TCEQ 2016 The 20152016 TNI Standard and the

TCEQ 2016 The 2015/2016 TNI Standard and the EPA MDL Update Richard Burrows, Ph. D. Test. America Inc. 1 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

A Revision to the Method Detection Limit EPA published a revision to the 40 CFR Part 136 MDL procedure in the Federal Register on Thursday February 19 th 2015 EPA expects that the rule will be finalized in late 2016 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved. 2

TNI Chemistry Standard Revisions to the Detection, Quantitation and Calibration language in TNI Volume 1 Module 4 (Quality Systems for Chemical Testing) are complete and will be incorporated into the TNI 2015/2016 Standard. The 2015/2016 TNI Standards should be adopted by the States in 2017/2018 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved. 3

MDL and TNI revisions!!? ? • What if they have different requirements? • What if they are implemented at different times? • What if different states adopt the new requirements at different times? • Will I have to have different detection limits for different states? 4 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

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Good News • The revised MDL and TNI Chemistry standards are fully compatible • If you are compliant with the revised EPA MDL then you are also compliant with the current MDL • One exception, discussed later • If you are compliant with the revised TNI Chemistry standard then you are compliant with the current TNI Chemistry standard 6 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Details of the Modifications Initial MDL 7 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

First, what stays the same? • Fundamental concept is unchanged • What is the lowest result that is qualitatively reliable, i. e. , the lowest result that reliably indicates the analyte is in the sample? • Fundamental approach is unchanged • Describe the distribution as Student’s t times the standard deviation of results 8 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

What is different? • Requires calculation of a MDL based on blanks as well as a MDL based on spikes (the higher of the two becomes the MDL) • Incorporates longer term variance • Includes checks for reasonableness • Works effectively with various quantitation limit concepts and procedures 9 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Details, details • Spiking level • 2 -10 times estimated MDL • Run spiked replicates in at least 3 separate preparation and analysis batches • Multiple instruments • At least 2 spike replicates on each instrument • If blanks give ND, MDLB does not apply • Addendum for MDL determined on a specific matrix • No 10 X rule • Use all method blanks unless batch was rejected 10 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Ongoing verification 11 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

MDL verification • Collect data quarterly • One spike per instrument (two if only one instrument) • Requirement is detection • Analyze data annually • Recalculate MDL based on last two years spike and blank data • If within factor of 3 (possibly 2) then lab option to leave unchanged or update • If outside that range then update 12 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

The TNI MDL Requirements 13 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

2009 TNI standard requirements for detection limit 2009 “appropriate and relevant for the intended use of the data “all sample processing and analysis steps shall be included” “Each target analyte” Verify with a spike at 1 -3 or 1 -4 X the LOD – detection above zero Verification is required on each instrument Verification is required annually Only required if reporting below the LOQ Repeat each time there is a change that affects the sensitivity of the analysis Copyright © 2015, Test. America Laboratories, Inc. All rights reserved. 14

What is the MDL? 15 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Lloyd Currie’s original concept LC • The lowest result that can be reliably distinguished from a blank Equals the MDL equals the TNI LOD LD • The lowest amount present in a sample that will reliably give a result that is above LC Not routinely used in environmental testing (included in the DOD QAPP) LQ • The lowest amount that gives quantitative results Conceptually, equals TNI LOQ, EPA ML and EPA LLOQ Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

MDL The method detection limit (MDL) is defined as the minimum measured concentration of a substance that can be reported with 99% confidence that the measured concentration is distinguishable from method blank results MDL 3. 14 x Standard Deviation 0 0 MDL LC 40 CFR Part 136 Currie’s Critical Level Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

What the MDL is (and is not): MDL = Lowest result that can be distinguished from blanks Or, lowest result that means there is actually something in the sample MDL ≠ Lowest amount in a sample that can be reliably detected 18 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

MDL and Currie’s LD is the minimum true concentration that is reliably detected (i. e. , gives a result above the MDL) MDL LD 1% chance of false negative 0 0 0 MDL LC DL -LD (LODDOD) 40 CFR Part 136 Currie’s Detection Level DOD Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

What does this mean regarding verification? • MDL can be verified by examining blank results • MDL cannot be verified with spiked samples • (Curries LD could be verified with spiked samples) 20 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

2015/16 TNI standard requirements for detection limit 2005/16 “reflect current operating conditions” “entire analytical process” “analytes of interest in each test method in quality systems matrix of interest” “Include data from low level spikes and routine method blanks” “Include evaluation of false positive rates” Verification includes a minimum of one low level spike and one blank per quarter per instrument Verification is only required if reporting below the LOQ One option is to follow the EPA MDL procedure Copyright © 2015, Test. America Laboratories, Inc. All rights reserved. 21

