D 14163

Quantitation of AFB₁ was carried out using HPLC (KNAUER D-14163 UV-VIS system, Germany). Briefly, 50 µl of each of the chloroformic extracts sample was injected into the HPLC C18 reverse phase column (TSKgel ODS-80TS; 4.6 mm ID × 150 mm, Tosoh Bioscience, Japan) and eluted at a flow rate of 1 ml/min by water-acetonitrile-methanol (60:25:15, v/v/v). The amount of AFB₁ was measured at a wavelength of 365 nm using a fluorescence detector. A standard curve of AFB₁ was plotted using various concentrations of AFB₁ standard (1, 5, and 10 mg), and the amounts of AFB₁ in unknown samples were calculated by comparison of the undercurved areas of the samples with authentic standards. The elution time of the samples was compared with pure AFB₁ retention time (12.25 min) and quantified based on the ratio of the peak area of samples to those of the standards [ 21 (link)
].

The DFX concentrations of different formulations were determined by HPLC (KNAUER D-14163; Berlin, Germany). Chromatographic separations were performed using a KNAUER C18 column (4.6 mm × 250 mm), UV detector set at 245 nm, and ChromGate Clint software version 3.1.7. The mobile phase consisted of acetonitrile : methanol : water (40 : 20 : 40), the volume of injection was 20 μl, and the flow rate was 0.7 ml/min. The standard curve for DFX was constructed over a range of 0.5–50 μg/ml [24 (link), 30 ].