Butane Biostimulation Technologies; AquaTane Technology

1. Global BioSciences, Inc.
“ Sustainable technologies for a better
environment ”

2. Global BioSciences, Inc.
Technologies Sold Exclusively Through
Business Partners:
• Environmental consulting, engineering
and remediation firms
• Business Partners receive generous
margins
• Several levels of partner participation &
margin

3. Business Partner Program
• GBI leases hardware and technology to
Authorized Business Partners (BPs)
• BPs add value by providing marketing,
sales, system design, project
management, O&M
• GBI provides technical support
• In addition to margins BPs project
referrals resulting from GBI marketing
• The number of BPs and current discounts
will be limited

4. Pricing Alternatives
• Panels provided exclusively on a
lease basis
• BPs provide air compressors,
butane, nitrogen, Aquatane
components, piping and installation
• Technology lease fees
• Monthly, fixed-fee or Pay-for-
Performance pricing available

5. Pilot Tests
• Site-specific Pilot Tests
recommended
• GBI support provided
• No technology lease fees for
pilot tests or bench scale
studies

6. Patents
22 U.S. patents
Many international patents*
*Remediation patents in Europe, Eurasia, Mexico, Brazil, Canada, Australia, New Zealand and Japan.

7. Global BioSciences (GBI)
Technologies for:
• Soil and groundwater remediation
• Wastewater treatment
• Odor Control
• Plant growth, nitrogen fixation
• Metal Recovery

8. GBI Remediation Technologies
Butane Biostimulation (BBT)
– Butane Biosparging
– Butane Bioventing
– Butane Biopiles
AquaTane
– Low molecular weight hydrocarbon
substrate designed for aerobic
treatment

9. GBI Remediation Technologies
• Easy to implement
• Low cost
– Up to 60% less than alternatives
• Field proven
• No residual impacts

10. Butane Biostimulation Technologies
Treats many types of contaminants
– Petroleum products
(aromatics and PAHs)
– Chlorinated solvents
– PCBs
– 1,4 - dioxane
– Others
Effective in soil and ground water

11. Why
Butane?
• Butane gas provides a sustainable carbon source to stimulate
the growth of indigenous microbial communities
• Many forms of bacteria exhibit accelerated growth under
butane stimulation
• Butane-grown bacteria also produce enzymes that rapidly
degrade toxic chemicals
• Butane is the heaviest and most soluble of the alkane gases
(propane, methane) – important for bioavailability

12. Why
Butane?
• As a gas, butane disperses and diffuses rapidly and broadly
in the subsurface – large radius of influence
• Gaseous nature makes butane uniquely suited to remediation
in low-permeability environments
• Contaminated soil in the unsaturated zone and capillary fringe
also treated
• Easily adaptable to biopiles

13. Why
Butane?
• Butane-utilizing bacteria have been shown to fix nitrogen -
able to produce their own nutrients
• Ability to treat contaminants in either aerobic or anaerobic
environments
• Simple infrastructure – easy to switch rapidly from aerobic to
anaerobic conditions at most sites
• Degradation of CVOC daughter products occurs
simultaneously

14. BBT Currently Accepted* by......
US EPA
Massachusetts - DEP
Florida - DEP
Maryland - MDE
Pennsylvania - DEP
Iowa - DNR
CETESB – Brazil
* never rejected

15. Butane Breakdown Products
CH3-CH2-CH2-CH3 (Butane)
CH3-CH2-CH2-CH2-OH (1-Butanol)
CH3CH2CH2CH=O (Butyraldehyde)
CH3CH2CH2COO- (Butyrate)
Further Metabolized

16. TCA/TCE Degradation Pathway Using
Butane Remediation
Chlorinated Ethenes (TCE Epoxidation)
CHCl=CCl2 + H2O = CHClOCCl2 + 2H+ + 2e-
Chlorinated Ethanes (1,1,1-TCA Hydroxylation)
CH3CCl3 (TCA) + H2O = CH3CCl2OH +2H+ + 2e-
Final products CO2, H2O, and chloride ions

17. MTBE Degradation Pathway Using
Butane Remediation
• MTBE = (CH3)3COCH3 / C5H12O
• Enzymes accelerate C5H12O
Demethylation
• MTBE degraded to form tert-butyl
alcohol (TBA) and formaldehyde
• These species quickly decompose
into H2O and CO2

18. Butane Safety
• Non-Toxic – Used in food processing
• Safe, UL engineered panel
• Concentrations generated in the
subsurface with the butane injector are
less than 0.01% of the LEL for butane
in air
• Excess butane in soil/groundwater
decomposes into CO2, H2O, and other
inert species in 2-3 days

