Determination of toxaphene residues in fish foodstuff by GC-MS.

A method for the determination of toxaphene residues in fish and fish-based baby foods has been developed. The cleanup of the fatty matrices was performed using an acid treatment on an Extrelut-NT3 and ENVI-Florisil SPE cartridges system, using light petroleum as eluent. Instrumental analysis was carried out by gaschromatography with mass spectrometry detector (GC/MS) in SIM mode. Recoveries from spiked samples were tested at 0.005 and 0.01 mg/kg per single congener and were in the range 82-104% while relative standard deviations (RSDs) were in the range 3.7-10.9%. Nineteen samples of both frozen fish food and fish-based baby foods representative of the Italian market were collected from local dealers and analysed.

Organochlorine pesticides and polychlorinated biphenyls in 12 edible marine organisms from the Adriatic Sea, Italy, Spring 1997.

Edible portions of 12 marine organisms from several areas of the Adriatic Sea, Italy, were collected during Spring 1997 and analysed for 32 organochlorine pesticides residues and 27 polychlorobiphenyl congeners. Only eight organochlorine pesticides - hexachlorobenzene (HCB), hexachlorocyclohexane (HCH) isomers, diphenyl-dichloro-trichloroethane (DDT) group, dieldrin - were determined at levels in the range <0.01-19.88 ng g(-1) wet weight, with 1,1'-dichloro-2,2'-bis(4-chlorophenyl)ethylene (DDE) being the more relevant single organochlorine. The contamination by organochlorine pesticides was comparable in organisms from the North, Centre and South Adriatic. Polychlorinated biphenyls (PCBs) were determined at levels in the range <0.05-14.46 ng g(-1), with CB 101, 118, 138, 153, 180 and 187 being more relevant (penta-, hexa- and hepta-chlorinated congeners). The sum of PCBs congeners determined were in the range 1.18-69.05 ng g(-1). The contamination by PCBs is more relevant in organisms from the North Adriatic Sea owing to the antropic discharge from major rivers such as Po and Adige that flow through highly industrialized and densely populated areas.

Estimation of dietary intake of pesticide residues by the Italian population during 1997.

In 1997 the Italian monitoring programme for pesticide residues in food comprised 7951 samples of 152 different foods for which a total of 275 pesticides were analysed. For 90 pesticides, the National Estimated Daily Intakes (NEDIs) were calculated by using the 90th percentile value and the food consumption data for the general Italian population. The calculated NEDIs were found to represent only a small portion of the respective Acceptable Daily Intake (ADI).

Determination of pyrethroid pesticide residues in fatty materials by solid-matrix dispersion partition, followed by mini-column size-exclusion chromatography.

The method studied uses a combination of a solid-matrix dispersion partition (SMDP) followed by high-performance size-exclusion chromatography on a minicolumn (HPmSEC) of 7.8 mm I.D. for the separation of pyrethroid (PYR) residues from fatty material. The solid-matrix dispersion extraction is carried out by absorbing a fat solution onto an Extrelut-3 cartridge (filled with a macroporous diatomaceous material) and extracting the PYR residues with acetonitrile. Up to 1 g of fatty material can be extracted with 15 ml acetonitrile. The small amount (mean +/- S.D. = 12.4 +/- 5.9 mg) of fatty material which is eluted into the acetonitrile is further removed by HPmSEC. PYR pesticide residues are collected in a 2-ml fraction between 7 and 9 ml, the column being washed up to 24 ml. The two techniques used in series allow a better removal of fat, a greater input of sample and a lower consumption of solvent compared to the sole SEC on macrocolumns, and a lower limit of determination compared to the sole SEC on minicolumns. Recoveries of 9 PYR out of the 14 investigated residues from soya oil were in the range 66-83% at spiking levels ranging 0.49-2.57 mg/kg, while for 6 PYR residues tested at spiking levels in the range 0.13-0.53 mg/kg the recoveries were in the range 80-111%. Recovery of fluvalinate and permethrin could not be calculated due to interferences from soya oil, while lambda-cyhalothrin, esfenvalerate and tralomethrin gave low recovery. The final extract contains small amount (mean +/- S.D. = 2.4 +/- 0.9 mg) of lipid residue and is not completely free from interferences.

Selective clean-up applicable to aqueous acetone extracts for the determination of carbendazim and thiabendazole in fruits and vegetables by high-performance liquid chromatography with UV detection.

