Influence of dietary menadione nicotinamide bisulphite (vitamin K3) and phylloquinone (vitamin K1) on Atlantic salmon (Salmo salar L.) tissue levels, determined by high-performance liquid chromatography with fluorescence detection

Abstract

The aim of the present study was to investigate the retention of menadione nicotinamide bisulphite (MNB; vitamin K3) and phylloquinone (vitamin K1) in Atlantic salmon (Salmo salar L.). Another objective was to find a reliable method for determination of menadione in fish feed, and to include and validate more matrices in the methods for phylloquinone and menaquinones (vitamin K2). Duplicate tanks of Atlantic salmon (∼93 g) were fed four levels (0–1000 mg menadione kg−1 feed) of MNB for 9 weeks. The concentration of menadione and phylloquinone in the feed and the concentration of phylloquinone and menaquinone‐4 (MK‐4) in the tissues were determined. The analysed concentration of dietary menadione found in feed indicated a substantial loss of MNB during feed production. This assumption was supported by screening 15 commercial fish feed samples which also revealed menadione concentrations far below the recommended level. MNB fed salmon showed only a minor increase in liver MK‐4 concentration, compared to salmon fed phylloquinone which had a considerably higher level of liver phylloquinone, indicating a higher retention of phylloquinone compared to menadione in Atlantic salmon. Due to highly varying stability and bioavailability of the different vitamin K derivatives, vitamin K supplementation in fish feed needs a revision.

Vitamin K in fish nutrition

“…It is not itself biologically active and is easily excreted, but can at least be partly alkylated enzymatically to MK‐4 in tissues when present in animal feeds (Dialameh et al. 1971; Udagawa 2000; Graff et al. 2002, 2010; Okano et al.…”

Vitamin K-dependent γ-glutamylcarboxylase in Atlantic salmon (Salmo salar L.)

“…This is in agreement with earlier studies by Sadowski et al (1976) and Buitenhuis et al (1990), which demonstrated that only vitamin K 1 and K 2 can work as a cofactor for GGCX. In farmed Atlantic salmon fed a standard commercial diet, Graff et al (2009) found *70 lg MK-4 kg -1 in heart, 45 lg MK-4 kg -1 in kidney and 30 lg MK-4 kg -1 in both liver and muscle. However, our results showed more than a two-fold higher activity of GGCX in liver compared to muscle.…”

Scientific Opinion on the safety and efficacy of vitamin K3 (menadione sodium bisulphite and menadione nicotinamide bisulphite) as a feed additive for all animal species

“…However, Graff et al (2010) added increasing amounts of MNB to salmon diets (10, 100 and 1 000 mg menadione/kg feed), which was produced according to commercial manufacturing procedures, and found recovery of only 3-5 % two months after production.…”

“…The real requirements may therefore be considerably lower. They require at least a new assessment (Graff et al, 2010).…”

“…No adverse effects on feed intake and laying performance were found. Graff et al (2010) supplemented salmon diets with MNB to provide 0, 10, 100 or 1 000 mg menadione/kg. The recovery in feed was only 3-5 % of the intended values.…”