## Method 8276 Technical Toxaphene and Toxaphene Co ngeners by Gas Chromatography – Negative Ion Chemical Ionization /Mass Spectroscopy (GC-NICI/MS)

U.S. EPA Office of Resource Conservation and Recovery Inter-laboratory Validation for Fish Tissue STATISTICAL ANALYSIS REPORT

August 2012

Submitted to: U. S. Environmental Protection Agency Office of Resource Conservation and Recovery 1200 Pennsylvania Avenue, NW Washington, DC 20460

Submitted by: TechLaw, Inc. 14500 Avion Parkway, Suite 300 Chantilly, VA 20151

EPA Contract No. GS-10F-0168J Task Order No. EP-G11S-00001 Work Order No. B-008

### TABLE OF CONTENTS
I. Introduction ............................................................................................................... .................1 . II. Inter-Laboratory Comparisons .............................................................................................. .....3 III. Intra-Laboratory Comparisons ............................................................................................. ......9 IV. Statistical Tests ......................................................................................................... ...............11 V. References ................................................................................................................ ................32 Appendix A – Normality Testing of Technica l Toxaphene and Toxaphene Congener Data Appendix B – Results and 3-Sigma Control Li mits for Terry Creek Fish Tissue Samples Appendix C – Results and 3-Sigma Control Charts for Quality Control Samples
TABLES Table II-1: Terry Creek 1 Fish Sample Results ........................................................................4 Table II-2: Terry Creek 2 Fish Sample Results ........................................................................4 Table II-3: Technical Toxaphene 2 nd 

### Source Standard and LCS Recoveries ...........................5 Table II-4: Toxaphene C ongeners LCS Recoveries .................................................................6 Table II-5: Technical Toxaphe ne Matrix Spike Recoveries .....................................................7 Table II-6: Toxaphene Congene rs Matrix Spike Recoveries ...................................................8 Table III-1: Intra-Laboratory Co mparison for Terry Creek Samples ........................................9 Table III-2: Intra-Laboratory Comparison for Laboratory Control Samples (LCS) ........................................................................................10 Table III-3: Intra-Laboratory Comparison for Matrix Spikes/Matrix Spike Duplicates (MS/MSD) ................................................................................10 Table IV-1: Individual Values Ou tside of 3-Sigma Control Limits ........................................13 Table IV-2: Laboratory Observations W ithin and Outside of Control Limits .........................14 Table IV-3: Method Blanks – Detect ions and Associated Laboratories ..................................15 Table IV-4: Source Fish – Detect ions and Associated Laboratories .......................................16

### TABLE OF CONTENTS (continued) TABLES (continued) Table IV-5: Second Source Standa rd – Toxaphene and Surrogates ........................................17 Table IV-6: Low-Level LCS Results – Toxaphene and Congeners ........................................17 Table IV-7: Mid-Level LCS Resu lts – Toxaphene and Congeners .........................................18 Table IV-8: Spiked Surrogates ............................................................................................... .18 Table IV-9: MS/MSD Results – To xaphene, Surrogates and Congeners ...............................19 Table IV-10: Means and Standard De viations Comparing Toxaphene Results Between Analysts – Fish 1 ...................................................................................20 Table IV-11: Means and Standard De viations Comparing Toxaphene Results Between Analysts – Fish 2 ...................................................................................20 Table IV-12: Means and Standard De viations Comparing Hx-Sed Results Between Analysts – Fish 1 ...................................................................................21 Table IV-13: Means and Standard Deviations Comparing Hx-Sed Results Between Analysts – Fish 2 ...................................................................................21 Table IV-14: Means and Standard Deviations Comparing Hp-Sed Results Between Analysts – Fish 1 ...................................................................................22 Table IV-15: Means and Standard Deviations Comparing Hp-Sed Results Between Analysts – Fish 2 ...................................................................................22 Table IV-16: Means and Standard Deviations Comparing P26 Results Between Analysts – Fish 1 ...................................................................................23 Table IV-17: Means and Standard Deviations Comparing P26 Results Between Analysts – Fish 2 ...................................................................................23 Table IV-18: Means and Standard Deviations Comparing P40 Results Between Analysts – Fish 1 ...................................................................................24 Table IV-19: Means and Standard Deviations Comparing P40 Results Between Analysts – Fish 2 ...................................................................................24 Table IV-20: Means and Standard Deviations Comparing P41 Results Between Analysts – Fish 1 ...................................................................................25 Table IV-21: Means and Standard Deviations Comparing P41 Results Between Analysts – Fish 2 ...................................................................................25

### TABLE OF CONTENTS (continued) TABLES (continued) Table IV-22: Means and Standard Deviations Comparing P44 Results Between Analysts – Fish 1 ...................................................................................26 Table IV-23: Means and Standard Deviations Comparing P44 Results Between Analysts – Fish 2 ...................................................................................26 Table IV-24: Means and Standard Deviations Comparing P50 Results Between Analysts – Fish 1 ...................................................................................27 Table IV-25: Means and Standard Deviations Comparing P50 Results Between Analysts – Fish 2 ...................................................................................27 Table IV-26: Means and Standard Deviations Comparing P62 Results Between Analysts – Fish 1 ...................................................................................28 Table IV-27: Means and Standard Deviations Comparing P62 Results Between Analysts – Fish 2 ...................................................................................28 Table IV-28: Z-Scores for Laboratory Quality Assurance Samples .........................................30

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 1 

### I. INTRODUCTION Objective Fish tissue extract and quality control sample data collected from Phase III of the SW-846 Method 8276 validation study was as sessed using statistical methodol ogies in order to determine the robustness and ruggedness of Method 8276 for use in the analysis of technical toxaphene and toxaphene congeners in fish tissu e. The method performance charac teristics used to evaluate the data included the accuracy/bias and precision of the data sets. Additionally, the data sets were subjected to distribution analysis to determine th e normality and overall variance. Data sets were assessed considering the entire population of data submitted by all participating laboratories (inter-laboratory data), with specific evaluations for inter-laboratory data sets as well as data sets which compared results between analysts from a single laboratory (i ntra-laboratory). Prior to conducting the statistical evaluation, data were first subject ed to validation to ensure all method and quality control parameter requirements were met for the analysis. Only valid data that were determined to be usable for the pur pose of evaluating the efficacy of Method 8276 for fish tissue analysis were included in the statisti cal evaluation. Data validation resulted in the elimination of two data sets. These two data sets were generated using a HP-5MS analyticalcolumn, which was allowed by Method 8276, but was not the recommended or preferred DBXLB column. Data generated using this column resulted in toxaphene congener results which did not conform to the distributi on of results reported by all othe r participating laboratories, or which were not reportable due to co-elution problems. All other data were deemed valid and usable for inclusion in the statistical evaluation. Method Performance Characteristics Bias/Trueness For the purposes of this study, trueness is a perf ormance characteristic th at represents the known systematic error in a result; therefore, bias is a measure of trueness. According to EPA guidance document QA/G-5, Guidance for Quality Assurance Project Plans , bias is the systematic or persistent distortion of a measur ement process and is measured as a function of the difference between a reported value, or the statistical m ean for a set of reported values, and the known amount of the analyte present. Usually, systematic error is caused by the e ffect a specific sample matrix has on the ability to recover and quantitat e a target analyte using a specific method. In the Phase III validation study for technical t oxaphene and toxaphene congeners, bias was evaluated through the analysis of fish tissue sample extracts prepared by EPA Region 4 Lab, matrix spikes, surrogate recoveries, and eval uation of the laboratory control samples.

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 2 

### Percent recovery provides a measure of the bi as in the results, and is calculated as follows:
Recovery 

### 100 ) (%       

 

n s m 

### Conc Conc Conc R

m 

### Conc = Concentration of target analyte meas ured in analysis of spiked sample

s 

### Conc = Concentration of target an alyte in the unspiked sample

n 

### Conc = Nominal concentration of the spiked aliquot Precision Precision is a measure of the magnitude of random e rror(s), which can be attributed to the degree of repeatability and reproducibility of the result s from a specific preparatory and/or analytical method. The precision of the technical toxa phene and toxaphene congeners results were determined using the statistical calculations fo r standard deviation (SD) and relative standard deviation (RSD) on the mean. Data values reported by each Phase III participan t were compiled to include each concentration of technical toxaphene and toxaphe ne congeners, surrogates in spiked samples, and each spiked QC sample. The reported values from the Phase III participants were summed for each concentration level reported. A mean value was calculated using the equation below. The arithmetic mean is the sum of the result s, divided by the number of measurements, as follows: 

# 

 

##  n

i i 

## x n x 1 

## 1



# x = mean

n = number of measurements

i 

### x = result The SD and RSD represent the amount of variability or statistical dispersion of the results from the mean value, and provide an indication of the magnitude of random error. Relative standard deviation (% RSD) is the absolute value of th e coefficient of variation and is determined by dividing the variance of the results by the average (mean) and multiplying by 100.

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 3 

### Standard Deviation (SD) measures the am ount of dispersion of the result, as follows:

ܵܦ ൌ ඪ ቌ 

# 

 n

i 1 

### ሺ x ୧ 

### െ x ሻ ቍ

ሺ ݊െ1 ሻ Percent relative standard de viation (%RSD) is a measure of variance, expressed as a percentage, as follows:
ൌ ܦ%ܴܵ ܦܵ
x 100	ݔ Precision can also be evaluated for sample pairs (duplicates) by calculating a percent difference (%D), also known as the relativ e percent difference (RPD), as follows: %	ܦ݂݂݅ ൌ ቮ ݔ ଵ 

### െ ݔ ଶ 

### ቀ ݔ ଵ 

### ݔ ൅ ଶ 

### 2 ቁ ቮ 	ܺ	100

II. INTER-LABORATORY COMPARISONS Ruggedness is a measure of the degree to which an analytical method results in accurate and reproducible results when performed multiple times by independent laboratories. The Toxaphene Phase III method validation study pr imarily focused on the evaluation of interlaboratory data generated by severa l laboratories that were invite d from inside and outside of EPA to join this Phase III study. This evaluati on included an assessment of the degree to which inter-laboratory results for fish tissue sample s and quality control samples met reasonable benchmarks for acceptable bias/trueness, and precision/variance. The measurement performance criteria for bias and variance, as well as contro l charts were developed, and are included in this report for the purpose of establishi ng the validity of the analyt ical protocol in Method 8276 for fish tissue analysis.

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 4 Table II-1 Terry Creek 1 Fish Sample Results Sample ID No. of Participants Grand Mean (ug/kg) Standard Deviation Reproducibility (%RSD) Toxaphene 

### 8 7030 1170 16.7

Hx-Sed 

### 8 160 10.7 6.65 Hp-Sed 

### 8 184 10.3 5.59

P26 

### 8 16.0 0.738 4.62 P40 

### 8 21.8 2.05 9.37 P41 

### 8 8.93 0.322 3.60 P44 

### 8 9.53 0.693 7.28 P50 

### 8 16.5 1.20 7.25 P62 

### 8 22.3 7.01 31.5

Table II-2 Terry Creek 2 Fish Sample Results Sample ID No. of Participants Grand Mean (ug/kg) Standard Deviation Reproducibility (%RSD) Toxaphene 

### 8 650 112 17.2

Hx-Sed 

### 8 10.8 0.838 7.75 Hp-Sed 

### 8 15.1 0.963 6.37

P26 

### 8 2.32 0.063 2.71 P40 

### 8 2.49 0.114 4.57 P41 

### 8 1.05 0.027 2.53 P44 

### 8 1.34 0.099 7.40 P50 

### 8 2.57 0.125 4.87 P62 

### 8 3.07 1.02 33.3

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 5 Technical Toxaphene 2 nd 

### Source Standard and LCS Recoveries Table II-3

2nd Source Standard Toxaphene
ID No. of Participants True Value (pg/uL) Grand Mean (pg/uL) % Recovery Std. Deviation Reproducibility (% RSD) Second Source Std. 

