Can any body please suggest TLC or Spectrophotometric determination of Ochratoxin?
I have worked on aflatoxin and used TLC for detection it. TLC only can show you that in comparison to standard loaded beside other samples which of them have the same Rf and maybe it is the same as standard, for instance if you load ochratoxin as standard you can find out which sample is ochratoxin. but if you want to know more detail for example the amount of toxin and some other details it is better to use another method fro detection the toxin.
TLC shows the presence or absence of mycotoxins (OTA ,AFLA B1, ...) in your samples
For detection and quantification of mycotoxins using your high-performance chromatography and fluorescence detection (HPLC FD), and ELISA immunoassay.
You can download and use Romer Labs method for TLC determination of ochratoxins if you have a dencitometer to quantify, however you will also need pure standards for quantification
The chromatoplaques are placed vertically in the migration tank filled to a height of 0.5 cm from the migration of solvent composed of (5v toluene / 4v acetate d'ethyl/1V formic acid), the constituents of the sample are eluted by the mobile phase, which migrates by capillary action upwardly from the plate, they can be identified by comparison with the simultaneous elution of the standard control reports comparing their front (Rf).
The chromatoplaques developed until the solvent reached the limit of the upper edge, then they will be promptly removed is dried in a drying cabinet.
Revelation chromatograms:
Reading TLC plates in a darkroom under UV lamp 366nm where the revelation of ochratoxin A that results in fluorescence and the presence of aflatoxin B1 that results in a blue fluorescence.
The TLC method is a technique qualitatife standard, but for more advanced analysis according to international standards for the detection and quantification of OTA study is carried out by reversed HPLC column with fluorescence detection. With a flow rate of 1mL/min, the mobile phase consisting of acetonitrile-water mixture, acetic acid, glacial (55: 43: 2, v / v / v). The detection of OTA is performed at wavelengths of excitation (333nm) and emission (460 nm). Quantification of OTA is performed from the surface of the absorption peak and the OTA rates are determined in the samples according to the calibration curve.
See as attachment our poster of "Development of planar chromatography methods for determination of 11 mycotoxins in biological materials combined with immunoaffinity chromatography clean-up step:
My best regards
Vladimir
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