The TNI LOQ Requirements Limit of Quantitation 22 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

2009 TNI Standard LOQ • 2009 standard • Include all sample processing and analysis steps • Verify with analysis of a standard at 1 -2 times LOQ • Verify annually, but not required if the LOD was determined or verified on that instrument No way to determine precision and accuracy 23 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

2015/16 TNI Standard LOQ requirements • 2016 • At least 3 X the MDL • All processing steps • Above lowest calibration standard Verification (initial) • At least 7 spikes at of below the LOQ • 3 batches on 3 separate days • Distribute across instruments, at least 2 per instrument • Results must meet identification criteria and be above zero and within the laboratory established recovery criteria 24 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

LOQ verification • Ongoing • One verification sample on each instrument each quarter • Must meet identification criteria and be above zero • Once per year compile the data and create a statement of precision and accuracy 25 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

How Does This All Fit Together? 26 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

TNI LOQ and EPA MDL 27 MDL LOQ Initial 7 spikes 3 separate batches 3 separate days 2 per instrument Ongoing One spike per inst. per quarter (2 if only one inst. ) Meet qualitative ID, above zero Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

TNI LOQ and EPA MDL Annual calculation requirement is different: MDL • Calculate MDL(s) as Students t x Standard Deviation LOQ • Create statement of precision and accuracy We will see how this works out in the next Section 28 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Why Do we need a MDL and a LOQ? 29 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Reason #1 that we need MDLs We need to make the Quantitation limit meaningful • Applies to MRL, LLOQ, or any quantitation limit • Without a MDL, a true concentration at or close to the LOQ is probably going to be reported as a false negative In other words, without a MDL, our quantitation limit is not only not quantitatively reliable, it is not qualitatively reliable either Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

90% recovery, 9% RSD Spike #1 #2 #3 #4 #5 #6 #7 Mean MDL 10 9 8. 3 9. 8 9. 3 8. 1 8. 6 10. 0 9. 0 2. 29 9 11 Spike True Value = LLOQ 10 9 Mean recovery Reported Concentration 8 7 6 5 4 3 MDL = 2. 29 Calculated MDL 2 1 0 0 1 2 3 4 5 6 7 Replicate Number Reported results (without MDL) ND ND Copyright © 2015, Test. America Laboratories, Inc. All rights reserved. ND ND 10

Bottom Line If you don’t report below the LOQ you have a lot of false negatives You can’t minimize false negatives and false positives at the same level With the LOQ and the MDL, false negatives are controlled at the LOQ and false positives are controlled at the MDL Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Reason #2 that we need MDLs are needed in risk assessment • Handling non-detects ~ Substitute a value such as ½ detection limit or detection limit ~ More sophisticated methods such as Maximum Likelihood estimation and Regression on Order statistics − These still benefit from a detection limit as low as possible If we do not have a detection limit, the Quantitation limit will become the new Detection limit Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