### 8 150 159 106 9.85 6.20 LCS 1 8 50 57.3 115 9.7 16.9 LCS 2 8 200 207 104 13.4 6.50 PCB 209 LCS1 8 100 95.0 95.0 1.9 2.00 eHCH LCS1 8 250 230.5 92.2 21.0 9.10 PCB 209 LCS2 8 100 90.2 90.2 5.4 6.00 eHCH LCS2 8 250 240 95.8 15.4 6.40

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 6 Toxaphene Congeners LCS Recoveries Table II-4 Hx-Sed
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD LCS1 

### 8 100 98.4 98.4 5.84 5.94 LCS2 

### 8 2 1.95 97.4 0.16 8.00 Hp-Sed
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD LCS1 

### 8 100 98.9 98.9 5.77 5.84 LCS2 

### 8 2 1.99 99.3 0.13 6.30 P26
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD LCS1 

### 8 100 97.9 97.9 5.27 5.39 LCS2 

### 8 2 1.83 91.4 0.36 19.7 P40
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD LCS1 

### 8 100 98.1 98.1 6.33 6.45 LCS2 

### 8 2 1.94 97.2 0.15 7.46 P41
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD LCS1 

### 8 100 98.6 98.6 6.31 6.40 LCS2 

### 8 2 2.03 102 0.17 8.32 P44
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD LCS1 

### 8 100 99.4 99.4 8.85 8.91 LCS2 

### 8 2 1.99 99.8 0.16 8.09 P50
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD LCS1 

### 8 100 96.9 96.9 7.47 7.71 LCS2 

### 8 2 2.01 101 0.07 3.55 P62
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD LCS1 

### 8 226 237 105 26.0 11.0 LCS2 

### 8 22.6 21.0 92.9 4.51 21.5

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 7 Technical Toxaphene Matrix Spike Recoveries Table II-5 ID No. of Participants True Value Grand Mean % Recovery Std. Deviation % RSD Matrix Spike 

### 8 200 186 92.8 23.0 12.4 Matrix Spike Duplicate 

### 8 200 182 90.9 21.3 11.7

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 8 Toxaphene Congeners Matrix Spike Recoveries Table II-6

Hx-Sed
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD MS 

### 8 100 92.6 92.6 6.29 6.79 MSD 

### 8 100 89.8 89.8 5.92 6.60 Hp-Sed
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD MS 

### 8 100 91.9 91.9 5.56 6.05 MSD 

### 8 100 89.2 89.2 4.78 5.36 P26
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD MS 

### 8 100 88.5 88.5 4.93 5.57 MSD 

### 8 100 83.3 83.3 4.23 5.08 P40
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD MS 

### 8 100 91.9 91.9 6.11 6.65 MSD 

### 8 100 88.9 88.9 4.66 5.24 P41
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD MS 

### 8 100 91.8 91.8 6.67 7.26 MSD 

### 8 100 88.0 88.0 6.00 6.81 P44
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD MS 

### 8 100 95.7 95.7 8.94 9.34 MSD 

### 8 100 91.2 91.2 7.63 8.37 P50
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD MS 

### 8 100 90.4 90.4 6.67 7.37 MSD 

### 8 100 85.1 85.1 6.54 7.68 P62
ID No. of Participants True Value Grand Mean % Recovery Std. Deviation %RSD MS 

### 8 226 227 101 33.5 14.7 MSD 

### 8 226 217 95.9 29.0 13.4

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 9 

### III. INTRA-LABORATORY COMPARISONS Robustness is a measure of the degree to which the method is unaffected by minor variations in operating conditions and/or analys ts and laboratories. Robustn ess was generally assessed by comparison of intra-laboratory an alyst precision. As part of th is evaluation, the precision or agreement between fish tissue and quality contro l sample results among analysts within each lab was assessed. The EPA Region 4 laboratory in At hens, GA reported data from three analysts. The intra-laboratory data was evaluated for analys t precision and therefor e was evaluated against the benchmarks of obtaining a percent relative standard deviation (%RSD ) of less than 30% for all measurements. This benchmark cr iteria was met for this evaluation.
Table III-1 Intra-Laboratory Comparison For Terry Creek Samples

ID % RSD Fish 1 % RSD Fish 2 EPA Region 4 Lab – 3 analysts Toxaphene 

### 7.78 5.17 Hx-Sed 

### 4.07 7.01 Hp-Sed 

### 4.47 7.61 P26 

### 4.48 6.74 P40 

### 2.19 4.60 P41 

### 2.58 5.07 P44 

### 6.68 7.37 P50 

### 3.51 4.85 P62 

### 13.9 11.5
Fish tissue Terry Creek 1 and Terry Creek 2 variance for technical toxaphene results

 %RSD < 7.78% for all three analysts from EPA Region 4 laboratory Fish tissue Terry Creek 1 and Terry Creek 2 variance for toxaphe ne congener results

 %RSD < 13.9% for all three analysts from EPA Region 4 laboratory

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 10 Table III-2 Intra-Laboratory Comparison For Laboratory Control Samples (LCS)

ID % RSD LCS 1 % RSD LCS 2 EPA Region 4 Lab – 3 analysts Toxaphene 

### 17.6 9.54 Hx-Sed 

### 4.15 5.69 Hp-Sed 

### 4.93 4.10 P26 

### 4.57 8.71 P40 

### 3.40 5.03 P41 

### 3.63 5.55 P44 

### 4.42 5.27 P50 

### 3.60 4.93 P62 

### 5.88 5.14
Laboratory Control Sample 1 (LCS 1) and LC S 2 variance for techni cal toxaphene results

 %RSD < 17.6% for all three analysts from EPA Region 4 laboratory LCS 1 and LCS 2 variance for toxaphene congener results

 %RSD < 8.71% for all three analysts from EPA Region 4 laboratory
Table III-3 Intra-Laboratory Comparison For Matrix Spikes/Matrix Spike Duplicates (MS/MSD)

ID % RSD MATRIX SPIKE % RSD MATRIX SPIKE

DUPLICATE EPA Region 4 Lab – 3 analysts Toxaphene 

### 4.92 4.57 Hx-Sed 

### 5.74 4.42 Hp-Sed 

### 6.18 4.74 P26 

### 5.84 3.93 P40 

### 4.66 3.26 P41 

### 4.76 3.33 P44 

### 6.36 4.67 P50 

### 4.46 3.10 P62 

### 7.06 6.53
Matrix Spike (MS) and Matrix Spike Duplicate (MSD) variance for tech nical toxaphene results

 %RSD < 4.92% for all three analysts from EPA Region 4 laboratory Matrix Spike (MS) and Matrix Spike Duplicate (MSD) variance for toxaphene congener results

 %RSD < 7.06% for all three analysts from EPA Region 4 laboratory

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 11 

### IV. STATISTICAL TESTS Normality Testing The distribution of the data generated duri ng Phase III of the SW-846 Method 8276 validation study is important to ascertain fo r several reasons: 1) it can i ndicate the need to transform a dataset prior to evaluation; 2) it can influe nce the decision to use parametric versus nonparametric statistical evaluati on methods (since some methods require data to be normally distributed); and 3) it can signal potential biases or ot her issues associated with the analytical method used to generate the data. For the purposes of this study, statistica l methods were used to determine if the results reporte d by Method 8276 on the fish tissue extracts and quality control samples met certain population characteristics for normality and were within reasonable agreement and/or specified variance limits. No rmality testing was perf ormed using the Minitab statistical software program. In order to de tect departures from normality present in the analytical results, the Anderson-Darling Test for Normality (ADT) was used. The ADT is considered one of the more robust statistical te sts to detect departures from normality and was applied to the Terry Creek analytical results fo r toxaphene and for each congener. The test was run independently for each fish (i.e., Fish 1 and Fish 2). ADT results are included in graphical form in Appendix A. Interpretation of the ADT output provided in Appendix A is relatively straightforward. The ADT returns what is termed an A-squared test sta tistic, as well as an associ ated p-value. The ‘A squared’ value is obtained from a probability distribution which defines whether to accept or reject the null hypothesis at the de fined level of significance, whic h in this case is 0.05 or 95% confidence level. When the associated p-value is less than or equal to 0.05, the hypothesis of normality is rejected at the 95% confidence level. In other words, when the p-value is less than or equal to 0.05, we can say with 95% certainty that the data are not normally distributed. Conversely, when the p-value is greater than 0.05 , it can be said that no significant deviation from normality was observed. For the Terry Creek results, no significant deviation from normality was observed (i.e., all p values were greater than 0.05), with the following exceptions:

 Fish 1, P40 (A-squared = 0.88, p = 0.019)

 Fish 2, Toxaphene (A-squared = 1.41, p < 0.005)

 Fish 2, P50 (A-squared = 0.96, p = 0.012) A lognormal transformation was performed for each of the above datasets to mitigate the observed deviations in normality. After transf ormation, the ADT was once again performed. In each case, the deviation from normality was not mitigated by lognormal transformation. ADT output for lognormal transformed data is provided in Appendix A. Given these results, it appears that in these limited cases, there is a significant deviation from normality.

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 12 

### In order to determine whether these departures from normality indicate a methodological bias or other systemic issue, the Phase III data were evaluated following the Inter-laboratory Validation for Fish Tissue Study Plan, dated September 2011, using the method performance characteristics and evaluation criteria. The data from th e Terry Creek fish tissue were assessed for variance/statistical dispersion by calculating the standard de viation and relative standard deviation (RSD) for the fish tissue Terry Creek samples, as well as for the quality control samples, including the LCS and Matrix Spikes. A 30% RSD was used as the benchmark, which would indicate variance was within an acceptabl e limit. However, since the benchmarks foracceptable precision (dispersion) of 30 % RPD and/or 30 % RSD were met for all interlaboratory and intra-laboratory Terry Creek 1 and 2 sample data and quality control sample data, the few exceptions to normality in the data dist ribution do not appear to indicate there is a problem with the method. Rather, it is likely the va riability in analytical technique or process. The variability in the analyses can sometimes be correlated to the quality control results. For the Technical Toxaphene analyses, vari ability was primarily noted in LCS and matrix spikes results. For the LCS results, variability was evident in the high recoveries reported for the Low-Level LCS toxaphene (LCS 1 spiked at 50 pg/uL) reported by NEIC (141% and 132%) and EPANERL/ORD-LV-2 (133% and 150%). As reported in the Analytical Data Report, one possible source of variability occurred as a result of the presence of contam inant peaks that eluted within the retention times/peak area assigned to quantita tion of technical toxaphene, as evidenced by blank contamination. Variability was also not ed in the LCS and matr ix spike results for congeners. The variability in LCS congeners resu lts stems from the low recoveries reported for Parlar 62 in the Low-Level LCS (LCS 2 spiked at 2 pg/uL) by SCCWRP (69.4%, 67.8%, for two of the three replicates); EPA-NERL/ORD-LV-2 (45.8% and 46.5%); and EPA-NERL/ORD-LV1 (59.8% and 74.6%). The low recoveries for th e LCS results reported by SCCWRP may be a result of variability in instrument response n ear the LLOQ. The low LC S recoveries reported by the EPA-NERL/ORD-LV-1 participant may be due to the method of quantitation used. The EPA-NERL/ORD-LV-1 participant used a linear re gression curve in the calibration rather than a relative response factor or qua dratic curve method. As noted in Method 8276, Section 11.4.5.4, the use of linear regression anal ysis has the potential to cause a significant bias in the lower portion of the calibration curve, while the rela tive percent difference and quadratic methods do not. Matrix spike recoveries were low for technical toxaphene results repor ted by all laboratories except for SCCWRP, while matrix spike results were slightly low for toxaphene congener results reported by all laboratories except for EPA-R4- 1. The low matrix spike results may indicate possible matrix interference due to the fish matrix. Other than the potential sources of variability, no trends or biases were noted in the Analytical Data Report for the congener data. Therefore, the deviations from normality demonstrated for the P40 and P50 data were most likely a result of normal statistical variability in the analytical process, and do not appear to indicate a si gnificant problem with the Method 8276 methodology.

