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Determination of folate monoglutamates in biofortified rice with UPLC-MS/MS

Veerle De Brouwer UGent, Serguei Storojenko UGent, Christophe Stove UGent, Dominique Van Der Straeten UGent and Willy Lambert (2009) PTERIDINES. 20(3). p.148-155
abstract
Folates are essential nutrients for human beings. To reduce folate deficiencies and consequences thereof, such as spina bifida, in developing areas like Northern China, biofortification of rice by means of genetic modification is one of the approaches. We successfully overexpressed two plant genes involved in the folate pathway in rice (1) and evaluated these interventions by analysing 6 monoglutamate folates present in rice matrix, by a validated ultra high performance chromatography-tandem mass spectrometry (UPLC-MS/MS) method. Rice was homogenized and heated, followed by a a tri-enzyme treatment (a-amylase, protease and deconjugase). Subsequently, the extract was centrifuged, ultrafiltrated, and injected on an Acquity UPLC (R) HSS T3 column (150mm x 2.1mm, 1.8-mu m particle size), coupled to tandem mass spectrometric detection. The method was validated based on linearity, sensitivity, precision, accuracy, recovery, stability and matrix effects. The limits of detection and the lower limits of quantification varied between 0.06 and 0.45 mu g/100g and 0.12 and 0.91 mu g/100g, respectively. Analysis of the distribution and levels of folates in wild-type and folate-biofortified rice showed up to 50-fold enrichment in biofortified rice, with total folate levels of up to 900 mu g/100g rice. 5-Methyl-THF was the dominant natural folate form (+/- 90% of total folate content for biofortified rice). This shows the successful implementation of this UPLC method for the rapid, quantitative determination of folates.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (proceedingsPaper)
publication status
published
subject
keyword
Folates, UPLC-MS/MS, Rice analysis, Biofortification, TANDEM MASS-SPECTROMETRY, NEURAL-TUBE DEFECTS, BIOANALYTICAL METHODS, FOLIC-ACID, EXTRACTION, SERUM, QUANTIFICATION, DISEASE
journal title
PTERIDINES
Pteridines
volume
20
issue
3
pages
148 - 155
conference name
14th International symposium on Pteridines and Folates
conference location
Jeju, South Korea
conference start
2009-06-07
conference end
2009-06-12
Web of Science type
Article; Proceedings Paper
Web of Science id
000284232100024
JCR category
BIOCHEMISTRY & MOLECULAR BIOLOGY
JCR impact factor
0.379 (2009)
JCR rank
271/281 (2009)
JCR quartile
4 (2009)
ISSN
0933-4807
language
English
UGent publication?
yes
classification
A1
id
838816
handle
http://hdl.handle.net/1854/LU-838816
alternative location
http://www.pteridines.sk/center.php?act=V&volId=12&type=abstract
date created
2010-01-26 10:53:40
date last changed
2017-09-14 09:14:44
@article{838816,
  abstract     = {Folates are essential nutrients for human beings. To reduce folate deficiencies and consequences thereof, such as spina bifida, in developing areas like Northern China, biofortification of rice by means of genetic modification is one of the approaches. We successfully overexpressed two plant genes involved in the folate pathway in rice (1) and evaluated these interventions by analysing 6 monoglutamate folates present in rice matrix, by a validated ultra high performance chromatography-tandem mass spectrometry (UPLC-MS/MS) method. 
Rice was homogenized and heated, followed by a a tri-enzyme treatment (a-amylase, protease and deconjugase). Subsequently, the extract was centrifuged, ultrafiltrated, and injected on an Acquity UPLC (R) HSS T3 column (150mm x 2.1mm, 1.8-mu m particle size), coupled to tandem mass spectrometric detection. 
The method was validated based on linearity, sensitivity, precision, accuracy, recovery, stability and matrix effects. The limits of detection and the lower limits of quantification varied between 0.06 and 0.45 mu g/100g and 0.12 and 0.91 mu g/100g, respectively. 
Analysis of the distribution and levels of folates in wild-type and folate-biofortified rice showed up to 50-fold enrichment in biofortified rice, with total folate levels of up to 900 mu g/100g rice. 5-Methyl-THF was the dominant natural folate form (+/- 90\% of total folate content for biofortified rice). This shows the successful implementation of this UPLC method for the rapid, quantitative determination of folates.},
  author       = {De Brouwer, Veerle and Storojenko, Serguei and Stove, Christophe and Van Der Straeten, Dominique and Lambert, Willy},
  issn         = {0933-4807},
  journal      = {PTERIDINES},
  keyword      = {Folates,UPLC-MS/MS,Rice analysis,Biofortification,TANDEM MASS-SPECTROMETRY,NEURAL-TUBE DEFECTS,BIOANALYTICAL METHODS,FOLIC-ACID,EXTRACTION,SERUM,QUANTIFICATION,DISEASE},
  language     = {eng},
  location     = {Jeju, South Korea},
  number       = {3},
  pages        = {148--155},
  title        = {Determination of folate monoglutamates in biofortified rice with UPLC-MS/MS},
  url          = {http://www.pteridines.sk/center.php?act=V\&volId=12\&type=abstract},
  volume       = {20},
  year         = {2009},
}

Chicago
De Brouwer, Veerle, Serguei Storojenko, Christophe Stove, Dominique Van Der Straeten, and Willy Lambert. 2009. “Determination of Folate Monoglutamates in Biofortified Rice with UPLC-MS/MS.” Pteridines 20 (3): 148–155.
APA
De Brouwer, V., Storojenko, S., Stove, C., Van Der Straeten, D., & Lambert, W. (2009). Determination of folate monoglutamates in biofortified rice with UPLC-MS/MS. PTERIDINES, 20(3), 148–155. Presented at the 14th International symposium on Pteridines and Folates.
Vancouver
1.
De Brouwer V, Storojenko S, Stove C, Van Der Straeten D, Lambert W. Determination of folate monoglutamates in biofortified rice with UPLC-MS/MS. PTERIDINES. 2009;20(3):148–55.
MLA
De Brouwer, Veerle, Serguei Storojenko, Christophe Stove, et al. “Determination of Folate Monoglutamates in Biofortified Rice with UPLC-MS/MS.” PTERIDINES 20.3 (2009): 148–155. Print.