Simultaneous determination of furanocoumarins in infusions and decoctions from "Carapiá" (dorstenia species) by high-performance liquid chromatography

The use of furanocoumarins, which are photosensitizing compounds, combined with exposure to UV-A radiation is a common treatment for vitiligo, psoriasis, and a number of other skin diseases. Although furanocoumarins plus UV-A treatment is highly effective, several studies have shown that exposure to high doses increases the risk to development of cutaneus carcinoma. Several Dorstenia species are used in folk medicine, mainly against skin diseases, because of the presence of biologically active compounds. We present here analysis of the chemical composition of furanocoumarins from infusion and decoction of "Carapiá" (Dorstenia species), which is used in Brazil against several diseases. We have employed high-performance liquid chromatography (HPLC) procedures for the quantitative determination of psoralen, bergapten, and isopimpinellin. The contents of furanocoumarins revealed an insignificant difference between infusion and decoction. Dorstenia tubicina and D. asaroides contained psoralen and bergapten only in the rhizomes, whereas D. vitifolia shows solely isopimpinellin in both rhizomes and aerial parts.     

Simultaneous detection of residues of 25 β₂-agonists and 23 β-blockers in animal foods by high-performance liquid chromatography coupled with linear ion trap mass spectrometry

A sensitive method has been developed for the simultaneous determination of residues of 25 β?-agonists and 23 β-blockers in animal foods by high-performance liquid chromatography coupled with linear ion trap mass spectrometry (HPLC-LIT-MS). This method is based on a new procedure of hydrolysis and extraction by 5% trichloracetic acid, and then cleaned up by mixed strong cation exchange (MCX) cartridges coupled with a novelty cleanup step by methanol. Methanol and 0.1% formic acid were used as mobile phases for gradient elution, while a Supelco Ascentis Express Rp-Amide column was used for LC separation. ESI positive ion scan mode was used with consecutive reaction monitoring (CRM, MS3). Nine β?-agonists labeled by the deuterium isotope were used as internal standards for quantification. The linear ranges of 48 analytes were from 5 to 200 μg/L; the coefficient of correlation was not less than 0.995. Blank pork muscle, blank liver, and blank kidney were selected as representative matrix for spiked standard recovery test. The recoveries of each compound were in the range of 46.6-118.9%, and the relative standard deviations were in the range of 1.9-28.2%. Decision limits (CCα, α = 0.01) of 48 analytes in muscles, liver, and kidney samples ranged from 0.05 to 0.49 μg/kg, and the detection capability (CCβ, β = 0.05) ranged from 0.13 to 1.64 μg/kg. This method was successfully applied to 110 real animal origin food samples including meat, liver, and kidney of pig and chicken samples.     

Modeling of urea degradation in white and rosé wines by acid urease

The specific activity of a whole cell acid urease preparation was tested in five wines manufactured in the Apulia region of Italy in the 2003 vintage at both short and long treatment times, thus confirming the validity of the pseudo-first-order kinetic model to describe urea removal in real wines. The ratio between the experimental pseudo-first-order kinetic rate constant (kIe) for any real wine tested and that (kI) referred to a model wine solution having the same composition and pH reduced from about 0.21 to 0.02 as the overall content of phenolic compounds (P) increased from 109 to 853 g m-3 of gallic acid equivalent (GAE). The specific inhibitory effect of such compounds was explained by accounting for the equilibrium constant (KP) of the reaction of polyphenols with acid urease, which was found to be about 21 g of GAE m-3 for the real wines tested, whereas it ranged from about 16 to 45 g of GAE m-3 when the model wine solution was enriched with tannins extracted from grape seeds or skins, respectively. A sequential experimental procedure consisting of accelerated acid urease tests at high doses of enzyme followed by accelerated ethyl carbamate tests on the resulting acid urease treated wine was recommended to assess preliminarily the technoeconomic feasibility of the acid urease hydrolytic process for the wine of concern. Keywords: Acid urease; real and model wines; phenolics; pseudo-first-order kinetic rate constant; inhibitory effect; urea degradation kinetics.     

