Hypoxanthine, uric acid and allantoin as indicators of in vivo free radical reactions. Description of a HPLC method and human brain microdialysis data.

A practical one-step high performance liquid chromatography (HPLC) method was developed for the simultaneous determination of hypoxanthine, uric acid and allantoin in small (4 microL) microdialysis samples. The rationale for this work was the current interest in hypoxanthine as a marker for energy perturbation in hypoxia/ ischemia, in uric acid as an endogenous antioxidant, and in allantoin as a marker for in vivo formation of reactive oxygen species (ROS). The method is based on ion pairing reversed phase chromatography and UV detection. The coefficient of variance for within and between run imprecision was generally below 5%, somewhat higher for concentrations close to the detection limit (0.4-1.0 pmoles). Recoveries ranged from 89 to 102% and linearity was observed in the concentration range from 0.25 to 25.0 mmol/L. There was an excellent correlation between the present method and available reference methods. The method was applied to cerebral microdialysis samples from a patient with severe subarachnoid haemorrhage complicated by secondary ischemia leading to cerebral infarction. The secondary ischemia was associated with an increase of hypoxanthine followed by increasing allantoin and uric acid levels. We submit that this pattern of chemical changes indicates increased ROS formation produced by secondary ischemia. The HPLC method appears to be a useful tool for studying changes of hypoxanthine, uric acid and allantoin levels in microdialysis samples.

The validity and accuracy of blood lactate measurements for prediction of maximal endurance running capacity. Dependency of analyzed blood media in combination with different designs of the exercise test.

The effect of using different blood lactate sampling sites in combination with different exercise test designs on the validity and accuracy for prediction of maximal endurance running velocity was investigated. Ten aerobically all-round trained firemen and nine aerobically endurance trained long-distance runners performed six differently designed treadmill running blood lactate accumulation tests. Each test consisted of five consecutive running periods on a treadmill of either 4, 6 or 8 min duration, with a mean increase in running velocity between each period of either 0.25 or 0.5 m.s-1. The corresponding treadmill running velocity to a lactate concentration of 4.0 mmol.l-1 in capillary and venous hemolysed blood and plasma for each running velocity. The mean running velocity from a maximal 12 km run for the firemen and a maximal 21 km run for the runners served as the reference of maximal endurance running velocity. There were both significant (p < 0.001) and similar relationships (r = 0.86-0.94) and no difference in mean prediction error between the predicted and measured maximal endurance running velocities with all tested protocols. However, there was a high risk of making both over- and underestimations (5% to -4%). The lowest risk of making an inaccurate prediction was found when a running duration of 8 min for each running period was used in combination with an increase in running velocity of 0.25 m.s-1, and the lactate measurements were performed in hemolysed capillary blood.

The effect of different blood sampling sites and analyses on the relationship between exercise intensity and 4.0 mmol.l-1 blood lactate concentration.

The aim of the study was to examine whether the difference in lactate concentration in different blood fractions is of practical importance when using blood lactate as a test variable of aerobic endurance capacity. Ten male firefighters performed submaximally graded exercise on a cycle ergometer for 20-25 min. Venous and capillary blood samples were taken every 5 min for determination of haematocrit and lactate concentrations in plasma, venous and capillary blood. At the same time, expired air was collected in Douglas bags for determination of the oxygen consumption. A lactate concentration of 4.0 mmol.l-1 was used as the reference value to compare the oxygen consumption and exercise intensity when different types of blood specimen and sampling sites were used for lactate analysis. At this concentration the exercise intensity was 17% lower (P less than 0.01) when plasma lactate was compared to venous blood lactate, and 12% lower (P less than 0.05) when capillary blood lactate was used. Similar discrepancies were seen in oxygen consumption. The results illustrated the importance of standardizing sampling and handling of blood specimens for lactate determination to enable direct comparisons to be made among results obtained in different studies.

Lactate concentration differences in plasma, whole blood, capillary finger blood and erythrocytes during submaximal graded exercise in humans.

The aim of the study was to investigate the distribution of lactate in plasma, whole blood, erythrocytes, and capillary finger blood, before and during submaximal exercise. Ten healthy male subjects performed submaximal graded cycle ergometer exercise for 20-25 min. Venous blood samples and capillary finger blood samples were taken before exercise and every 5th min during exercise for lactate determination. The plasma lactate concentration was significantly higher (P less than 0.001, approximately 50%) than in the erythrocytes. This difference was not altered by the venous blood lactate concentration or exercise intensity. A significant difference (P less than 0.01) in lactate concentration was also found between capillary whole blood and venous whole blood. It was concluded that direct comparisons between lactate in capillary finger blood, venous whole blood and plasma could not be made.