Headquarters U.S.

Air Force
Integrity - Service - Excellence

Sampling for Perchlorate

in Biota
Doris (Andy) Anders, PhD
William H. Batschelet, PhD
HQ AFCEE/ERC

Mike Wantland
Clayton Group Services

1
 Overview
 Sources of perchlorate
 Primary Oxidizer in Solid
Rockets
 Titan III and IV, Polaris,
Minuteman, Peacekeeper,
Hawk, Space Shuttles
 USA, USN, USAF, NASA

 Also occurs naturally

 Integrated Approach
 Analytical
 Health Effects
 Treatment Technology
 Ecological

I nt eg ri ty - Se rvic e - Exc el len c e 2

 Sampling

 Matrices
 Soil
 Water
 Biota

 Results used
for human and
ecological
risk assess-
ments and
cleanup

I nt eg ri ty - Se rvic e - Exc el len c e 3

 Overview
• Background
• Introduction
• Sample Preparation (water and tissue)
• Sample Clean-up
• Water Methodology
• Frog Tissue Methodology
• Validation
• Conclusions
 Background
• Bio-uptake of perchlorate in food chain
• Current data gaps tied to reporting limits
• Current tissue methods have insufficiently
low reporting limits (RL)
• Lower RL required to access “risk” to
human health and environment
 Introduction
• Develop and validate an IC/MS/MS method for
tissue that produces a lower detection/reporting
limit (RL=20 ppb in tissues)
• Existing IC Methods
– solids
– vegetation
– tissue
– water
 Sample Preparation
• Water
– Filter (Acrodisc) and analyze

• Frog tissue (whole body)

– Size reduced using knife
– Immersed in liquid nitrogen and homogenized
using commercial stainless steel blender
 Tissue Sample Cleanup

– Extract tissue aliquot in DI water

– Centrifuge
– Expose to Alumina Oxide
– Centrifuge
– Dilute sample with DI water
– Analyze
 Instrumentation
• Micromass Quattro LC
– ElectroSpray Ionization
– Instrument run in negative ion mode
 Water Methodology
• Determine levels of perchlorate (ClO4) and
other ions
– Screen on IC
– If ClO4 level <10 ppb, analyze by IC/MS/MS
Water Chromatogram
Comparison
Ion Chromatograph - 12 ppb

IC/MS/MS - 0.2 ppb

 Tissue Methodology
• Frog tissues - whole body
• Clean-up
• Analyze by IC/MS/MS
• IC/MS/MS conditions
– Utilizes Dionex AG-16 column
– Monitors Transition from 99-83 m/z
– Run time 17 minutes
Tissue Chromatogram
Comparison
Ion Chromatograph - 40 ppb

IC/MS/MS - 20 ppb
 IC/MS/MS Validation Data
• Linear range -- 0.1 to 10 ppb (r2: >0.999)
• Water Matrix Verification
– Average Recovery: 102.6%
– MDL: 0.0118 ppb
– RSD: 3.64
• Tissue Matrix Verification
– Average Recovery: 79.6%
– MDL: 1.19 µg/kg
– RSD: 2.46
 Water Verification Data
Spike ID Target Replicate Analyzed Recovery
Conc. Conc. (%)
(ppb) (ppb)
1 x LOQ 0.20 1 0.204 102.2
2 0.216 108.2
3 0.205 102.4
2.5 x LOQ 0.50 1 0.511 102.3
2 0.508 101.5
3 0.503 100.6
5 x LOQ 1.0 1 1.008 100.8
2 1.021 102.1
3 1.038 103.8
 Tissue Validation Data
Spike ID Target Replicate Anayzed Recovery
Conc. Conc (%)
(ug/kg) (ug/kg)
1 x LOQ 20 1 16.1 80.5
2 15.5 77.5
3 16.2 81.0
2 x LOQ 40 1 32.0 80.0
2 32.0 80.0
3 32.9 82.3
5 x LOQ 100 1 80.0 80.0
2 78.2 78.2
3 77.3 77.3
 Conclusions
• IC/MS/MS method is accurate, sensitive, easy
to use and capable of high sample throughput
• Instrument detection level
– IC/MS/MS: 0.020 ppb
– IC: 0.4 ppb
• Verify the extent of contamination in the low
part per billion range in various ecosystems
and specific inhabitant species
• Achieves levels meaningful to environmental
risk assessment activities
For Technical Questions Call

Mr. Kem Charron

or
Dr. Kristine Kurtz
(248) 344-1770

For General Questions Call

Mr. Mike Wantland
800-996-2050