                              EPA Office of Water
                          New Method Application Form
                             For Chemical Analytes
                                                    Applicant Name and Address:
Hach Company
5600 Lindbergh Drive
Loveland, Colorado 80539
Attn: Cary B. Jackson
970 669-3050
cjackson@hach.com
                                 EPA Use Only
Case No.
                                                    Date Application Submitted:
February 2012
                                                       Method Number and Title:
Hach Method 10206  -  Hach Method 10206 TNTplus 835/836 Spectrophotometric Measurement of Nitrate in Water and Wastewater	
                                                                   Analytes(s):
Nitrate, Nitrite/Nitrate
                                                          Type (WW, DW, WW/DW):
WW Tier 3
                                                                  Level of Use:
                                                                     (LU or NW)
NW
Attachments:
Validation Study Report/Justification for New Method
Method in standard EPA format
Digital Attachments

                                       


Revised Report of Hach Method 10206 Spectrophotometric Measurement of Nitrate in Water and Wastewater
                               October 12, 2012 
                   
                   
                                       


Acknowledgments

	This report was prepared under the direction of Dr. Cary B. Jackson, Director of Regulatory Affairs for Hach Company.  Contributors to this study include Dave MacDonald, Scott Tucker, and Tom Haukebo of Hach Company; Lemuel Walker, USEPA Engineering and Analysis Division; and Dr. Richard Burrows et al. of Test America laboratories.

Questions or comments concerning this report should be addressed to:

Dr. C. B. Jackson
Director of Regulatory Affairs
Hach Company
5600 Lindbergh Drive
Loveland, Colorado 80539
970 669-3050
cjackson@hach.com

Preface

This document is a revision (October 12, 2012) to the Report of the Hach Method 10206 Spectrophotometric Measurement of Nitrate in Water and Wastewater, which was submitted on December 16, 2011 to Mr. Lemuel Walker, Clean Water Act ATP Coordinator, United Sates Environmental Protection Agency, Engineering and Analysis Division.  During the EPA review of the ATP application and Report, it was determined that several statistical calculations were detected.  This revision report contains the corrections to the statistical calculations, with all other document- content remaining identical to the original report submission.

Document Changes From Original Report

Section 2.7.3, paragraph #1, sentence #3, - Change from: "5.3". Change to: "5.4".

Section 2.7.3, paragraph #1, sentence #4, - Change from: "11.7". Change to: "11.4".

Table 5, Standard Deviation of Means % Recovery across Labs  -  Change from: "5.3" Change to: "5.4".

Table 5, Standard Deviation of Means % Recovery across Labs  -  Change from: "11.7" Change to: "11.3".



Table of Contents																							Page
Report												1

Section 1	Introduction									2
Section 2	Multi-laboratory Validation Study						3
	
Section 2.1	Study Objectives								3
Section 2.2	Sample Source								3
Section 2.3	Sample Collection and Handling Activities					4
Section 2.4	Analytical Study Design							4

Section 2.4.1	MDL and ML Test Description						4
Section 2.4.2	IPR Test Description								5
Section 2.4.3 	Matrix Spike and Matrix Spike Duplicate Test Description			5

Section 2.5	Data Validation Description							5
Section 2.6	Analytical Results								5
Section 2.7	Discussion of Results								6

Section 2.7.1	MDL and ML Test								6
Section 2.7.2	IPR Test									6
Section 2.7.3	MS and MSD Test Results							6

Section 2.8	Validation Conclusions							7

Tables												8

Table 1 and 2	Method Detection Limit and Method Limit					9
Table 3 and 4	Initial Precision and Recovery						10
Table 5 and 6	Matrix Spike and Matrix Spike Duplicate Recovery				11
Table 7	Pooled Multi-laboratory Quality Control Performance Results		12

Appendices											13

Appendix A	Wastewater Source								14
Appendix B	Testing laboratories								15
Appendix C	Screening Results								16
Appendix D	Hach Method 10206  -  EPA Format						17
Appendix E	Hach Method 10206- Hach Procedure					18
Appendix F	Laboratory Standard Operating Procedure					19

 
Appendix G	New Method Data Form and Laboratory Bench Sheets			20Report

Section 1
Introduction 
	 
	Hach Method 10206, TNTplus 835/836(TM) Nitrate using dimethylphenol chemistry is a green chemistry alternative to that of EPA Reference EPA Method 353.2, 300.1 and the approved alternative methods of SM4500-NO3[-] E.  The TNTplus Nitrate chemistry follows classical electrophillic aromatic substitution in that nitrate in the presence of sulfuric acid yields a nitronium ion ([+]NO2) and HSO4[-].  Nitronium ions are electrophiles that attack the aromatic ring of the dimethylphenol reagent to form intermediate nitro-carbonium ions.  The basic HSO4[-] ion then extracts a hydrogen ion from the nitro-carbonium intermediate to yield a stable substitution product (o, or p-nitro-dimethylphenol). The nitrodimethylphenol product is a highly colored (directly related to the nitro functional group), quantifiable by its visible absorption spectra.
























