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.     

Second derivative spectrophotometric determination of acetaminophen and phenacetin in presence of their degradation products

A rapid and accurate method for determining acetaminophen and phenacetin in presence of their degradation products is presented. Solutions of these drugs in 0.1N HCl were analyzed by measuring their second derivative spectral response at 295 nm where the degradation products do not interfere. The mean percent recoveries for mixtures of acetaminophen and/or phenacetin with the corresponding degradation products were 100.2 +/- 0.6 and 100.6 +/- 1.1, respectively. The method can be used for assessing the stability of the 2 drugs. The proposed method is also applied to the determination of acetaminophen in tablets and syrups.     

Pyrolysis of lignin in the presence of tetramethylammonium hydroxide (TMAH): products stemming from beta-5 substructures

Lignin model compounds, synthetic lignins, and cedar wood have been analyzed by pyrolysis-gas chromatography(-mass spectrometry) in the presence of tetramethylammonium hydroxide (TMAH) to examine the behavior of beta-5 substructures specifically under these conditions. Two model compounds contained a beta-5 linkage and a gamma-CH2OH group. The phenolic model compound produced stilbene products by way of a formaldehyde elimination of the gamma-CH2OH. The nonphenolic model compound underwent dehydration to give arylbenzofuran products. Dehydrogenation polymers of coniferyl alcohol gave a large amount of stilbene products in TMAH/pyrolysis. TMAH/pyrolysis of a Japanese cedar (Cryptomeria japonica) wood yielded a very small amount of stilbene products. The results demonstrated that synthetic lignins are rich in terminal beta-5 substructures, but cedar (a softwood) contains a paucity of the terminal beta-5 substructures.     

纤维素超低酸水解产物的分析

基于木质纤维素类生物质水解为人类提供乙醇等能源、化工产品的重要性，该文以定量滤纸模拟生物质的主要组分－纤维素，以其超低酸水解试验得到的最佳工况下液体产物和固体残渣为研究对象，通过高效液相色谱(HPLC)结合KS-802糖柱对产物质中糖的种类进行了划分，以GC-MS对副产物进行定性，并通过扫描电镜(SEM)及热重和工业分析元素分析对固体残渣作了从表观到内部的研究，发现纤维素超低酸水解主要生成纤维四糖、三糖和二糖等低聚糖和葡萄糖及果糖，反应副产物有糠醛、羟甲基糠醛、乙酰丙酸及一些小分子酮、醛类和酸类极性化合物，水解残渣已完全改变了原始形貌，热裂解活性增强，残留物中碳和灰分含量有较大增加，最后根据分析结果探讨了纤维素超低酸水解的反应途径，为木质纤维素类生物质水解技术的规模化利用打下基础。     

Analysis of free and esterified ergosterol in tomato products

A new extraction and chromatographic procedure to quantify free and esterified ergosterol in tomato products was devised. The extraction solution was composed of a dichloromethane/methanol mixture in a 2:1 (v/v) ratio. This extraction solvent allowed for higher ergosterol recovery from tomato products (an average of 25% more) compared to hexane, which is frequently employed for ergosterol extraction. Both free and esterified ergosterol were determined by HPLC reverse-phase chromatography employing a Nova-Pak C-18 column (300 x 3.9 mm), filled with 4 mm average particle size and a guard column of the same material. The elution was performed at a flow rate of 1 mL. min(-1) with a linear gradient of solvent A (methanol/water, 80:20, v/v) and solvent B (dichloromethane). The gradient, starting at sample injection, was from 0 to 50% B for 20 min for the free ergosterol analysis and additional 15 min at 50% B to analyze the ergosterol esters. This technique has proven to be more sensitive for ergosterol determination than other reported chromatographic procedures. Moreover, ergosterol esters, extracted from various fungal sources, separated well and were easily quantified.     

Ameliorative effects of proanthocyanidin on oxidative stress and inflammation in streptozotocin-induced diabetic rats

