Simultaneous determination of purine metabolites, creatinine and pseudouridine in ruminant urine by reversed-phase high-performance liquid chromatography.

Determination of purine metabolites, pseudouridine and creatinine in both bovine and ovine urine using high-performance liquid chromatography (HPLC) is described. Following dilution and filtration, urine samples were analysed directly. Separation and quantification was achieved using a Spherisorb ODS II C18 column (250x4.6 mm I.D.) under isocratic conditions. The mobile phase contained 7.5 mM ammonium dihydrogen phosphate, 10 mM sodium 1-heptane sulphonic acid and 1.0 mM triethylamine at pH 3.0. Chromatography was achieved at a flow-rate of 1.0 ml/min and monitoring column effluent at 218 nm. Total analysis time was 60 min. Recovery of all compound standards added to urine was above 96%. In all cases, close spectral matches of compound standards and corresponding identified peaks in ovine and bovine urine were obtained. Lowest detectable concentrations of allantoin, uric acid, xanthine, hypoxanthine, creatinine and pseudouridine were 1.1, 1.0, 1.0, 3.0 and 0.4 micromol/l, respectively. Advantages of simultaneous determination of purine metabolites, creatinine and pseudouridine in ruminant urine collected from both sheep and cattle exist over current methods.

Determination of allantoin in bovine milk by high-performance liquid chromatography.

Determination of allantoin in bovine milk based on high-performance liquid chromatography (HPLC) is described. Following dilution and filtration, milk samples were analysed directly. Separation and quantification of allantoin was achieved using a Spherisorb 5 NH2 column (250 x 4.6 mm ID), acetonitrile-water (90:10, v/v) mobile phase at a flow-rate of 2.0 ml min(-1), temperature 20 degrees C and monitoring the effluent at 214 nm. Total analysis time was 10 min. Recovery of allantoin added to milk was 97 (+/-3.7, n = 30)%. Lowest detectable concentration was 1 micromol l(-1). Within-day and between-day variability were less than 3%. Advantages of improved retention and separation of allantoin, and less complicated sample preparation exist over current methods.

The absorption of magnesium from the forestomachs, stomach and small intestine of sheep.

The passage of magnesium from the mouth to the proximal duodenum and from the proximal duodenum to the terminal ileum was measured continuously in two rams for 24 and 19 days. The results indicate a net disappearance of 25 per cent and 34 per cent of ingested magnesium from the forestomachs and stomach, and no set absorption from the small intestine.

The effect of glucose infusion on the plasma free amino acids in sheep.

1. Glucose in solution in saline, or saline alone, was administered to a group of twenty ewes during late pregnancy and again after lambing. Sequential blood samples were taken before and after the infusion and the concentration of plasma free amino acids was determined. 2. The effect of glucose was to reduce the concentrations of all amino acids except alanine. The reduction was greatest for tryptophan in the pregnant sheep, but this amino acid showed no significant change in the lactating animals. 3. An attempt to rank the amino acids on the basis of their response to glucose infusion indicated that, with the exception of tryptophan for the preparturient ewes, groups of essential amino acids could not be distinguished from each other. These groups were, for the preparturient sheep, valine, leucine, phenylalanine and isoleucine, and for the postparturient animals, isoleucine, lysine, methionine, valine and phenylalanine.