Validation of Alternative Methods for the Analysis of Drinking Water and Their Application to Escherichia coli

Abstract

ABSTRACT In Europe, the Drinking Water Directive of the European Commission indicates which methods (most of which are CEN/ISO-standardized methods) should be used for the analysis of microbiological parameters (European Commission, Environment, Council Directive 98/83/EC of 3 November 1998). According to the Directive, alternative methods “may be used, providing it can be demonstrated that the results obtained are at least as reliable as those produced by the methods specified.” The prerequisite for the routine use of any alternative method is to provide evidence that this method performs equivalently to the corresponding reference method. In this respect, the ISO 16140 standard (ISO, ISO 16140. Microbiology of Food and Animal Feeding Stuffs—Protocol for the Validation of Alternative Methods , 2003) represents a key issue in generating such a procedure based on an interlaboratory study. A new statistical tool, called the accuracy profile, has been developed to better interpret the data. The study presented here is based upon the enumeration of Escherichia coli bacteria in water. The reference method may require up to 72 h to provide a confirmed result. The aim of this publication is to present data for an alternative method by which results can be obtained in 18 h (Colilert-18/Quanti-Tray) based upon defined substrate technology (DST). The accuracy profile is a statistical and graphical decision-making tool and consists of simultaneously combining, in a single graphic, β expectation tolerance intervals (β-ETIs) and acceptability limits (λ). The study presents the validation criteria calculated at the three levels of contamination used in the trial for a β equal to 80% and a λ equal to ±0.3 and combines the accuracy profiles of Escherichia coli for a λ of ±0.3 log 10 unit/100 ml, a λ of ±0.4 log 10 unit/100 ml, and a β of 80% or 90%. Several interesting conclusions can be drawn from these data. The accuracy profile method has been applied to the validation of the Colilert-18/Quanti-Tray method against reference method ISO 9308-1 (ISO, ISO 9308 - 1. Water Quality—Detection and Enumeration of Escherichia coli and Coliform Bacteria. Part 1. Membrane Filtration Method , 2000), using a β of 80% and a λ of 0.4; the alternative method can be validated between 1.00 and 2.05 log 10 units/100 ml, equivalent to 10 to 112 CFU/100 ml.