Validation of liquid chromatographic method for assay of calcitriol and alfacalcidol in capsule formulations

The validation of a liquid chromatographic procedure suitable for the determination of calcitriol and alfacalcidol in their respective formulations labeled to contain at least 0.25 micrograms drug per unit is described. The capsule content is diluted and chromatographed in 15-20 min on silica columns (5 micron) with a mobile phase of hexane-tetrahydrofuran-methylene dichloride-isopropanol (72 + 12 + 12 + 4, v/v) with detection at 254 nm. The calibration curve is linear. Recoveries of "spikes" averaged 101% with a standard deviation of 2%. Precision was better than 1.5%.     

Effect of Pasteurella multocida and Haemophilus pleuropneumoniae toxins on swine alveolar macrophages

Supernatants were obtained from 18 hr. broth cultures of Pasteurella multocida strains D82 and D62 (serotype D, toxigenic), Kobe 6 (type D, non-toxigenic), A50 and X73 (type A, non-toxigenic), Haemophilus pleuropneumoniae serotypes 1, 5 and 6, Haemophilus sp. taxon "minor group" (2 strains) and an avirulent serotype 1 strain of H. pleuropneumoniae. The supernates were filtered, pH-neutralized and tested for cytotoxicity after incubation for 18 hours in the presence of swine alveolar macrophage monolayers. Supernatants from H. pleuropneumoniae serotypes 1, 5 and 6 were cytocidal.