Mercury As An Indicator Of Autism Spectrum Disorder

Mercury As An Indicator Of Autism Mercury As An Indicator Of Autism Spectrum DisorderSpectrum Disorder

Charles LamarCharles LamarEnvironmental Testing Laboratories/Environmental Testing Laboratories/

Freeport High School Freeport High School

Patty ElsPatty ElsEnvironmental Testing LaboratoriesEnvironmental Testing Laboratories

Edward IrwinEdward IrwinFreeport High SchoolFreeport High School

AABSTRACTBSTRACT

This study was initiated to examine if levels of excreted This study was initiated to examine if levels of excreted mercury in head hair could be used as accurate indicator of mercury in head hair could be used as accurate indicator of autistism spectrum disorder. Cold vapor atomic absorption was autistism spectrum disorder. Cold vapor atomic absorption was used to quantitatively measure the level of mercury excreted in used to quantitatively measure the level of mercury excreted in head hair. This process varies from traditional techniques in that head hair. This process varies from traditional techniques in that a flame is not used to vaporize the sample and the Perkin-Elmer a flame is not used to vaporize the sample and the Perkin-Elmer Flow Injection Mercury system used in this study is a dedicated Flow Injection Mercury system used in this study is a dedicated mercury analysis system.mercury analysis system.

AABSTRACTBSTRACT

Hair samples were taken form both autistic and non-Hair samples were taken form both autistic and non-autistic volunteers. The parent of each autistic volunteer cut a autistic volunteers. The parent of each autistic volunteer cut a small locket of hair from the back of the head. This was small locket of hair from the back of the head. This was accomplished in the student’s home during a regularly accomplished in the student’s home during a regularly scheduled hair cut without any disturbance in the child’s scheduled hair cut without any disturbance in the child’s environment or routine. In addition, individual volunteers will environment or routine. In addition, individual volunteers will be followed for mercury excretion over a period of time. be followed for mercury excretion over a period of time.

GGOALOAL

One of the major goals of this study is One of the major goals of this study is to design a new method of diagnosing to design a new method of diagnosing Autism Spectrum Disorder.Autism Spectrum Disorder.

PPROBLEMSROBLEMS

There are three primary research questions being There are three primary research questions being addressed by this study:addressed by this study:

1) Is cold vapor atomic absorption capable of quantitatively 1) Is cold vapor atomic absorption capable of quantitatively measuring the level of mercury excreted in head hair?measuring the level of mercury excreted in head hair?

2) Can levels of excreted mercury in head hair be used as an 2) Can levels of excreted mercury in head hair be used as an accurate indicator of autism spectrum disorder?accurate indicator of autism spectrum disorder?

3) Does the excretion of mercury through head hair fluctuate? 3) Does the excretion of mercury through head hair fluctuate? Furthermore, does it fluctuate in a cyclical or seasonal pattern?Furthermore, does it fluctuate in a cyclical or seasonal pattern?

IINTRODUCTIONNTRODUCTION

Elemental mercury is Elemental mercury is found as a liquid at room found as a liquid at room temperature. Itemperature. It’s compounds and t’s compounds and vapors are considered highly vapors are considered highly toxic. It is often used in a number toxic. It is often used in a number of industrial processes including of industrial processes including the production of batteries, the production of batteries, thermometers, florescent lamps, thermometers, florescent lamps, barometers, and thermostats. barometers, and thermostats. However, due to widespread However, due to widespread health concerns, toxic use health concerns, toxic use reduction efforts are attempting reduction efforts are attempting to cut back or eliminate mercury to cut back or eliminate mercury in said products.in said products.


Ethylmercury is a Ethylmercury is a bioaccumulative organic bioaccumulative organic compound. It is composed of an compound. It is composed of an ethyl group and a mercury ethyl group and a mercury atom.atom. Ethylmercury constitutes Ethylmercury constitutes 49% of thimerosal, a 49% of thimerosal, a preservative formerly used in preservative formerly used in vaccines. In infants it has been vaccines. In infants it has been suggested that thimerosal in suggested that thimerosal in vaccines could contribute to the vaccines could contribute to the cause neurodevelopmental cause neurodevelopmental disorders, most notably, Autism disorders, most notably, Autism Spectrum Disorder. Spectrum Disorder.


MMethylmercuryethylmercury is is composed of a methyl group composed of a methyl group bonded to a mercury atom. It is bonded to a mercury atom. It is introduced into the environment introduced into the environment mainly through the burning of mainly through the burning of fossil fuels. Large fish like fossil fuels. Large fish like marlin, swordfish and tuna tend marlin, swordfish and tuna tend to have a high concentration to have a high concentration and are the largest dietary and are the largest dietary source. Once ingested by source. Once ingested by humans, the chemical is readily humans, the chemical is readily absorbed by the gastrointestinal absorbed by the gastrointestinal tract. tract.


