4/3/2014
®
We will start momentarily at 2pm ET
All recordings will be available to only ACS Members
http://acswebinars.org/detecting-toxins Contact ACS Webinars ® at
[email protected] 1
Have Questions? Type Questions Box! “Why am I muted?” Don’t worry. Everyone is muted except the presenter and host. Thank you and enjoy the show.
Or tweet using #acswebinars All recordings will be available to only ACS Members
http://acswebinars.org/detecting-toxins Contact ACS Webinars ® at
[email protected] 2
1
4/3/2014
®
®
If you enjoy ACS Webinars every Thursday… please support the program!
www.join.acs.org ACS Network (search for group acswebinars) www.communities.acs.org
Find the many benefits of ACS membership! 3
www.facebook.com/acswebinars 4
2
4/3/2014
ACS Sci-MindTM has an incredible group of experts ready and willing to help you solve your technical challenges. Check out what Sci-MindTM has to offer at www.sci-mind.org
5
Q: “Hungry for a brain snack?” A:
on TM
“ACS Webinets are 2 minute segments that bring you valuable snippets from some of our most popular full length ACS Webinars® ” TM
See all of our ACS Webinets on YouTube at http://bit.ly/acswebinets 6
3
4/3/2014
How has ACS Webinars benefited you?
®
“ACS Webinars give me a justifiable outlet for my mind to wander while I'm at work! Sometimes the best ideas come to you while you let your mind run free.”
Fan of the Week Trisha H. Miller, Ph.D. Systems Research and Analysis Sandia National Laboratories
Be a featured fan on an upcoming webinar! Write to us @
[email protected] 7
®
Beginning in 2014 all recordings of ACS Webinars will be available only to current ACS members. We appreciate your patience while we work to complete the migration of past and current episodes, which we hope to have available as soon as possible. Live weekly ACS Webinars will continue to be available to the general public. Contact ACS Webinars ® at
[email protected] 8
4
4/3/2014
Upcoming ACS Webinars® www.acs.org/acswebinars. Thursday, April 10, 2014
“The Chemistry of Cocktails: Bruising and Louching and Fire Oh My!” Darcy J. Gentleman, Ph.D., Science Communicator, ACS Office of Public Affairs Kathryn Verona, ACS Office of Public Affairs
Thursday, April 17, 2014
“Red and White Beer for St. George’s Day” Dr. Charlie Bamforth, UC Davis Bill Courtney, Cheese-ology Macaroni & Cheese
Contact ACS Webinars ® at
[email protected] 9
®
Detecting Human Exposure to Toxins
Elizabeth Hamelin Centers for Disease Control and Prevention
Lucas Zarwell DC Office of the Chief Medical Examiner
Recordings will be available to only ACS Members
http://acswebinars.org/detecting-toxins Contact ACS Webinars ® at
[email protected] 10
5
4/3/2014
?
Chronic Exposure
Acute Exposure
Symptoms
dose
effect
Case Study: Symptomology
Case 1 Severe gastrointestinal illness
Case 3 Severe vomiting
Case 2 Dizziness Tingling of lips Floating sensation
Case 4 Large blisters on arm
6
Observations
4/3/2014
Unique marker Only a few sources
Visual cues Questions & answers
Diagnosis
General marker Impacted by many sources
Why do we need a specific test? Identify toxin Evaluate long-term health effects Determine exposed vs. unexposed Determine geographical distribution
7
4/3/2014
Case Study: Toxins Saxitoxin
Ricin
Abrin
Sulfur Mustard, toxicant
Absorption Ingestion Inhalation Injection Absorption Toxication
Distribution Reabsorption
Elimination Distribution Excretion Detoxification 16
8
4/3/2014
Toxicant Excretion Specific enzyme
10 Concentration (arbitrary)
In vitro
Animal studies
In vivo
Toxic Compound or Marker
8
Metabolite
6 4 Adduct
2 0
Exposure events
Human exposure
0
50 100 Time (hours)
150
200
What? * Where? * How much? * How long?
