1. OSBI Toxicology Lab Current Technologies for Blood Testing
Robert G. Weston
Forensic Toxicologist

2. We are… Seven full-time toxicologists One part-time technician
Combined 90 years of tox experience

3. What law enforcement officers do…

4. Recent DUI caseload DUI cases submitted: 2011 – 3987 cases

5. Unlike CSI, we…
must separate the drugs from the blood through sample preparation.
set up the analysis and wait for the data to be collected.
review and interpret the data.
do not know everything about every drug seen in the blood.
determine the identity of substances we haven’t seen before.
do not have a universal test that is effective for all drugs.

6. Everyone knows how drugs get in…

7. …but how do we get them out???

8. Composition of blood Red blood cells (40-50% of blood volume)
White blood cells (1 % of blood volume)
Platelets
Plasma
Water (>92%)
Sugars
Lipids
Vitamins
Minerals
Hormones
Enzymes
Antibodies
Proteins

9. Volatiles analysis Blood placed into a vial Internal standard is added
Vial is heated
Only volatile components
evaporate into the headspace
Alcohols (ethanol, methanol, isopropyl)
Acetone
Toluene
Difluoroethane (from spray dusters)

10. Volatiles analysis A portion of the headspace gas is introduced into a
gas chromatograph
Comparison is made between
the sample and known standards
Quantification of ethanol
Identification of other volatiles

11. Enzyme-linked immunosorbent assay
ELISA
Enzyme-linked immunosorbent assay

12. ELISA Presumptive for classes of compounds
Eight classes currently tested for are
Barbiturates
Benzodiazepines
Carisoprodol/meprobamate
Cocaine/benzoylecgonine
Phencyclidine
Phenethylamines (meth)
Opiates
Tetrahydrocannabinols (marijauna)
Results must be confirmed by a chromatography/mass spectrometry method

13. At OSBI, all confirmatory testing in toxicology involves…
CHROMATOGRAPHY

15. The separation process
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62. Alkaline drug analysis
2 milliliters of blood
+internal standards
+borate buffer
+n-butyl chloride
+mixing
+centrifuging
DRUGS MOVE

63. Alkaline drug analysis
Transfer top layer to new tube
Add hydrochloric acid
Mix
Centrifuge
DRUGS MOVE

64. Alkaline drug analysis
Dispose of top layer
Add sodium hydroxide
Mix
Add chloroform
Centrifuge
Transfer chloroform to vial

65. Gas chromatography mass spectrometry

66. Why negative results? Not all drugs extract well
Drug may be present in an amount less than what our methods can detect
Some drugs metabolize very quickly
We don’t have methods to test for every drug

67. New technology for OSBI

68. LC/MS/MS Advantages Less sample needed Less sample prep (generally)
Lower limits of detection
Now capable of detecting previously unseen drugs
2.0 milliliters of blood for GC/MS testing
0.2 milliliters of blood for LC/MS/MS testing

69. Blank blood

70. New analysis capability - SPICE
Seven synthetic cannabinoids
0.5 ng/mL
AM-2201
JWH-018
JWH-073
JWH-081
JWH-122
JWH-210
JWH-250
3.0 minutes!!

71. A case sample – AM-2201 found on driver and in blood
Small amounts of other “spice” compounds.

72. What else can we do? 12 benzodiazepines Under 4 minutes
Limit of detection: 5 ng/mL

73. An real case example.

74. ELISA positive for benzos, but none found
Temazepam would be found here.

75. Now we can see…
temazepam
oxazepam

76. Why lower limits of detection?
Fewer steps in sample prep – fewer opportunities to “lose” drugs.
We can program the instrument to “see” only the drugs we are interested in.
Other drugs/chemicals present do not cause a response.

77. High concentrations of other drugs
Blank Opiates Mix

78. High concentrations of other drugs
Blank THCs Mix

79. High concentrations of other drugs
Blank Spice Mix

80. LC/MS/MS limitations Limited to analyzing what we have validated
Method validation time consuming
Not currently set up for unknown screening
Different methods for different compounds – multiple analyses required for multiple compound types
We will be able to analyze for most drugs with LC/MS/MS – it is just a matter of time

81. Upcoming projects Continue to expand capacities Quantitations Opiates
More synthetic cannabinoids
“Bath salts”
Cocaine
Quantitations