Aluminum tolerances of two snapbean cultivars related to organic acid content evaluated by high‐performance liquid chromatography

High‐performance liquid chromatography (HPLC) was used to determine aluminum (Al)‐induced changes in organic acid (OA) concentrations of Al‐tolerant ‘Dade’ and Al‐sensitive ‘Romano’ snapbean cultivars. Two week old ‘Dade’ and ‘Romano’ snapbean were grown in 1/5‐strength Steinberg nutrient solution for 10 days and then subjected to 0, 2, 4, 6, and 8 mg L^‐1 Al treatments at pH 4.5 for an additional 3–15 days. Current studies confirmed earlier findings that the Dade cultivar was significantly more tolerant to Al than the Romano variety. Organic acid analyses were performed on extracts of root and leaf, and on stem exudates. The organic acids were separated on an ion exclusion column using a mobile phase of 0.01 N H₃PO₄. Individual OA were quantified with a variable wavelength detector operating at 210 nm. Aluminum stress tended to reduce the concentrations of citric, malonlc, malic, glycolic, fumaric, and acetic acids in the roots and increased the OA concentrations in stem exudates. In the presence or absence of Al stress, the Al tolerant Dade cultivar contained higher OA concentrations than did the Al‐sensitlve Romano. Aluminum stress reduced total OA levels in root extracts from Al‐sensltive Romano plants to a greater extent than in those of the Al‐tolerant Dade. Malic and citric acid concentrations were decreased more than those of the other organic acids examined. Results indicate that the Al‐tolerant Dade snapbean cultivar has a higher potential for Al‐chelation and detoxification than does the Al‐sensitive Romano. Hence, an Al‐chelation mechansism may be involved in differential Al tolerance within this species.     

Phytotoxicity of organic acids as influenced by montmorillonite,hydroxy‐Al montmorillonite and phosphate fertilization

Abstract

The germination and growth of wheat seedlings were studied at pH 5.5 in liquid growth media with organic acids in concentrations ranging from 800 to 1200 ppm, before and after interaction of the organic acids with montmorillonite and hydroxy‐Al montmorillonite. Germination was not affected by the organic acids, but subsequent growth was dependent on the kind and concentration of organic acid in the growth medium. Acetic acid was more inhibitory than citric acid. Gallic acid polymerized at pH 5.5 and was phytotoxic at 1200 ppm. Interaction of organic acids with montmorillonite and hydroxy‐Al montmorillonite reduced the concentration of organic acids in solution by adsorption. Despite this reduction in concentration the phytotoxicity of the growth media was enhanced after the interaction. This was caused by the dissolution of surface Al and the effect was more pronounced in cases where hydroxy‐Al interlayered montmorillonite was the adsorbent. Additions of P as KH₂PO₄ alleviated the phytotoxicity of organic acids but not the associated Al toxicity. The amount of P lost by adsorption or precipitation was dependent on the form of Al in the growth media.     

Formation of volatile organic products in soils under anaerobiosis—II: Metabolism of amino acids

Volatile organic compounds generated microbiologically under anaerobic conditions from various soils amended with glucose and amino acids were identified by combined gas chromatographymass spectrometry. Volatile compounds were usually found up to 4 weeks and sometimes longer. Acetaldehyde, acetone, methyl ethyl ketone, methyl propyl ketone, diacetyl, ethanol, n- and iso-propanol, n-, 2- and iso-butanol, acetic acid, butyric acid, ethyl acetate, ethyl butyrate, butyl acetate, butyl butyrate, methane thiol, dimethysulfide, dimethyldisulfide and methyl thioacetate were detected at various time intervals. Volatile organic sulfur compounds were evolved from soils amended with glucose and methionine but not with glucose and cysteine. Volatile nitrogen containing organic compounds were not detected with any of the amended soils.     

Extraction and reverse‐phase high‐pressure liquid chromatography of gibberellic acid in soil

Abstract

Several aqueous extractants buffered above pH 6.5 were evaluated for their ability to extract gibberellic acid from a Xerollic Calciorthid. More than 90% of the GA₃ added to soil was recovered with 0.01 M KH₂PO₄, pH 7.4. Gibberellic acid in soil extracts concentrated under vacuum was determined by reverse‐phase high‐pressure liquid chromatography (HPLC) by comparing the reproducible GA₃ peak areas with those of standards prepared in the same background solution.     

Determination of total organic‐C in soils by an improved chromic acid digestion and spectrophotometric procedure

Abstract

An improvement to the Walkley‐Black wet digestion method for the rapid determination of organic carbon over the range 0.2–5.5% in air‐dry soil is described. It permits total recovery of the organic‐C in finely ground soil samples digested with the heat of dilution from mixing N K₂ Cr₂ O₇ with concentrated H₂SO₄. in test tubes followed by external heating from a hot‐plate digestor. The organic‐C concentrations are determined directly, as the Cr product in diluted soil digests, by absorptiometry at 600 nm with calibration against similarly treated sucrose standards in solution. For the soils tested, there were negligible interferences from carbonates, wood charcoal, coke, Fe⁺² and readily reducible Mn; Cl does not interfere with the organic‐C assay in non‐saline soils but for saline soils a correction based on 1/12 Cl^‐ assay of the soil is necessary. The present method is compared with Tabatabai and Bremner's dry combustion procedure and Allison's manometric adaptation for calcareous soils. The procedure described here does not require carbonate to be determined and is therefore simpler. In addition it is cheaper, faster and more effective in controlling interferences than dry combustion procedures.     

