Quantitation and confirmation of sulfamethazine residues in swine muscle and liver by LC and GC/MS

The present paper describes a liquid chromatographic (LC) method for purification of crude swine tissue extracts before gas chromatographic/mass spectrometric (GC/MS) quantitation and confirmation of sulfamethazine at low ppb levels. Fractions corresponding to sulfamethazine were collected, evaporated to dryness, N-methylated with diazomethane, concentrated, and analyzed by GC/MS. A mass spectrometer was set to selected ion monitoring (SIM) mode. Ions 233, 227, 228, and 92 m/z were detected. Ratio 227/233 m/z (sulfamethazine/internal standard, [phenyl 13C6] sulfamethazine) was used for quantitation, while ratios 228/227 and 92/227 m/z, respectively, were used for confirmation. Quantitation in spiked blank muscle tissue was tested from 100 to 1 ppb and found acceptable at all concentrations studied; coefficients of variations ranged from 4.9 to 14.4%. Similar results were obtained for liver tissue from 5 to 20 ppb; coefficients of variation ranged from 1.2 to 9.1%.     

速冻莲藕片贮藏过程中品质变化动力学模型

为了探寻贮藏温度对速冻莲藕片品质的影响及预测其货架期,该文研究在贮藏温度?5、?15和?25℃条件下,速冻莲藕片维生素C、色泽和硬度随着贮藏时间的变化,建立了3个指标变化动力学模型。结果表明:随着贮藏温度的升高,速冻莲藕片维生素C、色泽和硬度的变化速率逐渐增加,?5℃条件下贮藏的速冻莲藕片品质快速下降,?15和?25℃条件下贮藏前期莲藕片品质下降不显著(P0.05),随着时间的延长,?25℃部分玻璃态贮藏的莲藕片品质最佳;不同贮藏温度下速冻莲藕片维生素C和硬度变化符合一级反应,总色差和亮度变化符合零级反应,速冻莲藕片维生素C、色泽和硬度反应速率常数符合Arrhenius方程,并建立莲藕片货架期动力学模型,通过对其验证发现模型拟合度良好(R20.9),能预测速冻莲藕片贮藏期各品质指标变化和不同温度下的货架期寿命。研究结果可为速冻莲藕片低温贮藏品质变化和货架寿命预测提供理论依据。     

Evidence for recycling of N from plants to soil during the growing season

In spite of the known below-ground biomass production of plant roots that concurrently introduce significant amounts of carbon and nitrogen into the soil, the effects of these inputs on N cycling in the soil–plant system are seldom considered. Here, we report on two field experiments carried out between 1995 and 1997 at the FAM Research Station Scheyern: (1) a N-turnover experiment to determine the N fluxes derived from ¹⁵N-labeled clover residues incorporated into the plough layer of defined plots, and (2) a root production experiment to assess the above (shoot) and below ground (gross and net root) biomass production of winter wheat in different fields, but nearby the ¹⁵N plots. An initial 50% decrease in soil organic ¹⁵N at 0–20-cm soil depth was recorded between fall, 1996 (incorporation of clover straw) and spring, 1997 (138 days after incorporation), which was then followed by a period of stability in ¹⁵N levels in the soil organic N until the harvest of winter wheat (286 days after incorporation). This stability may be explained in two ways: (a) actual stability of clover-derived ¹⁵N remaining in the second phase, e.g., due to recalcitrant compounds or microbial immobilization; or (b) apparent stability, e.g., because the actual mineralization of clover-derived ¹⁵N in the soil was compensated by secondary inputs of organic ¹⁵N (recycling). Further results showed that the first explanation was unlikely, as (1) between 138 and 286 days after clover incorporation, the mean ¹⁵N signature in soil mineral N was 2.1 at.%, indicating a persistent mineralization of clover residues; and (2) a decrease in soil microbial biomass ¹⁵N occurred in the second phase, indicating a continued N turnover in the soil. The amount of clover-derived ¹⁵N accumulated below the plough layer at 20–110-cm soil depth (11.5%) between early spring and the harvest of wheat also corroborated the return of mineralized ¹⁵N into the soil being due to the root N inputs by winter wheat. Based on the depth distribution of winter wheat net root biomass (root production experiment) and on soil organic ¹⁵N depth distribution (¹⁵N-turnover experiment), the root N input into soil was estimated to be 282 kg ha⁻¹, equivalent to 54% of total net N assimilation of winter wheat. Thus, the results of this study give substantial evidence for a N loop between soil and growing plants, whereby a part of the net mineralized N taken up by plants is continuously returned into the soil by their roots. The implications of this N loop for the interpretation of ¹⁵N experiments and for plant nutrition are discussed.     

