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

WhyCommon 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

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

This is your liver on mycotoxin

Farm Related Benefits

Environmental Farm Contaminant

Storage Contaminant

Field Practices

Shipping commodities

Disease Prevention

Still on WHYChemical 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’tbelieve 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 Stachybotrysalternans strainsAccording to Gajdusek (1953), cases of human stachybotryotoxicosishave been found mainly in regions where the equine disease has alsobeen reported. Forgacs et al. (1958) studied the toxicity of 40 strainsThe 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

Glove BoxThe 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.

AJ-1 Aranizer

Cup & Tubing

Equipment DataAJ-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

Sampling2 - 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

minutesAfter another 11 hours 1 – 10 mL sample was taken labeled 12 hours

Sampling ScheduleTest start time 1:16 AM 80 mL Sample at 1:46 AM 70 mL (1:49AM)Sample at 2:19 AM 60 mL (2:22 AM) Sample at 2:22 PM Last sample

Reduction of Mycotoxin using Allotropic Oxygen

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Aranization for 30 minutes

Reduction of Mycotoxin using Allotropic Oxygen

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Aranization for 60 minutes

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Aranization for 12 hours

30 Minute Aran TreatmentAnalytical resultsChemicalMycotoxins Time Aran ™ Aflatoxin B1 Initial – ½ hr 2525 mL/hr

Starting Measurable ng/mL: 26,000 ng/mLEnding Measurable 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 TreatmentAnalytical results

Chemical

Mycotoxins Time Aran ™

Aflatoxin B1 Initial – 1 hour 2525 mL/hr

Starting Measurable ng/mL: 26,000 ng/mL

Ending Measurable 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

Analytical resultsChemicalMycotoxins Time Aran ™

Aflatoxin B1 Initial – 12 hr 2525 mL/hr

Starting Measurable ng/mL: 26,000 ng/mLEnding Measurable 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

12 hour Aran Treatment

References1. Source: World of Forensic Science, ©2006 Gale Cengage2. Herman Schuette Octozone Patent Jan. 12 19433. Aflatoxin Handbook USDA March 20024. Report on Carcinogens, Twelfth Edition (2011) Aflatoxins CAS No. 1402-68-25. Multimolecular Oxygen 2 Dr. Basil Earle Wainwright 20006. Molecular Variants of Oxygen Atoms by Basil E. Wainwright 10-23-20007. Code of practice for the prevention and reduction of mycotoxin contamination in cereals, including annexes on

ochratoxin a, zearalenone,fumonisins, trichothecenes CAC/RCP 51-20038. The chemistry and biology of aflatoxin b1: from mutational spectrometry to carcinogenesis Carcinogenesis Vol.22

no.4 pgp. 535-545, 20019. Diener et al., 1987 and(Maren A. Klich USDA 200210. Vet. Med. – Czech, 46, 2001 (6): 169–174 Potential Hazard of Simultaneous occurence of aflatoxin B1 and

ochratoxin A11. Review of the Toxicology of Aflatoxin by W.H. Butler MRC Toxicology Unit Surrey England12. 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 200313. 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 Aldrich16. Cayman Chemical17. 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 Hazards20. Worldwide regulations for mycotoxins in food and feed in 200321. FDA and USDA materials