Transcript Mycotoxin Testing with Aran Gas

Aflatoxin B1 Mycotoxin Destruction using Allotropic
Oxygen, O4, O5, O6
By
Mary McPherson, Microbiologist
Scientific Analytical Institute
June 21 2012
Aran is…
Molecular Oxygen Plasma or what we call
Polyatomic Oxygen Polymer - “POP” called
AranTM
Oxygen commonly exists as an O2
molecule. The AranizerTM creates, through
energy, higher allotropes of oxygen, O4,
O5, O6, O8… These oxygen molecules
have oxidizing properties far greater than
ozone and a much longer half-life. Thus we
have a powerful oxidizer.
Reaction
5 molecules of aran
to one molecule of
mycotoxin will be
enough aran to
theoretically
breakdown the
Aflatoxin B1
Theoretical Formulation
5O5 + C17H12O6 
CO2 (vapor) + H2O (vapor)
Basic theoretical formula
Tested Agent
 7-18-05 AJ-1
Aflatoxin B1 Aspergillus flavus mycotoxin
 Chemical Formula C17H12O6 Formula Weight 312.3
The Aflatoxin B1 compound was bought from Sigma Aldrich
Company. It is a standard reagent grade mycotoxin, catalog #
A6636 neat powder.
Aspergillus species capable of producing aflatoxins
Species







Mycotoxins produced
AFB AFG CPA*
Major sources Geographical distribution
A. flavus
+
–
–
All kinds of foods Ubiquitous in warmer latitudes
A. parasiticus
+
+
–
Peanuts Specific areas
A. nomius
+
+
–
Bees USA, Thailand
A. pseudotamarii
+
–
+
Soil Japan
A. bombycis
+
+
–
Silkworm frass Japan, Indonesia
A. ochraceoroseus
+
–
–
Soil Africa
A. australis
+
+
+
Soil, peanuts Southern hemisphere
*AFB, B aflatoxins; AFG, G aflatoxins; CPA, cyclopiazonic acid
IARC MONOGRAPHS VOLUME 82
Why
 Common Health Problem – “listing was revised to known to
be human carcinogens in the Sixth Annual Report on Carcinogens in
1991.” National Toxicology Program, Department of Health and
Human Services
This is your
liver on
mycotoxin
Liver Cancer
Occurrence in foods
cereals, oilseeds, spices and tree nuts, maize, groundnuts (peanuts), pistachios, brazils,
chilies, black pepper, dried fruit and figs, milk, nonfat dry milk, cheese, yogurt, peanut
butter, eggs, meat, and just about any other food substance that is grown in warm
moist climates
Farm Related
Benefits
Field Practices
Environmental Farm Contaminant
Shipping commodities
Storage Contaminant
Disease Prevention
Still on WHY
 Chemical Formula - C17H12O6 important because this has
benzene ring and bonds that are in many other compounds that the
allotropic oxygen can breakdown such as solvents, formaldehyde, and
other biological agents.
Aflatoxin B1 History
 Aflatoxins were first identified in England in 1960’s the shorthand of its
causative agent: A. fla.10,000 turkeys and ducks died within a few months called Turkey X disease
Cause - Aspergillus flavus contamination of peanut meal
Previous to this:
Khrushchev in 1939 was faced with horses dying on farms all over
the western parts of the Ukraine. Khrushchev wrote, ‘‘I can’t
believe that science is absolutely helpless here…” ‘‘We had won more
than just a victory for our agriculture. It was a moral and political victory
as well. But how many collective farm chairmen, cattle raisers,
agronomists, animal husbandry specialists, and scientists had lost their
heads as saboteurs before I stepped in and took charge of the
situation.’’
Korneev (1948) fed white mice with oats infested with Stachybotrys
alternans strains
According to Gajdusek (1953), cases of human stachybotryotoxicosis
have been found mainly in regions where the equine disease has also
been reported.
Forgacs et al. (1958) studied the toxicity of 40 strains
The Role of Stachybotrys in the Phenomenon Known as Sick Building Syndrome EEVA-LIISA HINTIKKA Finnish Institute of Occupational Health
Uusimaa Regional Institute of Occupational Health Arinatie 3A, FIN-00370Helsinki, Finland
AJ-1
Aranizer
Glove Box
The final setup is:
an unknown
portion of
Aflatoxin B1
placed in a clean
(sterile) 250 mL
Erlenmeyer flask.
A 100 mL aliquot
of distilled water
was added to the
Erlenmeyer flask.
