# Presentation: 036336

## Alternate Matrices: II. Analyzing for Opiates

- John M. Mitchell, Ph.D.
- Drug Testing Advisory Board Meeting
- 11 March 2003

## Source of Analytical Values

- 2 Urine Maintenance PT Cycles for all labs currently certified in the NLCP
- 4 Hair Pilot PT Cycles
- The 6 labs that completed all 4 cycles
- 3 Oral Fluids Pilot PT Cycles
- The 9 labs that completed all 3 cycles

## Evaluation of Analytical Values Reported for Morphine and its Metabolites

- Confirmatory testing
-  Urine,
- GC/MS
- quantitations are in ng/mL (ppm-ppb)
- Hair,
- mostly GC/MS (2 by MS/MS)
- quantitations are in pg/mg (ppb)
- for Oral Fluid
-  mostly GC/MS (2 by MS/MS)
- quantitations are in ng/mL (ppb)

## Relative Sensitivity of Assays at Cutoffs for Morphine/Codeine

**Notes:**
Urine- All labs are required to demonstrate required sensitivity prior to certification.

Hair- based on sensitivity required in one lab.  All labs may not have the same level of sensitivity.  Each lab sent 100 mg.  Some evidence that some labs may have utilized more than 10 mg for confirmational analysis.

Urine-   based on sensitivity required in one lab.  All labs may not have the same level of sensitivity.  Each lab sent 2 mL.  

All Matrices:  Sensitivities do not account for extraction efficiencies

## Relative Sensitivity of Assays at Cutoffs for 6-Acetylmorphine

**Notes:**
Better means of comparison
Comparing theoretical amount need for analysis at cutoffs
Ignoring extraction efficiency
Ignoring the amount of material placed on the MS instrument for analysis.
Using parameter from one lab from each matrix 

## Performance around cutoffs 
Graphs of reported values from each lab versus the mean value containing 
+20% and +50% quantitation ranges 
relative position of cutoff 
distribution below the cutoff 
Variance
Industry agreed-upon cutoffs
Means of reported values 
Calculated for samples with three or more values
Annotated above each histogram bar
Percent coefficient of variation
Mathematical indicator of the variation

- Graphs of reported values from each lab versus the mean value containing
- _+_20% and _+_50% quantitation ranges
- relative position of cutoff
- distribution below the cutoff
- Variance
- Industry agreed-upon cutoffs
- Means of reported values
- Calculated for samples with three or more values
- Annotated above each histogram bar
- Percent coefficient of variation
- Mathematical indicator of the variation

- Evaluation

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## Cutoff

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## Analysis of Opiate Analytes in Hair: Recommended Cutoffs

- Initial Test
- Opiate Metabolites – 200 pg/mg
- Confirmatory Test
- Morphine/Codeine – 200 pg/mg
- 6-Acetylmorphine  – 200 pg/mg

## Confirmatory Analysis of Opiates in Hair 

- NLCP Pilot PT Program Cycles 1-4
- 6 laboratories -  4 MS & 2 MS/MS
- 6 Laboratories – Morphine/Codeine/6-AM
- Hair Preparation
- Prewash + Digestion 	2 Laboratories
- Prewash + Powder    	1 Laboratory
- Prewash + Small Pieces  1 Laboratory
- Powder 			2 Laboratories
- Samples-
- Challenges in three cycles (1, 2 & 4)

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## Cutoff

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## Cutoff

## Analysis of Opiate Analytes in OralFluids: Recommended Cutoffs

- Initial Test
- Opiate Metabolites – 40 ng/mL
- Confirmatory Test
- Morphine/Codeine – 40 ng/mL
- 6-Acetylmorphine  – 4 ng/mL

## Collection Devices Utilized by Participating Laboratories

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## Cutoff

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## Urine Summary

- Confirmatory procedures provide sufficient sensitivity and precision for present analytes

## Hair Summary

- Wash procedures appear to lower opiate concentration in hair PT samples and appeared to decrease the variability for 6-acetyl morphine determinations.
- Efforts to demonstrate analytical sensitivity, precision and accuracy and to evaluate the appropriateness of cutoffs have been hampered by the limited number and variability of analytical values.
- Future PT cycles to assess inter and intra laboratory variability

**Notes:**
Unable to investigate. Insufficient values for both treatments from laboratories.

## Oral Fluid Summary

- Values obtained for neat and device treatments were equivalent.
- Most laboratories demonstrate sensitivity below the suggested opiate cutoffs
- In PT cycle 3, the variability was usually  less than 40% of the group mean for Morphine, and 30% or less for 6-AM and and 20% or less for codeine.
- Future sets to investigate issues associated with collection devices and inter and intra lab variability.