EPA Method Detection Limit Procedure

APPENDIX B TO PART 136 – DEFINITION AND PROCEDURE FOR THE DETERMINATION OF THE METHOD DETECTION LIMIT – REVISION 1.11

Definition

The method detection limit (MDL) is defined as the minimum concentration of a substance that can be measured and reported with 99% confidence that the analyte concentration is greater than zero and is determined from analysis of a sample in a given matrix containing the analyte.

Scope and Application

This procedure is designed for applicability to a wide variety of sample types ranging from reagent (blank) water containing analyte to wastewater containing analyte. The MDL for an analytical procedure may vary as a function of sample type. The procedure requires a complete, specific, and well defined analytical method. It is essential that all sample processing steps of the analytical method be included in the determination of the method detection limit.

The MDL obtained by this procedure is used to judge the significance of a single measurement of a future sample.

The MDL procedure was designed for applicability to a broad variety of physical and chemical methods. To accomplish this, the procedure was made device- or instrument-independent.

Procedure

1. Make an estimate of the detection limit using one of the following:
   1. The concentration value that corresponds to an instrument signal/noise in the range of 2.5 to 5.
   2. The concentration equivalent of three times the standard deviation of replicate instrumental measurements of the analyte in reagent water.
   3. That region of the standard curve where there is a significant change in sensitivity, i.e., a break in the slope of the standard curve.
   4. Instrumental limitations.

   It is recognized that the experience of the analyst is important to this process. However, the
   analyst must include the above considerations in the initial estimate of the detection limit.

2. Prepare reagent (blank) water that is as free of analyte as possible. Reagent or interference free water is defined as a water sample in which analyte and interferant concentrations are not detected at the method detection limit of each analyte of interest. Interferences are defined as systematic errors in the measured analytical signal of an established procedure caused by the presence of interfering species (interferant). The interferant concentration is presupposed to be normally distributed in representative samples of a give matrix.
3. 1. If the MDL is to be determined in reagent (blank) water, prepare a laboratory standard (analyte in reagent water) at a concentration which is at least equal to or in the same concentration range as the estimated detection limit. (Recommend between 1 and 5 times the estimated detection limit.) Proceed to Step 4.
   2. If the MDL is to be determined in another sample matrix, analyze the sample. If the measured level of the analyte is in the recommended range of one to five times the estimated detection limit, proceed to Step 4.
      If the measured level of analyte is less than the estimated detection limit, add a known amount of analyte to bring the level of analyte between one and five times the estimated detection limit.
      If the measured level of analyte is greater than five times the estimated detection limit, there are two options.
      1. Obtain another sample with a lower level of analyte in the same matrix if possible.
      2. This sample may be used as is for determining the method detection limit if the analyte level does not exceed 10 times the MDL of the analyte in reagent water. The variance of the analytical method changes as the analyte concentration increases from the MDL, hence the MDL determined under these circumstances may not truly reflect method variance at lower analyte concentrations.
4. 1. Take a minimum of seven aliquots of the sample to be used to calculate the method detection limit and process each through the entire analytical method. Make all computations according to the defined method with final results in the method reporting units. If a blank measurement is required to calculate the measured level of analyte, obtain a separate blank measurement for each sample aliquot analyzed. The average blank measurement is subtracted from the respective sample measurements.
   2. It may be economically and technically desirable to evaluate the estimated method detection limit before proceeding with 4a. This will: (1) Prevent repeating this entire procedure when the costs of analyses are high and (2) insure that the procedure is being conducted at the correct concentration. It is quite possible that an inflated MDL will be calculated from data obtained at many times the real MDL even though the level of analyte is less than five times the calculated method detection limit. To insure that the estimate of the method detection limit is a good estimate, it is necessary to determine that a lower concentration of analyte will not result in a significantly lower method detection limit. Take two aliquots of the sample to be used to calculate the method detection limit and process each through the entire method, including
      blank measurements as described above in 4a. Evaluate these data:
      1. If these measurements indicate the sample is in desirable range for determination of the MDL, take five additional aliquots and proceed. Use all seven measurements for calculation of the MDL.
      2. If these measurements indicate the sample is not in correct range, reestimate the MDL, obtain new sample as in 3 and repeat either 4a or 4b.
5. Calculate the standard deviation (s) of the replicate measurements.
6. Compute the MDL,
   MDL = t_{(n−1, 99%)}(s)

   where: t_{(n-1, 99%)} = Students’ t value for a 99% confidence level for a one-tailed distribution and n−1 degrees of freedom (See Note 2 below.) s = standard deviation of the replicate analyses.

Reporting

The analytical method used must be specifically identified by number of title and the MDL for each analyte expressed in the appropriate method reporting units. If the analytical method permits options which affect the method detection limit, these conditions must be specified with the MDL value. The sample matrix used to determine the MDL must also be identified with MDL value. Report the mean analyte level with the MDL. If a laboratory standard or a sample that contained a known amount analyte was used for this determination, also report the mean recovery.

If the level of analyte in the sample was below the determined MDL or exceeds 10 times the MDL of the analyte in reagent water, do not report a value for the MDL.

Notes:

1. This document was adapted by Dr. Zelewski from the Electronic Code of Federal Regulations Title 40: Protection of Environment, Part 136: Guidelines for Establishing Test Procedures for the Analysis of Pollutants, Appendix B: Definition and Procedure for the Determination of the Method Detection Limit – Revision 1.11 (link)

2. The t-table in the textbook, Exploring Chemical Analysis, by Harris is for a two-tailed distribution. That is, a 99% confidence interval means that 0.5 % of the results are outside the 99% interval on the low side (or tail) and 0.5% results are outside the 99% interval on the high side (or tail). Similarly, a 98% confidence interval means that 1% of the results are outside the 98% interval on the low side (or tail) and 1% results are outside the 98% interval on the high side (or tail).

For MDL, one only cares about the tail on the high side. That is, the MDL specifies that there is at most a 1% chance of incorrectly concluding that a blank has analyte above the detection limit (a false positive). One does not care about the tail on the low side as it is well below the detection limit. The t-values for a 99% one-tailed distribution are equal to the t-values for a 98% two-tailed distribution. So, if a t table for a one-tailed distribution is not available, one can use the t values for the 98% confidence level of a two-tailed distribution.