	The quality control (QC) requirements incorporated into Hach Method 10206 exceed and improve upon that of the currently approved 40 CFR part 136 nitrate methods.  The QC acceptance criteria in this method were derived from the results of an interlaboratory study conducted by 9 laboratories and 9 different matrices.  This report gives the details of the validation study and associated results.  

Section 2
Multi-laboratory Validation Study 

	This section of the report provides the details of the multi-laboratory validation study conducted by Hach Company for the determination of comparability of measurement of nitrate-N and nitrite/nitrate-N with Hach Method 10206 to EPA Reference Methods 353.2, 300.0, and SM4500-NO3 E.

2.1	Study Objectives

	The purposes of the multi-laboratory study were to:

   *          Validate the Hach Method 10206 according to the Protocol for EPA Approval of New Methods for Organic and Inorganic Analytes in Wastewater and Drinking Water (USEPA, 1999).
   *          Submission of validation study report to USEPA for ultimate approval and use in 40 CFR part 136 NPDES program monitoring of nitrate and nitrite/nitrate nitrogen.

2.2	Sample Source 

	The multi-laboratory validation study required collection of a variety of industrial, municipal, and treated drinking water samples in order to assess the effect of sample type on the dimethylphenol chemistry of the Hach Nitrate Method.  When selecting these sites, various factors were considered and included the following criteria:

   * The expected range of nitrate-N concentrations, with a range of 0.20  -  35 mg/L

   * Industrial categories with existing nitrate effluent limitations

   * Waste streams that have measurable quantities of nitrate and nitrite 

   * Accessibility of waste streams

	Representatives of candidate facilities were initially contacted to request cooperation, verify plant and waste stream characteristics, and to coordinate sampling schedules.  Facilities volunteering to participate were selected from EPA Region VIII for industrial samples, Regions IV, V, and X for drinking water samples, and Regions II, VIII, and IX for municipal samples.  A summary of the wastewater sources is presented in Appendix A, which shows the facility types, industrial categories, and waste streams included in the study.


2.3	Sample Collection and Handling Activities 

	At each site, sample material was collected in clean polyethylene containers. Industrial and municipal waste samples were collected, preserved, and shipped to the study sponsor facility (Hach Company) according to the specifications in 40 CFR part 136, Table II.  Aliquots were preserved on-site with H2SO4 to a pH of less than 2, shipped overnight to the sponsor facility at a temperature not greater than 6[o] C., but not less than 1[o] C. Drinking Water sample matrices were collected at the testing laboratory tap, then immediately preserved.  The testing laboratories for this validation study are listed in Appendix B.

	Upon receipt of collected industrial and municipal wastes samples to the sponsor facility, each matrix was screened by the Hach 10206 procedure to determine the background concentration of nitrate.  Where concentrations of screened analytes exceeded the range of the test procedure, a dilution was prepared and re-preserved to a pH of less than 2, then shipped overnight to the validation study laboratory at a temperature not greater than 6[o] C., but not less than 1[o] C.  Screening results and dilution preparation information are presented in Appendix C and D.

2.4	Analytical Study Design

	The multi-laboratory validation study consisted of three parts tests, the method detection limit (MDL) and minimum limit (ML) tests, an initial precision and recovery  (IPR) test, and a matrix and matrix spike duplicate test.  The participating validation laboratories are to perform these tests for the proposed Hach Method 10206 and either of the EPA Reference Methods 353.2, 300.0 or SN 4500--NO3[-] E.  The Validation Study Protocol is described in Appendix E

2.4.1	MDL and ML Test Discription

	The MDL is defined as the minimum concentration of a substance that can be measured and reported with 99 percent confidence that the analyte concentration is greater than zero.  To determine the MDL, the laboratories were required to follow the procedure in Appendix B to 40 CFR part 136. This procedure consists of the analysis of seven aliquots of reagent water that are spiked with nitrate-N. The MDL is calculated by multiplying the standard deviation of the seven replicate analyses by the student's t value for (n-1) degrees of freedom, where n equals the number of replicates.  

