VALIDATION PROTOCOL FOR DETERMINATION OF FUMONISINS IN CORN
Date Issued
2022-09-22
Author(s)
Biljana Stojanovska-Dimzoska
Angelevska, Aleksandra
Abstract
Fumonisins are a group of at least 15 closely related mycotoxins produced principally
by Fusarium verticillioides and Fusarium proliferatum. Fumonisin B1 is the most
frequently found in food products (especially corn) representing 70–80% of the total
of fumonisin content and, together with FB2 and FB3, seem to be the major fumonisin
due to its toxic properties. It is most important in veterinary medicine as a cause of
leukoencephalomalacia in horses, liver cancer in rats, and porcine pulmonary edema. In
areas of high maize consumption fumonisins may be responsible for esophageal cancer in
humans. FB1 is classifi ed as possibly carcinogenic to humans (IARC group 2B). As an
animal and human health threat, fumonisins are regulated by legislation worldwide.
Maximum limits for the total content of fumonisins have been established in the EU in
maize and maize based products (EC No 1126/2007). Among various analytical methods
for the determination of mycotoxins, ELISA method is still the method of choice for
screening purposes. The aim of this paper was to test and validate a commercial ELISA kit
for determination of fumonisins. The validation procedure was performed in compliance
with Commission Decision 2002/657/EC. For linearity test, six standard solutions were
used in a concentration range of 0 – 2 mg/kg and a satisfactory coefficient of correlation
was found. The LOD was accomplished from the measurement of the background response
from 20 blank corn samples and it was found to be 50 μg/kg. The determination of trueness
was performed by means of an analysis of six replicates of the CRM and the obtained
value was 83.5%. Recovery, the other accuracy parameter, was achieved by fortifying
blank samples at level of one-half of MRL and the obtained value was 112.6%. The both
values (trueness and recovery) were in accordance to the performance criteria. CCβ was
established by analyzing at least 20 blank samples fortified at level one-half of MRL and
the achieved value was 0.63 ± 0.31 mg/kg. Repeatability was estimated using the data from
the recovery and the RSDr was 17.33%. The within-laboratory reproducibility (RSDR) of
the method was 27.8% which is in accordance to the EU validation criteria. The realised
validation protocol shows that this rapid ELISA method is simple, because it employs staff
with lesser technical training; easier, it saves time and costs, saves investments in complex
instruments and it is accurate. It can be implemented for routine analysis of fumonisins.
by Fusarium verticillioides and Fusarium proliferatum. Fumonisin B1 is the most
frequently found in food products (especially corn) representing 70–80% of the total
of fumonisin content and, together with FB2 and FB3, seem to be the major fumonisin
due to its toxic properties. It is most important in veterinary medicine as a cause of
leukoencephalomalacia in horses, liver cancer in rats, and porcine pulmonary edema. In
areas of high maize consumption fumonisins may be responsible for esophageal cancer in
humans. FB1 is classifi ed as possibly carcinogenic to humans (IARC group 2B). As an
animal and human health threat, fumonisins are regulated by legislation worldwide.
Maximum limits for the total content of fumonisins have been established in the EU in
maize and maize based products (EC No 1126/2007). Among various analytical methods
for the determination of mycotoxins, ELISA method is still the method of choice for
screening purposes. The aim of this paper was to test and validate a commercial ELISA kit
for determination of fumonisins. The validation procedure was performed in compliance
with Commission Decision 2002/657/EC. For linearity test, six standard solutions were
used in a concentration range of 0 – 2 mg/kg and a satisfactory coefficient of correlation
was found. The LOD was accomplished from the measurement of the background response
from 20 blank corn samples and it was found to be 50 μg/kg. The determination of trueness
was performed by means of an analysis of six replicates of the CRM and the obtained
value was 83.5%. Recovery, the other accuracy parameter, was achieved by fortifying
blank samples at level of one-half of MRL and the obtained value was 112.6%. The both
values (trueness and recovery) were in accordance to the performance criteria. CCβ was
established by analyzing at least 20 blank samples fortified at level one-half of MRL and
the achieved value was 0.63 ± 0.31 mg/kg. Repeatability was estimated using the data from
the recovery and the RSDr was 17.33%. The within-laboratory reproducibility (RSDR) of
the method was 27.8% which is in accordance to the EU validation criteria. The realised
validation protocol shows that this rapid ELISA method is simple, because it employs staff
with lesser technical training; easier, it saves time and costs, saves investments in complex
instruments and it is accurate. It can be implemented for routine analysis of fumonisins.
Subjects
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