The use of gas‐liquid chromatography to determine the proportions of inositol isomers present as pentakis‐ and hexakisphosphates in alkaline extracts of soils

Abstract

A method for determining the relative proportions of inositol isomers present in soil as their pentakis‐ and hexakisphosphate derivatives (IP₅ + IP₆) is presented. The method involves determination of the isomers as their hexakis‐O‐acetyl derivatives by gas‐liquid chromatography. The only isomers detected in extracts of 4 soils were chiro‐inositol, neo‐inositol, myo‐inositol and soyllo‐to‐inositol. Little variation in the relative proportions of the isomers was found between the 4 soils but the relative proportion of the neo‐isomer was considerably greater than previously published estimates. The method also makes possible the estimation of the (IP₅ + IP₆) fraction, using anion‐exchange chromatography on a aliquot of the extract.     

The use of comprehensive two-dimensional gas chromatography and structure–activity modeling for screening and preliminary risk assessment of organic contaminants in soil, sediment, and surface water

Purpose

This article aims to investigate the use and benefits of using comprehensive two-dimensional gas chromatography (GC?×?GC) and structure?Cactivity relationship modeling for screening and prioritization of organic contaminants in complex matrices. The benefit of applying comprehensive screening techniques to samples with high organic contaminant content is primarily that compounds with diverse physicochemical properties can be analyzed simultaneously. Here, a heavily contaminated industrial area was surveyed for organic pollutants by analyzing soil, sediment, and surface water samples. The hazard of the pollutants were ranked using SARs.

Material and methods

The water samples were liquid?Cliquid extracted using dichloromethane and directly analyzed by GC?× GC?Ctime-of-flight mass spectrometry (GC?×?GC?CTofMS). Soil and sediment samples were extracted with dichloromethane in an ultrasonic bath and subjected to gel permeation chromatography to eliminate lipids and humic matter. The low molecular weight fraction was then analyzed with GC?×?GC?CTofMS.

Results and discussion

More than 10,000 components were found in each sample, of which ca. 300 individual compounds were unambiguously identified using the National Institute of Standards and Technology mass spectra library and authentic reference standards. Alkanes, polycyclic aromatic hydrocarbons, and phthalates were generally the most abundant and were found in all matrices. In contrast, chlorinated compounds such as chlorophenols, biphenyls, and chlorinated pesticides were only detected in samples from a few hotspot regions. The toxicities of the most frequently detected compounds and of the compounds detected at the highest concentrations in samples from hotspot regions were estimated by ecological structure?Cactivity relationships. The ratio of the measured concentration to the predicted toxicity level was then calculated for each compound and used for an initial risk assessment in order to prioritize compounds for further transport and fate modeling, complementary measurements, and more advanced risk assessments.

Conclusions

The advantage of using of GC?×?GC?CTofMS for preliminary screenings of contaminated areas was evaluated at a polluted area in northern Sweden. The area was found to carry organic pollutants such as polyaromatic hydrocarbons, aliphatic hydrocarbons, polychlorinated biphenyls, phthalic compounds, and many chlorinated pesticides. Preliminary risk assessments indicate which compounds to use for subsequent remediation experiments based on their availability on the site or toxicity.     

Chromatographic quantitation of free amino acids: S‐methylmethionine,methionine and lysine in corn

The Pickering lithium gradient system was used with a modified Hewlett‐Packard HPLC and post‐column ninhydrin derivatization to separate and visualize the free amino acids and other ninhydrin‐positive substances in corn (Zea mays). About 50 major and minor components were discernible, with generally very good separation. Analytical focus was on S‐methylmethionine, principal precursor of the volatile flavor component dimethyl sulfide, in several sweet corn genotypes. The standard Pickering gradient program gave excellent separation of S‐methylmethionine from the other basic amino acids, permitting detection of as little as 20 pmoles. Quantitation was highly reproducible when canavanine was used as internal standard. Free methionine and lysine were also measured.     