	The ML is defined as the level at which the entire analytical system produces a recognizable signal and an acceptable calibration point, and is determined by multiplying the MDL by 3.18 and rounding the result to the nearest factor of 10 multiple of 1, 2, or 5.  The value 3.18 represents the ration between the student t multiplier used to determine the MDL and the 10 times multiplier used in the American Chemical Society (ACS) limit of quantity (LOQ) calculations (i.e. 10 / 3.143 = 3.18).  For example, if the calculated MDL is 5.8, the resulting ML will be equal to 5.8 times 3.18, or 18.4.  Rounding to the nearest factor of 10 multiple of 1, 2, or 5 then establishes the ML at 20.
2.4.2	IPR Test Description

	The IPR test is used for initial demonstration of a laboratory's capability to produce results that are at least as precise and accurate as results from practice of the method by other laboratories. The IPR test also is used to demonstrate that a new method or method modification will produce results that are as precise and accurate as the reference method.  The IPR test consists of analyzing at least four replicate aliquots of a reference matrix spiked with the analytes of interest and with either surrogate compounds or, for isotope dilution analysis, labeled compounds. The concentration of the target analytes in the spike solution may vary between one and five times the concentration used to establish the lowest calibration point (e.g., one to five times the ML). The spiked aliquots are carried through the entire analytical process.  Specifications are developed for the permissible range of recovery for each analyte and for an upper limit on the standard deviation or RSD of recovery.

2.4.3 	Matrix, Matrix Spike and Matrix Spike Duplicate Test

	The matrix and matrix spike/matrix spike duplicate (MS/MSD) test is used to assess method performance in the sample matrix. Analytes of interest are added to a field sample aliquot that is then thoroughly homogenized and split into two spiked replicate aliquots.  One of these replicates is identified as the matrix spike sample and the other is identified as the matrix spike duplicate sample. The recoveries of the analytes, relative to the spike, are determined in each sample.  The precision of the determinations also is assessed by measuring the relative percent difference (RPD) between the analyte concentrations measured in the MS and MSD. 

2.5	Data Validation

	Data generated from this study were reviewed and validated according to the previously referenced USEPA validation protocol (Protocol for EPA Approval of , 1999 Methods for Organic and Inorganic Analytes in Wastewater and Drinking Water).  Final results, including all calculations, were verified from the raw data and bench sheets provided.  All quality control requirements in the study plan and draft version of Hach Method 10206 and USEPA Reference Methods 353.2, 300.0 or SM 4500-NO3 E were met, and all data reviewed were determined to be of acceptable quality.

2.6 	Results

	Nine laboratories submitted results for nitrate determination for each of the three performance tests (MDL/ML, IPR, Matrix and MS/MSD), by the proposed Hach Method 10206 and USEPA Reference Method 353.2, 300.0, or SM 4500-NO E.  For each reference matrix (reagent water) and field sample, the analytical results were subjected to statistical evaluation as described in section 2.4 above.  The statistical performance calculations of analytical results from each laboratory are summarized in Tables 1 through 6.    The pooled multi-laboratory validation results are summarized in Table 7.

2.7	Discussions 
	
2.7.1	 MDL and ML Test 

	All nine laboratories were successful with the analysis of the MDL samples. The overall pooled MDL with Hach Method 10206 was determined to be 0.05 mg/L and 0.02 mg/L for the USEPA Reference Methods.  Likewise, the pooled and rounded ML for each method was determined to be 0.20 and 0.10, respectively mg/L.  

	This similarity in MDL and ML results indicates Hach Method 10206 is equally effective and comparable to USEAP Method 353.2, 300.0, and SM4500-NO3 E with respect to method detection in the standard reference matrix of reagent water.

2.7.2	IPR Test

	All nine laboratories were successful with the analysis of IPR samples.  There were no outliers and all data points were used to determine precision and accuracy in the standard reference matrix of reagent water.  The mean percent recovery and standard deviation of mean percent recoveries across laboratories with Hach Method 10206 was determined to be 98.6 and 1.3, respectively.  The mean percent recovery and standard deviation of mean percent recoveries across laboratories with USEPA Reference Methods was determined to be 101 and 4.7, respectively.  

	The Hach Method 10206 results of mean percent recovery, standard deviation of mean percent recoveries, further indicates equal effectiveness and comparability to USEPA Reference Methods in the standard reference matrix of reagent water.

