Bone-protective studies are beginning to show the positive effects of dried prune consumption on bone health in young women using hormonal contraceptives. Dried plums are known to be sources of boron, potassium, and phenolic compounds have shown inhibitory effects on bone resorption and stimulate bone formation. The effects can be visualized through the analysis of the biomarkers chlorogenic acid, neochlorogenic acid, and cryptochlorogenic acid. Untargeted metabolomics was employed for broader analysis to explore all possible metabolites found in both serum and fecal samples, with a particular interest in the previously mentioned biomarkers. Subjects were divided into a control group administered oral contraceptives with no dried plum or an experimental group administered oral contraceptives with dried plums. Fecal and plasma samples were collected on day 1 and day 21 of the trial. After samples were collected, they were flash frozen with liquid nitrogen to halt any metabolic processes. Fecal samples were pulverized, slurried with LCMS grade water, then mixed with a solution of 50:50 acetonitrile:methanol and sterile filtered. Small volumes of plasma samples were treated with 50:50 acetonitrile:methanol, precipitated, centrifuged and its supernatant extracted. All the samples went through a nitrogen gas dry down and were reconstituted with 50:50 acetonitrile:water. They were then analyzed using liquid chromatography-high resolution mass spectrometry in reverse phase to determine the effectiveness of metabolites crossing into the bloodstream and if any became modified through glucuronidation or sulfonation during the digestive process. The metabolomics bioinformatics platform XCMS Online was used to perform feature detection, retention time alignment, and statistical analysis to identify dysregulated metabolites in fecal and serum samples.

Identification of bone-protective biomarkers in human fecal and plasma using high resolution mass spectrometry

ACS Spring 2022

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