Toxicity of Cu and Cd in Soils and Detoxication by Additives

Results from laboratory experiments indicated that the concentrations and toxicities of both water-soluble and 0.1 M HCl-extractable Cu and Cd from soils were in the order of red soil> yellow brown earth> black earth. The toxicity of soil varied with the concentrations of metals. The form, concentration and toxicity of Cu and Cd in soils were determined by cation exchange capacity, content of organic matter and composition of clay minerals in the soil. Addition of CaCO3 could significantly decrease the concentration and toxicity of water-soluble and 0.1 M HCl-extractable Cu or Cd from the red soil, and could notably transform the Cu and Cd from the water-soluble or exchangeable form into the organic, free oxides-occluded or sulfic form.     

Input and migration of dissolved organic carbon in soils of forest ecosystems in the subzone of deciduous–coniferous forests

The results of four-year monitoring of natural waters in forest ecosystems of the Zvenigorod Biological Station (Moscow State University, Moscow oblast) show regular changes in the concentrations and fluxes of dissolved organic carbon (DOC) within the atmospheric precipitation–throughfall–soil waters system. Precipitation passing through the tree canopy is enriched with DOC (2–3 and 9–24 mg/L). The average carbon concentration in soil waters reaches 100–110 mg/L in complex spruce and pine–spruce forests and does not exceed 40–60 mg/L in spruce–birch forests.     

Nitrogen and organic matter of volcanic ash soils of Santa Bárbara Association in Chile

Santa Bárbara association is a group of Andosols widely distributed on the western foothills of the Andes Mountains in the Central Valley of Chile from about 36° to 40° south latitude. The soils of this association may be divided into two groups—south and north—by the characteristics of soil organic matter, although they are much alike in morphology. The line of demarcation between the groups lies at about 38° south latitude.

The potential nitrogen fertility or , the content of easily decomposable organic nitrogen is greater in the south soils than in the north soils of which humic matter is at a more advanced stage of humification, as compared with that of the south soils. Dry summers have caused the north soils to lessen in their potential nitrogen fertility and to stimulate humification of organic matter, altering the organic matter in quantity as well as in nature. It is evident that the analysis of soil organic matter is an effectual measure for the characterization of Andosols, and hence for the classification of the soils.

It is very probable that a considerable water erosion has occurred in the soils of this association, transporting the fine soil particles from the mid-slope to the foot of the hills.     

Soil organic carbon as affected by direct drilling and mulching in sugar beet – wheat rotations

The effect of conventional ploughing, mulching, and direct drilling on the soil organic C (soil C_org) contents through the soil profile and on total soil C_org stocks (0–45 cm) was investigated at five different German sites. All sites showed similar results: after 10–13 years, soil C_org contents in the surface soil (0–10 cm) were 15–71% and 33–42% higher under direct drilling and mulching, respectively, than under ploughing (8–18 g kg^?1). Under ploughing, the soil C_org contents were distributed homogenously through the soil profile. Either mulching or direct drilling resulted in 3–28% higher soil C_org stocks than ploughing (49–116 t ha^?1). However, the tillage management was no significant factor since the sites showed the effects to different extents but were the mathematical replications. Five to six years later, trends and values were similar. We concluded that the main effect of mulching or direct drilling was the stratification with higher soil C_org contents in the surface soil. Since this is a positive means for soil protection, we suggested that the use of mulching or direct drilling can contribute to a sustainable soil management in crop rotations with sugar beet which are characterized by a strong physical impact on the soil during harvest.     

Hazard identification of contaminated sites—ranking potential toxicity of organic sediment extracts in crustacean and fish