2.7.3	Matrix Spike Matrix Spike Duplicate Test

	All nine laboratories were successful with the analysis of field matrix spike and matrix spike duplicate samples.  There were no outliers and all data points were used to determine precision and accuracy in the standard reference matrix of reagent water.  The mean percent recovery and standard deviation of mean percent recoveries across laboratories with Hach Method 10206 was determined to be 95.6 and 5.4, respectively.  The mean percent recovery and standard deviation of mean percent recoveries across laboratories with USEPA Reference Methods was determined to be 102 and 11.3, respectively.  

	The Hach Method 10206 results of mean percent recovery, standard deviation of mean percent recoveries across labs further indicates equal effectiveness and comparability to USEPA Reference Methods in various matrices of drinking water, treated wastewater from POTWs, and industrial treated wastewater effluents.


2.8 	Validation Conclusions

	A nation-wide multi-laboratory validation study was successfully completed using nine highly-centralized commercial testing laboratories, each laboratory analyzing a different matrix with Hach Method 10206 and EPA Reference Method 353.2, 300.0, or SM 4500-NO3 E, for the measurement of total nitrate (nitrite-nitrate nitrogen).   Based on the comparative side-by-side results of all laboratories and all matrices, Hach Method 10206, for the measurement of nitrate nitrogen equaled or provided better performance to that of the EPA Reference Methods. 

Tables


                                    Table 1
                      Multi-laboratory Validation Results
                       Method Detection and Method Limit
                               Hach Method 10206
                               Nitrate-N (mg/L)

                                       
                                    Lab #1
                                    Lab #2
                                    Lab #3
                                    Lab #4
                                    Lab #5
                                    Lab #6
                                    Lab #7
                                    Lab #8
                                    Lab #9
                                    Rep #1
                                     0.325
                                     0.322
                                     0.225
                                     0.299
                                     0.261
                                     0.218
                                     0.244
                                     0.276
                                     0.239
                                    Rep #2
                                     0.320
                                     0.341
                                     0.225
                                     0.345
                                     0.233
                                     0.225
                                     0.254
                                     0.239
                                     0.272
                                    Rep #3
                                     0.271
                                     0.269
                                     0.246
                                     0.264
                                     0.230
                                     0.237
                                     0.262
                                     0.270
                                     0.247
                                    Rep #4
                                     0.280
                                     0.266
                                     0.259
                                     0.260
                                     0.236
                                     0.241
                                     0.246
                                     0.331
                                     0.240
                                    Rep #5
                                     0.285
                                     0.284
                                     0.244
                                     0.260
                                     0.227
                                     0.230
                                     0.248
                                     0.254
                                     0.232
                                    Rep #6
                                     0.253
                                     0.324
                                     0.244
                                     0.284
                                     0.238
                                     0.239
                                     0.251
                                     0.231
                                     0.231
                                    Rep #7
                                     0.260
                                     0.313
                                     0.239
                                     0.288
                                     0.245
                                     0.226
                                     0.248
                                     0.252
                                     0.239
                              Standard Deviation
                                     0.028
                                     0.030
                                     0.012
                                     0.030
                                     0.011
                                     0.008
                                     0.006
                                     0.033
                                     0.014
                                      MDL
                                     0.088
                                     0.093
                                     0.038
                                     0.095
                                     0.036
                                     0.027
                                     0.019
                                     0.102
                                     0.045
                                      ML
                                     0.279
                                     0.295
                                     0.121
                                     0.302
                                     0.115
                                     0.085
                                     0.060
                                     0.325
                                     0.142
                                  ML (Rounded
                                     0.28
                                     0.30
                                     0.12
                                     0.30
                                     0.12
                                     0.09
                                     0.070
                                     0.33
                                     0.14

                                    Table 2
                      Multi-laboratory Validation Results
                       Method Detection and Method Limit
                    USEPA Method 353.2/300.0/SM 4500-NO3 E
                               Nitrate-N (mg/L)
                                       