Competitive direct enzyme-linked immunosorbent assay for detection of sulfamethazine residues in swine urine and muscle tissue

A sensitive assay for the detection of sulfamethazine in swine urine and muscle tissue using a direct competitive enzyme-linked immunosorbent assay (ELISA) has been developed. Undiluted urine or a phosphate-buffered saline extract of pork muscle tissue is mixed with an enzyme-labeled conjugate of sulfamethazine and horseradish peroxidase. The mixture is added to wells of a microtiter plate coated with antibody to sulfamethazine. After the test system is incubated, washed, and re-incubated with substrate and the reaction is stopped, the absorbance is measured at 405 nm. Levels of sulfamethazine as low as 20 ng sulfamethazine/g muscle tissue and 10 ng sulfamethazine/mL swine urine were detected and estimated.     

Lipid oxidation in fillets of herring (Clupea harengus) during frozen storage. Influence of prefreezing storage.

Fillets of herring (Clupea harengus) were kept on ice for 0, 3, 6, and 9 days prior to storage at -18 degrees C for 0, 21, 42, 63, and 84 days. At each storage point, peroxide value (PV), absorbance at 268 nm (A(268)), fluorescent products (FP), alpha-tocopherol, glutathione peroxidase (GSH-px) activity, and ascorbic acid were measured. As shown by regression analyses, samples held for 6 days on ice formed oxidation products at the highest rate during frozen storage, followed by, for PV and FP, the 9-day samples. These data indicate that severe changes that negatively affect the oxidation process took place in the herring muscle between 3 and 6 days after catch. Both the initial antioxidant levels and the rate of antioxidant loss at -18 degrees C decreased with increased prefreezing holding time, the latter being most obvious for GSH-px activity and ascorbic acid. alpha-Tocopherol showed the largest losses and had disappeared entirely from the 6- and 9-day samples at the end of the frozen storage. Partial least-squares regression analysis of the data showed that ice storage had a greater effect than frozen storage on changes in PV, A(268), FP, alpha-tocopherol, and ascorbic acid. For GSH-px activity, frozen storage had the greatest effect.     

A sensitive method for detection of sulfamethazine and N4-acetylsulfamethazine residues in environmental samples using solid phase immunoextraction coupled with MALDI-TOF MS

Sulfamethazine (SMT) and its major metabolite, N(4)-acetylsulfamethazine (NA-SMT), were each recovered from spiked water (0.1 ppb) and 10% (w/v) aqueous suspensions of soil (1 ppb) or composted manure (1 ppb), by using a three-stage solid phase immunoextraction (SPIE) system, followed by detection with matrix-assisted laser/desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Sulfonamide recovery rates are reported for separate stages of the SPIE system and for trace-level sulfonamide SPIE extraction from the environmental samples. SPIE MALDI-TOF MS is a rapid and definitive technique with potentially better efficiency relative to other established trace-level sulfonamide analytical methods. SPIE MALDI-TOF MS required 1.5 h per batch (8-24 samples/batch) for sample enrichment, 5 min per batch for probe preparation, and 5 min per sample to acquire and process the spectrum. This is the first time MALDI-TOF MS has been reported as a potential means of detecting trace-level drug residues in complex environmental samples.     

煤矿区生态储存估算及其对土地利用的综合响应评价

为对煤矿区生态储存估算及其对土地利用的综合响应进行评价,以山西省位于长河流域的晋煤集团11个集中连片煤矿区为例,在生态服务功能理论的基础上,通过收集研究区10a的土地利用类型以及矿区与非矿区的相关数据,分析各类土地利用类型的面积、分布和变化方向,并建立评价指标体系以估计不同时期的生态储存,进而得出煤矿区基于煤炭开采的生态储存的响应。研究结果:研究区整体的生态储存呈下降趋势,除了天安圣华,其他煤矿区的生态都出现不同程度的退化,且矿区的退化速度远高于非矿区;12个区域中(11个煤矿区和1个非煤矿区)有一半以上区域的生态储存等级为第3级,研究区11个煤矿区的生态储存能力普遍较高,生态改善的可能性较大,可为煤矿区生态保护规划以及相应的管理提供基础数据和应用借鉴。     

Changes in volatile compounds of carrots (Daucus carota L.) during refrigerated and frozen storage

