Determination of atrazine metabolites in human urine: Development of a biomarker of exposure

Abstract

Enzyme-linked immunosorbent assays (ELISAs) are reported for the detection of atrazine and its principle metabolite in human urine. The ELISAs can be used with crude urine or following extraction and partial purification by methods described in this report. GC, MS, and HPLC techniques were used to confirm and complement the ELISA methods for qualitative and quantitative detection of urinary metabolites. A series of samples from workers applying this herbicide confirmed a mercapturic acid conjugate of atrazine as a major urinary metabolite. The mercapturate was found in concentrations at least 10 times that of any of the N-dealkylated products or the parent compound. Atrazine mercapturic acid was isolated from urine using affinity extraction based upon a polyclonal antibody for hydroxy-s-triazines and yielded products sufficiently pure for structure confirmation by MS/MS. In a pilot study monitoring applicators, a relationship between cumulative dermal and inhalation exposure and total amount of atrazine equivalents excreted over a 10-day period was observed. On the basis of these data, we propose that an ELISA for the mercapturate of atrazine could be developed as a useful marker of exposure.

Biomonitoring of Human Exposure to Triazine Herbicides

“…Za određivanje izvornih triazinskih spojeva i njihovih mono-i didealkiliranih metabolita u urinu ljudi najčešće se rabi plinskokromatografska analiza uz detektor selektivan za spojeve dušika i fosfora (engl. Gas Chromatography-Nitrogen Phosphorus Detector, GC-NPD) (34,49,50) ili uz detekciju spektrometrom masa (engl. Gas ChromatographyMass Spectrometry, GC-MS) (50)(51)(52).…”

“…Gas Chromatography-Nitrogen Phosphorus Detector, GC-NPD) (34,49,50) ili uz detekciju spektrometrom masa (engl. Gas ChromatographyMass Spectrometry, GC-MS) (50)(51)(52). Za određivanje polarnijih metabolita, primjerice konjugata s merkapturnom kiselinom, prikladnija je tekućinska kromatografi ja visoke djelotvornosti uz UV-detektor (engl.…”

Two-dimensional high performance liquid chromatography separation and tandem mass spectrometry detection of atrazine and its metabolic and hydrolysis products in urine

“…ATZ also can be metabolized to the corresponding hydroxyl derivatives (ATZ-OH, DEA-OH, DIA-OH and DAA-OH) in plants and soil bacteria [4]. From ATZ-OH, liver enzymes can produce other desalkyl OH analogs of desethyl hydroxy atrazine (DEA-OH), desisopropyl hydroxy atrazine (DIA-OH), and diamino hydroxy atrazine (DAA-OH) [8].…”

“…Developing a selective and sensitive method to measure simultaneously trace concentrations of Cl, Mer, and OH ATZ metabolites in a biological matrix is an analytical chemistry challenge. Concentrations of the relatively volatile Cl-derivatives can be quantified by gas chromatography/mass spectrometry (GC/MS) detection [8][9][10]. However, the GC/MS methodology for measuring DAA in biological matrices is not selective enough to eliminate signal interferences from urinary components [11].…”

“…By contrast, simultaneous determination of Cl, Mer, and OH derivatives has been more successful with high performance liquid chromatography (HPLC) coupled with atmospheric pressure chemical ionization (APCI) tandem mass spectrometry (MS/MS) detection [8,[12][13][14]. In these studies, ATZ and its metabolites are separated from each other and from other matrix components either by reversed-phase (RP) [8,12,13] or strong cation exchange (SCX) chromatography [13,14]. Neither of these HPLC modes by itself allows the separation of the full panel of ATZ metabolites that includes DAA, DAAMer and DAA-OH.…”