                                       
                                    Lab #1
                                    Lab #2
                                    Lab #3
                                    Lab #4
                                    Lab #5
                                    Lab #6
                                    Lab #7
                                    Lab #8
                                    Lab #9
                                    Rep #1
                                     0.279
                                     0.069
                                     0.577
                                     0.097
                                     0.108
                                     0.078
                                     0.019
                                     0.031
                                     0.020
                                    Rep #2
                                     0.285
                                     0.072
                                     0.585
                                     0.036
                                     0.110
                                     0.072
                                     0.019
                                     0.039
                                     0.019
                                    Rep #3
                                     0.274
                                     0.074
                                     0.588
                                     0.048
                                     0.109
                                     0.071
                                     0.019
                                     0.039
                                     0.020
                                    Rep #4
                                     0.279
                                     0.074
                                     0.576
                                     0.050
                                     0.109
                                     0.075
                                     0.018
                                     0.037
                                     0.018
                                    Rep #5
                                     0.278
                                     0.072
                                     0.576
                                     0.050
                                     0.107
                                     0.078
                                     0.018
                                     0.035
                                     0.020
                                    Rep #6
                                     0.287
                                     0.073
                                     0.586
                                     0.051
                                     0.107
                                     0.076
                                     0.019
                                     0.041
                                     0.023
                                    Rep #7
                                     0.257
                                     0.071
                                     0.586
                                     0.049
                                     0.106
                                     0.082
                                     0.018
                                     0.035
                                     0.023
                              Standard Deviation
                                     0.010
                                     0.002
                                     0.005
                                     0.019
                                     0.001
                                     0.004
                                     0.001
                                     0.003
                                     0.002
                                      MDL
                                     0.031
                                     0.005
                                     0.017
                                     0.061
                                     0.003
                                     0.012
                                     0.002
                                     0.011
                                     0.006
                                      ML
                                     0.098
                                     0.017
                                     0.054
                                     0.195
                                     0.011
                                     0.038
                                     0.007
                                     0.034
                                     0.019
                                  ML (Rounded
                                     0.10
                                     0.02
                                     0.05
                                     0.20
                                     0.01
                                     0.40
                                     0.01
                                     0.03
                                     0.02
                                       

                                    Table 3
                      Multi-laboratory Validation Results
                        Initial Precision and Recovery
                               Hach Method 10206
                          Nitrate-N Percent Recovery
                                       
                                       
                                    Rep #1
                                    Rep #2
                                    Rep #3
                                    Rep #4
                           Average Percent Recovery
                                      Std
                                   Deviation
                                    Lab #1
                                     99.6
                                     97.8
                                     98.2
                                     98.4
                                     98.5
                                      0.8
                                    Lab #2
                                     99.6
                                     98.6
                                     98.2
                                     99.8
                                     99.1
                                      0.8
                                    Lab #3
                                     98.8
                                     98.6
                                     98.6
                                     98.2
                                     98.6
                                      0.3
                                    Lab #4
                                      100
                                     99.4
                                     99.2
                                     99.0
                                     99.5
                                      0.6
                                    Lab #5
                                     99.0
                                     98.6
                                     98.4
                                     99.6
                                     98.9
                                      0.5
                                    Lab #6
                                     96.0
                                     96.4
                                     97.0
                                     94.4
                                     96.0
                                      1.1
                                    Lab #7
                                      100
                                     99.6
                                     99.0
                                     99.2
                                     99.5
                                      0.4
                                    Lab #8
                                      100
                                      100
                                     99.6
                                      100
                                      100
                                      0.3
                                    Lab #9
                                     97.2
                                     97.0
                                     97.4
                                     96.6
                                     97.1
                                      0.3
                                       
Mean % Recovery across Labs				       98.6
				  		
Std. Deviation of Mean % Recoveries across Labs	        			1.3	

                                    Table 4
                      Multi-laboratory Validation Results
                        Initial Precision and Recovery
                    USEPA Method 353.3/300.0/SM 4500-NO3 E
                          Nitrate-N Percent Recovery
                                       
                                       
                                    Rep #1
                                    Rep #2
                                    Rep #3
                                    Rep #4
                           Average Percent Recovery
                                      Std
                                   Deviation
                                    Lab #1
                                      100
                                     99.4
                                     99.2
                                      101
                                      100
                                      0.8
                                    Lab #2
                                     91.2
                                      102
                                      102
                                      102
                                     99.0
                                      5.2
                                    Lab #3
                                      104
                                      104
                                      104
                                      104
                                      104
                                      0.0
                                    Lab #4
                                      108
                                      107
                                      107
                                      108
                                      108
                                      0.7
                                    Lab #5
                                      107
                                      106
                                      105
                                      103
                                      105
                                      1.5
                                    Lab #6
                                     94.9
                                     95.9
                                     95.2
                                     96.5
                                     95.6
                                      0.7
                                    Lab #7
                                      101
                                      103
                                      103
                                      103
                                      103
                                      1.1
                                    Lab #8
                                     92.5
                                     92.7
                                     92.5
                                     93.1
                                     92.7
                                      0.3
                                    Lab #9
                                     98.6
                                      102
                                     99.6
                                     99.7
                                      100
                                      1.5

Mean % Recovery across Labs				       101
Std. Deviation of Mean % Recoveries across Labs	       			4.7


