Enzymatic release of sequestered cows' milk fluoride for analysis by the hexamethyldisiloxane microdiffusion method.

The concentrations of diffusible and total fluoride in cows' milk samples from areas with widely different fluoride levels in drinking water were determined using a fluoride electrode. The diffusible fluoride was determined by direct hexamethyldisiloxane microdiffusion while for total fluoride, samples were subjected to either open ashing or digestion with proteolytic enzymes before microdiffusion. Magnesium nitrate was studied as a new fixative for milk during open ashing and compared with magnesium acetate. Diffusible fluoride ranged from 0.024 to 0.28 microgram ml-1 while total fluoride ranged from 0.05 to 0.31 microgram ml-1. The use of proteolytic enzymes before microdiffusion resulted in total fluoride measurement. It was concluded that all fluoride in milk is inorganic in nature with the bound fluoride being physically or chemically sequestered in the milk proteins. The proposed method is convenient for total fluoride analysis in milk.