Recent evidence strongly suggests that oxidative stress due to redox imbalance is causally associated with inflammatory processes and various diseases including diabetes. We examined the effects of proanthocyanidin from persimmon peel, using both oligomers and polymers, against oxidative stress with elucidation of the underlying mechanisms in streptozotocin-induced diabetic rats. The elevation of lipid peroxidation in the kidney and serum under the diabetic condition was decreased by the administration of proanthocyanidin. The suppression of reactive oxygen species generation and elevation of the reduced glutathione/oxidized glutathione ratio were observed in the groups administered proanthocyanidin. These results support the protective role of proanthocyanidin from oxidative stress induced by diabetes. Moreover, proanthocyanidin, especially its oligomeric form, affected the inflammatory process with regulation of related protein expression, inducible nitric oxide synthase, cyclooxygenase-2, and upstream regulators, nuclear factor kappaB, and inhibitor-binding protein kappaB-alpha. Proanthocyanidin ameliorated the diabetic condition by decreases of serum glucose, glycosylated protein, serum urea nitrogen, urinary protein, and renal advanced glycation endproducts. In particular, oligomeric proanthocyanidin exerted a stronger protective activity than the polymeric form. This suggests that the polymerization of proanthocyanidin has an effect on its protective effect against diabetes. The present study supports the beneficial effect of proanthocyanidin against diabetes and oxidative stress-related inflammatory processes.     

Carbon-14 biolabeling of (+)-catechin and proanthocyanidin oligomers in willow tree cuttings.

Proanthocyanidin polymers, oligomers, and the structurally related monomer (+)-catechin were labeled by incorporation of radioactive precursors in shoots of willow tree (Salix caprea L.). [1-(14)C]-Acetate and [U-(14)C]-phenylalanine precursors were fed through the cut stems or petioles of leaves. Optimization of several parameters such as the nature and origin of the plant material, leaf maturity, nature, and quantity of radioactive precursor applied and the duration of metabolism led to incorporation yields of 3.2% and to specific activities of 500 microCi/g. Detailed characterization of the products (polymerization degree, procyanidin/prodelphinidin ratio, specific activities) and purification by chromatography are reported. Some sugars bound to radiolabeled proanthocyanidin polymers were removed by enzymic treatment with a mixture of glycosidases. A radioactive purity close to 100% and specific activities suitable for bioavailability studies were obtained.     

Antioxidant activities of phenolic,proanthocyanidin, and flavonoid components in extracts of Cassia fistula

Cassia fistula L., a semi-wild Indian Labernum, is widely cultivated in Mauritius as an ornamental tree for its beautiful bunches of yellow flowers and also used in traditional medicine for several indications. The total phenolic, proanthocyanidin, and flavonoid contents, and the antioxidant activities, of fresh vegetative and reproductive organs of Cassia fistula harvested at different stages of growth were determined using the Trolox equivalent antioxidant capacity (TEAC) and ferric-reducing antioxidant power (FRAP) assays. The antioxidant activities were strongly correlated with total phenols (TEAC r = 0.989; FRAP r = 0.951) in all organs studied, and with proanthocyanidins (TEAC r = 0.980; FRAP r = 0.899) in reproductive organs including fruits. The antioxidant activities of reproductive parts were higher than those of the vegetative organs, with the pods having highest total phenolic, proanthocyanidin, and flavonoid contents and antioxidant potentials (TEAC = 992 +/- 0.4 micromol/g dry weight; FRAP = 811 +/- 23 micromol/g dry weight).     

Analysis of coffee for the presence of acrylamide by LC-MS/MS

A variety of popular instant, ground, and brewed coffees were analyzed using a modified liquid chromatography-tandem mass spectrometry (LC-MS/MS) method specifically developed for the determination of acrylamide in foods. Coffee test portions were spiked with 13C3-labeled acrylamide as an internal standard prior to their extraction and cleanup. Ground coffees (1 g) and instant coffees (0.5 g) were extracted by shaking with 9 mL of water for 20 min. Brewed coffee test portions (9 mL) were taken through the cleanup procedure without further dilution with extraction solvent. Coffee test portions were cleaned up by passing 1.5 mL first through an Oasis HLB (hydrophilic/lipophilic copolymer sorbent) solid phase extraction (SPE) cartridge and then a Bond Elut-Accucat (cation and anion exchange sorbent) SPE cartridge. The cleaned up extracts were analyzed by positive ion electrospray LC-MS/MS. The MS/MS data was used to detect, confirm, and quantitate acrylamide. The limit of quantitation of the method was 10 ng/g for ground and instant coffees and 1.0 ng/mL for brewed coffee. The levels of acrylamide ranged from 45 to 374 ng/g in unbrewed coffee grounds, from 172 to 539 ng/g in instant coffee crystals, and from 6 to 16 ng/mL in brewed coffee.     