Carrots (Daucus carota L.) of cv. Bolero and cv. Carlo were processed into shreds and stored for up to 4 months at -24 degrees C (frozen storage), or the roots were stored for up to 4 months at 1 degrees C (refrigerated storage) followed by processing into shreds. Volatiles from the carrot shreds were collected by dynamic headspace technique and analyzed by GC-FID, GC-MS, GC-MS/MS, and GC-O to determine the volatile composition and aroma active components of carrots stored under different temperature conditions. A total of 52 compounds were quantified, of which mono- and sesquiterpenes accounted for approximately 99% of the total volatile mass. Major volatile compounds were (-)-alpha-pinene, beta-myrcene, (-)-limonene, (+)-limonene, (+)-sabinene, gamma-terpinene, p-cymene, terpinolene, beta-caryophyllene, alpha-humulene, and (E)- and (Z)-gamma-bisabolene. A considerable increase in the concentration of mono- and sesquiterpenes was observed during refrigerated storage, whereas the concentration of terpenoids was around the same level during frozen storage. GC-O revealed that the major volatiles together with (+)-alpha-pinene, (-)-beta-pinene, (+)-beta-pinene, 6-methyl-5-hepten-2-one, (-)-beta-bisabolene, beta-ionone, and myristicin had an odor sensation, which included notes of "carrot top", "terpene-like", "green", "earthy", "fruity", "citrus-like", "spicy", "woody", and "sweet".     

Evidence for acclimation of N cycling to episodic N inputs in anthropogenically-affected intertidal salt marsh sediments

Nitrate removal was compared in anthropogenically-impacted and unimpacted salt marsh sediments in microcosms using the acetylene block technique and ¹⁵N natural abundance measurements. Potential denitrification rates were greater at the impacted site than the unimpacted site at all added NO₃⁻ concentrations (233, 467, or 700 μg N g dw⁻¹). Although the change in concentration of NH₄⁺ over time was small (69-104 μg N g dw⁻¹), the δ¹⁵N of accumulated NH₄⁺ increased significantly (0.26-13.22‰), and was more enriched for all NO₃⁻ treatments in the impacted sediments than the unimpacted site. The impacted site may be acclimated to episodic N inputs, and based on concentrations and ¹⁵N natural abundance had greater denitrification and N cycling than the unimpacted site.     

Raman spectroscopic study of structural changes in Hake (Merluccius merluccius L.) muscle proteins during frozen storage

This paper examines changes in the structure and functionality of fish muscle proteins at frozen storage temperatures known to render very different practical storage lives (-10 and -30 degrees C). Apparent viscosity and dimethylamine (DMA) content showed drastic temperature-related differences during storage. Raman spectroscopy revealed the occurrence of some structural changes involving secondary and tertiary protein structures. The changes in secondary structure were quantified, showing an increase of beta-sheet at the expense of alpha-helix structure. The nuC-H stretching band near 2935 cm(-)(1) increased in intensity, indicating denaturation of the muscle proteins through the exposure of aliphatic hydrophobic groups to the solvent. These structural changes were more pronounced at -10 degrees C but occurred at both storage temperatures, whereas changes in apparent viscosity and DMA only occurred in storage at -10 degrees C. The possible utility of these structural changes for quality assessment is discussed.     

Phenolic compounds in olive oils intended for refining: formation of 4-ethylphenol during olive paste storage

The phenolic composition of "lampante olive oil", "crude olive pomace oil", and "second centrifugation olive oil" was characterized by high-performance liquid chromatography with UV, fluorescence, and mass spectrometry detection. The phenolic profile of these olive oils intended for refining was rather similar to that previously reported for virgin olive oil. However, a new compound was found in these oils, which is mainly responsible of their foul odor. It was identified as 4-ethylphenol by comparison of its UV and mass spectra with those of a commercial standard. Although 4-ethylphenol was discovered in all oils intended for refining, its presence was particularly significant in "second centrifugation olive oils", its concentration increasing with time of olive paste storage. Similar trends were observed for hydroxytyrosol, hydroxytyrosol acetate, tyrosol, and catechol, the concentration of these substances reaching values of up to 600 mg/kg of oil, which makes their recovery for food, cosmetic, or pharmaceutical purposes attractive.     