Fungicide residues in vegetables (benomyl, carbendazim, thiabendazole) are analyzed through a clean-up procedure that uses a portion of the aqueous acetone extract prepared for multiresidue methodology. A portion of the aqueous acetone extract (equivalent to 5 g of vegetables) is loaded onto an Extrelut-20 cartridge (the cartridge is filled with a coarse, large-pore diatomaceous material). Then, acetone is partially removed by an upward stream of nitrogen at 2l/min for 30 min. Benzimidazolic fungicides are recovered by percolating the cartridge with 100 ml of 0.1 M phosphoric acid solution, which also serves to convert benomyl to carbendazim. The percolating acid solution is drained on-line through a strong cation-exchange (SCX) solid-phase extraction cartridge with the aid of a slight vacuum. Benzimidazolic fungicides are retained on the SCX cartridge. The phosphoric acid solution is discarded together with the washings of the SCX cartridge, i.e., water followed by methanol-water (75:25), that remove unwanted coextractives. Finally, benzimidazolic fungicides are recovered by eluting the SCX cartridge with methanol-ammonium formate buffer (75:25). The final extract is then analyzed by reversed-phase HPLC with UV detection. Recoveries from crops such as apples, lettuce, strawberries and citrus fruits are generally greater than 80% and no interferences were observed. The clean-up is simple and straightforward, requires only disposable items, water solutions and a few milliliters of solvent and a minimum number of manipulations, and does not require concentration steps or electrical equipment.

Clean-up of aqueous acetone vegetable extracts by solid-matrix partition for pyrethroid residue determination by gas chromatography-electron-capture detection.

Disposable, ready-to-use cartridges filled with macroporous diatomaceous material are used to carry out a partition clean-up that, in a single step, is capable of transferring pesticide residues from aqueous acetone extracts into light petroleum-dichloromethane (75:25, v/v). This procedure takes the place of some functions (such as separatory-funnel partition, drying over anhydrous sodium sulphate and partial adsorption clean-up) usually performed by separate steps in classical schemes. Fourteen pyrethroid pesticides, including tefluthrin, tetramethrin, cyphenothrin, cyfluthrin, flucythrinate, tau-fluvalinate, deltamethrin, bioallethrin, fenpropathrin, lambda-cyhalothrin, permethrin, alpha-cypermethrin, esfenvalerate and tralomethrin were determined using the described procedure with satisfactory recoveries for most of them, at spiking levels ranging from 0.08 to 0.82 mg/kg for the different compounds. Crops subjected to the described procedure included strawberry, apple, and orange gave extracts containing a mass of co-extractives that was between 5 and 30 mg. Compared with classical schemes, the described procedure is simple, less labour intensive, allows parallel handling of several extracts and does not require the preparation and maintenance of equipment. Troublesome emulsions such as those frequently observed in separation funnel partitioning do not occur.

Selective extraction of pyrethroid pesticide residues from milk by solid-matrix dispersion.

A rapid procedure has been developed that allows a single-step, selective extraction and clean-up of pyrethroid (PYR) pesticide residues from milk dispersed on solid-matrix diatomaceous material filled into disposable cartridges and eluted by means of light petroleum saturated with acetonitrile and ethanol. The extract was cleaned up by high-performance size-exclusion chromatography. Determinations were carried out by gas chromatography with electron-capture detection. Recovery experiments were carried out on homogenized commercial milk (3.6% fat content) that was spiked with solutions of 14 PYR pesticides, viz., tefluthrin, tetramethrin, cyphenothrin, cyfluthrin, flucythrinate, fluvalinate, deltamethrin, bioallethrin, fenpropathrin, lambda-cyhalothrin, permethrin, alpha-cypermethrin, esfenvalerate and tralomethrin, at levels ranging from 0.04 to 0.41 mg/kg for the different PYR pesticides. Average recoveries were in the range 60-119% for the different PYR pesticides, with relative standard deviations from ca. 2.5 to 14.4%. Coextracted fatty material amounted to an average of ca. 5 mg/ml of milk. The sole extraction step requires about 30 min. The main advantages of the procedure are that extraction of PYR pesticides (with a minimum carry over of fat) is performed in a single step, emulsions do not occur, several samples can be run in parallel by a single operator, reusable glassware is not needed and simple operations are required.

Selective, solid-matrix dispersion extraction of organophosphate pesticide residues from milk.