Method Application
3.0e6
Intensity, cps
2.4e6
Symptomatic Identify compound in poisoning scenarios
1.8e6
Asymptomatic
1.2e6
Differentiate between exposed/unexposed
6.0e5
0.0
2.8
3.0
3.2
3.4
3.6
3.8
4.0
4.2
4.4
4.6
4.8
Time, min
9
4/3/2014
Analytical method development Sample preparation
Matrix
Separation
Detection
Dilute and shoot
IS
SPE
Evaporation
Reconstitution
HPLC Matrix
MS
IS
Online SPE Matrix IS
Challenges: Potential False Positives
Similar compounds 2-CHA metabolite from riot control agent CS 4-CHA metabolite from pharmaceutical
Matrix components
Analyte
Urinary component at same transition
Matrix
10
4/3/2014
Challenges: Potential False Negatives
Extraction efficiency
Average Extraction Efficiencies
120% 100% Strata X
80%
WAX 60%
SAX ZrO2
40%
Si
20% 0% Water
Matrix effects
Urine
Serum
3.5e5
In Solvent
3.0e5
In Matrix
2.5e5 Intensity, cps
2.0e5 1.5e5 1.0e5 5.0e4 0.0 0.0 0.2
0.4
0.6 0.8
1.0 1.2
1.4 1.6 1.8 Time, min
2.0
2.2 2.4
2.6 2.8
Audience Trivia Question What is the source of the toxin ricin?
Tree Bark Rosary Peas Castor Beans Bamboo Rice Silver Beatles
11
4/3/2014
Case 1: Ricin Suicide Attempt 2005, California 58-year-old homeless man, chewed 6 castor seeds
LD50 Inhalation or injection: 5-10 mg/kg Higher for ingestion
Ricin
Ricinine
Ricinine method % Ricin + % RCA % Ricinine -
5% of castor bean 20% of ricin content 1% of castor bean
Ricin toxicity = 5.0 mg/kg Method Performance QL
QH
Precision
14%
0.78%
Accuracy
106%
102%
Lowest Reportable
0.03 ng/mL
12
4/3/2014
Castor Bean Ingestion Analysis of Ricin Biomarker in urine 800
ug Ricinine/g Creatinine
~ 6 Castor Seeds 700 600 500 400 300 200 100 0
20
40
60
80
Time after ingestion (hrs)
Ricinine: Other Sources
Present in cosmetics, oils, and pharmaceuticals 12 positive of 989 urine samples (1.2%)
Ricinine (ng/mL)
6 4 2 0 1
3
5
Sample
7
9
11
Results ranged from 0.186-4.15 ng/mL
Must take analytical results in context with symptomology
13
4/3/2014
Case 2: Saxitoxin poisoning from shellfish 2011, Alaska Two persons present with dizziness, shortness of breath, nausea, floating sensation following ingestion of mussels, which they had harvested in the wild
Happy Clams
Red Tide w/Toxins
Clams w/Toxins
Captured Clams
Happy People Consume Clams
Unhappy People w/Toxins
Artwork by Ken Hsuan Liu
Saxitoxin in Urine XIC of +MRM (5 pairs): 300.100/282.100 Da from Sample 13 (Curve 6_RR_13) of Curve 6.wiff (Turbo Spray)
Max. 4.3e5 cps.
3.26
Saxitoxin
4.2e5 4.0e5 3.8e5 3.6e5 3.4e5 3.2e5 In te n s ity , c p s
3.0e5 2.8e5 2.6e5 2.4e5 2.2e5 2.0e5 1.8e5 1.6e5 1.4e5
Internal Standard
1.2e5 1.0e5 8.0e4 6.0e4 4.0e4 2.0e4 0.0
2.2
2.4
2.6
2.8
3.0
3.2
3.4
3.6 Time, min
3.8
4.0
4.2
4.4
4.6
4.8
QL
QH
Precision
8.0%
5.5%
Accuracy
99.3%
99.6%
Lowest Reportable
4.8 ng/mL
14
4/3/2014
Audience Trivia Question
About how much water is in urine? 73% 80% 84% 92% 95%
Southeast Alaska Outbreak May – June 2011 Patient
A B C D E F
Symptoms Ataxia, difficulty swallowing, dizziness, difficulty moving, floating sensation, nausea, paresthesia, shortness of breath, weaknes Paresthesia Paresthesia Ataxia, dysphagia, floating sensation, paresthesia, weakness Paresthesia Ataxia, dysphagia, floating sensation, paresthesia, shortness of breath, weakness
Time from consumption Type of shellfish to symptom consumed onset
Toxin level in shellfish Toxin level consumed (µg in urine Hospitalized saxitoxins/100 g of (ng/mL) meat*)
3.5--4 hrs
Cockles
528
N/A
Yes (ICU)
10--15 min 2.5--3 hrs
Cockles Cockles
528 528
N/A N/A
No No
