Residues of azoxystrobin from grapes to raisins

Azoxystrobin, a fungicide of the strobilurin group, has an European Union maximum residue level (MRL) of 2 mg/kg for grapes. This work aimed to assess residues on fresh and washed grapes and on raisins following processing with (i) alkali treatment and sun drying and (ii) sun drying only. QUADRIS 25% SC was applied according to good agricultural practice for two consecutive years on a typical cv. Thomson seedless and a seed-producing clone. Samples were collected 0, 15, and 21 days postapplication and analyzed using gas chromatography/electron capture detection; recoveries were 86 +/- 12% for grapes and 99 +/- 15% for raisins. Residues on grapes were 0.49-1.84 mg/kg, and washing removed 75% of the residue. Residues in raisins produced from seedless grapes were 0.51-1.49 (treatment 1) and 1.42-2.08 mg/kg (treatment 2), with residue transfer factors sometimes >1, even following alkali treatment, which reduced residues considerably. To avoid trade problems, a higher MRL for raisins is necessary.     

Gas chromatographic determination of captan, folpet, and captafol residues in tomatoes, cucumbers, and apples using a wide-bore capillary column: interlaboratory study.

An interlaboratory study of the determination of captan, folpet, and captafol in tomatoes, cucumbers, and apples was conducted by 4 laboratories using wide-bore capillary column gas chromatography with electron capture detection. The 3 fungicides were determined using the Luke et al. multiresidue method modified to include additional solvent elution in the optional Florisil column cleanup step used with this method. The crops were fortified with each fungicide at 3 levels per crop. Mean recoveries ranged from 86.2% for a 25.1 ppm level of captan in apples to 115.4% for a 0.288 ppm level of captafol in apples. Interlaboratory coefficients of variation ranged from 3.4% (24.7 ppm folpet) to 9.7% (0.243 ppm captafol) for tomatoes; from 2.8% (2.0 ppm captafol) to 8.2% (24.8 ppm captan) for cucumbers; and from 1.5% (0.234 ppm folpet) to 22.1% (0.266 ppm captafol) for apples.     

Residues of spiroxamine in grapes following field application and their fate from vine to wine

Dissipation of the fungicide spiroxamine in grapes of two vine varieties, Roditis and Cabernet Sauvignon, exposed to field treatments was evaluated. Vines of a grape vineyard located in central Greece were sprayed once or twice with a commercial formulation of the fungicide at 30 g a.i./hL. Residues in grapes, must, and wine were determined by gas chromatography/IT-MS after extraction with cyclohexane-dichloromethane (9:1), with a limit of quantitation 0.02 mg/kg in grapes and 0.012 mg/kg in wine. Under field conditions, spiroxamine dissipation on grapes was faster during the first 2 weeks and then slower to the sixth week. About 7 days after application, half of the initial spiroxamine concentration remained on the grapes; the respective proportion at 42 days was about 10%. At 14 and 35 days, residues were lower than 0.44 and 0.22 mg/kg, respectively, values below the maximum residue levels set by the European Union (1 mg/kg). Spiroxamine residues transferred from grapes into the must and through the vinification process into the wine were also studied. Mean transfer factors of 0.26 and 0.55 were found from grapes into wine for the wines obtained without maceration and with maceration, respectively. Residues in wine, prepared from grapes with a spiroxamine content of 0.11-0.20 mg/kg, varied from <0.026 to 0.09 mg/kg. Spiroxamine diastereomer B was found to dissipate slower than diastereomer A in the field as well as during the vinification process.     

Bacterial degradation of fungicide captan

The phthalimide fungicide captan has been widely used to control plant pathogenic fungi. A strain of Bacillus circulans utilized the fungicide captan as sole source of carbon and energy. The organism degraded captan by a pathway involving its initial hydrolysis to yield cis-1,2,3,6-tetrahydrophthalimide, a compound without fungicidal activity. The formation of this compound was confirmed by HPLC, IR, NMR, and mass spectral analysis. The results also revealed that cis-1,2,3,6-tetrahydrophthalimide was further degraded to o-phthalic acid by a protocatechuate pathway. These findings indicated that there was a complete mineralization of fungicide captan by B. circulans.     

