The influence of haemolysis on the determination of vitamin E in cattle

Abstract

The study deals with the influence of haemolysis on the results of vitamin E determination in plasma (serum) in cattle. Although nowadays specific High Performance Liquid Chromatography (HPLC) techniques are used almost exclusively for the determination of vitamin E, this indicator is also influenced by haemolysis. This occurs in the pre-analytical phase due to the fact that iron contained in haemoglobin is able to catalyze peroxidation reactions. Subsequently, changes occur mainly in polyunsaturated fatty acids in the lipoprotein components of serum/plasma. The vitamin E present inhibits this process, as a result of which its concentration is reduced. The experiment was performed by preparing model samples with a defined degree of haemolysis by adding haemolysate to the centrifuged plasma in the range of ca 0–12 g/l, i.e. mild to severe haemolysis. After 4 h of incubation at room temperature, the vitamin E concentration was determined by the HPLC method. Haemolysis was found to reduce the test result; mild one (approximately up to 2 g/l) non-significantly, medium and severe haemolysis by up to tens of percent, which warrants repeated sampling. False reductions in results will not endanger the patient’s health, but economic losses may occur due to unnecessary check-ups and increased vitamin E supplementation.