Background, aim, and scope  It is well known that contaminated sediments represent a potential long-term source of pollutants to the aquatic environment. To protect human and ecosystem health, it is becoming common to remediate contaminated sites. However, the great cost associated with, e.g., dredging in combination with the large numbers of contaminated sites makes it crucial to pinpoint those sites that are in greatest need of remediation. In most European countries, this prioritization process has almost exclusively been based on chemical analyses of known substances; only seldom toxicity data has been considered. The main objective of the current study was therefore to develop a tool for hazard identification of sediment by ranking potential toxicity of organic sediment extracts in a crustacean and a fish. A secondary objective was to investigate the difference in potential toxicity between compounds with different polarities. Materials and methods  Early life stages of the crustacean Nitocra spinipes and the fish Oncorhynchus mykiss, which represent organisms from different trophic levels (primary and secondary consumer) and with different routes of exposure (i.e., ingestion through food, diffusive uptake, and maternal transfer), were exposed to hexane and acetone fractions (semi-polar compounds) of sediment from five locations, ranging from heavily to low contaminated. Preliminary tests showed that the extracts were non-bioavailable to the crustacean when exposed via water, and the extracts were therefore loaded on silica gel. Rainbow trout embryos were exposed using nano-injection technique. Results and discussion  Clear concentration–response relationships of both mortality and larval development were observed in all tests with N. spinipes. Also for rainbow trout, the observed effects (e.g., abnormality, hemorrhage, asymmetric yolk sac) followed a dose-related pattern. Interestingly, our results indicate that some of the locations contained toxic semi-polar compounds, which are normally not considered in risk assessment of sediment since they are focused on compounds isolated in the hexane fraction. Conclusions  The ranking of the five sediments followed the expected pattern of potential toxicity in both organisms, i.e., sediments with known pollution history caused major effects while reference sediments caused minor effects in the two test systems. Silica gel turned out to be an excellent carrier for exposure of N. spinipes to very hydrophobic and otherwise non-bioavailable sediment extracts. Recommendations and perspectives  Since both test systems demonstrated that a substantial part of the potential toxicity was caused by semi-polar compounds in the acetone fractions, this study enlightens our poor understanding of which compounds are causing adverse effects in environmental samples. Therefore, by investigating potential toxicity (i.e., hazard identification) as a first screening step in prioritizing processes, these implications could be avoided. For proper sediment risk assessment, we however recommend whole sediment toxicity tests to be used for selected sites at following tiers.     

Similarities in the aroma chemistry of Gewürztraminer variety wines and lychee (Litchi chinesis sonn.) fruit

GC/O analysis of canned lychees indicated that cis-rose oxide, linalool, ethyl isohexanoate, geraniol, furaneol, vanillin, (E)-2-nonenal, beta-damascenone, isovaleric acid, and (E)-furan linalool oxide were the most odor potent compounds detected in the fruit extracts. However, on the basis of calculated odor activity values (OAVs), cis-rose oxide, beta-damascenone, linalool, furaneol, ethyl isobutyrate, (E)-2-nonenal, ethyl isohexanoate, geraniol, and delta-decalactone were determined to be the main contributors of canned lychee aroma. When these results were compared with GC/O results of fresh lychees and Gewürztraminer wine, 12 common odor-active volatile compounds were found in all three products. These included cis-rose oxide, ethyl hexanoate/ethyl isohexanoate, beta-damascenone, linalool, ethyl isobutyrate, geraniol, ethyl 2-methylbutyrate, 2-phenylethanol, furaneol, vanillin, citronellol, and phenethyl acetate. On the basis of OAVs, cis-rose oxide had the highest values among the common odorants in the three products, indicating its importance to the aroma of both lychee fruit and Gewürztraminer wines. Other compounds that had significant OAVs included beta-damascenone, linalool, furaneol, ethyl hexanoate, and geraniol. This indicated that while differences exist in the aroma profile of lychee and Gewürztraminer, the common odorants detected in both fruit and wine, particularly cis-rose oxide, were responsible for the lychee aroma in Gewürztraminer wine. When headspace SPME was used as a rapid analytical tool to detect the levels of selected aroma compounds deemed important to lychee aroma in Gewürztraminer-type wines, cis-rose oxide, linalool, and geraniol were found to be at relatively higher levels in Gewürztraminers. No cis-rose oxide was detected in the control wines (Chardonnay and Riesling), while lower levels were detected in the Gewürztraminer-hybrid wine Traminette. Gewürztraminers produced in the Alsace region showed differences in the levels of the 3 monoterpenes when compared to those from New York State, which could be attributed to differences in viticultural and enological practices between regions.     

Sugar analysis with the shaffer‐somogyi micro‐analysis,high performance liquid chromatography and enzymatic analysis in crop samples

Abstract

To evaluate the reliability of the Shaffer‐Somogyi (SS) micro‐analysis of reducing sugars, extracts of 14 dried crop samples were analyzed before and after hydrolysis in 0.05 N ^H ₂SO₄with this method, with High Performance Liquid Chromatography (HPLC) and with an enzymatic glucose and fructose assay. The values, obtained with the SS micro‐analysis were for many samples higher than those, obtained with HPLC, suggesting that other compounds than sugars, present in certain plant tissues, respond in this non‐specific method. Enzymatic analysis tended to give lower values for sugar content than HPLC. It is recommended, that routine analysis of crop samples with the SS micro‐analysis is preceeded by analysis with HPLC to assess the contribution of non‐sugars to the outcome of the former.