GC-ITMS determination and degradation of captan during winemaking

Captan and its metabolite tetrahydrophthalimide (THPI) were determined in grapes, must, and wine by GC-ITMS. Pesticides were extracted with acetone/petroleum ether (50:50 v/v). Because of the high selectivity of the ITMS detector, no interferent was found and cleanup was not necessary. Recoveries from fortified grapes, must, and wines ranged between 90 and 113% with a maximum coefficient of variation of 11%. Limits of quantitation were 0.01 mg/kg for both compounds. In model systems, captan and its metabolites, THPI, cis-4-cyclohexene-1,2-dicarboxylic acid, and 1,2,3,6-tetrahydrophthalamic acid, were determined by HPLC. The degradation of captan during winemaking was studied. Captan degraded in must, giving 100% THPI, and at the end of fermentation, only THPI was found in wine. The degradation of captan to THPI was due to the acidity in must and wine. This metabolite was present at low levels on grapes, and, unlike captan, it had no negative effect on the fermentative process. Model systems showed that the mechanism of disappearance of captan in grapes was due to photodegradation and codistillation.     

Antioxidant and antiproliferative activities of strawberries

Strawberries contain high levels of antioxidants, which have been correlated with a decreased risk of chronic disease. To more fully characterize the antioxidant profiles and possible associated health benefits of this fruit, the total free and bound phenolic, total flavonoid, and total anthocyanin contents of eight strawberry cultivars (Earliglow, Annapolis, Evangeline, Allstar, Sable, Sparkle, Jewel, and Mesabi) were measured. Cultivar effects on phenolic contents were compared with antioxidant capacities, as measured by the total oxyradical scavenging capacity (TOSC) assay, and to antiproliferative activities, as measured by inhibition of HepG(2) human liver cancer cell proliferation in vitro. Free phenolic contents differed by 65% between the highest (Earliglow) and the lowest (Allstar) ranked cultivars. The water soluble bound and ethyl acetate soluble bound phenolic contents averaged 5% of the total phenolic content of the cultivars. The total flavonoid content of Annapolis was 2-fold higher than that of Allstar, which had the lowest content. The anthocyanin content of the highest ranked cultivar, Evangeline, was more than double that of the lowest ranked cultivar, Allstar. Overall, free phenolic content was weakly correlated with total antioxidant activity, and flavonoid and anthocyanin content did not correlate with total antioxidant activity. The proliferation of HepG(2) human liver cancer cells was significantly inhibited in a dose-dependent manner after exposure to all strawberry cultivar extracts, with Earliglow exhibiting the highest antiproliferative activity and Annapolis exhibiting the lowest. No relationship was found between antiproliferative activity and antioxidant content.     

Gas-liquid chromatographic determination of captan and two metabolites in milk and meat

A gas-liquid chromatographic (GLC) method has been developed for the determination of captan (N-trichloromethylthio-4-cyclohexene-1,2-dicarboximide) and 2 metabolites, tetra-hydrophthalimide (THPI) and tetrahydrophthalamic acid (THPMA), in milk and meat. The sample is extracted with ethyl acetate and is cleaned up by acetonitrile partition and silica gel chromatography where captan, THPI, and THPMA are separated. Captan is directly determined by GLC. THPI and THPMA are separately derivatized in an acetone solution of pentafluorobenzyl bromide. The resultant derivatives are purified separately on an Al2O3 column and quantitated by GLC, using an electron capture detector. Recoveries from milk samples fortified at 0.02-10 ppm ranged from 71 to 102%; recoveries from meat samples fortified at 0.04-10 ppm ranged from 75 to 99%.     

