Isolation, Purification and Production of Aflatoxin B1 And T-2 Toxin Reference Mycotoxin Standards for Estimation of Mycotoxins in Poultry (Animal) Feeds.
KrishiKosh
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Title |
Isolation, Purification and Production of Aflatoxin B1 And T-2 Toxin Reference Mycotoxin Standards for Estimation of Mycotoxins in Poultry (Animal) Feeds.
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Creator |
Krishnamurthy, T. N.
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Contributor |
Devegowda, G.
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Subject |
mycotoxins, toxins, productivity, rice, poultry equipment, extraction, purity, diseases, wheats, yields
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Description |
An in vitro trial was conducted in the Department of Avian Production and Management to develop the technology for production, isolation, purification and standardization of aflatoxin B1and T-2 toxin reference standards. Aflatoxin B1 was produced on different substrates like broken wheat, broken maize and broken rice at different moisture levels, different incubation periods and at different temperature levels by inoculating with A. parasiticus (MTCC 411) using BOD incubator. The maximum yield of aflatoxin B1 was obtained at 7 days of incubation in broken rice with a moisture level of 35% and temperature maintained at 30 0c. T-2 toxin was produced on different substrates viz., whole wheat, whole maize and whole rice at different moisture levels, different incubation periods and at different temperature levels by inoculating with F. sporotrichoides (MTCC 1894) incubated in BOD incubator. The maximum yield of T-2 toxin was obtained at 21 days of incubation in whole wheat with a moisture level of 35% and at 200c. Aflatoxins were extracted using Acetone: water (85: 15), shaken for one hour, then passed in column for removing fat and other impurities then toxins were eluted with chloroform. Trichothecenes were extracted using Methanol: water (50: 50), shaken for one hour, then passed in separating funnels for removing fat and other impurities then toxins were eluted with diethyl ether. The aflatoxin B1 and T-2 toxins were isolated with preparative thin layer chromatography (TLC) of 1.5 to 2.0 mm plate by repeating the same procedure for four times. The purity was tested by chromatographic and spectrphotometric methods and purity of 99.00 per cent was achieved for aflatoxin B1 and 96.45 per cent for T-2 toxin reference standards. Aflatoxin B1 and T- 2 toxin reference standards were labeled and packed, distributed to various feed industries, Quality Control Laboratories, State and Central Research Laboratories on complimentary basis with brochure containing the information like mode of use, solvent used, etc. From the results we can conclude that: The maximum yield of aflatoxin B1 can be produced by inoculating with A. parasiticus on broken rice at 7 days of incubation with a moisture level of 35% and temperature maintained at 300c. The maximum yield of T-2 toxin can be produced by inoculating with F. sporotrichoides on whole wheat at 21 days of incubation with a moisture level of 35% and temperature maintained at 200c. Acetone : water (85:15l) was the solvent of choice for extraction of aflatoxins and methanol : water (50 : 50) for the trichothecenes. Preparative thin layer chromatography was found to be economical, method of choice for isolation and purification, and purity to the extent of 99.00 per cent for aflatoxin B1 and 96.45 per cent for T-2 toxin could be achieved. Establishment of linear curve at different concentration in UV spectroscopy was the indicator of purity. The molar absorptivity value at different concentrations should be within the limits of 19,800 in benzene- acetonitrile (98:2) at wavelength of 348nm (long UV-wave) for aflatoxin B1 and for T- 2 toxin it should be within the limits of 6,800 in ethyl acetate at wavelength of 333 nm. The UV–spectrum of aflatoxin B1 reference standard should show the maximum absorbance at the wavelength of 275 nm and for T- 2 toxin at the wavelength of 255nm. |
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Date |
2016-06-22T15:51:14Z
2016-06-22T15:51:14Z 2005-09-24 |
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Type |
Thesis
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Identifier |
http://krishikosh.egranth.ac.in/handle/1/67755
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Language |
en
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Format |
application/pdf
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Publisher |
Karnataka Veterinary, Animal and Fisheries Sciences University, Bidar
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