Characterization of the tolerance to excess manganese in four maize varieties

Abstract

Manganese (Mn) is an essential micronutrient in all organisms, but may become toxic when present in excess. Four maize (Zea mays L.) varieties, Kneja 605, Kneja 434, Kneja 509 and Kneja 537, were studied with respect to their responses to excess Mn in hydroponic solution. In the varieties Kneja 605, Kneja 509 and Kneja 537, increasing Mn concentrations in the nutrient solution negatively affected biomass accumulation, photosynthetic rate, transpiration, stomatal conductance and chlorophyll content. In addition, these varieties showed increased electrolyte leakage and lipid peroxidation (malondialdehyde [MDA] content). Increased Mn leaf concentrations, higher contents of chlorophyll a and chlorophyll b, higher photosynthetic rate and transpiration, lower concentrations of MDA and insignificant changes in the electrolyte leakage in the leaves were found in var. Kneja 434 compared with the other maize varieties studied. This variety appeared to possess a stronger ability to cope with Mn phytotoxicity, suggesting high potential for Mn detoxification and var. Kneja 434 could be a good candidate for improving maize productivity on acid soils under non-tropical conditions.     

Analysis of combustion products using time-of-flight mass spectrometry

Time of flight mass spectrometry (TOF-MS) combined with resonant two-photon ionization is applied to detect aromatic hydrocarbons in combustion products. The sample is introduced through a heated capillary into the supersonic jet of a TOF-MS. For ionization, a tunable, frequency-doubled dye-laser is used. The inlet system allows a direct measurement of samples at atmospheric pressure. The resonant ionization can distinguish structural isomers without preceding chromatography. The short response time of the system allows for example on-line monitoring of combustion emissions. This is demonstrated by thermodesorption experiments of anthracene from activated charcoal and from diesel soot. A significant higher desorption temperature from charcoal than from diesel soot is found. In addition, examples showing selective measurements of benzene, toluene and three dimethyl-benzene isomers in the exhaust gas of a gasoline engine are presented.     

Influence of alternative liquid chromatography techniques on the chemical complexity and bioactivity of isolated proanthocyanidin mixtures

Extracts rich in proanthocyanidins, which are implicated in multiple human health benefits, were comparatively separated using alternative separation methods [vacuum or open column liquid chromatography], separation matrices [Toyopearl, Sephadex, or silica gel], and degrees of subfractionation [8 or 12 subfraction series], to evaluate the influence of separation technique on the resolution of the chemical composition and the biological activity of separated proanthocyanidin mixtures in individual subfractions. Bioactivity was assessed using a DNA human topoisomerase II bioassay and structural composition by acid thiolysis (average degree of polymerization, DP) and HPLC-ESI/MS. The amount of parent fraction needed to inhibit 50% of topoisomerase II was 3.38 ng/mL with an average DP of 25.5. A 2(3) factorial analysis revealed that the vacuum and open column strategies for separation, when individually considered, did not yield significantly different results in terms of mass recovery, DP, or bioactivity; however, interactions with other factors such as matrix or subfraction series resulted in distinctive shifts in fraction profiles and biological activity. In general, Sephadex as a matrix permitted elution and separation of discrete, polymerized subfractions with potent inhibition against human topoisomerase II. Sephadex vacuum chromatography, Toyopearl open column chromatography, and Toyopearl vacuum chromatography separation techniques eluted highly polymerized proanthocyanidin mixtures, but the inhibitory bioactivity was attenuated as compared to the parent fraction, whereas Sephadex open column chromatography eluted highly polymerized subfraction mixtures that retained bioactive potential.     

Effects of proanthocyanidin preparations on hyperlipidemia and other biomarkers in mouse model of type 2 diabetes

The protective effect of proanthocyanidins from persimmon peel, using both oligomers and polymers, was investigated in a db/db type 2 diabetes model. Male db/db mice were divided into three groups: control (vehicle), polymer-, or oligomer- (10 mg/(kg body weight x day x p.o.)) administered mice. Age-matched nondiabetic m/m mice were used as a normal group. The administration of proanthocyanidins reduced hyperglycemia in db/db mice through a decline in the serum level of glucose and glycosylated protein. In addition, it had a strong effect on hyperlipidemia through lowering levels of triglyceride, total cholesterol, and nonesterified fatty acids. The protective effect against hyperglycemia and hyperlipidemia was greater in the groups administered the oligomeric rather than polymeric form. The increased oxidative stress in db/db mice was attenuated by the administration of oligomers through inhibiting the generation of reactive oxygen species and lipid peroxidation and elevating the reduced glutathione/oxidized glutathione ratio. On the other hand, polymers did not show such an effect. Moreover, expressions in the liver of sterol regulatory element binding protein (SREBP)-1 and SREBP-2 were downregulated by the administration of proanthocyanidins, especially the oligomeric form. Oligomers caused a slight elevation in the expression of peroxisome proliferator-activated receptors alpha. Furthermore, oligomeric proanthocyanidin regulated the expression of nuclear factor kappaB in db/db type 2 diabetes via the activation of inhibitor protein kappaB-alpha. It also attenuated the protein expressions of cyclooxygenase-2 and inducible nitric oxide synthase. This suggests that oligomers would act as a regulator in inflammatory reactions associated with oxidative stress in type 2 diabetes. The present study results suggest that proanthocyanidin administration, especially the oligomeric form, may improve oxidative stress via the regulation of hyperlipidemia than hyperglycemia in type 2 diabetes.     

