Validation of quantativemethod for thiaminedetection in animalfeed, tissue and plasma
Project Number1U18FD008346-01
Contact PI/Project LeaderSNEKVIK, KEVIN ROY Other PIs
Awardee OrganizationWASHINGTON STATE UNIVERSITY
Description
Abstract Text
1 Project Summary
2 The Thiamine HPLC-FLD quantification project is proposed by the WADDL-ASL laboratories to
3 fill the analytical gap in the Vet-LIRN network by developing a standardized HPLC-FLD
4 (fluorescence) method to quantify the concentration of thiamine, thiamine
5 pyrophosphate(diphosphate), and thiamine monophosphate in animalfeeds, tissues, and blood
6 that can be implemented by all in-network laboratories with HPLC-FLD capabilities. Thiamine is
7 an essential vitamin that must be supplied in the diet of many animal species and severe
8 disease can quickly develop in deficient diets. The current gold standard for diagnosis relies on
9 recognition of clinical signs and response to treatment as thiamine testing in tissue and blood
10 samples is not readily available in veterinary diagnostic laboratories despite being routine in
11 human clinical laboratories. A method utilizing HPLC-FLD instrumentation will be extrapolated
12 from an HPLC method validated in fish tissues and roe into an HPLC-FLD method for increased
13 sensitivity and the matrices expanded to include animal and pet feed and animal (non-fish)
14 tissues (liver, brain, and muscle) and blood. A Level 2 FDA validation will be done in the lead
15 laboratory using prepared and verified homogeneous test materials including animal and pet
16 feed, animaltissues (liver, brain, and muscle), blood, and three species of fish roe. A Level 3
17 FDA validation will then be done by collaborators at the Cornell University Animal Health
18 Diagnostic Center and supply additional recommendations for optimization of the method. After
19 completion of the validation, the SOP for the Thiamine HPLC-FLD quantification method will be
20 made available to Vet-LIRN network laboratories and added to the literature in a peer-reviewed
21 journal. Oral and/or poster presentations at appropriate conferences will be made to increase
22 visibility and awareness of the new standardized SOP available for use and implementation.
23 Additionally, method development will be completed to expand the method to an HPLC-UV/VIS
24 to increase accessibility in Vet-LIRN laboratories without HPLC-FLD testing capabilities in case
25 of an event requiring significant surge analysis capacity.
Public Health Relevance Statement
Project Narrative
The proposed Thiamine HPLC-FLD quantification method will fill a diagnostic gap in the
laboratories of the FDA Vet-LIRN network by standardizing a method for accurate quantification
of thiamine, thiamine pyrophosphate(diphosphate), and thiamine monophosphate in animalfeeds, tissues, and blood. This standardized method will facilitate implementation at laboratories
with HPLC-FLD testing abilities, improving the availability of monitoring pet feeds to provide
early warnings for mis-formulations and create surge capacity in emergent cases that might
require significant feed recalls. The extension of this method into tissues and blood will allow
confirmation of thiamine deficiency in suspect cases, facilitating treatment of any affected
animals and providing evidence for any necessary related feed recalls.
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