Journal of AOAC International

Determination of dialkyl phosphates as breakdown products of organophosphorus insecticides in fruit juices by HPTLC with fluorescence detection.

PMID 19610356


Dialkyl phosphates (DAP) are common degradation products of organophosphorus pesticides that are used as urinary biomarkers for human exposure. An HPTLC method was developed for the quantitative determination of DAP in fruit juices, i.e., dimethyl phosphate (DMP), dimethyl thiophosphate (DMTP), diethyl phosphate (DEP), and diethyl thiophosphate (DETP). Dibutyl phosphate (DBP) was used as an internal standard. The method was based on precipitation of fruit acids in the presence of barium chloride and acetonitrile and liquid-liquid extraction with acetonitrile-diethyl ether. Extracted DAP were derivatized with 1-(bromoacetyl)pyrene (BAP), and the BAP derivatives separated on HPTLC amino plates with dichloromethane as the mobile phase. Densitometry was performed by measurement of fluorescence at 366/>400 nm. The limit of quantification (LOQ) values were between 0.8 and 1.4 ng/zone. Fluorescence enhancement was achieved by dipping the plate into a paraffin oil solution, increasing the sensitivity and resulting in an LOQ of 0.5-0.6 ng/zone. Repeatabilities with relative standard deviations of < or = 3.5% (n = 5, at 15-20 ng/zone) and coefficients of correlation of 0.9999 were highly satisfactory for rapid trace analysis of DAP in the fruit juices by HPTLC. The mean recoveries from apple juice spiked at 0.5 mg/L were 74, 83, 70, and 57% for DMP, DEP, DMTP, and DETP, respectively. If an application volume of 5 microL of apple juice extract was applied, the LOQ in apple juice was 300 microg/L. However, this can be lowered by application of higher volumes (up to 50 microL) or a more concentrated derivatization batch.

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1-(Bromoacetyl)pyrene, 97%