的安全。采用气相色谱法（GC-NPD）测定毒死蜱残留量,研究了硝酸稀土对菠菜中毒死蜱残留动态的影响。结果表明,不论是喷施农药之前2d还是喷施农药之后2d喷施硝酸稀土,不同的硝酸稀土对菠菜中的毒死蜱残留都有不同程度的降解作用,且随着喷药后时间的延长,毒死蜱在菠菜中的残留量逐渐减少。不同时间喷施硝酸稀土,对菠菜中毒死蜱残留降解的效果存在差异,药后喷施的效果优于药前喷施。在硝酸稀土种类的选择上,首先选择对毒死蜱降解效果好的硝酸铈和硝酸钕,其次选择常乐益植素和硝酸镧。根据稀土农用的安全性分析,参考植物性食品中稀土最大残留限量标准,选择硝酸稀土作为农药残留降解制剂用于蔬菜安全生产,在技术上是可行的,人类食用是安全的。     

Evolution of volatile sulfur compounds from soils treated with sulfur-containing organic materials

Release of volatile S compounds from soils treated with S-containing organic materials was studied by sensitive gas chromatographic techniques. Methyl mercaptan, dimethyl sulfide, dimethyl disulfide, carbonyl sulfide and carbon disulfide were identified as gaseous products of decomposition of animal manures, sewage sludges and plant materials in soils under aerobic or waterlogged conditions. No release of hydrogen sulfide was detected. Most of the S volatilized from soils treated with sludges was in the form of dimethyl sulfide and dimethyl disulfide. whereas most of the S volatilized from soils treated with manures and plant materials was in the form of methyl mercaptan and dimethyl sulfide. More S compounds were released, and more S was volatilized, by decomposition of manures, sludges or plant materials in soils under waterlogged conditions than by decomposition under aerobic conditions. When calculated as a percentage of the S added as organic material, the average amount of S volatilized under aerobic or waterlogged conditions was < 0.2% < 0.5% and < 3.4% for the sludges, manures and plant materials, respectively. The five volatile S compounds produced by decomposition of manures, sludges and plant materials in soils under aerobic and waterlogged conditions also were produced by decomposition of plant proteins (zein, gluten and gliadin). It is concluded that the volatile S compounds released by decomposition of the above organic materials in soils are largely, if not entirely, produced by microbial degradation of methionine and cystine in these materials.     

Nonenzymatic antioxidant activity of four organosulfur compounds derived from garlic

The nonenzymatic antioxidant activity of diallyl sulfide (DAS), diallyl disulfide (DADS), S-ethyl cysteine (SEC), and N-acetyl cysteine (NAC) in the liposome system was examined. The antioxidant protection from these organosulfur agents was concentration dependent (p < 0.05). SEC and NAC showed significantly lower lipophilicity and greater reducing power than DAS and DADS (p < 0.05). Greater antioxidant protection was presented in the combinations of alpha-tocopherol with four organosulfur agents than alpha-tocopherol treatment alone (p < 0.05), and SEC and NAC showed greater sparing effects on alpha-tocopherol (p < 0.05). Four organosulfur agents lost antioxidant activity when the temperature was 65 degrees C (p < 0.05). At pH 2.5 and 10, DAS and DADS still showed antioxidant activity (p < 0.05). On the basis of the observed nonenzymatic antioxidant protection, these organosulfur compounds are potent agents for enhancing lipid stability.     

Comparative studies of the quantification of genetically modified organisms in foods processed from maize and soy using trial producing

Seven types of processed foods, namely, cornstarch, cornmeal, corn puffs, corn chips, tofu, soy milk, and boiled beans, were trial produced from 1 and 5% (w/w) genetically modified (GM) mixed raw materials. In this report, insect resistant maize (MON810) and herbicide tolerant soy (Roundup Ready soy, 40-3-2) were used as representatives of GM maize and soy, respectively. Deoxyribonucleic acid (DNA) was extracted from the raw materials and the trial-produced processed food using two types of methods, i.e., the silica membrane method and the anion exchange method. The GM% values of these samples were quantified, and the significant differences between the raw materials and the trial-produced processed foods were statistically confirmed. There were some significant differences in the comparisons of all processed foods. However, our quantitative methods could be applied as a screening assay to tofu and soy milk because the differences in GM% between the trial-produced processed foods and their raw materials were lower than 13 and 23%, respectively. In addition, when quantitating with two primer pairs (SSIIb 3, 114 bp; SSIIb 4, 83 bp for maize and Le1n02, 118 bp; Le1n03, 89 bp for soy), which were targeted within the same taxon specific DNA sequence with different amplicon sizes, the ratios of the copy numbers of the two primer pairs (SSIIb 3/4 and Le1n02/03) decreased with time in a heat-treated processing model using an autoclave. In this report, we suggest that the degradation level of DNA in processed foods could be estimated from these ratios, and the probability of GM quantification could be experimentally predicted from the results of the trial producing.     

Food and Drug Administration pesticide residue monitoring of foods: 1978-1982

Pesticide residues in foods are reported for the 5-year period 1978-1982 [fiscal years (FY) 78-82]. Results were compiled from the 2 complementary elements that comprise the Food and Drug Administration's (FDA) program for monitoring pesticide residues in foods. Under regulatory monitoring, which focuses on residues in raw agricultural commodities, a total of 49,877 samples (30,361 domestic and 19,516 import) that included fresh fruits and vegetables, grains, milk and dairy products, seafoods, and a variety of processed foods were analyzed. No residues were found in about 55 and 44% of the domestic and import samples, respectively. About 3% of the domestic and 7% of the import samples were classed as violative. Data from the Total Diet Study, which is conducted to determine dietary intakes of a variety of chemicals, showed that residues of 42 pesticides were found in 1044 composites of table-ready foods. Results of FDA's monitoring for FY78-82 demonstrate that pesticide residue levels in the U.S. food supply were generally well below regulatory limits, and dietary intakes were manyfold lower than the Acceptable Daily Intakes established by international agencies.     

Antioxidative and antiglycative effects of six organosulfur compounds in low-density lipoprotein and plasma

Low-density lipoprotein (LDL) and plasma were isolated from patients with non-insulin-dependent diabetes. The protective effects of six organosulfur compounds (DAS, diallyl sulfide; DADS, diallyl disulfide; SAC, S-allylcysteine; SEC, S-ethylcysteine; SMC, S-methylcysteine; SPC, S-propylcysteine) against further oxidation and glycation in these already partially oxidized and glycated samples were studied. DAS and DADS showed significantly greater oxidative-delaying effects than four cysteine-containing compounds in both partially oxidized LDL and plasma samples (P < 0.05). However, cysteine-containing agents were superior to DAS and DADS in delaying glycative deterioration in already partially glycated LDL (P < 0.05). The observed delays of oxidative and glycative effects from each agent were significantly concentration-dependent (P < 0.05). Furthermore, six organosulfur agents significantly decreased the loss of catalase and glutathione peroxidase activities in plasma and increased alpha-tocopherol retention in LDL and plasma (P < 0.05). These results suggested that the use of these organosulfur agents derived from garlic at these concentrations could protect partially oxidized and glycated LDL or plasma against further oxidative and glycative deterioration, which might benefit patients with diabetic-related vascular diseases.