Different responses of soil CO2 and N2O emissions to simulated N deposition in forests with divergent N transformation characteristics

CO₂ and N₂O are important greenhouse gases that are related to soil mineralization–immobilization turnover and nitrification. To explore the responses of CO₂ and N₂O emissions to N deposition in forests with different N transformation characteristics, CO₂ and N₂O fluxes were measured in two NH₄NO₃ fertilized plots. One plot was in a temperate pine plantation in Heilongjiang Liangshui National Nature Reserve (LS) with slow and minimally coupled mineralization–immobilization turnover and a high nitrification rate. The other plot was in a subtropical bamboo forest in the Fujian Daiyun Mountain National Nature Reserve (DY) in China with rapid and coupled mineralization–immobilization turnover but a low nitrification rate. The results showed that CO₂ emissions in the DY with a high mineralization rate were greater than those in the LS. Cumulative CO₂ emissions were significantly enhanced by N addition in DY, but in LS, they were not affected. The mean N₂O fluxes in the control were 0.010 and 0.008 mg N m^?2 hr^?1 for LS and DY, respectively. High N addition stimulated N₂O emissions in both LS and DY, but the response ratio for N₂O flux in LS (8.6) was larger than that in DY (2.9). These results suggested that soils with rapid and coupled mineralization–immobilization turnover are beneficial to CO₂ emissions and their positive response to N deposition. A high nitrification rate contributed to high N₂O emissions and the sensitive response of N₂O emissions to N deposition.     

N transformation mechanisms and N dynamics of organic fertilisers as partial substitutes for chemical fertilisers in paddy soils

Journal of Soils and Sediments - Knowledge of nitrogen (N) dynamics of organic fertilisers as partial substitutes for chemical fertilisers could improve the retention of mineral N and optimise...     

Effect of milk protein concentrate on lipid oxidation and formation of fishy volatiles in herring mince (Clupea harengus) during frozen storage

The effect of milk protein concentrate (MPC) at 0, 2, 4, and 6% on lipid oxidation and volatile formation in frozen stored herring mince (-18 degrees C) was evaluated by analyzing samples at 0, 2, and 4 months for fatty acid composition, volatiles, and thiobarbituric acid reactive substances (TBARS). Sensory evaluation was also conducted to assess the intensity of fishy odor, and the volatiles were analyzed using static headspace gas chromatography-mass spectrometry (SHGC-MS). The addition of 4 and 6% MPC to herring mince resulted in a 33% and 50% reduction of TBARS, respectively, at month 4 and lessened the intensity of fishy odor throughout storage. However, MPC did not protect fatty acids from enzymatic degradation unless it was added immediately after mincing. Volatile analysis using SHGC-MS showed that 4% MPC was able to reduce headspace volatiles associated with fishy odor. MPC is most effective for reducing 4-heptenal, 3-methyl-1-butanol, 2-hexenal, and 1-penten-3-ol, which are known to be potent odorants associated with lipid oxidation.     

Structural changes in natural actomyosin and surimi from ling cod (Ophiodon elongatus) during frozen storage in the absence or presence of cryoprotectants

Surimi and natural actomyosin (NAM) from ling cod (Ophiodon elongatus) were subjected to frozen storage in the absence or presence of cryoprotectants (sorbitol, sucrose, lactitol, and Litesse, either individually or in combination). Effects of frozen storage were studied for NAM frozen at -10 degrees C for 10 days and for surimi after eight freeze-thaw cycles. A commercial blend cryoprotectant (4% sucrose and 4% sorbitol), individual cryoprotectants at 8%, and optimal blends at 4, 5.5, 6, and 8%, were effective in maintaining the gel strength of surimi and NAM gels. Surimi or NAM frozen in the absence of cryoprotectants or with only 4% individual cryoprotectants, showed increased percent alpha-helical content by Raman analysis. Increased disulfide content was also observed in the treatment without cryoprotectants by the Raman SS stretching band and by chemical determination. Tyrosine residues were in a buried environment before and after freezing for all treatments, and surface hydrophobicity measured by 1-anilinonaphthalene-8-sulfonate decreased after frozen storage in the absence of cryoprotectants.     

Gross N transformation rates and net N mineralisation rates related to the C and N contents of soil organic matter fractions in grassland soils of different age

