Abstract
2,4-Dichlorophenoxyacetic acid (2,4-D) is a chlorophenoxy herbicide used worldwide. We describe a high-performance liquid chromatography (HPLC) method with UV detection for the determination of 2,4-D in female and male rat serum. This allows to observe the change of serum 2,4-D concentration in rats with time and its pharmacokinetics characteristics with a simple, rapid, optimized and validated method. The serum samples are pretreated and introduced into the HPLC system. The analytes are separated in a XDB-C18 column with a mobile phase of acetonitrile (solvent A) and 0.02 M ammonium acetate (containing 0.1% formic acid) (solvent B) using a gradient elution at a flow rate of 1.0 mL/min. The wavelength for UV detection was set at 230 nm. Calibration curve for 2,4-D was constructed over a range of 0.1–400 mg/L. The method was successfully applied to study the pharmacokinetics of 2,4-D in rats in this study. After oral administration of 300 mg/kg and 60 mg/kg 2,4-D, the mean Cmax values were 601.9 and 218.4 mg/L, the AUC0→∞ values were 23,722 and 4,127 mg×h/L and the clearance (Cl) were 1.10 and 0.02 L/(h×kg), respectively. The developed method was found to be specific, precise, reproducible and rapid.
Highlights
The widely-used herbicide 2,4-Dichlorophenoxyacetic acid (2,4-D; CAS No.94-75-7) was first synthesized in 1941 [1, 2]
All other reagents used in this study were of analytical or high-performance liquid chromatography (HPLC) grade
Representative chromatograms for blank rat serum and blank rat serum spiked with 2,4-D are shown in Fig 1A and 1B (Fig 1)
Summary
The widely-used herbicide 2,4-Dichlorophenoxyacetic acid (2,4-D; CAS No.94-75-7) was first synthesized in 1941 [1, 2]. Skin contact and inhalation are the three main pathways of human exposure to 2,4-D herbicides [3]. The overuse of 2,4-D by both the farmers and the manufacturers in the tropics has greatly damaged the health of the local ecosystem because it is deleterious to both terrestrial and aquatic living organisms [4]. The subchronic exposure of 2,4-D has shown toxic effects on the kidneys (increased kidney weight, histopathological lesions) and on the liver (increased liver weight, increased liver enzymes) [5]. Chronic exposure in rats is manifested by decreased body-weight gain, altered organ weights and hematological parameters and other biochemical changes [6].
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