                                    Table 5
                      Multi-laboratory Validation Results
               Matrix Spike and Matrix Spike Duplicate Recovery
                               Hach Method 10206
                          Nitrate-N Percent Recovery

                                       
                                 Matrix Spike
                            Matrix Spike Duplicate
                             Mean Percent Recovery
                                      Std
                                  Deviation 
                                    Lab #1
                                     92.2
                                     91.6
                                     91.9
                                      0.4
                                    Lab #2
                                     88.8
                                     93.4
                                     91.1
                                      3.3
                                    Lab #3
                                     96.3
                                     93.9
                                     95.1
                                      1.7
                                    Lab #4
                                     96.2
                                     95.8
                                     96.0
                                      0.3
                                    Lab #5
                                     99.0
                                     99.0
                                     99.0
                                      0.0
                                    Lab #6
                                     99.2
                                     99.0
                                     99.1
                                      0.1
                                    Lab #7
                                     84.4
                                     82.4
                                     83.4
                                      1.4
                                    Lab #8
                                      102
                                      102
                                      102
                                      0.0
                                    Lab #9
                                     97.0
                                     99.2
                                     98.1
                                      1.6

Mean % Recovery across Labs		               95.1 
Std. Dev. of Mean % Recoveries across Labs              		5.6	
		  		
	
                                    Table 6
                      Multi-laboratory Validation Results
               Matrix Spike and Matrix Spike Duplicate Recovery
                    USEPA Method 353.2/300.0/SM 4500-NO3 E
                          Nitrate-N Percent Recovery
                                       
                                       
                                 Matrix Spike
                            Matrix Spike Duplicate
                           Average Percent Recovery
                                      Std
                                   Deviation
                                    Lab #1
                                     96.8
                                     96.8
                                     96.8
                                      0.0
                                    Lab #2
                                     93.0
                                      112
                                      102
                                     13.3
                                    Lab #3
                                      102
                                      102
                                      102
                                      0.0
                                    Lab #4
                                      109
                                      109
                                      109
                                      0.0
                                    Lab #5
                                     95.4
                                     89.6
                                     92.5
                                      4.1
                                    Lab #6
                                     81.9
                                     83.7
                                     82.8
                                      1.3
                                    Lab #7
                                      100
                                      108
                                      104
                                      5.4
                                    Lab #8
                                      123
                                      126
                                      124
                                      2.3
                                    Lab #9
                                     97.2
                                      106
                                      102
                                      6.5

Mean % Recovery across Labs		               102 
Std. Dev. of Mean % Recoveries across Labs               		11.3 	
		

                                    Table 7
          Pooled Multi-laboratory Quality Control Performance Results
                                       
                           Quality Control Parameter
                               Hach Method 10206
                      EPA Method 353.2/300.0/SM4500-NO3 E
Pooled MDL (mg/L)
                                     0.05
                                     0.02
Pooled ML (mg/L)
                                     0.17
                                     0.06
Rounded ML (mg/L)
                                     0.20
                                     0.10
IPR RSDMax
                                     0.011
                                     0.033
IPR Limit of Recovery Range (%)
                                 95.4  -  102
                                 89.4  -  112
Matrix Spike RPDMax
                                     0.048
                                     0.173
Matrix Spike Limit of Recovery Range (%)
                                 90.5  -  101
                                  89.6 - 114

 

Appendices
	

Appendix A
Wastewater Sources

Matrix Category - Public Owned Treatment Works
Colorado, California, New York

Matrix Category  -  Industrial
Food Processing, Dairy processing, Beef Slaughter and Processing

Matrix Category  -  Treated Drinking Water
Florida, Indiana, Oregon,


Appendix B
Testing Laboratories/Matrices

Hach Company  -  Screening Laboratory

Lab #1:  Hach Company  -  Validation Laboratory/DSRWWTP
Lab #2:  Test America Cedar Fall  -  Validation Laboratory/DW
Lab #3:  Test America Phoenix  -  Validation Laboratory/DW
Lab #4:  Test America Watertown  -  Validation Laboratory/DW
Lab #5:  Test America Portland  -  Validation Laboratory/FTCWWTP
Lab #6:  Test America Valparaiso  -  Validation Laboratory/LPWWTP
Lab #7:  Test America Tallahassee  -  Validation Laboratory/Industrial KFE
Lab #8:  Test America Houston  -  Validation Laboratory/Industrial DFAE
Lab #9:  Test America Dayton  -  Validation Laboratory/CWWTP

