Validation and application of Biocrates AbsoluteIDQ® p180 targeted metabolomics kit using human milk

Authors: HAMPEL, DANIELA - University Of California, Davis; Shahab-Ferdows, Setti; HOSSAIN, MUTTAQUINA - International Centre For Diarrhoeal Disease Research; ISLAM, MUNIRUL - International Centre For Diarrhoeal Disease Research; AHMED, TAHMEED - International Centre For Diarrhoeal Disease Research; Allen, Lindsay - A

Submitted to: Nutrients
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/17/2019
Publication Date: 7/26/2019
Citation: Hampel, D., Shahab-Ferdows, S., Hossain, M., Islam, M., Ahmed, T., Allen, L.H. 2019. Validation and application of Biocrates AbsoluteIDQ® p180 targeted metabolomics kit using human milk. Nutrients. 11(8):1733. https://doi.org/10.3390/nu11081733.
DOI: https://doi.org/10.3390/nu11081733

Interpretive Summary: We tested a commercially available targeted metabolomics kit (BIOCRATES AbsoluteIDQ® p180; 40 acylcarnitines, 42 amino acids/biogenic amines, 91 phospholipids, 15 sphingolipids, and sum of hexoses) for its suitability for human milk analysis using the ABSciex 5500QTRAP mass-spectrometer in liquid chromatography-tandem mass-spectrometry (LC-MS/MS) and flow-injection analysis (FIA) mode. In addition, we analyzed milk (<6mo lactation) from A) Bangladeshi apparently healthy mothers (BMI> 18.5; n=12) and B) Bangladeshi mothers of stunted infants (HAZ-score<-2; n=13) to cover a wide range of metabolite concentrations. We detected 123 compounds of possible 188 in a pooled human milk sample. New internal standards and adjusted calibrator levels were beneficial to accuracy of the analysis. Metabolite recoveries were between 43-120% (CV: 2.4-24.1%, 6 replicates). Milk consumed by stunted infants vs. milk from healthy mothers was lower in 6 amino acids/biogenic amines and higher in isovalerylcarnitine, two phospholipids, and one sphingomyelin. The AbsoluteIDQ® p180 kit is a rapid and useful tool for human milk analysis.

Technical Abstract: Human milk targeted metabolomics analysis offers novel insights into milk composition and relationships with maternal and infant phenotypes and nutritional status. The Biocrates AbsoluteIDQ® p180 kit, targeting 40 acylcarnitines, 42 amino acids/biogenic amines, 91 phospholipids, 15 sphingolipids, and sum of hexoses, was evaluated for human milk using the ABSciex 5500QTRAP mass-spectrometer in liquid chromatography-tandem mass-spectrometry (LC-MS/MS) and flow-injection analysis (FIA) mode. Milk (<6mo lactation) from A) Bangladeshi apparently healthy mothers (BMI> 18.5; n=12) and B) Bangladeshi mothers of stunted infants (HAZ-score<-2; n=13) was analyzed. 123 of the possible 188 metabolites were detected in milk. New internal standards and adjusted calibrator levels were used for improved precision and concentration ranges for milk metabolites. Recoveries ranged between 43-120% (CV: 2.4-24.1%, 6 replicates). Milk consumed by stunted infants vs. that from healthy mothers was lower in 6 amino acids/biogenic amines but higher in isovalerylcarnitine, two phospholipids, and one sphingomyelin (p<0.05 for all). Associations between milk metabolites differed between groups. The AbsoluteIDQ® p180 kit is a rapid analysis tool, suitable for human milk analysis and reduces analytical bias by allowing the same technique for different specimens. More research is needed to examine milk metabolite relationships with maternal and infant phenotypes.