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 13 

### Given this, as well as the fact that the deviat ions from normality were limited to only three analytes across both sets of fish samples, the data will be evaluated in its raw (i.e., untransformed) state using standard parametric analytical techniques. Bar Graphs and 3-sigma Control Limits – Terry Creek In order to statistically evaluate the data ge nerated during this study, uppe r control limits (UCL) and lower control limits (LCL) were established using what is termed a 3-sigma approach (a standard approach for generating statistical quality control charts ). For ease of display, bar graphs depicting analytical re sults were constructed for toxa phene and each congener for the Terry Creek data and are included in Appendix B. The bar graphs represent individual analytical results (for each replicate). A center line representing the Grand M ean is indicated on the graphs, along with lines repres enting the UCL and LCL. The UCLs and LCLs represent three standard deviations (or 3-sigma) above (UCL) and below (LCL) the Grand Mean. These control levels were selected because in a normal distri bution, 99.7% of observations will fall within three standard deviations of the mean. Th erefore, if a process is in a state of statistical control, all data points should fall within the 3-sigma UCL and LCL. Data points that fall outside of the control limits may be indicative of a process problem (such as the nature of the media, method of sample preparation, and/or analytical te chnique) that may warrant furt her investigation. Table IV-1 shows the number of individual an alytical results outside of the 3-sigma control limits. Table IV-2 shows the total number of observations for each analyst, and the number of those observations that fell outsi de of the control limits.
Table IV-1 Individual Values Outside of 3-sigma Control Limits * Analyte Number of Individual Values Outside of Control Limits - Fish 1 Number of Individual Values Outside of Control Limits - Fish 2 Toxaphene 0 (18) 0 (18) Hx-Sed 0 (18) 0 (18) Hp-Sed 0 (18) 0 (18) P26 0 (18) 0 (18) P40 0 (18) 0 (18) P41 0 (18) 0 (18) P44 0 (18) 2 (18) P50 0 (18) 1 (18) P62 0 (16) 0 (18) Total 0 (160) 3 (162) *Total number of individual values is included in parentheses.

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 14 

### Table IV-2 Laboratory Observations Within and Outside of Control Limits Laboratory (Analyst) Total Number of Observations Number of Observations Outside

of Control Limits Percentage of Observations Outside

of Control Limits Ashland 36 1 2.8 EPA NEIC 34 0 0.0 EPA R4 1 54 0 0.0 EPA R4 2 36 0 0.0 EPA R4 3 36 0 0.0 SCCWRP 54 0 0.0 EPA ORD-LV 1 36 0 0.0 EPA ORD-LV 2 36 2 5.6 Total 322 3 0.9 As shown in Table IV-1, none of the 160 individual measurements for Fish 1 and only three (two for P44 and one for P50) of the 162 individual meas urements (< 2%) for Fish 2 fall outside of the 3-sigma control limits. In each of these cases, the analytical value outside of the 3-sigma control limits was one of two replicates, with the other fall ing within the control lim its. If an average of replicates is taken to generate a single analytical result for th e analyst (as was done to generate the Grand Mean), then the average of each of th ese replicates falls within the control limits. Based on this evaluation, it appears that the analytical process is in a state of statistical control. Control Charts – Laboratory Quality Control Samples Individual control charts (I-charts) were constructed for laboratory quality control samples and are included in Appendix C. Charts were develo ped for the following quality control samples:

 Blank Samples
o Method Blanks
o Source Fish

 Spiked Samples
o Second Source Standard
o LCS (low-level and mid-level)
o Surrogates
o Matrix Spikes/Matrix Spike Duplicates I-charts developed for the quality control samp les do not employ a standard 3-sigma UCL and LCL approach as was done for the Terry Creek samp les. Rather, alternative reference lines have been provided to assist in eval uating laboratory performance.

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 15 

### For the method blanks and source fish, the followi ng reference lines are in cluded on the I-charts:

 zero concentration line (this in cludes “non-detects” as well as laboratory results reported as “<”)

 mean concentration line

 calibration standard lines (set at 50% of the lo west calibration standard [25 pg/uL] for technical toxaphene, and at 50-100% of the lowest calibra tion standard for toxaphene congeners) For spiked samples, the following reference lines are included:

 mean concentration line

 true value line (actual spiked concentrat ion of the second source standard, LCS, surrogates, and MS/MSDs)

 true value reference lines (+/- 20% and +/- 30% of true value for second source standard; +/- 10%, +/- 15% and +/- 20% of true value for mid-level LCS; +/- 20%, +/- 30% and +/- 50% of true value for low-level LCS; +/- 20 % and +/- 30% of true value for surrogates and MS/MSDs) A discussion of laboratory quality cont rol sample results is provided below. Blank Samples Tables IV-3 and IV-4 show the di stribution of analytical results compared to the reference points discussed above for method blanks and source fi sh, as well as the analytical laboratories associated with analyte detections. Table IV-3 Method Blanks – Detections and Associated Laboratories * Analyte Number of Detections Number of Detections ≤ 50% of the lowest

calibration

standard Number of Detections Between 50%- 100% of the lowest calibration standard Number of Detections >100% of the lowest calibration standard Toxaphene 12 (18) 12 (18) 0 (18) 0 (18) Hx-Sed 3 (18) 3 (18) 0 (18) 0 (18) Hp-Sed 3 (18) 3 (18) 0 (18) 0 (18) P26 2 (18) 0 (18) 2 (18) 0 (18) P40 2 (18) 0 (18) 2 (18) 0 (18) P41 2 (18) 0 (18) 2 (18) 0 (18) P44 2 (18) 0 (18) 2 (18) 0 (18) P50 2 (18) 0 (18) 2 (18) 0 (18) P62 2 (18) 2 (18) 0 (18) 0 (18) Total 30 (162) 20 (162) 10 (18) 0 (18) *Total number of individual values is included in parentheses.

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 16 

### Table IV-4 Source Fish – Detections and Associated Laboratories * Analyte Number of Detections Number of Detections ≤ 50% of the lowest

calibration

standard Number of Detections Between 50-100% of the lowest calibration standard Number of Detections >100% of the lowest calibration standard Toxaphene 17 (18) 13 (18) 4 (18) 0 (18) Hx-Sed 5 (18) 5 (18) 0 (18) 0 (18) Hp-Sed 5 (18) 5 (18) 0 (18) 0 (18) P26 9 (18) 7 (18) 1 (18) 1 (18) P40 5 (18) 3 (18) 2 (18) 0 (18) P41 6 (18) 4 (18) 2 (18) 0 (18) P44 6 (18) 4 (18) 2 (18) 0 (18) P50 10 (18) 6 (18) 4 (18) 0 (18) P62 5(18) 5 (18) 0 (18) 0 (18) Total 68 (162) 52 (162) 15 (162) 1 (162) *Total number of individual values is included in parentheses. 

### As shown in Table IV-3, and Appendix C-1 to C-5, laboratories reported method blank detections in 30 (19%) of 162 individual measur ements. Of those 30 detections, 20 fell below 50% of the lowest calibration standard referen ce line, with the remain ing 10 falling between 50% and 100% of the lowest calibration standard re ference line. As shown in Table IV-4, and Appendix C-6 to C-10, laboratories reported source fi sh detections in 68 (42%) of 162 individual measurements. Of those 68 detections, 52 fell below 50% of the lowest calibration standard reference line; 15, fell between 50% and 100%; and one, above 100% of the lowest calibration standard reference line.

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 17 

### Spiked Samples Tables IV-5 through IV-9 show the distribution of analytical results compared to the reference points discussed above for the following spiked sa mples: second source standard, low-level LCS, mid-level LCS, spiked surrogates and MS/MSDs.
Table IV-5 Second Source Standard – Toxaphene and Surrogates * Analyte Number of Observations +/- 20% of True Value Between +/- 20% and +/- 30% of True Value > +/- 30% of True Value Toxaphene 18 (18) 0 (18) 0 (18) PCB 209 18 (18) 0 (18) 0 (18) ε -HCH 17 (18) 1 (18) 0 (18) Total 53 (54) 1 (54) 0 (54) *Total number of individual values is included in parentheses. 

### As shown in Table IV-5 and Appendix C-11 to C- 12 of the 54 individual results for the second source standard samples, 53 (98%) were within +/ - 20% of the true value. One result (2%) fell between +/- 20% and +/- 30% of the true value, with none exceeding +/- 30%.
Table IV-6 Low-Level LCS Results – Toxaphene and Congeners * Analyte Number of Observations +/- 20% of True Value Between +/- 20% and +/- 30% of

True Value Between +/- 30% and +/- 50% of

True Value > +/- 50% of True Value Toxaphene 10 (18) 2 (18) 6 (18) 0 (18) Hx-Sed 17 (18) 1 (18) 0 (18) 0 (18) Hp-Sed 17 (18) 1 (18) 0 (18) 0 (18) P26 15 (18) 0 (18) 2 (18) 1 (18) P40 17 (18) 0 (18) 1 (18) 0 (18) P41 17 (18) 1 (18) 0 (18) 0 (18) P44 17 (18) 0 (18) 1 (18) 0 (18) P50 18 (18) 0 (18) 0 (18) 0 (18) P62 14 (18) 1 (18) 3 (18) 0 (18) Total 143 (162) 5 (162 ) 13 (162) 1 (162) *Total number of individual values is included in parentheses. 

### As shown in Table IV-6 and Appendix C-13, and C-15 to C-22, of the 162 individual results for the low-level LCS, 143 (88%) were within +/- 20% of the true value. Five measurements (3%) fell between +/- 20% and +/- 30% of the true value, and 13 (8%) between +/- 30% and +/- 50%. Only one result (1%) was more than 50% lower than the true spiked value.

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 18 

### Table IV-7 Mid-Level LCS Results – Toxaphene and Congeners * Analyte Number of Observations +/- 10% of True Value Between +/- 10% and +/- 15% of

True Value Between +/- 15% and +/- 20% of

True Value > +/- 20% of True Value Toxaphene 13 (18) 4 (18) 1 (18) 0 (18) Hx-Sed 17 (18) 1 (18) 0 (18) 0 (18) Hp-Sed 17 (18) 1 (18) 0 (18) 0 (18) P26 17 (18) 0 (18) 1 (18) 0 (18) P40 15 (18) 3 (18) 0 (18) 0 (18) P41 16 (18) 2 (18) 0 (18) 0 (18) P44 14 (18) 2 (18) 1 (18) 1 (18) P50 16 (18) 0 (18) 2 (18) 0 (18) P62 16 (18) 0 (18) 2 (18) 0 (18) Total 141(162) 13 (162) 7 (162) 1(162) *Total number of individual values is included in parentheses. 