A rapid procedure has been developed that allows a single-step, selective extraction and cleanup of organophosphate (OP) pesticide residues from milk dispersed on solid-matrix diatomaceous material filled into disposable cartridges by means of light petroleum saturated with acetonitrile and ethanol. Recovery experiments were carried out on homogenized commercial milk (3.6% fat content) spiked with ethanolic solutions of 24 OP pesticides, viz., ethoprophos, diazinon, dimethoate, chlorpyrifos-methyl, parathion-methyl, chlorpyrifos-ethyl, malathion, isofenphos, quinalphos, ethion, pyrazophos, azinphosethyl, heptenophos, omethoate, fonofos, pirimiphos-methyl, fenitrothion, parathion, chlorfenvinphos, phenthoate, methidathion, triazophos, phosalone, azinphos-methyl, at levels ranging for the different OP pesticides from 0.02 mg/kg to 1.11 mg/kg. Average recoveries of four replicates were in the range 72-109% for the different OP pesticides, with relative standard deviations (R.S.D.) from ca. 1 to 19%, while dimethoate and omethoate were not recovered. Coextracted fatty material amounted to an average of about 4.0 mg/ml of milk. The extraction procedure requires about 30 min. The main advantages are that extraction and cleanup are carried out in a single step, emulsions do not occur, several samples can be run in parallel by a single operator, reusable glassware is not needed and simple operations are required.

Application of solid-phase partition cartridges in the determination of fungicide residues in vegetable samples.

Disposable, ready-to-use cartridges filled with a macroporous diatomaceous material are used to extract in a single step fungicide residues with dichloromethane from aqueous acetone extracts of vegetables. This procedure takes the place of some functions (such as separating funnel partition, drying over anhydrous sodium sulphate and clean-up) usually performed by separate steps in classical schemes. Fourteen fungicides (dichloran, vinclozolin, chlorthalonil, triadimefon, dichlofluanide, procymidone, hexaconazole, captan, folpet, ditalimfos, iprodione, captafol, pyrazophos and fenarimol) were determined using the described procedure with recoveries between 83 and 107% at spiking levels ranging for the different compounds from 0.04 to 0.40 mg/kg. Crops subjected to the described procedure included lettuce, strawberry, apple, yellow pepper and peach, and gave extracts containing a mass of co-extractives between 5 and 30 mg. Compared with classical schemes, the described procedure is simple, less labour intensive, allows parallel handling of several extracts and does not require preparation and maintenance of equipment. Troublesome emulsions such as those frequently observed in separating funnel partitioning do not occur.

Pesticide residue control in the years 1988-1989 in Italy.

The paper reports the monitoring for pesticide residue contamination of foodstuffs carried out in the years 1988-1989 by the Italian peripheral laboratories officially commissioned for the task. The tables show the analytical data for each class of substrate versus the limits in force at the time. 2506 food samples and 555 samples from the environment were analysed and a total of 16,259 determinations made. The residues were absent in 90% of the cases; the percentage of irregularities was 1.52%. These results confirm the data obtained in previous years, which is particularly significant considering the much larger number of analytical determinations performed this time. It has been deemed convenient to survey specific types of foodstuffs (for example, cereals and animal products) by examining a larger number of samples.

Quantitative relationship between ethylated DNA bases and gene mutation at two loci in CHO cells.

In a previous study we showed that the formation of O6-ethylguanine (O6-EtGua) in the DNA of CHO cells in culture correlated with mutations induced by ethylnitrosourea (ENU) and diethylsulfate (DES) at the hypoxanthine-guanine-phosphoribosyltransferase (hprt) locus but not at the Na, K-ATPase locus. This study was extended to another ethylating agent, ethyl methanesulfonate (EMS). DNA adduct formation and induction of mutation at the two gene loci were determined simultaneously in CHO cells after EMS exposure. The extent of ethylation at the N7 and O6 positions of guanine and at the N3 site of adenine were measured and the possible correlations with 6-thioguanine resistance (6-TGr) and ouabain resistance (ouar) mutations were investigated. A good correlation between the levels of ethylation at O6 guanine and mutation frequency at hprt gene by all three ethylating agents was observed. In the case of the ouar locus, the frequency of O6-EtGua adducts correlated with mutation induction by EMS and ENU but not by DES. Although both EMS and DES have similar reaction mechanisms, these results highlight differences in their mutational specificity. The comparison of this type of analysis with mutational spectra revealed that correlation studies may be inadequate to analyse multicomponent phenomena like mutation formation.

Pesticide residue control in the years 1986-1987.