Autism or, Autism Autism or, Autism Spectrum Disorder (ASD), is a Spectrum Disorder (ASD), is a disability resulting from a disability resulting from a neurological disorder that affects neurological disorder that affects the normal development of the the normal development of the brain. ASD is one of the five brain. ASD is one of the five Pervasive Developmental Pervasive Developmental Disorders that are characterized Disorders that are characterized by “severe and pervasive by “severe and pervasive impairment in several areas of impairment in several areas of development”. ASD is the most development”. ASD is the most prevalent of the five disorders prevalent of the five disorders occurring in approximately 1 in occurring in approximately 1 in 166 births. 166 births.


The most common The most common symptoms of ASD include symptoms of ASD include difficulties in the areas of difficulties in the areas of verbal communication, verbal communication, social interactionsocial interaction. . Repetitive motion and self-Repetitive motion and self-injurious behavior are also injurious behavior are also common.common.


Most cases of ASD Most cases of ASD are treated with are treated with Methylphenidate. Methylphenidate. Methylphenidate is a mild Methylphenidate is a mild nervous system stimulant nervous system stimulant sold under the brand name sold under the brand name Ritalin in North America. Ritalin in North America.


Chelation therapy has Chelation therapy has also been used in the treatment also been used in the treatment of ASD. Chelation therapy is a of ASD. Chelation therapy is a series of intravenous infusions series of intravenous infusions containing EDTA containing EDTA (ethylenediaminetetraacetic) (ethylenediaminetetraacetic) and various other substances. and various other substances. Heavy metals are bound and Heavy metals are bound and removed from the bloodstream. removed from the bloodstream. Clinical trials thus far have Clinical trials thus far have yielded mixed results. yielded mixed results.


Recent scientific developments suggest that there Recent scientific developments suggest that there exists a direct link between the levels of mercury exists a direct link between the levels of mercury exposure a person receives during early development, exposure a person receives during early development, and their chances of developing ASD. It is believed, and their chances of developing ASD. It is believed, that when mercury enters the body, it binds to that when mercury enters the body, it binds to sulfhydryl groups on enzymes and other proteins. sulfhydryl groups on enzymes and other proteins. When the mercury enters the brain and spinal chord, it When the mercury enters the brain and spinal chord, it affects the uptake of dopamine, serotonin, affects the uptake of dopamine, serotonin, acetylcholine, and norepinephrine. This is what is acetylcholine, and norepinephrine. This is what is believed to cause the lack of motor coordination and believed to cause the lack of motor coordination and difficulty with communication that is seen in autistic difficulty with communication that is seen in autistic patients. patients.

PPROCEDUREROCEDURE

MEHTOD ONE: Sample CollectionMEHTOD ONE: Sample CollectionInformed consent is obtained from a Informed consent is obtained from a legal guardian. The hair samples from legal guardian. The hair samples from the autistic volunteers are collected the autistic volunteers are collected during a regularly scheduled haircut during a regularly scheduled haircut without any disturbance to the child’s without any disturbance to the child’s environment or routine. A locket of environment or routine. A locket of hair is cut, not pulled, from the back hair is cut, not pulled, from the back of the head. Data regarding the of the head. Data regarding the participant’s age, gender, and any participant’s age, gender, and any medical treatment is recorded. The medical treatment is recorded. The hair is stored in a Ziploc bag to reduce hair is stored in a Ziploc bag to reduce the risk of contamination. Digestion is the risk of contamination. Digestion is typically conducted within seven days. typically conducted within seven days.

PPROCEDUREROCEDUREMETHOD TWO: Sample DigestionMETHOD TWO: Sample Digestion

0.6g Sample0.6g Sample

↓↓+5mL Reagent Grade Water+5mL Reagent Grade Water

+3mL HCl+3mL HCl+2mL HNO+2mL HNO33

2min at 95°C2min at 95°CCool to Room TemperatureCool to Room Temperature

↓↓+50mL Reagent Grade Water+50mL Reagent Grade Water

+15mL 5% KMnO+15mL 5% KMnO44

30 min at 95°C30 min at 95°CCool to Room TemperatureCool to Room Temperature

↓↓+6mL Sodium+6mL Sodium

ChlorideChlorideHydroxylamine Hydroxylamine

SulfateSulfate

↓↓Analysis Within 28 DaysAnalysis Within 28 Days

PPROCEDUREROCEDURE

METHOD THREE: Mercury METHOD THREE: Mercury Analysis Analysis Mercury concentration is Mercury concentration is measured in parts per million by measured in parts per million by atomic spectroscopy using the atomic spectroscopy using the Perkin Elmer FIMS. 3% HCl is Perkin Elmer FIMS. 3% HCl is used as a carrier solution. Argon used as a carrier solution. Argon is used as a carrier gas. is used as a carrier gas. Measurements are reported in Measurements are reported in μg/g using a formula that takes μg/g using a formula that takes into account the mass, volume, into account the mass, volume, and absorbance reading of the and absorbance reading of the sample.sample.

RRESULTSESULTS

Question: Can Cold Vapor Atomic (CVAA) Question: Can Cold Vapor Atomic (CVAA) Absorption be used to quantitatively measure the Absorption be used to quantitatively measure the level of mercury in head hair?level of mercury in head hair?