### As shown in Table IV-7 and Appendix C-13, and C-15 to C-22, of the 162 individual results for the mid-level LCS, 141 (87%) were within +/- 10% of the true spiked value. Thirteen measurements (8%) fell between +/- 10% and +/- 15% of the true value, and 7 (4%) between +/- 15% and +/- 20%. Only one result (1%) was 20% lower than the true spiked value.
Table IV-8 Spiked Surrogates* Analyte Number of Observations +/- 20% of True Value Between +/- 20% and +/- 30% of True Value > +/- 30% of True Value PCB 209 52 (54) 2 (54) 0 (54) ε -HCH 52 (54) 2 (54) 0 (54) Total 104 (108) 4 (108) 0 (108) *Total number of individual values is included in parentheses. 

### As shown in Table IV-8 and Appendix C-14, of the 108 individual results for the spiked surrogates, 104 (96%) were within +/- 20% of the true spiked va lue. Four measurements (4%) fell between +/- 20% and +/- 30% and no results ex ceeded +/- 30% of the true spiked value.

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 19 

### Table IV-9 MS/MSD Results – Toxaphene, Surrogates and Congeners * Analyte Number of Observations +/- 20% of True Value Between +/- 20% and +/- 30% of True Value > +/- 30% of True Value Toxaphene 29 (36) 6 (36) 1 (36) PCB 209 0 (36) 11 (36) 25 (36) ε -HCH 18 (36) 18 (36) 0 (36) Hx-Sed 36 (36) 0 (36) 0 (36) Hp-Sed 36 (36) 0 (36) 0 (36) P26 32 (36) 4 (36) 0 (36) P40 32 (36) 4 (36) 0 (36) P41 32 (36) 4 (36) 0 (36) P44 32 (36) 4 (36) 0 (36) P50 32 (36) 4 (36) 0 (36) P62 32 (36) 4 (36) 0 (36) Total 311 (396) 59 (396) 26 (396) *Total number of individual values is included in parentheses. 

### As shown in Table IV-9 and Appendix C-23 to C-28, of the 396 individual results for the MS/MSDs, 311 (79%) were within +/- 20% of the true value. Fifty-nine measurements (15%) fell between +/- 20% and +/- 30% of the true va lue, and 26 (7%) exceeded +/- 30% of the true spiked value. Analytical Means Testing In order to determine if there are significan t differences in mean analyte concentrations (toxaphene and associated congeners) between laboratories (specifi cally among laboratory analysts), One-way Analysis of Variance (ANOVA) was performed. Results of the ANOVA yield what is termed an F-statistic, as well as an associated p-value. When the associated p-value is less than or equal to 0.05, it can be said wi th 95% confidence that there is a significant difference in analytical results between laborator ies (and/or analysts). Conversely, when the pvalue is greater than 0.05, no signifi cant difference (at the 95% confid ence level) is observed. In cases where a significant difference between laboratories was observed, post-hoc Tukey’s comparisons were performed to elucidate specific differences in mean concentrations. In the tables below, means that do not share a letter in the “Grouping” colu mn are significantly different from one another (based on 95% confid ence). Results of this analysis are included below. Note that as discussed above, there is a significant deviation from normality in the distribution of data for three analytes. However, ANOVA is r obust against the assumption of normality, and therefore is an appr opriate test for evaluating differe nces in mean concentrations in this case.

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 20 

### Toxaphene A statistically significant diffe rence was found among laboratory an alysts on toxaphene results for Fish 1 (F(7,10) = 176.54, p= < 0.001) and Fi sh 2 (F(7,10) = 151.77, p = < 0.001). Tables IV10 and IV-11 show mean toxaphene concentrations for each laboratory (by analyst) for each fish. Means that do not share a letter in the “Grouping” column are si gnificantly different from one another (based on 95% confidence).
Table IV-10 Means and Standard Deviations Comparing To xaphene Results between Analysts – Fish 1 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA ORD-LV 2 2 475 7.85 A SCCWRP 3 424 1.15 B EPA R4 1 3 368 14.2 C Ashland 2 355 1.70 CD EPA R4 3 2 338 4.07 DE EPA R4 2 2 312 3.05 E EPA ORD-LV 1 2 275 11.5 F EPA NEIC 2 263 1.91 F
Table IV-11 Means and Standard Deviations Comparing To xaphene Results between Analysts – Fish 2 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA ORD-LV 2 2 492 3.96 A Ashland 2 327 14.6 B EPA NEIC 2 326 1.58 B EPA ORD-LV 1 2 324 0.421 B SCCWRP 3 314 10.1 B EPA R4 1 3 264 12.6 C EPA R4 2 2 245 9.13 C EPA R4 3 2 245 4.12 C

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 21 

### Hx-Sed A statistically significant diffe rence was found among laboratory analysts on Hx-Sed results for Fish 1 (F(7,10) = 71.35, p = < 0.001) and Fish 2 (F(7,10) = 15.80, p = < 0.001). Tables IV-12 and IV-13 show mean Hx-Sed concentrations for each laboratory (by analyst) for each fish. Means that do not share a letter in the “Grouping” column are si gnificantly different from one another (based on 95% confidence).
Table IV-12 Means and Standard Deviations Comparing Hx -Sed Results Between Analysts – Fish 1 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA R4 1 3 87.9 1.19 A Ashland 2 86.6 0.072 AB EPA R4 2 2 82.9 0.521 BC EPA R4 3 2 80.8 0.421 CD SCCWRP 3 80.4 1.74 CD EPA NEIC 2 77.8 0.312 D EPA ORD-LV 2 2 73.3 0.141 E EPA ORD-LV 1 2 72.6 0.642 E
Table IV-13 Means and Standard Deviations Comparing Hx -Sed Results Between Analysts – Fish 2 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA R4 1 3 61.3 2.88 A Ashland 2 60.0 2.90 AB EPA R4 2 2 55.6 0.522 ABC EPA R4 3 2 53.6 0.322 BCD SCCWRP 3 52.5 1.26 CD EPA ORD-LV 1 2 51.4 0.571 CD EPA NEIC 2 51.4 0.424 CD EPA ORD-LV 2 2 48.1 1.04 D

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 22 

### Hp-Sed A statistically significant diffe rence was found among laboratory analysts on Hp-Sed results for Fish 1 (F(7,10) = 14.09, p = < 0.001) and Fish 2 (F(7,10) =8.48, p = < 0.002). Tables IV-14 and IV-15 show mean Hp-Sed concentrations for each laboratory (by analyst) for each fish. Means that do not share a letter in the “Grouping” colu mn are significantly different from one another (based on 95% confidence).
Table IV-14 Means and Standard Deviations Comparing Hp -Sed Results Between Analysts – Fish 1 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA R4 1 3 99.9 1.27 A Ashland 2 97.5 1.13 AB SCCWRP 3 94.1 2.69 AB EPA R4 3 2 92.2 0.983 ABC EPA R4 2 2 92.0 1.07 BC EPA NEIC 2 90.2 0.201 BCD EPA ORD-LV 2 2 85.0 1.71 CD EPA ORD-LV 1 2 83.4 5.30 D
Table IV-15 Means and Standard Deviations Comparing Hp -Sed Results Between Analysts – Fish 2 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA R4 1 3 85.3 4.54 A Ashland 2 81.5 6.22 AB SCCWRP 3 76.3 2.15 ABC EPA R4 2 2 75.7 0.452 ABC EPA R4 3 2 74.5 0.453 BC EPA NEIC 2 72.9 1.05 BC EPA ORD-LV 1 2 71.9 0.142 BC EPA ORD-LV 2 2 66.4 1.84 C

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 23 

### P26 No statistically significant differences were found among laboratory analysts on P26 results for Fish 1 (F(7,10) = 2.24, p = 0.12) or Fish 2 (F(7 ,10) = 1.9, p = 0.17). Ta bles IV-16 and IV-17 show P26 mean concentrations for each laborator y (by analyst) for each fish. Note that all means are in the same grouping since, base d on a 95% confidence le vel, they are not significantly different.
Table IV-16 Means and Standard Deviations Comparing P26 Results Between Analysts – Fish 1 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA R4 1 3 8.58 0.142 A SCCWRP 3 8.47 0.424 A EPA ORD-LV 1 2 8.35 0.503 A EPA R4 3 2 8.09 0.072 A EPA NEIC 2 8.00 0.133 A EPA R4 2 2 7.84 0.231 A EPA ORD-LV 2 2 7.67 0.840 A Ashland 2 7.45 0.504 A
Table IV-17 Means and Standard Deviations Comparing P26 Results Between Analysts – Fish 2 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA R4 1 3 12.8 0.493 A SCCWRP 3 11.8 0.402 A EPA NEIC 2 11.8 0.522 A EPA R4 2 2 11.5 0.280 A EPA R4 3 2 11.4 0.541 A EPA ORD-LV 1 2 11.3 0.214 A Ashland 2 11.3 1.34 A EPA ORD-LV 2 2 11.2 0.662 A

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 24 

### P40 A statistically significant differe nce was found among laboratory anal ysts on P40 results for Fish 1 (F(7,10) = 4.39, p = 0.018) and Fish 2 (F(7,10) =6.69, p = 0.004). Tables IV-18 and IV-19 show mean P40 concentrations for each laboratory (by analyst) fo r each fish. Means that do not share a letter in the “Grouping” column are si gnificantly different from one another (based on 95% confidence).
Table IV-18 Means and Standard Deviations Comparing P40 Results Between Analysts – Fish 1 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA R4 1 3 12.2 0.333 A EPA R4 2 2 12.1 0.144 AB EPA R4 3 2 11.8 0.051 AB EPA NEIC 2 11.7 0.042 AB SCCWRP 3 11.3 0.701 AB EPA ORD-LV 2 2 10.8 0.742 AB EPA ORD-LV 1 2 9.75 1.06 AB Ashland 2 9.55 1.49 B
Table IV-19 Means and Standard Deviations Comparing P40 Results Between Analysts – Fish 2 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA R4 1 3 13.7 0.582 A EPA ORD-LV 2 2 13.1 0.152 AB EPA NEIC 2 12.8 0.011 AB EPA R4 2 2 12.8 0.092 AB EPA R4 3 2 12.8 0.153 AB SCCWRP 3 12.4 0.481 B EPA ORD-LV 1 2 11.7 0.781 B Ashland 2 11.6 0.144 B

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 25 

### P41 A statistically significant differe nce was found among laboratory anal ysts on P41 results for Fish 1 (F(7,10) = 4.71, p = 0.014). No significant differe nces were found among laboratory analysts for Fish 2 (F(7,10) =3.2, p = 0.051). Tables IV-20 and IV-21 show mean P41 concentrations for each laboratory (by analyst) for each fish. Means that do not share a letter in the “Grouping” column are significantly different from one another (based on 95% confidence). Note that for Fish 2, all means are in the same grouping since, based on a 95% confidence level, they are not significantly different.
Table IV-20 Means and Standard Deviations Comparing P41 Results Between Analysts – Fish 1 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA R4 1 3 4.71 0.082 A SCCWRP 3 4.63 0.231 AB EPA ORD-LV 1 2 4.60 0.000 ABC EPA NEIC 2 4.56 0.082 ABC EPA R4 2 2 4.53 0.062 ABC EPA R4 3 2 4.51 0.013 ABC Ashland 2 4.25 0.213 BC EPA ORD-LV 2 2 4.15 0.044 C
Table IV-21 Means and Standard Deviations Comparing P41 Results Between Analysts – Fish 2 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA R4 1 3 5.66 0.212 A EPA ORD-LV 1 2 5.40 0.141 A SCCWRP 3 5.37 0.314 A EPA NEIC 2 5.33 0.162 A EPA R4 2 2 5.30 0.013 A EPA R4 3 2 5.14 0.012 A Ashland 2 5.05 0.071 A EPA ORD-LV 2 2 5.05 0.064 A