Cup & Tubing
Equipment Data
 AJ-1 Aranizer output 2525 mg/hr Aran gas average
molecular weight of O6 previous similar models
tested by Campbell and Associates
 Spectrophotometer 2401 Schimadzu UVProbe
190 nm – 900 nm peak analyses peaking below 190 nm
- O4 peaks at 151 nm
 PK Laboratory Analyzed the series of samples and
blanks using HPLC
Sampling
 2 - Blanks taken from the dilution water not treated
 1 aliquot of dilution water was Aranized for comparison
 2 - 10 mL aliquots were taken from the original test matrix – labeled
“Start Point”
 NOTE the Aflatoxin did not dissolve well– therefore the sampling was
done while the sample was being stirred to ensure well mixed
samples
 After 30 minutes 1 – 10 mL
 After another 30 minutes 1 – 10 mL sample was taken labeled 60
minutes
 After another 11 hours 1 – 10 mL sample was taken labeled 12 hours
Sampling Schedule
 Test start time 1:16 AM 80 mL
 Sample at
 Sample at
 Sample at
1:46 AM 70 mL (1:49AM)
2:19 AM 60 mL (2:22 AM)
2:22 PM Last sample
Mycotoxin Test Results ng/mL
30000
25000
20000
15000
Mycotoxin Test
Results ng/mL
10000
5000
0
Aranization for 30 minutes
Mycotoxin Test Results ng/mL
30000
25000
20000
15000
10000
Mycotoxin Test
Results ng/mL
5000
0
Aranization for 60 minutes
Mycotoxin Test Results ng/mL
30000
25000
20000
15000
Mycotoxin Test
Results ng/mL
10000
5000
0
Aranization for 12 hours
30 Minute Aran Treatment
Analytical results
Chemical
Mycotoxins
Aflatoxin B1
Time
Initial – ½ hr
Aran ™
2525 mL/hr
Starting Measurable ng/mL:
Ending Measurable ng/mL:
26,000 ng/mL
11,000 ng/mL
Reduced amount of “Aflatoxin B1” ng/mL
15,000 ng/mL
A reduction of about 58% in 30 minutes time.
Mary McPherson
60 minute Aran Treatment
Analytical results
Chemical
Mycotoxins
Aflatoxin B1
Time
Initial – 1 hour
Aran ™
2525 mL/hr
Starting Measurable ng/mL:
Ending Measurable ng/mL:
26,000 ng/mL
4,000 ng/mL
Reduced amount of “Aflatoxin B1” ng/mL
22,000 ng/mL
A reduction of about 85% in 60 minutes time.
Mary McPherson
12 hour Aran Treatment
Analytical results
Chemical
Mycotoxins
Time
Aran ™
Aflatoxin B1
Initial – 12 hr
2525 mL/hr
Starting Measurable ng/mL:
Ending Measurable ng/mL:
26,000 ng/mL
520 ng/mL
Reduced amount of “Aflatoxin B1” ng/mL
25,480 ng/mL
A reduction of about 98% in 720 minutes time.
Mary McPherson
References
1. Source: World of Forensic Science, ©2006 Gale Cengage
2. Herman Schuette Octozone Patent Jan. 12 1943
3. Aflatoxin Handbook USDA March 2002
4. Report on Carcinogens, Twelfth Edition (2011) Aflatoxins CAS No. 1402-68-2
5. Multimolecular Oxygen 2 Dr. Basil Earle Wainwright 2000
6. Molecular Variants of Oxygen Atoms by Basil E. Wainwright 10-23-2000
7. Code of practice for the prevention and reduction of mycotoxin contamination in cereals, including annexes on
ochratoxin a, zearalenone,fumonisins, trichothecenes CAC/RCP 51-2003
8. The chemistry and biology of aflatoxin b1: from mutational spectrometry to carcinogenesis Carcinogenesis Vol.22
no.4 pgp. 535-545, 2001
9. Diener et al., 1987 and(Maren A. Klich USDA 2002
10. Vet. Med. – Czech, 46, 2001 (6): 169–174 Potential Hazard of Simultaneous occurence of aflatoxin B1 and
ochratoxin A
11. Review of the Toxicology of Aflatoxin by W.H. Butler MRC Toxicology Unit Surrey England
12. African Journal of Biotechnology Vol. 2 (9), pp. 254-263, September 2003 Mycotoxins in food in West Africa: current
situation and possibilities of controlling it by S.A. BANKOLE* and A. ADEBANJO Accepted 29 August 2003
13. Identification of common Aspergillus species by Maren A. Klich
14. First OIE/FAO-APHCA Regional Workshop on Feed Safety - Feed borne Disease Prevention (Tokyo, Japan, 12-15
July 2010)
15. Sigma Aldrich
16. Cayman Chemical
17. IMPORTANCE OF AFLATOXINS IN HUMAN AND LIVESTOCK HEALTH Ananth S Bommakanti, and Farid
Waliyar.
18. Journal of Applied Sciences Research, 8(2): 719-726, 2012 ISSN 1819-544X Work - Related Airborne Fungi and
the Biological Levels of Mycotoxin in Textile Workers by 1Amal Saad-Hussein, 1Safaa Elserougy, 1Safia Beshir,
2Mohamed I.M. Ibrahim, 3Abdel-Hameed Awad, and 2Mosaad A. Abdel-Wahhab,
19. Food Borne Aflatoxin Hazards
20. Worldwide regulations for mycotoxins in food and feed in 2003
21. FDA and USDA materials