We investigated the relationship between soil organic matter (SOM) content and N dynamics in three grassland soils (0-10 and 10-20 cm depth) of different age (6, 14 and 50 y-old) with sandy loam textures. To study the distribution of the total C and N content the SOM was fractionated into light, intermediate and heavy density fractions of particulate macro-organic matter (150-2000 μm) and the 50-150 μm and <50 μm size fractions. The potential gross N transformation rates (mineralisation, nitrification, NH₄⁺ and NO₃⁻ immobilization) were determined by means of short-term, fully mirrored ¹⁵N isotope dilution experiments (7-d incubations). The long-term potential net N mineralisation and gross N immobilization rates were measured in 70-d incubations. The total C and N contents mainly tended to increase in the 0-10 cm layer with increasing age of the grassland soils. Significant differences in total SOM storage were detected for the long-term (50 y-old) conversion from arable land to permanent grassland. The largest relative increase in C and N contents had occurred in the heavy density fraction of the macro-organic matter, followed by the 50-150 and <50 μm fractions. Our results suggest that the heavy density fraction of the macro-organic matter could serve as a good indicator of early SOM accumulation, induced by converting arable land to permanent grassland. Gross N mineralisation, nitrification, and (long-term) gross N immobilization rates tended to increase with increasing age of the grasslands, and showed strong, positive correlations with the total C and N contents. The calculated gross N mineralisation rates (7-d incubations) and net N mineralisation rates (70-d incubations) corresponded with a gross N mineralisation of 643, 982 and 1876 kg N ha⁻¹ y⁻¹, and a net N mineralisation of 195, 208 and 274 kg N ha⁻¹ y⁻¹ in the upper 20 cm of the 6, 14 and 50 y-old grassland soils, respectively. Linear regression analysis showed that 93% of the variability of the gross N mineralisation rates could be explained by variation in the total N contents, whereas total N contents together with the C-to-N ratios of the <50 μm fraction explained 84% of the variability of the net N mineralisation rates. The relationship between long-term net N mineralisation rates and gross N mineralisation rates could be fitted by means of a logarithmic equation (net m=0.24Ln(gross m)+0.23, R²=0.69, P<0.05), which reflects that the ratio of gross N immobilization-to-gross N mineralisation tended to increase with increasing SOM contents. Microbial demand for N tended to increase with increasing SOM content in the grassland soils, indicating that potential N retention in soils through microbial N immobilization tends to be limited by C availability.     

Changes in the fauna and its contribution to mass loss and N release during leaf litter decomposition in two deciduous forests

Investigations on the mass loss of leaf litter were carried out between 1992 and 1994 using litter bags of 0.02 mm and 5 mm mesh sizes in a beech and a mixed forest in northern Germany. The two forests on moder humus differed in soil faunal composition, vegetation type, and nutrient supply. Mass loss and N and C concentrations were determined from the litter at bimonthly intervals. From subsamples macrofauna were sorted by hand and mesofauna was extracted by heat. The biomass and N content of the litter bag fauna was estimated. Mass loss, particularly that attributed to the fauna, was different between the two sites with highest rates in the mixed forest and lowest at the beech site. A significantly higher rate of N release was found for the litter extracted from 5 mm mesh size litter bags in the mixed forest but not in the beech forest. Collembola and Cryptostigmata changed in numbers during litter breakdown. Collembola reached high numbers in the beginning, whereas Cryptostigmata dominated later. The diversity of Cryptostigmata increased at both sites during litter breakdown, whereas collembolan diversity only increased in the beech forest and remained at the same level in the mixed forest. Several species of Collembola and Cryptostigmata occurred earlier in the mixed forest than in the beech forest. Mass loss rate attributable to the fauna did not correspond to total faunal biomass. Only Isopoda, Diplopoda and Cryptostigmata appeared to affect the mass loss positively, whereas the biomass of Lumbricidae was negatively correlated with mass loss, particularly in the beech forest. On the other hand, the release of N attributable to the fauna was positively correlated with the total faunal biomass in the beech forest and Lumbricidae in particular were positively correlated with N-release at both sites.     

Evolution of phenolic composition of red wine during vinification and storage and its contribution to wine sensory properties and antioxidant activity

The objective of this work was to study the evolution of the phenolic composition of red wine during vinification and storage and its relationship with some sensory properties (astringency and bitterness) and antioxidant activities. Thus, red wine was made by a classic vinification method with Castela?o and Tinta Miu?da grapes (Vitis vinifera L.) harvested at maturity (3:2; w/w). Samples were taken at 2 and 7 days of maceration, at second racking, at the time of bottling and at 6 and 14 months after bottling. The total polyphenols extract (TPx) in each sample was isolated by column chromatography. The phenolic composition (anthocyanins and proanthocyanidins), in vitro antioxidant activity, and sensory property (astringency, bitterness) of the isolated TPx from different winemaking stages were evaluated through high-performance liquid chromatography-diode array detection, 1,1-diphenyl-2-picrylhidrazyl radical test, ferric reducing antioxidant power assay, total phenolic index, MWI (polyphenol molecular weight index), TSA (tannin specific activity), and sensory panel tasting. The results showed that the phenolic composition of red wine varied significantly during winemaking. The intensity of astringency (IA) and the intensity bitterness (IB) of the isolated TPx from different winemaking stages increased from 2 days of maceration until second racking and then decreased. Furthermore, MWI and TSA are positively correlated with IA and IB. The in vitro antioxidant activity of the isolated TPx from different winemaking stages maintained unchanged after alcoholic fermentation, which was independent of the variation of phenolic composition and sensory properties.