A method for the simultaneous quantification of eight metabolites of synthetic pyrethroids in urine of the general population using gas chromatography-tandem mass spectrometry.

Synthetic pyrethroids are highly effective, widespread insecticides applied worldwide for different purposes. Among the possible sources of exposure for the general population, pyrethroid residues in food and their prominent use for the conservation of wool carpets or in indoor pest control might play a major role. On the basis of previous works, we have developed and validated a highly sensitive and specific GC/MS/MS-method to simultaneously quantify the metabolites of the most common synthetic pyrethroids in urine, namely cis- and trans-(2,2-dichlorovinyl)-2,2-dimethylcyclopropanecarboxylic acid (DCCA), cis-(2,2-dibromovinyl)-2,2-dimethylcyclopropanecarboxylic acid (DBCA), 4-fluoro-3-phenoxybenzoic acid (F-PBA), 3-phenoxybenzoic acid (3-PBA) as well as the metabolites cis-3-(2-chloro-3,3,3-trifluoroprop-1-enyl)-2,2-dimethyl-cyclopropanecarboxylic acid (ClF3CA, λ-cyhalothrin/bifenthrin), 4-chloro-α-isopropylbenzene acetic acid (CPBA, esfenvalerate), and 2-methyl-3-phenylbenzoic acid (MPB, bifenthrin). After acidic hydrolysis to cleave conjugates in urine, the analytes are subjected to a pH-controlled extraction into n-hexane. After concentration, the analytes are derivatised using MTBSTFA and finally quantified by GC/MS/MS in EI-mode using d6-trans-DCCA and (13)C6-3-PBA as internal standards. The limit of quantification for these metabolites was 0.01 µg/L urine. Precision within and between series was determined to range between 1.6 and 10.7 % using a native quality control sample as well as a urine sample spiked with 0.3 µg/L of the analytes. To investigate possible background excretions, we analysed spot urine samples of 38 persons of the general population in a pilot study. cis- and trans-DCCA as well as 3-PBA could be quantified in every urine sample investigated, while MPB and F-PBA could only be detected in two samples. The median levels for excretion of cis-DCCA, trans-DCCA, 3-PBA, ClF3CA, DBCA, CPBA, F-PBA and MPA were 0.08, 0.17, 0.22, 0.04, 0.04, <0.01, <0.01 and < 0.01 µg/L urine, respectively. The excretion of metabolites revealed excellent correlations between cyclopropane carboxylic acids and 3-PBA. Our method is highly suitable for human biomonitoring of exposures to synthetic pyrethroids in environmental medicine. Remarkable are the high detection rates for the metabolites ClF3CA (90 %) and CPBA (40 %), proving that their parent pyrethroids have entered the market in Germany.

Sensitive and accurate analyses of free 3-nitrotyrosine in exhaled breath condensate by LC-MS/MS.

The quantitative determination of 3-nitro-l-tyrosine, a biological marker for inflammatory processes, in exhaled breath condensate (EBC) is described. The clean-up and preconcentration was performed by solid phase extraction (SPE). After liquid chromatography the specific detection was performed by tandem mass spectrometry using electron spray ionisation and selected reaction monitoring (SRM). 13C9-3-nitrotyrosine was used as an internal standard. For reliability, tests for the precision of the method, the losses during preparation, a test for nitrating artifacts and the comparibility of calibrants in EBC and buffer solution were performed. The calibration of the method was linear over a range of 10-500 pg/mL. The within-run coefficients of variation (CV) of the samples were found to be 8.4% at 25 pg/mL and 8.3% at 250 pg/mL. The day-to-day CV was found to be 11.2%. The limit of quantification was 3.9 pg/mL. The losses during preparation were 15%. The discrepancy between the calibration with EBC and buffer solution was below 10%. No artificial production of 3-nitrotyrosine was observed during the procedure. The application of the method on the EBC samples of healthy smokers (N=10) and non-smokers (N=10) showed no difference between the two groups. The concentration of 3-nitrotyrosine ranged between the limit of quantification and 184 pg/mL and was distinctly lower than data detected by an immunoassay procedure. The procedure was proven to be accurate, sensitive and in contrast to GC methods less elaborate and is recommended for the determination of 3-nitrotyrosine in exhaled breath condensate.