I have isolated Aspergillus ochraceus strains. I would like to quantify the ochratoxin-A produced by those cultures. I am growing the cultures in 250ml of broth medium. Now I would like to quantify the Ochratoxin-A of those cultures. How can I extract Ochratoxin from those liquid cultures?How can I test the presence of OTA through TLC? Later how can I quantify OTA through HPLC? What is the best detector (UV/Florescent) for detection of OTA? What are safe levels of OTA in foods and feeds especially in Poultry feeds.
HPLC is the best way to quantify any molecules. First we need to optimize solvent system for standard then maintain the same experimental condition for your crude sample also. If you get the peak at the same region (Rt) as per standard so we can say that peak is that sample. Then calculate the quantity of sample present in your crude sample by using area under that peak.
Dear Akarapa, the other option is to use an ELISA based method for estimation of Ochratoxin A. If you are interested in this, you would need an ELISA plate reader, and pure Ochratoxin for standardisation, Antisera for the toxin and BSA conjugate. the other necessary reagents are easily obtainable and a sample would cost you less than $5 when the procedure is optimised. All should be available in India if you are interested in following up on this. It is relatively much cheaper and provides the accuracy up to less than 1ppb. More recently, it has been more difficult (and thus slightly more expensive) to move pure toxin due to fears of bio-warfare but still easily accessable. Alternatively, you could ship you samples to ICRISAT hyderabad for analysis. Good luck.
Dear Acarapu, I send you our publication in Analytical Letters : "DETERMINATION OF OCHRATOXIN A IN FOOD BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY"
as attachment.
Best regards
Vladimir
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