How to do a LOQ / MDL study 34 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Select LOQ • Choose your LOQ • Must be at or above low calibration standard • If blanks are non zero, then calculate mean + 3 X standard deviation. This will approximate the MDL and spiking level should be at least 2 -3 times higher • Spike ideally in the range 0. 5 -1 x LOQ • 35 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Run the spiked and blank samples • Spikes • Minimum 7 spikes • Minimum 3 separate batches and three separate days ~ Prep and analytical • Minimum 2 on each instrument • Blanks • Use existing • If new method, minimum 7, at least 3 batches, at least 3 days, at least 2 per instrument 36 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Initial evaluation • Do the spike results meet qualitative identification criteria in the method? • Calculate precision and accuracy, and the MDL • Adjust the LOQ if necessary 37 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Good precision, good recovery Spike 1 10 9. 5 2 9. 8 3 4 10. 2 10. 6 MEAN STD. DEV MDL S 9. 9 0. 4 1. 3 Blanks ND ND ND MEAN STD. DEV MDL B 0. 0 3 X MDL LOQ MDL 1. 3 3. 9 10. 0 ND 5 6 7 9. 4 9. 7 9. 9 ND ND ND Success! Spikes work for calculation of the MDL And Verification of the LOQ Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Good precision, moderate recovery Spike 1 2 3 4 5 6 7 10 7 7. 3 6. 9 8. 1 7. 7 7. 3 7. 9 ND ND MEAN STD. DEV MDL S Blanks 7. 5 0. 5 1. 4 ND ND ND MEAN STD. DEV MDL B 0. 0 3 X MDL LOQ MDL 1. 4 4. 3 10. 0 Success! Spikes work for calculation of the MDL And Verification of the LOQ Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Poor/Moderate precision and recovery Spike 1 2 3 4 5 6 7 10 6 7. 3 7. 6 5. 7 7. 2 7. 9 5. 3 MEAN STD. DEV MDL S 6. 7 1. 0 3. 2 ND ND MEAN STD. DEV MDL B 0. 0 MDL 3 X MDL LOQ Blanks 3. 2 9. 7 10. 0 Success! Spikes work for calculation of the MDL And Verification of the LOQ Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Poor precision, poor recovery Spike 1 2 3 4 5 6 7 10 5 7. 1 3. 2 6. 5 7. 4 3 3. 3 ND ND MEAN STD. DEV MDL S Blanks 5. 1 1. 9 6. 1 ND ND ND MEAN STD. DEV MDL B 0. 0 MDL 3 X MDL LOQ 6. 1 18. 2 18. 0 Marginal! Spikes work for calculation of the MDL And Verification of the LOQ But LOQ must be elevated above the spiking level Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Poor precision, poor recovery Spike at ½ desired LOQ Spike 1 2 3 4 5 6 7 5 2. 5 3. 55 1. 6 3. 25 3. 7 1. 5 1. 65 ND ND MEAN STD. DEV MDL S 2. 54 0. 97 3. 04 Blanks ND ND ND MEAN STD. DEV MDL B 0. 0 MDL 3 X MDL LOQ 3. 04 9. 12 10 Success! Spikes work for calculation of the MDL And Verification of the LOQ Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Good precision, elevated blanks Spike 10 1 2 3 4 5 6 7 13. 5 12. 8 14. 2 14. 6 13. 4 13. 7 13. 9 MEAN STD. DEV MDL S Blanks 13. 7 0. 6 1. 8 3. 1 4. 2 3. 9 4. 4 3. 8 3. 2 4 MEAN STD. DEV MDL B 3. 8 0. 5 5. 3 MDL 3 X MDL LOQ 5. 3 16. 0 Marginal! Spikes work for calculation of the MDL And Verification of the LOQ But LOQ must be elevated above the spiking level Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Determine the LOQ – with 10% RSD 1. 2. 3. 4. 5. Spike at: If RSD is 10%, the standard deviation is: So MDL would be about: CHECK: Is spike concentration at least 3 x MDL? YES: MDL (3) x 3 = 9, and 10 is greater than 9. 10 1 3 Spike conc. 10 MDL Standard Deviation=1 c 0 1 2 3 4 5 6 7 8 9 10 11 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved. 12 13 14 15 16

Determine the LOQ – with 20% RSD 1. 2. 3. 4. 5. Spike at: If RSD is 20%, the standard deviation is: So MDL would be about: CHECK: Is spike concentration at least 3 x MDL? NO: MDL (6) x 3 = 18. Ten is not greater than 18. LOQ needs to be at least 18. 10 2 6 Spike conc. 10 MDL 0 1 2 3 4 5 Standard Deviation= 2 6 7 8 9 10 11 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved. 12 13 14 15 16

What do we expect if the true value is at the LOQ? Known precision? Known accuracy? Ability to detect and report? • Freedom from false negatives? Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

What do we expect from a true value at the MDL? Known accuracy? Ability to detect and report? Freedom from false negatives? We do expect freedom from false positives (99%) when the analyte is not present Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

What does this mean to labs? • • Clear requirements Sensible MDLs Level playing field Low transition costs since existing data can be used • Note – labs should start complying with 3 batch rule right now • Some additional organizational requirements 49 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

What does this mean to data users? • MDLs that make sense • Much lower rate of false positives, especially for ICP, ICPMS and some general chemistry tests • Easier to compare labs • In general, more reliable data = better decision making 50 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

How much will MDLs change? • Analytes with minimal or no detects in blanks, eg most GC/MS analytes at normal levels: Not Much • Analytes with frequent detects in blanks, eg, metals, very low level PAH, some general chemistry tests: Depends • If the lab is currently adjusting MDLs to avoid excessive false positives, not much • If the lab has been pushing MDLs below levels justified by the blanks, potentially quite a bit 51 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.

Questions? 52 Copyright © 2015, Test. America Laboratories, Inc. All rights reserved.