Determination of ethylenethiourea in grapes and wine

Residues of ethylenethiourea (ETU) in grapes and wine were determined by capillary gas chromatography and paper chromatography, without a cleanup step, and after derivatization to S-benzyl-ETU. The detection limit was 0.0002 mg/kg for flame ionization detection, 0.008 mg/kg for paper chromatography with photodensitometric evaluation of the detected spot. Results were compared with a generally used GC method specifying electron capture detection of trifluoroacetylated S-benzyl-ETU. The recoveries of ETU in grapes and wine at different concentration levels were determined. ETU residues were determined in treated grapes but no residues were detected in wine.     

Distribution of folpet on the grape surface after treatment

Field trials were carried out to evaluate whether folpet sprayed on grapevines penetrated the epicuticular wax and cell walls of grapes. Folpet showed poor penetration into the epicuticular wax; it was found almost totally on the surface. Despite its low solubility in water, perhaps due to the presence of adjuvants, its residues showed such a high resistance to washing that the action of rain was negligible in decreasing residues.     

Investigation on fungicide residues in greenhouse-grown strawberries

Maximum residue limits (MRL's) for different agricultural food products in Norway are harmonized with EU standards. In field-grown strawberries in Norway, tolylfluanid has a 7 day quarantine from last application to harvest, while other approved fungicides have 14 days quarantine. Greenhouse production of strawberries is newly introduced to the country. Residue levels in strawberries of the cultivar Korona grown in a commercial greenhouse were investigated 4, 7, and 14 days after application of eight different fungicides at rates recommended by the manufacturers and at half rates. Iprodione, tolylfluanid, and vinclozolin were tested in two experiments, while chinomethionat, chlorothalonil, imazalil, penconazole, and triadimefon were tested once. For chinomethionat, imazalil, iprodione, penconazole, and vinclozolin, the residue levels were below MRL 2 weeks after application. Application of triadimefon in normal rate gave residues below MRL 14 days after application. However, its metabolite, triadimenol, was above MRL at the same time. Tolylfluanid gave very high residue levels, and except from half concentration in the second experiment, all other residue levels were above MRL. Seven days after application, residues in both experiments were approximately 3 times higher than MRL when normal rate of tolylfluanid was applied. For chlorothalonil at the recommended rate, the residue level was above MRL at any sampling time, while half rate gave residues below MRL 14 days after treatment. In view of the present results, tolylfluanid, chlorothalonil, and triadimefon will need longer time from last application to harvest and/or reduced application rates in greenhouse-grown compared to field-grown strawberries. In addition or as an alternative, recommended rates could be lowered.     

Seasonal patterns of growth and tissue nutrient content in strawberries

At monthly intervals during the establishment year, 90 strawberry plants were excavated and separated into component parts, dried, weighed, and nutrient analyses were performed on the tissues. When growth resumed in spring, biomass and foliar nutrient levels were again measured until fruiting. Individual plant growth was linear during the establishment year, with leaves accounting for the highest percentage of accumulated biomass. In general, nutrient concentrations during the establishment year either remained relatively constant, or declined. The percent increase in biomass accumulation was always greater than the percent decline in nutrient concentration, indicating that strawberry plants have a continuing demand for nutrients during the establishment year. About 50% of the biomass was lost during the winter, mainly in foliage. However, within six weeks of resuming growth in spring, plant size was equivalent to that of the previous autumn. The root system was the only tissue to exhibit a reduction in biomass during fruiting. During this time, foliar nutrients displayed very different concentration patterns. Although this period of fluctuating nutrient levels is generally avoided for leaf sampling and analysis, it is likely that this is when nutrient limitations will have their most pronounced effects.     

Determination of sulfur dioxide in grapes and wines

Reasons for using SO2 in wine and the theoretical aspects of this use are discussed. Data are summarized on the amounts of SO2 found in wines in California and around the world. Methods of analysis are discussed.     

Optimization of the analysis of flavor volatile compounds by liquid-liquid microextraction (LLME). Application to the aroma analysis of melons, peaches, grapes, strawberries, and tomatoes

A fast method based on liquid-liquid microextraction (LLME) has been developed for the analysis of volatile compounds in fruit and vegetable juices. The method was tested in an aqueous solution containing 49 common flavor compounds typically found in fruit aroma. Influence on extraction yield of the salts used, their levels, and the time of extraction was investigated. The efficiency of n-propyl gallate to inhibit the formation of secondary compounds from lipids during the crushing of fruit tissues was also tested. The proposed method was then applied to several authentic samples such as melons, peaches, grapes, strawberries, and tomatoes. The advantages and limitations of LLME are discussed.     

