Collaborative evaluation of a fluorometric method for measuring alkaline phosphatase activity in cow's, sheep's, and goat's milk

J Food Prot. 2005 May;68(5):1047-53. doi: 10.4315/0362-028x-68.5.1047.


Pasteurization of raw milk was introduced to extend product shelf life and destroy pathogens. The measurement of alkaline phosphatase (ALP) activity has been used as an indicator of proper pasteurization in dairy products for more than 65 years. This study was undertaken to evaluate six different fluid dairy products at lower phosphatase levels than previously verified using the Fluorophos Test System, a sensitive and precise method for ALP activity detection. Thirteen laboratories participated in this collaborative, international study to evaluate the fluorometric test at 20, 40, 100, 350, and 500 mU/liter and extend the scope of the method to include milk from not only cows but also goats and sheep. Initially, the statutory level of ALP measured fluorometrically was set to equivalent levels of colorimetric test standards (500 mU/liter). The European Union recently announced its intention of lowering the legal limit from 500 to 350 mU/liter and, in addition, setting a target value of 100 mU/liter, which if exceeded would trigger an investigation into the pasteurizer plant performance. At 500 mU/liter of ALP, this trial generated relative standard deviation of repeatability values of 6.48, 5.69, and 1.74% and relative standard deviation of reproducibility values of 14.66, 13.30, and 5.33% for all cow's, sheep's, and goat's milk samples, respectively. Data from this study are comparable to data from previous studies and indicate the suitability of the Fluorophos Test System method for measuring ALP activity in milk from cows, sheep, and goats not only at the current European statutory level of 500 mU/liter but also at much lower levels.

MeSH terms

  • Alkaline Phosphatase / metabolism*
  • Animals
  • Cattle
  • Consumer Product Safety
  • Fluorometry / methods*
  • Food Contamination / analysis
  • Food-Processing Industry / standards*
  • Goats
  • Milk / enzymology*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Sheep


  • Alkaline Phosphatase