Results of analytical surveys on pesticide residue levels in foods for the years 1986-1987 are presented. Analytical data obtained from regional laboratories are tabulated in terms of various commodity groups (vegetables, fruit, cereals, processed or derived foodstuffs, other commodities, legumes, environmental samples, others) and are compared with maximum legal tolerances established by the ministerial ordinance (Italian Ministry of Health) 6.6. 1985. On a total number of 4715 determinations of classes of compounds or of single compounds made on 1601 commodities samples, only N 53 (1.1%) violative results appeared. No residue was found at levels exceeding regulatory limits or found in commodities where a tolerance had not been established for a specific pesticide/commodity combination.

Single-step multi-cartridge clean-up for organophosphate pesticide residue determination in vegetable oil extracts by gas chromatography.

A multi-cartridge system has been developed which, in a single step, performs the extraction and clean-up of organophosphate (OP) pesticide residues from oils and fatty extracts. A solution in hexane containing up to 1.8 g of lipidic material is loaded on to an Extrelut-3 column to which a silica-gel cartridge and a C18 silica cartridge have been connected in series. The OP pesticide residues are eluted with 15 ml of acetonitrile. Carry-over of fatty material is in the range 2-5 mg per 1.8 g of different oils, which makes the final solution amenable to capillary gas chromatography. Recoveries of 23 OP pesticides were in the range 82-111%. The whole procedure takes ca. 20 min and compares favourably with current procedures.

Tolerance to O6-methylguanine and 6-thioguanine cytotoxic effects: a cross-resistant phenotype in N-methylnitrosourea-resistant Chinese hamster ovary cells.

The biochemical and genetic characteristics of a clone of Chinese hamster ovary cells displaying resistance to N-methyl-N-nitrosourea (MNU) and 6-thioguanine (6-TG) were analyzed. The initial level of 7-methylguanine, 3-methyladenine, and O6-methylguanine formation and the repair rates for these alkylated bases were the same in the resistant and in the parental cell line, indicating that the resistance to alkylation damage is not due to differences in DNA alkylation. After exposure for 24 or 48 h to 6-TG (0.6 micrograms/ml) in culture medium, the resistant clone in contrast to them, was able to replicate the DNA containing the base analogue during the following 24 h. These data are in agreement with the hypothesis that resistant cells tolerate both O6-methylguanine and 6-TG present in DNA. The tolerance to MNU and 6-TG also included chromosomal damage induced by these two agents, and MNU-resistant cells incurred less sister chromatid exchanges after treatment with either MNU or 6-TG. 6-TG-resistant cells, selected by growth in 6-TG, exhibited cross-resistance to MNU but not to methyl methanesulfonate, confirming that a common pathway of tolerance is responsible for resistance to 6-TG and O6-methylguanine.

Evaluation of p,p'-DDE, p,p'-DDT and polychlorobiphenyls (PCBs) levels in samples of human milk from Rome, Florence and the surrounding areas.

Sixty-five samples of human milk obtained from individual donors living in Rome and surrounding areas between 1982 and 1984, and 28 samples from Florence and surrounding areas obtained during 1985 were analyzed for residues of p,p'-DDE, p,p'-DDT and PCBs. Levels of p,p'-DDE were between 5 and 126 ppb (micrograms/kg of milk), with an average value of 45 ppb (median 34). Levels of p,p'-DDT ranged from 1 to 79 ppb, with an average value of 10 ppb (median 7). PCBs were found at levels ranging from 7 to 304 ppb, with an average value of 74 ppb (median 66). As to the p,p'-DDT, when compared to the data previously obtained in Italy during 1975-77, the present findings show a decrease of the average value and a lower incidence of samples with higher values. These effects are less pronounced for the p,p'-DDE. No relevant variation was observed in the levels of PCBs in comparison with the data obtained in Italy during 1981-82.

Pesticide residue control in the years 1983-1985.

Levels of organochlorinated, organophosphate, dithiocarbamic, benzimidazolic, and phthalimidic compounds, as well as some other pesticide residues obtained from Regional Laboratories in the years 1983-1985, were assembled and presented as a part of a study aimed at evaluating food contamination. Approximately 2000 samples of vegetables, fruits, cereals, and food of animal origin were analyzed by local laboratories during the years 1983, 1984, and 1985. Samples were randomly obtained from the market, at production levels, or sometimes at custom sites. Approximately 10,000 determinations of single compounds or classes of compounds were carried out. The analytical results are presented and compared with the Italian maximum residue limits. In the majority of the samples analyzed, either there were no residues or the residues were below the permitted level for the specific pesticide in each commodity. High levels of residues were present in a few cases, mainly with respect to some fungicides in fruits and vegetables and chlorinated compounds in some animal products.