Cultural system affects fruit quality and antioxidant capacity in strawberries

Cultural system [hill plasticulture (HC) versus matted row (MR)] and genotype interactions affected strawberry fruit quality. In general, fruit soluble solids content, total sugar, fructose, glucose, ascorbic acid, titratable acid, and citric acid contents were increased in the HC system. Fruit from HC also had higher flavonoid contents and antioxidant capacities. Strawberry fruit contains flavonols as well as other phenolic compounds such as anthocyanins and phenolic acids. Pelargonidin-based anthocyanins such as pelargonidin 3-glucoside, pelargonidin 3-rutinoside, and pelargonidin 3-glucoside-succinate were the predominant anthocyanins in strawberry fruit. The content of cyanidin-based anthocyanins, cyanidin 3-glucoside and cyanidin 3-glucoside-succinate, was much lower than that of pelargonidin-based anthocyanins in either system. Strawberry fruit from the HC system had significantly higher amounts of p-coumaroylglucose, dihydroflavonol, quercetin 3-glucoside, quercetin 3-glucuronide, kaempferol 3-glucoside, kaempferol 3-glucuronide, cyanidin 3-glucoside, pelargonidin 3-glucoside, pelargonidin 3-rutinoside, cyanidin 3-glucoside-succinate, and pelargonidin 3-glucoside-succinate. Fruits from plants grown in the MR system generally had the lowest contents of phenolic acids, flavonols, and anthocyanins. Strawberry fruit grown under HC conditions had significantly higher peroxyl radicals (ROO*) absorbance capacity (ORAC).     

Survey of polyphenol constituents in grapes and grape-derived products

A rapid and comprehensive qualitative method has been developed to characterize the different classes of polyphenols, such as anthocyanins, flavonols, phenolic acids, and flavanols/proanthocyanidins, in grape products. The detection was achieved by two runs with the same LC gradient in different MS ionization modes and mobile phase modifiers (positive ionization mode and 0.4% trifluoroacetic acid for anthocyanins and flavonols; negative ionization mode and 0.1% formic acid for phenolic acids and flavanols). From an analysis of the MS and UV data and in comparison with the authenticated standards, a total of 53 compounds were identified, including 33 anthocyanins, 12 flavonols, 4 phenolic acids, and 4 flavanols/proanthocyanidins. With the method developed, a survey was then conducted to qualitatively assess the composition of polyphenols among 29 different grape products including original grape, grape juice, grape wine, and grape-derived dietary supplements, and their chemical profiles were systematically compared. This method provided a comprehensive qualitative insight into the composition of polyphenols in grape-derived products.     

Influence of processing on quality parameters of strawberries

To determine the effects of different processing steps, such as enzymatic treatment of the mash and pasteurization, on selected quality parameters, strawberries were processed to juices and purees. To identify the processing steps causing the highest losses, samples were taken after each step, and ascorbic acid, total phenols, anthocyanins, and antioxidant capacity were analyzed. To assess the antioxidant capacity, three different methods were applied: the trolox equivalent antioxidant capacity (TEAC), the ferric reducing antioxidant power (FRAP), and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, showing correlation coefficients of 0.889 to 0.948. The antioxidant capacity decreased with processing steps except heat treatment, which partly caused an increase due to the formation of antioxidant active products. The content of ascorbic acid, in comparison to that in the frozen strawberries, decreased significantly during the processing of the fruit to puree by 77%. In the pressed cloudy juices, the loss of ascorbic acid was 37%. The decline of phenolic compounds, measured as total polyphenols and anthocyanins, was smaller (between 30-40%). Pressing and pasteurization were the most critical steps for the decrease of these compounds. The enzymatic treatment of the mash within 90 min supported the release of secondary plant metabolites, while ascorbic acid is reduced up to 20%.