Appendix C
Industrial and Municipal Nitrate-N Screening Results




Appendix D
Hach Method 10206  -  EPA Format

Appendix E
Hach Method 10206 Procedure

Appendix F
Laboratory Standard Operating Procedure

Appendix G
New Method Data Form/Laboratory Bench Sheets

                             New Method Data Form

Method Title:	Hach Method 10206 Spectrophotometric Measurement of Nitrate in Water and Wastewater	

Validation Laboratory:	Hach Company
Matrix:  DSRWWTP

                       Method Detection Limit (MDL) Data
                                       
Spiking Concentration:  0.30 mg/L 
MDL Data:  0.325, 0.320, 0.271, 0.280, 0.285, 0.253, 0.260

                      Initial Precision and Recovery Data

Spiking Concentration: 5.0 mg/L
IPR Data:  4.98, 4.89, 4.91, 4.92

               Matrix Spike/Matrix Spike Duplicate (MS/MSD) Data

Spiking Concentration:  5.0 mg/L
MS Concentration: 2.49
MSD Concentration:  7.10
Background Concentration:  7.07

                        New Method Acceptance Criteria

IPR Recovery:  95.4  -  102
Precision:  0.011

MS/MSD Recovery:  90.5  -  101
RPD:  0.048

                             New Method Data Form

Method Title:	Hach Method 10206 Spectrophotometric Measurement of Nitrate in Water and Wastewater	

Validation Laboratory:  Test America Cedar Falls
Matrix:  Drinking Water

                       Method Detection Limit (MDL) Data
                                       
Spiking Concentration:  0.30 mg/L 
MDL Data:  0.322, 0.341, 0.269, 0.266, 0.284, 0.324, 0.313

                      Initial Precision and Recovery Data

Spiking Concentration: 5.0 mg/L
IPR Data:  4.98, 4.93, 4.91, 4.99

               Matrix Spike/Matrix Spike Duplicate (MS/MSD) Data

Spiking Concentration:  5.0 mg/L
MS Concentration:  4.90
MSD Concentration:  5.13
Background Concentration: 0.46

                        New Method Acceptance Criteria

IPR Recovery:  95.4  -  102
Precision:  0.011

MS/MSD Recovery:  90.5  -  101
RPD:  0.048

                             New Method Data Form

Method Title:	Hach Method 10206 Spectrophotometric Measurement of Nitrate in Water and Wastewater	

Validation Laboratory:  Test America Phoenix
Matrix:  Drinking Water

                       Method Detection Limit (MDL) Data
                                       
Spiking Concentration:  0.30 mg/L 
MDL Data:  0.225, 0.225, 0.246, 0.259, 0.244, 0.244, 0.239

                      Initial Precision and Recovery Data

Spiking Concentration: 5.0 mg/L
IPR Data:  4.98, 4.93, 4.91, 4.99

               Matrix Spike/Matrix Spike Duplicate (MS/MSD) Data

Spiking Concentration:  5.0 mg/L
MS Concentration:  5.12
MSD Concentration:  5.00
Background Concentration:  0.307

                        New Method Acceptance Criteria

IPR Recovery:  95.4  -  102
Precision:  0.011

MS/MSD Recovery:  90.5  -  101
RPD:  0.048

                             New Method Data Form

Method Title:	Hach Method 10206 Spectrophotometric Measurement of Nitrate in Water and Wastewater	

Validation Laboratory:  Test America Watertown
Matrix:  Drinking Water

                       Method Detection Limit (MDL) Data
                                       
Spiking Concentration:  0.30 mg/L 
MDL Data:  0.299, 0.345, 0.264, 0.260, 0.260, 0.284, 0.288

                      Initial Precision and Recovery Data

Spiking Concentration: 5.0 mg/L
IPR Data:  5.02, 4.97, 4.96, 4.95

               Matrix Spike/Matrix Spike Duplicate (MS/MSD) Data

Spiking Concentration:  5.0 mg/L
MS Concentration:  4.81
MSD Concentration:  4.79
Background Concentration:  non detect

                        New Method Acceptance Criteria

IPR Recovery:  95.4  -  102
Precision:  0.011

MS/MSD Recovery:  90.5  -  101
RPD:  0.048

                             New Method Data Form

Method Title:	Hach Method 10206 Spectrophotometric Measurement of Nitrate in Water and Wastewater	

Validation Laboratory:  Test America Portland
Matrix:  FTCWWTP

                       Method Detection Limit (MDL) Data
                                       
Spiking Concentration:  0.30 mg/L 
MDL Data:  0.261, 0.233, 0.230, 0.236, 0.227, 0.238, 0.245

