Micro-analytical techniques for the determination of calcium, phosphate, and pH in a small volume (less than 0.25 microliter) of plaque fluid are described and evaluated. The accuracy and the precision of the techniques were compared with those for standard macrotechniques applied to a large pooled plaque fluid sample. The results obtained for the micro-analysis of pooled plaque fluid were in excellent agreement with those obtained by macromethods. The described techniques were also used to analyze plaque fluid obtained from single quadrants of the oral cavities of five individuals. In this fashion, it was determined that, although a significant variation in plaque fluid composition exists between the quadrants, a greater variation exists between subjects. Analyses of plaque fluid obtained from six individuals, following sucrose exposure, were also conducted. The pH value of the fluid changed with time, following a typical Stephan curve, with a minimum value occurring between 15 and 30 minutes; following this, the pH increased to a value near that for resting plaque. An inverse relationship between pH and calcium and phosphorus concentrations was observed. It is noted that the described techniques are sensitive enough to carry out the above analyses both accurately and precisely using plaque obtained from a single quadrant.

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