Polyphosphorylated L-ascorbic Acid: A Stable Form of Vitamin C for Aquaculture Feeds

A protected form of vitamin C, L-ascorbyl-2-poly phosphate (APP), was tested for safety and efficacy in fishes. Phosphorylation of the 2-hydroxyl group protected the 2,3-enediol of L-ascorbic acid against oxidation. When APP was added to feed, about 15% of its vitamin C activity was lost during milling and storing 5–7 d at ambient temperature. Stability of APP in pelleted feeds at 25 or 40 C was up to 83 times or 45 times greater than that of ascorbic acid (AA), respectively. In later tests, trout feed mash, steam-pelleted in triplicate, frozen, and shipped on dry ice before analysis, had 46% loss of AA and no apparent loss of APP. With 16% milling loss (highest detected in trout feeds assayed in initial tests) and 22% storage loss after 90 d, conservative APP mix rates were estimated. Mixing 153 g L-ascorbate-equivalent as APP per metric ton of airdry ingredients (153 ppm) would give 128 ppm in the finished feed after steam-pelleting and 100 ppm after 60–90 d storage at 40 C. Low-temperature extrusion of Oregon-Moist salmon feed reduced AA slightly. Ascorbic acid oxidized in frozen storage and while thawing and, after 12 h at room temperature, only trace amounts remained. Ascorbyl-2-polyphosphate at 0.05 to 0.2 times that of equimolar AA in casein-base or practical feeds supported long-term growth and development in rainbow trout and other fishes. Trout feeding trials with practical feeds in actual production regimens documented APP activity at growth rates approaching the maximum for trout farming. Corrected for greater stability, APP antiscorbutic activity appeared to be comparable to that of AA on a molar-equivalent basis. Similarly, apparent equimolar activity was inferred from estimated equilibrium levels of liver, kidney, and whole-body AA and L-ascorbyl-2-sulfate (AS) after prolonged APP use in trout feeds. Fathead minnow growth and reproduction in egg-to-egg life-cycle tests also confirmed APP safety and efficacy as a stabilized source of vitamin C for feeds. None of the typical lesions of scoliosis, malformed gill lamellae, and crenulated opercles, nor conditions of poor health, anorexia, and eventual torpor seen in vitamin C-deficient controls occurred in trout fed APP. No liver pathology was evident in trout fed APP for 252 d at 1.7 to 3.3 times the National Research Council's recommended vitamin C (equivalent) rates.