An LC/MS/MS method for quantitation of chemopreventive sphingadienes in food products and biological samples

Area of Science:

• Nutritional Science
• Cancer Research
• Analytical Chemistry

Background:

• Colorectal cancer (CRC) is a major cause of cancer mortality, with diet playing a significant role in its risk.
• Dietary sphingolipids, specifically sphingadienes (SDs) from plant-based foods, exhibit cytotoxic and chemopreventive properties against colon cancer cells.
• Identifying and quantifying dietary compounds like SDs is crucial for developing strategies to reduce CRC incidence and promote gut health.

Purpose of the Study:

• To develop and validate a sensitive and robust ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method.
• To quantify sphingadienes (SDs) in various food products and biological matrices.
• To investigate the pharmacokinetic behavior of SDs in vivo.

Main Methods:

• Ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) was employed for quantification.
• Method validation included linearity, sensitivity (limit of detection), and post-extraction stability assessments.
• In vivo studies involved gavage delivery of SDs to mice, with subsequent plasma and gut sample analysis.

Main Results:

• The UHPLC-MS/MS assay demonstrated linearity from 80 nM to 50 μM with a limit of detection of 3.3 nM.
• Post-extraction stability of SDs was 100% over 24 hours, and soy oils contained approximately 10 nM SDs.
• SDs were detected transiently in mouse plasma within 10 minutes post-gavage, peaking in the duodenum within 1 hour, and declining to baseline by 1 hour.

Conclusions:

• A validated UHPLC-MS/MS method enables sensitive quantification of SDs in food and biological samples.
• SDs are present in soy oil and exhibit transient absorption and distribution in vivo.
• The rapid disappearance of SDs in the lower gastrointestinal tract suggests potential metabolism or export, warranting further investigation.