Studies on the Method to Quantify PPD and Its Adsorption onto Glass-Surface.
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The potency of PPD is very stable when kept as a lyophilized preparation or as asolution of high concentrations, but, as is well known, when diluted to the practical concentrations such as used in skin-testing for human, its potency diminishes rather rapidly. Ithas been considered that such decrease in potency is not due to denaturation of PPD, butdue to its adsorption onto the surface of glass- or plastic-container. The adsorption of PPD had been studied by several authors with radiolabeled PPD or by the potency test inexperimental animals or human. However, either method is not satisfactory because theyare not quantitative and not reproducible.<BR>Consequently, we developed a new method basing on ELISA to measure such low concentrations of PPD as used in skin-testing in human. Using this new method, we analyzed theadsorption of PPD onto glass-surface. The major characteristics of the PPD adsorptionrevealed by the present studies are as follows; (1) The adsorption of PPD takes place veryrapidly immediately after the distribution of PPD solution to a new container and thenproceeds in slower rates. (2) The rate of adsorption is influenced by both temperature andconcentration of PPD solution. The rates of adsorption are almost the same at -80°C andat 6°C, much higher at 22°C and further higher at 37°C at any concentration tested. At80°C and 6°C, there is no difference among the adsorption rates at three differentconcentrations, but at 22°C and 37°C, the rate of adsorption at the lowest concentration (0.25 μg/ml) is much higher than those at higher concentrations. (3) The amounts of PPDadsorbed are determined by the surface area of glass in contact with PPD solution. (4) Thelarger the volume of PPD solution distributed, the bigger the absolute amount of PPDadsorbed, but the smaller the rate of adsorption. (5) So-called "volume effects" areevidenced.
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