A Method to Estimate the Dietary α-Linolenic Acid Requirement Using Nonesterified DHA and Arachidonic Acid Oxylipins and Fatty Acids

The dietary requirement for α-linolenic acid (ALA) remains unclear, as evidenced by the absence of a Recommended Dietary Allowance (RDA) for this essential fatty acid (FA). In previous studies, we observed that the amount of dietary ALA required to maximize nonesterified (NE) DHA oxylipins appears to be higher than the amount required to maximize tissue esterified DHA, which have classically been used to estimate the ALA requirement. Further, we observed that dietary ALA reduces esterified arachidonic acid (ARA) and its NE oxylipins. Since NE oxylipins and FA mediate the biological activities of FA, we examined whether these DHA and ARA pools could be used to determine the dietary ALA requirement. Nine groups of 4-wk-old male Sprague-Dawley rats (n = 5) and 10 groups of male and female CD1 mice (n = 6) were provided 0.1–2.5 g ALA and 2 g of linoleic acid per 100 g of AIN93G-based diets. NE DHA and ARA and their oxylipins in serum, liver, kidney, and brain homogenates underwent solid phase extraction and were quantified by HPLC-MS/MS. Breakpoint analysis of transitions from increase to plateau was conducted using piecewise regression. In response to increasing dietary ALA, NE DHA oxylipins, and DHA in serum, liver, and kidney (but not the brain) initially increased rapidly and then reached a plateau whereas ARA oxylipins and ARA tended to decrease before reaching a plateau. Thus, breakpoints were calculated for the ratios of DHA/ARA and hydroxy-DHA/hydroxy-ARA (DHAOH/ARAOH), which consisted of oxylipins synthesized via pathways common to both FA. In serum, liver, and kidney, the highest estimated breakpoint indicated an ALA requirement of ∼0.7 g/100 g diet (1.7% energy), approximately twice that of previous estimations. This study supports the use of NE DHAOH/ARAOH or DHA/ARA as biochemical indicators of the ALA requirement. Applying this method in rats and mice indicates that the requirement is higher than previously estimated using esterified DHA alone.