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 26 

### P44 No statistically significant differences were found among laboratory analysts on P44 results for Fish 1 (F(7,10) = 3.13, p = 0.05) or Fish 2 (F(7 ,10) = 1.92, p = 0.17). Tables IV-22 and IV-23 show mean P44 concentrations fo r each laboratory (by analyst) for each fish. Note that all means are in the same grouping since, based on 95% confidence, they are not significantly different.
Table IV-22 Means and Standard Deviations Comparing P44 Results Between Analysts – Fish 1 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA NEIC 2 5.28 0.012 A EPA R4 1 3 5.16 0.254 A SCCWRP 3 4.90 0.101 A EPA R4 2 2 4.70 0.340 A EPA R4 3 2 4.65 0.042 A EPA ORD-LV 1 2 4.55 0.641 A EPA ORD-LV 2 2 4.43 0.113 A Ashland 2 4.40 0.280 A
Table IV-23 Means and Standard Deviations Comparing P44 Results Between Analysts – Fish 2 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
Ashland 2 7.55 1.06 A EPA R4 1 3 7.04 0.293 A EPA ORD-LV 2 2 6.74 0.383 A SCCWRP 3 6.67 0.354 A EPA NEIC 2 6.61 0.221 A EPA R4 2 2 6.32 0.151 A EPA R4 3 2 6.16 0.302 A EPA ORD-LV 1 2 5.75 1.20 A

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 27 

### P50 A statistically significant differe nce was found among laboratory anal ysts on P50 results for Fish 1 (F(7,10) = 15.97, p < 0.001) and Fish 2 (F(7,10 ) =6.50, p = 0.004). Tables IV-24 and IV-25 show mean P50 concentrations for each laboratory (by analyst) fo r each fish. Means that do not share a letter in the “Grouping” column are si gnificantly different from one another (based on 95% confidence).
Table IV-24 Means and Standard Deviations Comparing P50 Results Between Analysts – Fish 1 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA R4 1 3 9.32 0.114 A EPA R4 2 2 8.79 0.103 AB EPA R4 3 2 8.73 0.124 AB SCCWRP 3 8.60 0.363 AB EPA NEIC 2 8.50 0.182 AB EPA ORD-LV 2 2 8.03 0.221 B Ashland 2 7.80 0.571 BC EPA ORD-LV 1 2 6.95 0.212 C
Table IV-25 Means and Standard Deviations Comparing P50 Results Between Analysts – Fish 2 Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
EPA R4 1 3 14.3 0.552 A EPA ORD-LV 2 2 13.4 0.143 A EPA R4 2 2 13.3 0.000 A EPA NEIC 2 13.3 0.301 A EPA R4 3 2 13.1 0.214 A SCCWRP 3 13.0 0.473 A Ashland 2 12.4 0.642 AB EPA ORD-LV 1 2 10.7 1.49 B

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 28 

### P62 A statistically significant differe nce was found among laboratory anal ysts on P62 results for Fish 1 (F(6,9) = 8.18, p = 0.003) and Fish 2 (F(6,9) = 6.24, p = 0.005). Tables IV-26 and IV-27 show mean P62 concentrations for each laboratory (by an alyst) for each fish. Means that do not share a letter in the “Grouping” column are significa ntly different from one another (based on 95% confidence).
Table IV-26 Means and Standard Deviations Comparing P62 Results Between Analysts – Fish 1

Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
SCCWRP 3 16.4 0.903 A EPA ORD-LV 1 2 11.0 0.991 AB EPA R4 1 3 10.7 0.394 B EPA ORD-LV 2 2 9.51 4.48 B Ashland 2 8.90 0.281 B EPA R4 2 2 8.47 0.493 B EPA R4 3 2 8.26 0.253 B EPA NEIC * * * * *No data reported for this laboratory.


### Table IV-27 Means and Standard Deviations Comparing P62 Results Between Analysts – Fish 2

Laboratory (Analyst) Number of Observations Mean Concentration (pg/uL) Standard Deviation Grouping
SCCWRP 3 20.1 3.20 A EPA NEIC 2 19.9 2.25 A EPA ORD-LV 2 2 17.0 4.56 AB EPA ORD-LV 1 2 14.7 2.69 AB EPA R4 1 3 14.1 0.473 AB EPA R4 3 2 13.1 2.21 AB EPA R4 2 2 11.8 1.34 AB Ashland 2 7.90 0.00 B

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 29 

### Z-Scores – Laboratory Quality Control Samples In addition to using modified I-Charts to eval uate laboratory performance during this study, Zscores were generated for each quality contro l sample and averaged for each laboratory participant. In accordance with Internationa l Union of Pure and Applied Chemistry (IUPAC 1993) guidance, Z-scores can be used as indicators of accuracy in laboratory inter-comparison studies and reflect the difference between a laborat ory result and a target value or performance criterion. Z-scores are calcula ted using the following formula: Z-score = (laboratory estimate)/(pe rformance criterion) = (x - X)/ σ where x is the individual laborato ry analytical result, X is th e “exercise assigned value,” and σ is the established target value or cr iterion. Z-scores for this study we re calculated using an exercise assigned value equal to the averag e recovery (for each replicate) for each quality control sample (i.e., MS, MSD, LCS1, LCS2 and Second Source St andard). A fixed performance criterion for each analyte for each laboratory was calculated using 25% of the exercise assigned value ( σ = 25% X), yielding the following: Z-score = (laboratory analytical result – av erage recovery) / (25% average recovery) Z-scores that have been calculated for this study can be interpreted in the following way:

 A value of +1 indicates the laboratory result is 25% higher than the exercise assigned value.

 A value of -1 indicates the laboratory result is 25% lower than the exercise assigned value.

 A value of +2 indicates the laboratory result is 50% higher than the exercise assigned value.

 A value of -2 indicates the laboratory result if 50% lower than the exercise assigned value. An accepted approach to classifying and categor izing Z-scores includes the following (IUPAC 1993):

 |z| ≤ 2  Satisfactory

 2< |z| < 3  Questionable

 |z| ≥ 3  Unsatisfactory Table IV-28 summarizes the Z-scores (based on a 25% X criterion) for each participant and each laboratory quality control sample.

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 30 

### Table IV-28 Z-Scores for Laboratory Quality Assurance Samples

LaboratoryASHLANDEPA_ NEICEPA_R4_1EPA_R4_2EPA_R4_3SCCWRPEPA_NERL/ORD_1EPA_NERL/ORD_2



### Analyte Second Source Standard Toxaphene -0.18 0.15 0.17 0.16 0.13 -0.27 -0.45 0.17 PCB 209 -0.02 0.02 0.09 0.10 0.07 -0.12 -0.12 -0.03 eHCH -0.19 -0.05 0.02 -0.11 -0.13 -0.16 0.40 0.21

LCS1 Toxaphene -0.04 0.50 0.23 0.01 -0.96 -0.83 0.15 0.98 PCB 209 -0.22 0.02 0.01 -0.07 -0.05 0.09 0.00 0.05 eHCH 0.63 0.45 0.02 -0.08 -0.05 -0.11 -0.63 -0.10 Hx-Sed 0.38 0.06 0.25 0.08 -0.08 -0.01 -0.28 -0.35 Hp-Sed 0.34 0.05 0.27 -0.02 -0.11 0.07 -0.28 -0.33 P26 0.12 0.10 0.26 -0.03 -0.07 0.09 -0.42 -0.21 P40 0.20 0.07 0.26 0.10 -0.03 0.09 -0.48 -0.32 P41 0.12 0.06 0.27 0.06 -0.03 0.14 -0.54 -0.23 P44 0.01 0.05 0.39 0.11 0.01 0.12 -0.82 -0.16 P50 0.01 0.06 0.32 0.12 0.01 0.16 -0.65 -0.29 P62 0.11 -0.17 0.63 0.23 0.11 0.01 -0.90 -0.21

LCS2 Toxaphene 0.54 -0.06 0.23 -0.12 -0.53 0.17 -0.08 -0.04 PCB 209 -0.27 0.09 -0.41 0.01 0.09 0.15 0.08 0.23 eHCH 0.40 0.29 0.20 -0.06 -0.09 -0.22 -0.37 -0.09 Hx-Sed 0.09 0.06 0.54 0.12 0.10 -0.30 -0.26 -0.45 Hp-Sed 0.63 -0.02 0.28 0.03 0.01 -0.17 -0.19 -0.31 P26 0.34 0.52 0.69 0.08 0.50 -0.14 -1.00 -1.35 P40 0.72 0.16 0.21 -0.07 -0.22 -0.09 -0.55 0.07 P41 0.33 -0.11 0.18 -0.19 -0.21 -0.18 0.71 -0.31 P44 0.79 0.00 0.31 -0.10 0.00 -0.05 -0.68 -0.02 P50 -0.23 0.15 0.20 -0.16 -0.09 -0.15 -0.08 0.14 P62 0.35 0.43 0.83 0.43 0.41 -0.33 -1.73 -0.70

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 31 LaboratoryASHLANDEPA_ NEICEPA_R4_1EPA_R4_2EPA_R4_3SCCWRPEPA_NERL/ORD_1EPA_NERL/ORD_2



### Analyte

MS Toxaphene -0.63 -0.62 0.25 -0.02 0.20 0.47 0.44 -0.46 PCB 209 -0.39 -0.02 -0.05 -0.06 -0.11 0.14 -0.10 0.37 eHCH 0.49 0.28 0.00 -0.13 -0.03 -0.19 -0.23 0.01 Hx-Sed 0.32 -0.01 0.39 0.06 -0.09 -0.09 -0.37 -0.19 Hp-Sed 0.29 0.02 0.36 -0.07 -0.10 -0.01 -0.30 -0.23 P26 0.04 0.08 0.28 -0.16 -0.08 0.02 -0.46 0.12 P40 0.12 0.02 0.30 0.01 -0.07 0.06 -0.62 0.05 P41 0.04 0.00 0.28 -0.05 -0.07 0.16 -0.68 0.12 P44 -0.05 -0.06 0.35 -0.10 -0.14 0.17 -0.80 0.37 P50 -0.03 -0.02 0.24 -0.06 -0.10 0.15 -0.65 0.28 P62 -0.30 -0.27 0.53 -0.03 -0.01 0.23 -1.21 0.66

MSD Toxaphene 0.06 -0.66 0.32 -0.08 0.19 0.32 0.51 -0.61 PCB 209 -0.27 -0.05 0.04 -0.07 -0.19 0.05 -0.08 0.40 eHCH 0.39 0.32 -0.01 -0.12 0.02 -0.23 -0.27 0.04 Hx-Sed 0.50 -0.01 0.31 0.02 -0.04 -0.07 -0.37 -0.21 Hp-Sed 0.45 0.00 0.26 -0.12 -0.04 -0.02 -0.17 -0.24 P26 0.18 0.09 0.16 -0.18 0.02 0.05 -0.45 0.06 P40 0.12 0.02 0.19 -0.02 -0.06 0.09 -0.49 0.07 P41 0.10 0.02 0.20 -0.05 -0.04 0.17 -0.66 0.11 P44 -0.12 -0.03 0.25 -0.10 -0.10 0.22 -0.72 0.32 P50 -0.14 -0.02 0.17 -0.03 -0.07 0.21 -0.67 0.31 P62 -0.50 -0.32 0.48 0.01 -0.06 0.24 -0.96 0.67 

### As Table IV-28 indicates, Z-scores for each partic ipant and analyte all fell into the satisfactory category (|z| ≤ 2), with all scores falling between -1.73 a nd 0.98. These results indicate that there is an acceptable level of consistency between la boratories within the confines of this study. Please note that the percent recoveries, as well as the percent relative, percent differences, and relative standard deviation values for each laboratory control sample and matrix spike sample for all participants can be found in the Draft Phase III Toxaphene Data Tables in Attachment 3 of the Analytical Data Report.

