High Performance Liquid Chromatography of Mycotoxin Metabolites in Human Urine

Abstract

Because mycotoxins occur worldwide in grain and grain products, evaluating their effects on the health of the population has become important. The development of a high performance liquid chromatographic (HPLC) procedure was investigated for the analysis of aflatoxin B1, citrinin, and ochratoxin A, from hydrolyzed human urine. Solid-phase extraction (SPE) was used for sample clean-up and concentration. Reversed-phase liquid chromatography (RPLC) was used with fluorescence detection for sample analysis. The presence of aflatoxin B1 was confirmed by converting it to the hemiacetal. With 10 mL of urine, the detection limit for aflatoxin B1 and ochratoxin A was in the high parts per trillion (ppt) and that for citrinin was about 10 ppb.

Estimation of ochratoxin A in portuguese population: New data on the occurrence in human urine by high performance liquid chromatography with fluorescence detection

“…The assay with b-glucuronidase was performed according to the Orti et al (1986) method. For hydrolysis, 5 mL of b-glucuronidase solution and 5 mL of 0.1 M Na 2 SO 4 were added to 10 mL of urine, and the mixture was incubated overnight ($17 h) at 37°C.…”

“…After incubation with b-glucuronidase OTA was detected in the samples in higher amounts: 0.290, 0.167, 0.231 and 0.195 ng/mL, respectively. The hydrolysis procedure described here includes the addition of sodium sulphate to the urine sample to prevent interferences from the mucopolysaccharides that compete for the enzyme, according to the Orti et al (1986) method. They included incubation with b-glucuronidase in their method for OTA determination in human urine, however, as naturally contaminated samples were not analyzed in this study, OTA conjugation with glucuronic acid in human urine could not be proven.…”

Use of high-performance liquid chromatography to assess airborne mycotoxins

“…Hemiacetal forms of AFB1 and AFG1 (fluorescence active substances) were obtained by addition of a mixture of 50 L of trifluoroacetic acid and 200 L of n-hexane at 40 • C; the solution was mechanically shaken for 15 min [18]. The solution containing all four aflatoxins in its fluorescent form was subsequently dried by evaporation under gentle nitrogen stream, redissolved with 200 L acetonitrile-deionised water (30:70, v/v) and it was finally injected in the HPLC system (Fig.…”

“…HPLC is a common method used for determination of aflatoxins in contaminated food [18,19]. To carry out this study, the method by Brera et al [7] was modified to estimate the levels of aflatoxins and ochratoxin A in the air sampled in a coffee factory during the production process.…”

Application of restricted-access media column in coupled-column RPLC with UV detection and electrospray mass spectrometry for determination of azole pesticides in urine

“…Even a combination of three cartridges Si, CN and C8 has been used for clean-up of human urine [14].…”