Standardization and validation of assay of selected omega-3 and omega-6 fatty acids from phospholipid fraction of red cell membrane using gas chromatography with flame ionization detector.
Docosahexaenoic acid
Gas chromatography
Phospholipid
Polyunsaturated fatty acids
Validation
Journal
Journal of analytical science and technology
ISSN: 2093-3134
Titre abrégé: J Anal Sci Technol
Pays: Germany
ID NLM: 101559472
Informations de publication
Date de publication:
2021
2021
Historique:
received:
22
03
2021
accepted:
22
07
2021
entrez:
1
11
2021
pubmed:
2
11
2021
medline:
2
11
2021
Statut:
ppublish
Résumé
Docosahexaenoic acid (DHA) is an important structural component of human brain and retina. Evidence exists linking nutritional status of pregnant mothers and cognitive functions of their born infants. The DHANI (Maternal DHA Supplementation and Offspring Neurodevelopment in India) trial was implemented to evaluate the effect of maternal supplementation with DHA during pregnancy and for 6 months following delivery on motor and mental development of infants at 1 and 12 months. We describe here the standardization and validation of an assay for measurement of selected omega-3 and omega-6 fatty acids from the phospholipid fraction of red blood cells to assess their status in mothers at baseline, delivery and 6 months post-delivery and for infants in cord blood and at 1 and 12 months of age. The validated method has been used for the analysis of samples for DHANI. Lipids were extracted from a pool of red blood cells, separated using thin layer chromatography. The phospholipid fraction was esterified, and fatty acids were separated by gas chromatography using a flame ionization detector. The method accuracy for DHA was between 97 - 98% and between 91 - 95% for arachidonic acid (AA) at three different concentrations. The intra-assay and inter-assay coefficient of variation for the fatty acids ranged from 1.19 to 5.7% and 0.78 to 13.0% respectively. Intraclass correlation (ICC), as a measure of reproducibility, ranged between 0.689 and 0.996. A good linearity was observed for all the fatty acids between concentrations of 0.2-4 μg/ml. The standardized and validated method is suitable for implementation in large epidemiological studies for evaluation of fatty acids and in nutritional trials for assessment of fatty acid content of various lipid classes.
Sections du résumé
BACKGROUND
BACKGROUND
Docosahexaenoic acid (DHA) is an important structural component of human brain and retina. Evidence exists linking nutritional status of pregnant mothers and cognitive functions of their born infants. The DHANI (Maternal DHA Supplementation and Offspring Neurodevelopment in India) trial was implemented to evaluate the effect of maternal supplementation with DHA during pregnancy and for 6 months following delivery on motor and mental development of infants at 1 and 12 months. We describe here the standardization and validation of an assay for measurement of selected omega-3 and omega-6 fatty acids from the phospholipid fraction of red blood cells to assess their status in mothers at baseline, delivery and 6 months post-delivery and for infants in cord blood and at 1 and 12 months of age. The validated method has been used for the analysis of samples for DHANI.
METHODS
METHODS
Lipids were extracted from a pool of red blood cells, separated using thin layer chromatography. The phospholipid fraction was esterified, and fatty acids were separated by gas chromatography using a flame ionization detector.
RESULT
RESULTS
The method accuracy for DHA was between 97 - 98% and between 91 - 95% for arachidonic acid (AA) at three different concentrations. The intra-assay and inter-assay coefficient of variation for the fatty acids ranged from 1.19 to 5.7% and 0.78 to 13.0% respectively. Intraclass correlation (ICC), as a measure of reproducibility, ranged between 0.689 and 0.996. A good linearity was observed for all the fatty acids between concentrations of 0.2-4 μg/ml.
CONCLUSION
CONCLUSIONS
The standardized and validated method is suitable for implementation in large epidemiological studies for evaluation of fatty acids and in nutritional trials for assessment of fatty acid content of various lipid classes.
Identifiants
pubmed: 34721932
doi: 10.1186/s40543-021-00287-1
pii: 287
pmc: PMC8549946
doi:
Types de publication
Journal Article
Langues
eng
Pagination
33Informations de copyright
© The Author(s) 2021.
Déclaration de conflit d'intérêts
Competing interestsAll authors declare that they have no competing interest.
Références
J Biol Chem. 1957 May;226(1):497-509
pubmed: 13428781
Nutrients. 2020 Oct 03;12(10):
pubmed: 33023067
Clin Chim Acta. 1966 Feb;13(2):235-40
pubmed: 5941741
Nutrients. 2021 Feb 25;13(3):
pubmed: 33668849
J Lipid Res. 1965 Jul;6:428-31
pubmed: 14336214
Biometrics. 1977 Mar;33(1):159-74
pubmed: 843571
Clin Chem. 2006 Dec;52(12):2265-72
pubmed: 17053155
Pediatrics. 2018 Feb;141(2):
pubmed: 29358479
J Lipid Res. 1984 Dec 1;25(12):1391-6
pubmed: 6530596
BMC Pediatr. 2018 Aug 4;18(1):261
pubmed: 30077178