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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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Title Isolation, Purification and Production of Aflatoxin B1 And T-2 Toxin Reference Mycotoxin Standards for Estimation of Mycotoxins in Poultry (Animal) Feeds.
 
Creator Krishnamurthy, T. N.
 
Contributor Devegowda, G.
 
Subject mycotoxins, toxins, productivity, rice, poultry equipment, extraction, purity, diseases, wheats, yields
 
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.
 
Date 2016-06-22T15:51:14Z
2016-06-22T15:51:14Z
2005-09-24
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/67755
 
Language en
 
Format application/pdf
 
Publisher Karnataka Veterinary, Animal and Fisheries Sciences University, Bidar