                      Initial Precision and Recovery Data

Spiking Concentration: 5.0 mg/L
IPR Data:  4.95, 4.93, 4.92, 4.98

               Matrix Spike/Matrix Spike Duplicate (MS/MSD) Data

Spiking Concentration:  5.0 mg/L
MS Concentration:  6.20
MSD Concentration:  6.20
Background Concentration:  1.25

                        New Method Acceptance Criteria

IPR Recovery:  95.4  -  102
Precision:  0.011

MS/MSD Recovery:  90.5  -  101
RPD:  0.048

                             New Method Data Form

Method Title:	Hach Method 10206 Spectrophotometric Measurement of Nitrate in Water and Wastewater	

Validation Laboratory:  Test America Valparaiso
Matrix:  LPWWTP

                       Method Detection Limit (MDL) Data
                                       
Spiking Concentration:  0.30 mg/L 
MDL Data:  0.218, 0.225, 0.237, 0.241, 0.230, 0.239, 0.226

                      Initial Precision and Recovery Data

Spiking Concentration: 5.0 mg/L
IPR Data:  4.80, 4.82, 4.85, 4.72

               Matrix Spike/Matrix Spike Duplicate (MS/MSD) Data

Spiking Concentration:  5.0 mg/L
MS Concentration:  4.99
MSD Concentration:  4.98
Background Concentration:  0.032

                        New Method Acceptance Criteria

IPR Recovery:  95.4  -  102
Precision:  0.011

MS/MSD Recovery:  90.5  -  101
RPD:  0.048


                             New Method Data Form

Method Title:	Hach Method 10206 Spectrophotometric Measurement of Nitrate in Water and Wastewater	

Validation Laboratory:  Test America Tallahassee
Matrix:  Industrial KFE

                       Method Detection Limit (MDL) Data
                                       
Spiking Concentration:  0.30 mg/L 
MDL Data:  0.244, 0.254, 0.262, 0.246, 0.248, 0.251, 0.248

                      Initial Precision and Recovery Data

Spiking Concentration: 5.0 mg/L
IPR Data:  5.00, 4.98, 4.95, 4.96

               Matrix Spike/Matrix Spike Duplicate (MS/MSD) Data

Spiking Concentration:  5.0 mg/L
MS Concentration:  11.4
MSD Concentration:  11.3
Background Concentration:  7.18

                        New Method Acceptance Criteria

IPR Recovery:  95.4  -  102
Precision:  0.011

MS/MSD Recovery:  90.5  -  101
RPD:  0.048

                             New Method Data Form

Method Title:	Hach Method 10206 Spectrophotometric Measurement of Nitrate in Water and Wastewater	

Validation Laboratory:  Test America Houston
Matrix:  Industrial DFAE

                       Method Detection Limit (MDL) Data
                                       
Spiking Concentration:  0.30 mg/L 
MDL Data:  0.276, 0.293, 0.270, 0.331, 0.254, 0.231, 0.252

                      Initial Precision and Recovery Data

Spiking Concentration: 5.0 mg/L
IPR Data:  5.02, 4.98, 4.95, 4.96

               Matrix Spike/Matrix Spike Duplicate (MS/MSD) Data

Spiking Concentration:  5.0 mg/L
MS Concentration:  7.88
MSD Concentration:  7.84
Background Concentration:  2.76

                        New Method Acceptance Criteria

IPR Recovery:  95.4  -  102
Precision:  0.011

MS/MSD Recovery:  90.5  -  101
RPD:  0.048

                             New Method Data Form

Method Title:	Hach Method 10206 Spectrophotometric Measurement of Nitrate in Water and Wastewater	

Validation Laboratory:  Test America Dayton
Matrix:  CWWTP

                       Method Detection Limit (MDL) Data
                                       
Spiking Concentration:  0.30 mg/L 
MDL Data:  0.239, 0.272, 0.247, 0.240, 0.232, 0.231, 0.239

                      Initial Precision and Recovery Data

Spiking Concentration: 5.0 mg/L
IPR Data:  4.86, 4.85, 4.87, 4.83

               Matrix Spike/Matrix Spike Duplicate (MS/MSD) Data

Spiking Concentration:  5.0 mg/L
MS Concentration:  7.06
MSD Concentration:  7.17
Background Concentration:  2.21

                        New Method Acceptance Criteria

IPR Recovery:  95.4  -  102
Precision:  0.011

MS/MSD Recovery:  90.5  -  101
RPD:  0.048