### Method 8276 U.S. EPA ORCR Inter ‐ laboratory Validation: August 2012 Phase III Statistical Analysis Report Page 32 

### V. REFERENCES 1. TechLaw Incorporated, “Inter-laboratory Validation for Fish Tissue Study Plan,” Submitted to U.S. Environmental Protection Agency, September 2011. 2. TechLaw Incorporated, “Inter -laboratory Validation for Fi sh Tissue Analytical Data Report,” Submitted to U.S. Environmental Protection Agency, May 2012. 3. TechLaw Incorporated, “Inter-laboratory Validation for Fish Tissue Data Validation Report,” Submitted to U.S. Environmental Protection Agency, May 2012. 4. International Union of Pure and Applied Chemistry (1993). The International Harmonized Protocol for the Proficiency Test ing of (Chemical) Analytical Laboratories , Pure & Applied Chem., 65 (9), 2123-2144.

### APPENDIX A Normality Testing of Technical Toxa phene and Toxaphene Congener Data

### Normality Testing - Toxaphene

### Normality Testing - Congeners

### Normality Testing

– Transformed Data

### APPENDIX B Results and 3-Sigma Control Limits for Terry Creek Fish Tissue Samples

### Results and 3-Sigma Control Limits for Toxaphene – Terry Creek – Fish 1

Results and 3-Sigma Control Limits for Congeners – Terry Creek – Fish 1 Laboratory

Replicate

E P A _

N

E R L / O

R

D _

L V _

2

( 2 )

E P A _

N

E R L / O

R

D _

L V _

1

( 2 ) S

C

C W

R P E P A _

R 4 _

3
E P A _

R 4 _

2
E P A _

R 4 _

1
E P A _

N

E I C A

S H

L A

N

D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 600 500 400 300 200 100 0Toxaphene Concentration (pg/uL) 567 (+ 3-sigma)

351 (Grand Mean)

135 (- 3-sigma) 

### Toxaphene Results and Control Limits - Terry Creek Fish = 1

Laboratory Rep

E P A _

N

E R

L / O

R D _

L V _

2 ( 2 )

E P A _

N

E R L / O

R D _

L V _

1

( 2 ) S C

C W

R P E P A _

R 4 _

3
E

P A _

R

4

_ 2
E P A _

R 4 _

1
E P A _

N

E I C A

S H

L A

N

D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 100

80
60
40
20

0Hx-Sed Concentration (pg/uL) 97.1 (+ 3-sigma) 80.3 (Grand Mean) 63.5 (- 3-sigma) 

### Hx-Sed Results and Control Limits - Terry Creek Fish = 1

### Results and 3-Sigma Control Limits for Congeners – Terry Creek – Fish 1

Laboratory Rep

E P A _

N

E R

L / O

R D _

L V _

2 ( 2 )

E P A _

N

E R L / O

R D _

L V _

1

( 2 ) S C

C W

R P E P A _

R 4 _

3
E

P A _

R

4

_ 2
E P A _

R 4 _

1
E P A _

N

E I C A

S H

L A

N

D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 120 100

80 60 40 20

0Hp-Sed concentration (pg/uL) 109 (+ 3-sigma) 91.8 (Grand Mean) 74.8 (- 3-sigma) 

### Hp-Sed Results and Control Limits - Terry Creek Fish = 1

Laboratory Rep

E P A _

N

E R

L / O

R D _

L V _

2 ( 2 )

E P A _

N

E R L / O

R D _

L V _

1

( 2 ) S C

C W

R P E P A _

R 4 _

3
E

P A _

R

4

_ 2
E P A _

R 4 _

1
E P A _

N

E I C A

S H

L A

N

D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 9 8 7 6 5 4 3 2 1 0P26 Concentration (pg/uL) 9.25 (+ 3-sigma) 8.05 (Grand Mean) 6.85 (- 3-sigma) 

### P26 Results and Control Limits - Terry Creek Fish = 1

### Results and 3-Sigma Control Limits for Congeners – Terry Creek – Fish 1

Laboratory Rep

E P A _

N

E R

L / O

R D _

L V _

2 ( 2 )

E P A _

N

E R L / O

R D _

L V _

1

( 2 ) S C

C W

R P E P A _

R 4 _

3
E

P A _

R

4

_ 2
E P A _

R 4 _

1
E P A _

N

E I C A

S H

L A

N

D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 14 12 10

8 6 4 2 0P40 Concentration (pg/uL) 14.1 (+ 3-sigma) 11.1 (Grand Mean) 8.04 (- 3-sigma) 

### P40 Results and Control Limits - Terry Creek Fish = 1

Laboratory Rep

E P A _

N

E R

L / O

R D _

L V _

2 ( 2 )

E P A _

N

E R L / O

R D _

L V _

1

( 2 ) S C

C W

R P E P A _

R 4 _

3
E

P A _

R

4

_ 2
E P A _

R 4 _

1
E P A _

N

E I C A

S H

L A

N

D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 5
4
3
2
1
0P41 Concentration (pg/uL) 5.06 (+ 3-sigma) 4.49 (Grand Mean) 3.92 (- 3-sigma) 

### P41 Results and Control Limits - Terry Creek Fish = 1

### Results and 3-Sigma Control Limits for Congeners – Terry Creek – Fish 1

Laboratory Rep

E P A _

N

E R

L / O

R D _

L V _

2 ( 2 )

E P A _

N

E R L / O

R D _

L V _

1

( 2 ) S C

C W

R P E P A _

R 4 _

3
E

P A _

R

4

_ 2
E P A _

R 4 _

1
E P A _

N

E I C A

S H

L A

N

D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 6
5
4 3 2
1 0P44 Concentration (pg/uL) 5.75 (+ 3-sigma) 4.76 (Grand Mean) 3.77 (- 3-sigma) 

### P44 Results and Control Limits - Terry Creek Fish = 1

Laboratory Rep

E P A _

N

E R

L / O

R D _

L V _

2 ( 2 )

E P A _

N

E R L / O

R D _

L V _

1

( 2 ) S C

C W

R P E P A _

R 4 _

3
E

P A _

R

4

_ 2
E P A _

R 4 _

1
E P A _

N

E I C A

S H

L A

N

D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 12 10

8
6 4
2 0P50 Concentration (pg/uL) 10.5 (+ 3-sigma) 8.34 (Grand Mean) 6.15 (- 3-sigma) 

### P50 Results and Control Limits - Terry Creek Fish = 1

### Results and 3-Sigma Control Limits for Congeners – Terry Creek – Fish 1

Laboratory Rep

E P A _

N

E R

L / O

R D _

L V _

2 ( 2 )

E P A _

N

E R L / O

R D _

L V _

1

( 2 ) S C

C W

R P E P A _

R 4 _

3
E

P A _

R

4

_ 2
E P A _

R 4 _

1
E P A _

N

E I C A

S H

L A

N

D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 20

15

10

5

0P62 Concentration (pg/uL) 18.9 (+ 3-sigma)

2.07 (- 3-sigma) 10.5 (Grand Mean) 

### P62 Results and Control Limits - Terry Creek Fish = 1

### Results and 3-Sigma Control Limits for Toxaphene – Terry Creek – Fish 2

Results and 3-Sigma Control Limits for Congeners – Terry Creek – Fish 2 Laboratory

Replicate

E P A _

N E R L / O R D _

L V _

2

( 2 )

E P A _

N E R L / O R D

_ L V _

1

( 2 ) S C C W

R P
E P A _

R 4 _

3
E P A _

R 4 _

2
E P A _

R 4 _

1
E P A _

N E I C A S H L A

N D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 600 500 400 300 200 100 0Toxaphene Concentration (pg/uL) 554 (+ 3-sigma)

316 (Grand Mean)

78.1 (- 3-sigma) 

### Toxaphene Results and Control Limits - Terry Creek Fish = 2

Laboratory Rep

E P A _

N E R L / O

R D _

L V _

2 ( 2 )

E P A _

N E R L / O

R D _

L V _

1

( 2 ) S C C W

R P
E P A _

R 4 _

3
E P A _

R 4

_ 2
E P A _

R 4 _

1
E P A _

N E I C A S H L A

N D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 70 60 50 40 30 20 10

0Hx-Sed Concentration (pg/uL) 67.7 (+ 3-sigma)

40.7 (- 3-sigma) 54.2 (Grand Mean) 

### Hx-Sed and Control Limits - Terry Creek Fish = 2

### Results and 3-Sigma Control Limits for Congeners – Terry Creek – Fish 2

Laboratory Rep

E P A _

N E R L / O

R D _

L V _

2 ( 2 )

E P A _

N E R L / O

R D _

L V _

1

( 2 ) S C C W

R P
E P A _

R 4 _

3
E P A _

R 4

_ 2
E P A _

R 4 _

1
E P A _

N E I C A S H L A

N D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 90 80 70 60 50 40 30 20 10

0Hp-Sed Concentration (pg/uL) 93.0 (+ 3-sigma)

58.2 (- 3-sigma) 75.6 (Grand Mean) 

### Hp-Sed and Control Limits - Terry Creek Fish = 2

Laboratory Rep

E P A _

N E R L / O

R D _

L V _

2 ( 2 )

E P A _

N E R L / O

R D _

L V _

1

( 2 ) S C C W

R P
E P A _

R 4 _

3
E P A _

R 4

_ 2
E P A _

R 4 _

1
E P A _

N E I C A S H L A

N D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 14 12 10

8 6 4 2 0P26 Concentration (pg/uL) 13.2 (+ 3-sigma) 10.0 (- 3-sigma) 11.6 (Grand Mean) 

### P26 Results and Control Limits - Terry Creek Fish = 2

### Results and 3-Sigma Control Limits for Congeners – Terry Creek – Fish 2

Laboratory Rep

E P A _

N E R L / O

R D _

L V _

2 ( 2 )

E P A _

N E R L / O

R D _

L V _

1

( 2 ) S C C W

R P
E P A _

R 4 _

3
E P A _

R 4

_ 2
E P A _

R 4 _

1
E P A _

N E I C A S H L A

N D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 16 14 12 10

8 6 4 2 0P40 Concentration (pg/uL) 14.7 (+ 3-sigma) 12.6 (Grand Mean) 10.5 (- s-sigma) 

### P40 Results and Control Limits - Terry Creek Fish = 2

Laboratory Rep

E P A _

N E R L / O

R D _

L V _

2 ( 2 )