RRESULTSESULTSSummary RobU / Control MaleMean = 0.769 Std Dev. = 0.1037

(Values are +/- 1 Std. Dev.)

0

0.2

0.4

0.6

0.8

1

1.2

1 2 3 4 5 6 7 8 9 10

Individual Extractions

ug

Hg

pe

r g

ram

of

ha

ir

RRESULTSESULTS

Conclusion: Results suggest that CVAA is capable Conclusion: Results suggest that CVAA is capable of quantitatively measuring mercury excreted in of quantitatively measuring mercury excreted in head hair.head hair.

RRESULTSESULTS

Question: Can the concentration of excreted Question: Can the concentration of excreted mercury in head hair be used as an accurate mercury in head hair be used as an accurate indicator of ASD?indicator of ASD?

RRESULTSESULTS

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

ROBU

DJI (Control Male)

KFI (Autistic Male)

RRESULTSESULTS

Conclusion: Preliminary results from one autistic Conclusion: Preliminary results from one autistic individual suggests that levels of excreted mercury individual suggests that levels of excreted mercury in head hair can be used as an accurate indicator of in head hair can be used as an accurate indicator of ASD.ASD.

RRESULTSESULTS

Question: Does the excretion of mercury through Question: Does the excretion of mercury through head hair fluctuate? Furthermore, does it fluctuate head hair fluctuate? Furthermore, does it fluctuate in a cyclical or seasonal pattern?in a cyclical or seasonal pattern?

RRESULTSESULTSMonthly Haircuts: ETI

Mean = 12.756 Std. Dev. = 2.281(values are +/- 1 std dev. )

024

68

101214

161820

Sept. 05 Oct. 05 Nov. 05 Dec.05 Feb. 06 Mar. 06 Apr. 06 Jun. 06 Jul. 06

Haircut Date

ug

Hg

per

gra

m o

f h

air

RRESULTSESULTS

Conclusion: In the further pursuit of this question, Conclusion: In the further pursuit of this question, additional samples will be collected and additional samples will be collected and participants will be followed for analysis over participants will be followed for analysis over longer periods of time.longer periods of time.

DDISCUSSIONISCUSSION

The results show that the control The results show that the control participants were better able to excrete participants were better able to excrete mercury through their hair follicles. This mercury through their hair follicles. This supports the hypothesis that persons with supports the hypothesis that persons with ASD developed the disorder not due level ASD developed the disorder not due level of exposure, but rather their inability to of exposure, but rather their inability to release the mercury. release the mercury.

FFUTURE UTURE WWORKORK

New samples will be collected and New samples will be collected and analyzed. Additional aspects of the study will be analyzed. Additional aspects of the study will be explored in order to answer the underlying explored in order to answer the underlying questions. Participants will be followed over a questions. Participants will be followed over a period of time to examine possible cyclical period of time to examine possible cyclical patterns. Ten male and ten female autistic patterns. Ten male and ten female autistic volunteers will be recruited for that purpose. This volunteers will be recruited for that purpose. This new data will be further examined to find a new data will be further examined to find a possible correlation to hormonal imbalances. possible correlation to hormonal imbalances.

RREFERENCESEFERENCES DiCicco-Bloom, Emanuel et al. DiCicco-Bloom, Emanuel et al. The Developmental The Developmental

Neurobiology of Autism Spectrum DisorderNeurobiology of Autism Spectrum Disorder. The Journal . The Journal of Neuroscience, Jun, 2006of Neuroscience, Jun, 2006

Moldin, Steven O. et al. Moldin, Steven O. et al. Can Autism Speak to Can Autism Speak to NeuroscienceNeuroscience? The Journal of Neuroscience, Jun, 2006? The Journal of Neuroscience, Jun, 2006

Boaventura GR et al. Boaventura GR et al. Multivessel System For Cold-vapor Multivessel System For Cold-vapor Mercury Generation. Determination Of Mercury In Hair Mercury Generation. Determination Of Mercury In Hair And FishAnd Fish. Biological Trace Element Research, Oct-Nov, . Biological Trace Element Research, Oct-Nov, 1997 1997

Pineau A et al. Pineau A et al. Determination Of Total Mercury In Human Determination Of Total Mercury In Human Hair Samples By Cold Vapor Atomic Absorption Hair Samples By Cold Vapor Atomic Absorption SpectrometrySpectrometry. Journal of Analytical Toxicology, Jul-Aug, . Journal of Analytical Toxicology, Jul-Aug, 19901990

AACKNOWLEDGMENTSCKNOWLEDGMENTS

Dr. Sat Bhattacharya Dr. Sat Bhattacharya Harlem Children SocietyHarlem Children Society MSKCCMSKCC Edward IrwinEdward Irwin Patty ElsPatty Els Juan CubaJuan Cuba Kristin CookKristin Cook Ruth LawrenceRuth Lawrence

THANK YOU!THANK YOU!

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Mercury As An Indicator Of Autism Spectrum Disorder