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Improved Simple Sample Pretreatment Method for Quantitation of Major Human Milk Oligosaccharides Using Ultrahigh Pressure Liquid Chromatography with Fluorescence Detection
- Source :
- Journal of Agricultural and Food Chemistry. 67:12237-12244
- Publication Year :
- 2019
- Publisher :
- American Chemical Society (ACS), 2019.
-
Abstract
- Human milk oligosaccharides (HMOs) maintain and promote infant health. Most of the current methods for HMOs quantitation require labor-intensive and time-consuming steps for sample preparation. This study presents two very simple and easy-to-operate pretreatment methods, requiring either ultrafiltration or centrifugation to separate free oligosaccharides from whole fat human milk and other milk matrix before oligosaccharides labeling for quantifying HMOs using ultrahigh pressure liquid chromatography with fluorescence detection. A single chromatography run quantified 15 sialylated and neutral HMOs with high sensitivity (with an LOD less than 8 pg for all HMOs tested: about 1 pg for 2'-fucosyllactose, 3-fucosyllactose, 4'-galactosyllactose, 3'-galactosyllactose, and 6'-galactosyllactose) and good linearity with coefficient of correlation above 0.999. Accuracy and precision were satisfactory for both pretreatment methods. Overall, the centrifugation pretreatment was efficient and reliable for samples with high levels of oligosaccharides, and the ultrafiltration pretreatment was especially suitable for samples with low oligosaccharide abundance.
- Subjects :
- 0106 biological sciences
Ultrafiltration
Oligosaccharides
Centrifugation
Pretreatment method
01 natural sciences
Matrix (chemical analysis)
Simple sample
Humans
Sample preparation
Chromatography, High Pressure Liquid
health care economics and organizations
chemistry.chemical_classification
Chromatography
Milk, Human
010401 analytical chemistry
General Chemistry
Oligosaccharide
Fluorescence
0104 chemical sciences
chemistry
Female
General Agricultural and Biological Sciences
010606 plant biology & botany
Subjects
Details
- ISSN :
- 15205118 and 00218561
- Volume :
- 67
- Database :
- OpenAIRE
- Journal :
- Journal of Agricultural and Food Chemistry
- Accession number :
- edsair.doi.dedup.....57da32ee8c51af977cca33e1a2363eb4