E P A _

N E R L / O

R D _

L V _

1

( 2 ) S C C W

R P
E P A _

R 4 _

3
E P A _

R 4

_ 2
E P A _

R 4 _

1
E P A _

N E I C A S H L A

N D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 6 5
4 3
2 1 0P41 Concentration (pg/uL) 5.92 (+ 3-sigma) 5.29 (Grand Mean) 4.66 (- 3-sigma) 

### P41 Results and Control Limits - Terry Creek Fish = 2

### Results and 3-Sigma Control Limits for Congeners – Terry Creek – Fish 2

Laboratory Rep

E P A _

N E R L / O

R D _

L V _

2 ( 2 )

E P A _

N E R L / O

R D _

L V _

1

( 2 ) S C C W

R P
E P A _

R 4 _

3
E P A _

R 4

_ 2
E P A _

R 4 _

1
E P A _

N E I C A S H L A

N D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 9 8 7 6 5 4 3 2 1 0P44 Concentration (pg/uL) 8.25 (+ 3-sigma) 6.60 (Grand Mean) 4.95 (- 3-sigma) 

### P44 Results and Control Limits - Terry Creek Fish = 2

Laboratory Rep

E P A _

N E R L / O

R D _

L V _

2 ( 2 )

E P A _

N E R L / O

R D _

L V _

1

( 2 ) S C C W

R P
E P A _

R 4 _

3
E P A _

R 4

_ 2
E P A _

R 4 _

1
E P A _

N E I C A S H L A

N D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 18 16 14 12 10

8 6 4 2 0P50 Concentration (pg/uL) 16.1 (+ 3-sigma) 12.9 (Grand Mean) 9.72 (- 3-sigma) 

### P50 Results and Control Limits - Terry Creek Fish = 2

### Results and 3-Sigma Control Limits for Congeners – Terry Creek – Fish 2

Laboratory Rep

E P A _

N E R L / O

R D _

L V _

2 ( 2 )

E P A _

N E R L / O

R D _

L V _

1

( 2 ) S C C W

R P
E P A _

R 4 _

3
E P A _

R 4

_ 2
E P A _

R 4 _

1
E P A _

N E I C A S H L A

N D 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 3 2 1 30 25 20
15 10

5 0P62 Concentration (pg/uL)

2.41 (- 3-sigma) 14.8 (Grand Mean) 27.2 (+ 3-sigma) 

### P62 Results and Control Limits - Terry Creek Fish = 2

### Appendix C Results and 3-Sigma Control Char ts for Quality Control Samples

### Control Chart for Toxaphene – Method Blank

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart. 1715 1311 975 3 1 25

20

15

10

5

0

ObservationIndividualValue(pg/uL)

_ X=3.39 25 pg/uL,

0 pg/uL 

### I Chart of Toxaphene - Method Blan k

50% of LLOQ

### Control Chart for Congeners – Method Blank

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart.

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart. 1715 13 11 975 3 1 1.0

0.8

0.6

0.4

0.2

0.0

ObservationIndividualValue(pg/uL)

_ X=0.042 0.5 pg/uL,

0 pg/uL 1 pg/uL, 

### I Chart of Hx-Sed - Method Blank

LLOQ

17 15 13 11 97531 1.0

0.8

0.6

0.4

0.2

0.0

ObservationIndividualValue(pg/uL)

_ X=0.035 1 pg/uL,

0.5 pg/uL,

0 pg/uL 

### I Chart of Hp-Sed - Method Blan k 50% of LLOQ

LLOQ

50% of LLOQ

### Control Chart for Congeners – Method Blank

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart.

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart. 17151311 975 3 1 1.0

0.8

0.6

0.4

0.2

0.0

ObservationIndividualValue(pg/uL)

_ X=0.092 0.5 pg/uL,

0 pg/uL 1 pg/uL, 

### I Chart of P26 - Method Blan k

50% of LLOQ

17151311 975 3 1 1.0

0.8

0.6

0.4

0.2

0.0

ObservationIndividualValue(pg/uL)

_ X=0.066 0.5 pg/uL,

0 pg/uL 1 pg/uL, 

### I Chart of P40 - Method Blan k

50% of LLOQ LLOQ

LLOQ

### Control Chart for Congeners – Method Blank

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart.

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart. 17 151311 975 3 1 1.0

0.8

0.6

0.4

0.2

0.0

ObservationIndividualValue(pg/uL)

_ X=0.073 0.5 pg/uL,

0 pg/uL 1 pg/uL, 

### I Chart of P41 - Method Blan k

50% of LLOQ LLOQ

17 151311 975 3 1 1.0

0.8

0.6

0.4

0.2

0.0

ObservationIndividualValue(pg/uL)

_ X=0.101 0.5 pg/uL,

0 pg/uL 1 pg/uL, 

### I Chart of P44 - Method Blan k

50% of LLOQ LLOQ

### Control Chart for Congeners – Method Blank

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart.

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart. 1715 13 11 975 3 1 1.0

0.8

0.6

0.4

0.2

0.0

ObservationIndividualValue(pg/uL)

_ X=0.074 0.5 pg/uL,

0 pg/uL 1 pg/uL, 

### I Chart of P50 - Method Blan k

50% of LLOQ LLOQ

17 15 13 11 9753 1 5

4

3

2

1

0

ObservationIndividualValue(pg/uL)

_ X=0.161 2.5 pg/uL,

0 pg/uL 5 pg/uL, 

### I Chart of P62 - Method Blan k

50% of LLOQ LLOQMethod	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation Source	Fish	Control	Charts Phase III Statistical Analysis Report Page	1 

### Control Chart for Toxaphene – Source Fish

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart. 17 15 13 11 975 31 50

40

30

20

10

0

ObservationIndividualValue(pg/uL)

_ X=16.54 50 pg/uL,

0 pg/uL 25 pg/uL, 

### I Chart of Toxaphene - Source Fish

50% of LLOQ LLOQMethod	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation Source	Fish	Control	Charts Phase III Statistical Analysis Report Page	2 

### Control Chart for Congeners – Source Fish

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart.

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart. 17151311 975 3 1 1.0

0.8

0.6

0.4

0.2

0.0

ObservationIndividualValue(pg/uL)

_ X=0.043 1 pg/uL,

0.5 pg/uL,

0 pg/uL 

### I Chart of Hx-Sed - Source Fish

50% of LLOQ LLOQ

17151311 975 3 1 1.0

0.8

0.6

0.4

0.2

0.0

ObservationIndividualValue(pg/uL)

_ X=0.039 1 pg/uL,

0.5 pg/uL,

0 pg/uL 

### I Chart of Hp Sed - Source Fish

50% of LLOQ LLOQMethod	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation Source	Fish	Control	Charts Phase III Statistical Analysis Report Page	3 

### Control Chart for Congeners – Source Fish

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart.

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart. 17151311 9753 1 1.4
1.2
1.0
0.8
0.6
0.4
0.2
0.0

ObservationIndividualValue(pg/uL)

_ X=0.217 1 pg/uL,

0.5 pg/uL,

0 pg/uL 

### I Chart of P26 - Source Fish

50% of LLOQ LLOQ

17151311 9753 1 1.0

0.8

0.6

0.4

0.2

0.0

ObservationIndividualValue(pg/uL)

_ X=0.07 1 pg/uL,

0.5 pg/uL,

0 pg/uL 

### I Chart of P40 - Source Fish

50% of LLOQ LLOQMethod	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation Source	Fish	Control	Charts Phase III Statistical Analysis Report Page	4 

### Control Chart for Congeners – Source Fish

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart.

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart. 17 151311 975 3 1 1.0

0.8

0.6

0.4

0.2

0.0

ObservationIndividualValue(pg/uL)

_ X=0.098 1 pg/uL,

0.5 pg/uL,

0 pg/uL 

### I Chart of P41 - Source Fish

50% of LLOQ LLOQ

17 151311 975 3 1 1.0

0.8

0.6

0.4

0.2

0.0

ObservationIndividualValue(pg/uL)

_ X=0.12 1 pg/uL,

0.5 pg/uL,

0 pg/uL 

### I Chart of P44 - Source Fish

50% of LLOQ LLOQMethod	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation Source	Fish	Control	Charts Phase III Statistical Analysis Report Page	5 

### Control Chart for Congeners – Source Fish

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart.

Notes: Non-detects and results reported as “< ” are indicated as “0” on the chart. 17151311 975 3 1 1.0

0.8

0.6

0.4

0.2

0.0

ObservationIndividualValue(pg/uL)

_ X=0.201 1 pg/uL,

0.5 pg/uL,

0 pg/uL 

### I Chart of P50 - Source Fish

50% of LLOQ LLOQ

17 15 13 11 975 3 1 5

4

3

2

1

0

ObservationIndividualValue(pg/uL) 5 pg/uL,

2.5 pg/uL,

0 pg/uL _ X=0.173 

### I Chart of P62 - Source Fish

50% of LLOQ LLOQ

### Control Charts for Toxaphene and Surrogates – Seco nd Source Standard (Toxaphene spiked at 150 pg/uL; PCB209, 100 pg/uL; and eHCH, 250 pg/uL )

17151311 97531 200

180

160

140

120

100
ObservationIndividual Value (pg/uL)

_ X=159.5 195 (+30%)

120 (-20%) 180 (+20%)

150 - True Value

105 (-30%) 

### I Chart of Toxaphene - Second Source Standard

17151311 97531 130

120

110

100

90

80

70

ObservationIndividual Value (pg/uL)

_ X=109.29 130 (+30%)

80 (-20%) 120 (+20%)

100 - True Value

70 (-30%) 

### I Chart of PCB209 - Second Source Standard

### Control Charts for Toxaphene and Surrogates – Seco nd Source Standard (Toxaphene spiked at 150 pg/uL; PCB209, 100 pg/uL; and eHCH, 250 pg/uL )

1715 13 11 97531 340 320 300 280 260 240 220 200 180 160
ObservationIndividual Value (pg/uL)

_ X=263.7 325 (+30%)

200 (-20%) 300 (+20%)

250 - True Value

175 (-30%) 

### I Chart of eHCH - Second Source StandardMethod	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: LCS	Control	Charts Phase III Statistical Analysis Report Page	1 

### Control Charts for Toxaphene – LCS-1 (Low-50 pg/uL, at LLOQ) and LCS-2 (Mid-200 pg/uL)

1715 1311 97531 80

70

60

50

40

30

20
ObservationIndividual Value (pg/uL)

_ X=56.74 75 (+50%)
65 (+30%) 60 (+20%)

25 (-50%) 35 (-30%) 40 (-20%) 50 - True Value 

### I Chart of Toxaphene - LCS-1

17151311 97531 240 230 220 210 200 190 180 170 160 150
ObservationIndividual Value (pg/uL)

_ X=208.66

160 (-20%) 170 (-15%) 180 (-10%) 240 (+20%) 230 (+15%) 220 (+10%)

200 - True Value 

### I Chart of Toxaphene - LCS-2Method	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: LCS	Control	Charts Phase III Statistical Analysis Report Page	2 

### Control Charts for Spiked Surrogates – PCB209 (100 pg/uL) and eHCH (250 pg/uL)

51 4641363126211611 61 130

120

110

100

90

80

70

ObservationIndividual Value (pg/uL)

_ X=96.07 130 (+30%)
120 (+20%)

100 - True Value

80 (-20%)
70 (-30%) 

### I Chart of PCB209 - Spiked Surrogate

51 4641363126211611 61 340 320 300 280 260 240 220 200 180 160
ObservationIndividual Value (pg/uL)

