Work

Optimizing the uptake of deuterated docosapentaenoic acid by salmonid liver cells using a uniform shell design and liquid chromatography triple-quadrupole mass spectrometry

Year: 2025

Type: article

Abstract: Stable isotope-labelled fatty acids are valuable tracers for studying lipid metabolism in cell models. However, published methods rarely provide guidance on selecting the optimal concentration and incubation time to ensure accurate and reproducible measurements of cellular uptake and metabolism that reflect physiological conditions. This study systematically evaluated the uptake of deuterated docosapentaenoic acid (DPA-d 5 ) in Atlantic salmon ( Salmo salar L.) liver cells using a uniform shell design methodology. Cultures were exposed to DPA-d 5 at concentrations ranging from 0 to 100 μM for 24, 48, or 72 h and the uptake was quantified in cell lysates by liquid chromatography–mass spectrometry. Results indicate that the concentration of DPA-d₅ has the strongest positive influence on signal intensity. Incubation time slightly reduces the signal, but when combined with higher concentration, it increases the uptake of DPA-d₅. Modelling of the data allowed selecting 60 μM DPA-d 5 and 48 h as the optimal culturing condition that provides a balance between achieving a strong signal and minimizing the risk of oversaturation or metabolic artifacts. This work provides a methodological foundation for future fatty acid tracer experiments in aquaculture research. • First systematic/symultaneous evaluation of both tracer concentration and incubation time required to optimize deuterated fatty acid uptake in salmon liver cells. • The selected optimal condition (60 μM DPA-d₅ and 48 h) provides the best balance between signal strength, reproducibility, and avoidance of metabolic artifacts. • A robust methodological foundation for future fatty acid tracer experiments is proposed. • Clear LC-MS/MS separation of DPA-d₅, DHA-d₅, and their endogenous analogs with no contamination or carryover, demonstrating analytical robustness applicable to other isotope-tracer lipidomic studies.

Source: Journal of Chromatography B

DOI: 10.1016/j.jchromb.2025.124863

Authors: Pedro Araujo, Zhenxiao Zhuang, Maren Hoff Austgulen, Bjørg Kristine Hundal

Institutions: Norwegian Institute of Marine Research, Universidad de Las Palmas de Gran Canaria

Language: English

Topic: Aquaculture Nutrition and Growth

Subfield: Aquatic Science

Field: Agricultural and Biological Sciences

Domain: Life Sciences

Open Access status: hybrid

Estimated APC paid: $3,640

Funder: Havforskningsinstituttet