Analysis of free fatty acids in whey products by solid-phase microextraction

To evaluate the impact of Cheddar cheese starter cultures on the level of free fatty acids in liquid whey, a solid-phase microextraction (SPME) technique was utilized. The determination of response factors relative to an internal standard and the verification of linearity over a wide concentration range allowed for the quantitation of free fatty acids in experimentally produced liquid whey and in a wide range of dry whey ingredients. Results indicated that whey produced with a Lactococcus lactis subsp. lactis starter culture contained the highest level of total free fatty acids with significantly higher levels of lauric, myristic, and palmitic acids. Significant declines in oleic, linoleic, and palmitic acid occurred during storage. Commercial whey ingredients demonstrated a linear increase in free fatty acids with increasing percent protein, except for whey protein isolate, which had the lowest concentration. The use of SPME for the rapid analysis of free fatty acids in whey products should allow for further research to determine the importance of these compounds on the quality and stability of whey products.     

Analysis of the microbial communities in soils of different ages following volcanic eruptions

Volcanism is a primary process of land formation.It provides a model for understanding soil-forming processes and the role of pioneer bacteria and/or archaea as early colonizers in those new environments.The objective of this study was to identify the microbial communities involved in soil formation.DNA was extracted from soil samples from the Llaima volcano in Chile at sites destroyed by lava in different centuries(1640,1751,and 1957).Bacterial and archaeal 16 S r RNA genes were analyzed using quantitative polymerase chain reaction(q PCR)and Illumina Mi Seq sequencing.Results showed that microbial diversity increased with soil age,particularly between the 1751 and 1640 soils.For archaeal communities,Thaumarchaeota was detected in similar abundances in all soils,but Euryarchaeota was rare in the older soils.The analysis of bacterial 16 S r RNA genes showed high abundances of Chloroflexi(37%),Planctomycetes(18%),and Verrucomicrobia(10%)in the youngest soil.Proteobacteria and Acidobacteria were highly abundant in the older soils(16%in 1640 and 15%in 1751 for Acidobacteria;38%in 1640 and 27%in 1751 for Proteobacteria).The microbial profiles in the youngest soils were unusual,with a high abundance of bacteria belonging to the order Ktedonobacterales(Chloroflexi)in the 1957 soil(37%)compared with the 1751(18%)and 1640(7%)soils.In this study,we show that there is a gradual establishment of the microbial community in volcanic soils following an eruption and that specific microbial groups can colonize during the early stages of recovery.     

挤压温度对大豆分离蛋白与原花青素复合物结构和功能特性的影响

为了探究不同挤压温度（40、60、80、100和120℃）对大豆分离蛋白（Soy Isolate Protein,SPI）与葡萄籽原花青素（Grape Seed Proanthocyanidin Extract,GSPE）复合物功能性质及结构特性的影响。该研究以溶解度、乳化性、乳化稳定性、ζ-电位、粒度为指标,利用荧光光谱、红外光谱分析该复合体系中大豆分离蛋白功能性质及结构的变化。结果表明：相较于挤压SPI,经过挤压处理的SPI-GSPE复合物的溶解度、乳化活性指数、乳化稳定性指数、ζ-电位绝对值及持水性均显著提高（P<0.05）,其表面疏水性、持油性显著下降（P<0.05）。随着挤压温度的升高,SPI-GSPE复合物的溶解度、持油性及乳化活性均先增大后减小且在80℃达到最大值,而其表面疏水性先减小后增大且最小值在80℃,ζ-电位绝对值、乳化稳定性及持水性均随温度的升高而降低。粒径分析结果表明,挤压处理后SPI与GSPE形成了更加致密的复合物;荧光光谱及红外光谱结果表明,与GSPE的复合及挤压处理使SPI氨基酸残基所处微环境发生变化,蛋白结构发生变化。以上结果表明挤压温度为80℃时SPI-GSPE复合物功能性质提高幅度最大,为GSPE与SPI复合提高SPI的功能性质提供参考。