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An optimized LC-MS/MS method as a pivotal tool to steer thiamine biofortification strategies in rice

(2021) TALANTA. 224.
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Abstract
In developing countries, people mainly depend on rice as their primary source of calories. However, the thiamine content of rice is below minimal requirements. Biofortification, via genetic engineering, is a cost-effective strategy to increase thiamine content in rice. We report on the optimization of a matrix-specific method, including extensive optimization of the sample preparation to ensure maximal sensitivity and stability. The LCMS/MS method was fully validated for the simultaneous quantification of thiamine, its precursors 4-methyl-5-(2-hydroxyethyl) thiazole (HET) and 4-amino-2-methyl-5-hydroxymethylpyrimidine (HMP) and its diphosphate derivative (TDP) in both polished and unpolished rice. Bias was below 9% for all analytes and total imprecision (CV%) was within pre-set acceptance criteria (<= 15%) for both QCs and real samples. Thiamine monophosphate (TMP), for which no labeled analogue was available at the time of analysis, was determined without internal standard. Although both accuracy and precision criteria were met (bias and CV < 12%), the determination of TMP was considered semi-quantitatively. Moreover, TMP was found to be only a minor thiamine form (<1% of total thiamine in all lines analyzed, both wild-type and genetically engineered), with measurable levels only present in unpolished rice. Finally, the validity and applicability of the procedure were demonstrated via its successful application on rice lines, genetically engineered to enhance thiamine content. Consequently, this method allows to evaluate the success of biofortification strategies in rice.
Keywords
Thiamine, Rice, Biofortification, Liquid chromatography, Tandem mass spectrometry, RIBOFLAVIN, VALIDATION, RAW

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Citation

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MLA
Verstraete, Jana, et al. “An Optimized LC-MS/MS Method as a Pivotal Tool to Steer Thiamine Biofortification Strategies in Rice.” TALANTA, vol. 224, 2021, doi:10.1016/j.talanta.2020.121905.
APA
Verstraete, J., Strobbe, S., Van Der Straeten, D., & Stove, C. (2021). An optimized LC-MS/MS method as a pivotal tool to steer thiamine biofortification strategies in rice. TALANTA, 224. https://doi.org/10.1016/j.talanta.2020.121905
Chicago author-date
Verstraete, Jana, Simon Strobbe, Dominique Van Der Straeten, and Christophe Stove. 2021. “An Optimized LC-MS/MS Method as a Pivotal Tool to Steer Thiamine Biofortification Strategies in Rice.” TALANTA 224. https://doi.org/10.1016/j.talanta.2020.121905.
Chicago author-date (all authors)
Verstraete, Jana, Simon Strobbe, Dominique Van Der Straeten, and Christophe Stove. 2021. “An Optimized LC-MS/MS Method as a Pivotal Tool to Steer Thiamine Biofortification Strategies in Rice.” TALANTA 224. doi:10.1016/j.talanta.2020.121905.
Vancouver
1.
Verstraete J, Strobbe S, Van Der Straeten D, Stove C. An optimized LC-MS/MS method as a pivotal tool to steer thiamine biofortification strategies in rice. TALANTA. 2021;224.
IEEE
[1]
J. Verstraete, S. Strobbe, D. Van Der Straeten, and C. Stove, “An optimized LC-MS/MS method as a pivotal tool to steer thiamine biofortification strategies in rice,” TALANTA, vol. 224, 2021.
@article{8681286,
  abstract     = {{In developing countries, people mainly depend on rice as their primary source of calories. However, the thiamine content of rice is below minimal requirements. Biofortification, via genetic engineering, is a cost-effective strategy to increase thiamine content in rice. We report on the optimization of a matrix-specific method, including extensive optimization of the sample preparation to ensure maximal sensitivity and stability. The LCMS/MS method was fully validated for the simultaneous quantification of thiamine, its precursors 4-methyl-5-(2-hydroxyethyl) thiazole (HET) and 4-amino-2-methyl-5-hydroxymethylpyrimidine (HMP) and its diphosphate derivative (TDP) in both polished and unpolished rice. Bias was below 9% for all analytes and total imprecision (CV%) was within pre-set acceptance criteria (<= 15%) for both QCs and real samples. Thiamine monophosphate (TMP), for which no labeled analogue was available at the time of analysis, was determined without internal standard. Although both accuracy and precision criteria were met (bias and CV < 12%), the determination of TMP was considered semi-quantitatively. Moreover, TMP was found to be only a minor thiamine form (<1% of total thiamine in all lines analyzed, both wild-type and genetically engineered), with measurable levels only present in unpolished rice. Finally, the validity and applicability of the procedure were demonstrated via its successful application on rice lines, genetically engineered to enhance thiamine content. Consequently, this method allows to evaluate the success of biofortification strategies in rice.}},
  articleno    = {{121905}},
  author       = {{Verstraete, Jana and Strobbe, Simon and Van Der Straeten, Dominique and Stove, Christophe}},
  issn         = {{0039-9140}},
  journal      = {{TALANTA}},
  keywords     = {{Thiamine,Rice,Biofortification,Liquid chromatography,Tandem mass spectrometry,RIBOFLAVIN,VALIDATION,RAW}},
  language     = {{eng}},
  pages        = {{9}},
  title        = {{An optimized LC-MS/MS method as a pivotal tool to steer thiamine biofortification strategies in rice}},
  url          = {{http://doi.org/10.1016/j.talanta.2020.121905}},
  volume       = {{224}},
  year         = {{2021}},
}

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