_ X=238.8 325 (+30%) 300 (+20%)

200 (-20%) 250 - True Value

175 (-30%) 

### I Chart of eHCH - Spiked SurrogateMethod	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: LCS	Control	Charts Phase III Statistical Analysis Report Page	3 

### Control Charts for Congeners – LCS-1 (Mid-100 pg/uL for all, except 226 pg/uL for P62)

1715 13 11 97531 120

110

100

90

80

ObservationIndividual Value (pg/uL)

_ X=98.77 120 (+20%) 115 (+15%)

85 (-15%) 110 (+10%)

90 (-10%) 100 - True Value

80 (-20%) 

### I Chart of Hx-Sed - LCS-1

1715 13 11 97531 120

110

100

90

80

ObservationIndividual Value (pg/uL)

_ X=99.46 120 (+20%)

80 (-20%) 115 (+15%)

85 (-15%) 110 (+10%)

90 (-10%) 100 - True Value 

### I Chart of Hp-Sed - LCS-1Method	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: LCS	Control	Charts Phase III Statistical Analysis Report Page	4 

### Control Charts for Congners – LCS-1 (Mid- 100 pg/uL for all, except 226 pg/uL for P62)

17 151311 97531 120

110

100

90

80

ObservationIndividual Value (pg/uL)

_ X=98.40 120 (+20%)

80 (-20%) 115 (+15%)

85 (-15%) 110 (+10%)

90 (-10%) 100 - True Value 

### I Chart of P26 - LCS-1

17 151311 97531 120

110

100

90

80

ObservationIndividual Value (pg/uL)

_ X=98.65 120 (+20%)

80 (-20%) 115 (+15%)

85 (-15%) 110 (+10%)

90 (-10%) 100 - True Value 

### I Chart of P40 - LCS-1Method	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: LCS	Control	Charts Phase III Statistical Analysis Report Page	5 

### Control Charts for Congners – LCS-1 (Mid- 100 pg/uL for all, except 226 pg/uL for P62)

1715 13 11 97531 120

110

100

90

80

ObservationIndividual Value (pg/uL)

_ X=99.20 120 (+20%) 115 (+15%)

85 (-15%) 110 (+10%)

90 (-10%) 100 - True Value

80 (-20%) 

### I Chart of P41 - LCS-1

1715 13 11 97531 120

110

100

90

80

ObservationIndividual Value (pg/uL)

_ X=100.17 120 (+20%) 115 (+15%)

85 (-15%) 110 (+10%)

90 (-10%) 100 - True Value

80 (-20%) 

### I Chart of P44 - LCS-1Method	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: LCS	Control	Charts Phase III Statistical Analysis Report Page	6 

### Control Charts for Congners – LCS-1 (Mid- 100 pg/uL for all, except 226 pg/uL for P62)

1715 13 11 97531 120

110

100

90

80

ObservationIndividual (pg/uL)

_ X=97.66 120 (+20%) 115 (+15%)

85 (-15%) 110 (+10%)

90 (-10%) 100 - True Value

80 (-20%) 

### I Chart of P50 - LCS-1

1715 13 11 97531 280

260

240

220

200

180

ObservationIndividual Value (pg/uL)

_ X=239.4 271.2 (+20%)

192.1 (-15%) 259.9 (+15%)

203.4 (-10%) 248.6 (+10%)

226 - True Value

180.8 (-20%) 

### I Chart of P62 - LCS-1Method	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: LCS	Control	Charts Phase III Statistical Analysis Report Page	7 

### Control Charts for Congners – LCS-2 (Low-2 pg/uL for all, except 22.6 pg/uL for P62)

17151311 97531 3.0

2.5

2.0

1.5

1.0

ObservationIndividual Value (pg/uL)

_ X=1.955 3 (+50%)

1.4 (-30%) 2.6 (+30%)

1.6 (-20%) 2.4 (+20%)

2 - True Value

1 (-50%) 

### I Chart of Hx-Sed - LCS-2

17151311 97531 3.0

2.5

2.0

1.5

1.0

ObservationIndividual Value (pg/uL)

_ X=1.989 3 (+50%)

1 (-50%) 1.4 (-30%) 2.6 (+30%)

1.6 (-20%) 2.4 (+20%)

2 - True Value 

### I Chart of Hp-Sed - LCS-2Method	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: LCS	Control	Charts Phase III Statistical Analysis Report Page	8 

### Control Charts for Congners – LCS-2 (Low-2 pg/uL for all, except 22.6 pg/uL for P62)

17 15 13 11 97531 3.0

2.5

2.0

1.5

1.0

ObservationIndividual Value (pg/uL)

_ X=1.844 3 (+50%)

1 (-50%) 1.4 (-30%) 2.6 (+30%)

1.6 (-20%) 2.4 (+20%)

2 - True Value 

### I Chart of P26 - LCS-2

17 15 13 11 97531 3.0

2.5

2.0

1.5

1.0

ObservationIndividual Value (pg/uL)

_ X=1.948 3 (+50%)

1.4 (-30%) 2.6 (+30%)

1.6 (-20%) 2.4 (+20%)

2 - True Value

1 (-50%) 

### I Chart of P40 - LCS-2Method	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: LCS	Control	Charts Phase III Statistical Analysis Report Page	9 

### Control Charts for Congners – LCS-2 (Low-2 pg/uL for all, except 22.6 pg/uL for P62)

17 15 13 11 97531 3.0

2.5

2.0

1.5

1.0

ObservationIndividual Value (pg/uL)

_ X=2.031 3 (+50%)

1.4 (-30%) 2.6 (+30%)

1.6 (-20%) 2.4 (+20%)

2 - True Value

1 (-50%) 

### I Chart of P41 - LCS-2

17 15 13 11 97531 3.0

2.5

2.0

1.5

1.0

ObservationIndividual Value (pg/uL)

_ X=2.003 3 (+50%)

1.4 (-30%) 2.6 (+30%)

1.6 (-20%) 2.4 (+20%)

2 - True Value

1 (-50%) 

### I Chart of P44 - LCS-2Method	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: LCS	Control	Charts Phase III Statistical Analysis Report Page	10 

### Control Charts for Congners – LCS-2 (Low-2 pg/uL for all, except 22.6 pg/uL for P62)

17 15 13 11 97531 3.0

2.5

2.0

1.5

1.0

ObservationIndividual Value (pg/uL)

_ X=2.014 3 (+50%)

1.4 (-30%) 2.6 (+30%)

1.6 (-20%) 2.4 (+20%)

2 - True Value

1 (-50%) 

### I Chart of P50 - LCS-2

1715 13 11 97531 35

30

25

20

15

10
ObservationIndividual Value (pg/uL)

_ X=21.16 33.9 (+50%)

15.82 (-30%) 29.38 (+30%)

18.08 (-20%) 27.12 (+20%)

22.6 - True Value

11.3 (-50%) 

### I Chart of P62 - LCS-2Method	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: Matrix	Spikes	Control	Charts Phase III Statistical Analysis Report Page	1 

### Control Charts for Toxaphene and Surrogates– MS/MSD (Toxaphene spiked at 200 pg/uL; PCB20, 100 pg/uL; and eHCH, 250 pg/uL)

3329 25 21 1713 951 275

250

225

200

175

150

ObservationIndividual Value (pg/uL)

_ X=185.7 260 (+30%)

160 (-20%) 240 (+20%)

200 - True Value

140 (-30%) 

### I Chart of Toxaphene - MS/MSD

3329 25 21 1713 951 130
120
110
100

90
80
70
60
50
ObservationIndividual Value (pg/uL)

_ X=68.29 130 (+30%)

80 (-20%) 120 (+20%)

100 - True Value

70 (-30%) 

### I Chart of PCB209 - MS/MSDMethod	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: Matrix	Spikes	Control	Charts Phase III Statistical Analysis Report Page	2 

### Control Charts for Toxaphene and Surrogates– MS/MSD (Toxaphene spiked at 200 pg/uL; PCB20, 100 pg/uL; and eHCH, 250 pg/uL)

33 2925211713 951 340 320 300 280 260 240 220 200 180 160
ObservationIndividual Value (pg/uL)

_ X=201.4 325 (+30%)

200 (-20%) 300 (+20%)

250 - True Value

175 (-30%) 

### I Chart of eHCH - MS/MSDMethod	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: Matrix	Spikes	Control	Charts Phase III Statistical Analysis Report Page	3 

### Control Charts for Congeners – MS/MSD (All congeners spiked at 100 pg/uL, except for P62 which is spiked at 226 pg/uL)

33 2925211713 951 130

120

110

100

90

80

70

ObservationIndividual Value (pg/uL)

_ X=91.61 130 (+30%)

80 (-20%) 120 (+20%)

100 - True Value

70 (-30%) 

### I Chart of Hx-Sed - MS/MSD

33

2925211713 951 130

120

110

100

90

80

70

ObservationIndividual Value (pg/uL)

_ X=90.93 130 (+30%)

80 (-20%) 120 (+20%)

100 - True Value

70 (-30%) 

### I Chart of Hp-Sed - MS/MSDMethod	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: Matrix	Spikes	Control	Charts Phase III Statistical Analysis Report Page	4 

### Control Charts for Congeners – MS/MSD (All congeners spiked at 100 pg/uL, except for P62 which is spiked at 226 pg/uL)

33 2925211713 951 130

120

110

100

90

80

70

ObservationIndividual Value (pg/uL)

_ X=90.80 130 (+30%)

80 (-20%) 120 (+20%)

100 - True Value

70 (-30%) 

### I Chart of P40 - MS/MSD 332925211713 9 5 1 130

120

110

100

90

80

70

ObservationIndividualValue(pg/uL)

_ X=86.22 130 (+30%)

80 (-20%) 120 (+20%)

100 - True Value

70 (-30%) 

### I Chart of P26 - MS/MSDMethod	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: Matrix	Spikes	Control	Charts Phase III Statistical Analysis Report Page	5 

### Control Charts for Congeners – MS/MSD (All congeners spiked at 100 pg/uL, except for P62 which is spiked at 226 pg/uL)

33 2925211713 951 130

120

110

100

90

80

70

ObservationIndividual Value (pg/uL)

_ X=90.48 130 (+30%)

80 (-20%) 120 (+20%)

100 - True Value

70 (-30%) 

### I Chart of P41 - MS/MSD

332925211713 951 130

120

110

100

90

80

70

ObservationIndividual Value (pg/uL)

_ X=94.18 130 (+30%)

80 (-20%) 120 (+20%)

100 - True Value

70 (-30%) 

### I Chart of P44 - MS/MSDMethod	8276	U.S.	EPA	ORCR	Inter‐laboratory	Validation: Matrix	Spikes	Control	Charts Phase III Statistical Analysis Report Page	6 

### Control Charts for Congeners – MS/MSD (All congeners spiked at 100 pg/uL, except for P62 which is spiked at 226 pg/uL)

332925211713 951 130

120

110

100

90

80

70

ObservationIndividual Value (pg/uL)

_ X=88.29 130 (+30%)

80 (-20%) 120 (+20%)

100 - True Value

70 (-30%) 

### I Chart of P50 - MS/MSD

332925211713 951 300

275

250

225

200

175

150
ObservationIndividual Value (pg/uL)

_ X=224.6

158.2 (-30%) 293.8 (+30%)

180.8 (-20%) 271.2 (+20%)

226 - True Value 

### I Chart of P62 - MS/MSD