19. Injection Hardware is Simple, Safe
And Code Compliant
Single injection
panel (6-14 wells)
covers ~ 1-2 acre
area
Skid built into shed to provide comfortable
work space for on-site workers

20. Butane Injection System
• Modular panel cost effective at
larger Sites
• Controlled & monitored remotely
via Internet, cellular or radio
communication
• Turn Key (shipped via UPS)
• Rugged & compact
New Model Butane Injection System

21. Delivery System Flow Diagram
Note: Activated carbon has been removed from process stream – large cost savings to client

22. Comparative Rating of
Remediation Technologies (USEPA)
Rating Codes:
Av – Average; NA – Not applicable
Butane
Biosparging™ &
Butane Chemical Soil Vapor Air Thermal Excavation
Bioventing™ Oxidation Extraction Sparging Treatment / Disposal
Low Capital Cost Yes Av Yes Yes No No
Low O&M Cost Yes No No No No NA
Fast Cleanup Time Yes Av Yes Yes Yes No
Can Treat Soil and Groundwater Yes No No No No No
Minimal Site Disturbance Yes Yes Yes Yes No No
Avoids Harsh Treatments Yes No Yes Yes No Yes
System is Highly Reliable / Easy to Maintain Yes Yes Av Yes Yes NA
Can Remediate Nonhalogenated VOCs Yes Av Yes Yes Yes Av
Can Remediate Halogenated VOCs Yes Av Yes Av Yes Av
Can Remediate Nonhalogenated SVOCs Yes No No Av Yes Av
Can Remediate Halogenated SVOCs Yes Av No Av Yes Av
Can Remediate Fuels Yes No Yes No Yes Av

23. Case Studies
• Chlorinated Solvents
• MTBE
• Petroleum Hydrocarbons

24. Butane Biostimulation of TCA/TCE
Pilot Test
Outside treatment
Inside treatment zone
zone
MW-1 MW-2 MW-7
Parameter Parameter
0 6-months 0 6-months 0 6-months
1,1,1-TCA 1,1,1-TCA
8,320 49 7,160 6.0 0 0
1,1-DCA 1,1-DCA 1,130 1,160
17,400 460 5,170 16.6
1,1-DCE
1,1-DCE 40 50
1,800 49 1,400 0.8
cis 1,2-DCE 240 230
cis 1,2-DCE 22,500 2,230 5,720 32.6
TCE 19 19
TCE 5,320 70 1,400 6.5
All concentrations in µg/L
Project in Sioux City, Iowa for General Motors Corporation

25. Butane Biostimulation of Chlorinated
Solvents
São Paulo, Brazil
CHEMICALS OF CONCERN
1,1-dichloroethane Tetrachloroethene (PCE)
toluene
1,1-dichloroethene
trans-1,2-dichloroethene
1,1,1-trichloroethane
Trichloroethene (TCE)
1,2-dichloroethane
vinyl chloride
benzene
carbon tetrachloride
chloroform
cis-1,2-dichloroethene

26. Brazil Pilot Test Data
CVOCs – Aerobic Degradation
PZ-01
Carbon tetrachloride
1000000
Total CT (Daughter
Products)
Tetracloroethene
100000
Total PCE (Daughter
Products)
Expon. (Carbon
tetrachloride)
10000
Expon. (Total CT
umol/L
(Daughter Products))
Expon. (Tetracloroethene)
1000 Expon. (Total PCE
(Daughter Products))
Expon. (Total Mass of
VOCs )
100
10
05
05
06
06
5
5
-05
5
-06
-06
-06
5
5
6
6
v-0
o-0
t-0
i-0
z-0
i-0
r- 0
t-
-
r-
-
ju l
ju l
ju n
fe v
ja n
ju n
ma
ma
se
ou
ab
ma
no
ag
de

27. Brazil Pilot Test Data
CVOCs – Aerobic Degradation
MW-G201
1000000
Carbon tetrachloride
Total CT (Daughter
100000 Products)
Tetracloroethene
Total PCE (Daughter
10000
Products)
Expon. (Carbon
tetrachloride)
umol/L
1000 Expon. (Total CT
(Daughter Products))
Expon. (Tetracloroethene)
100
Expon. (Total PCE
(Daughter Products))
Expon. (Total Mass of
10 VOCs )
1
05
06
06
05
5
5
-06
-05
5
-06
-06
6
5
5
6
v-0
o-0
t-0
i-0
i-0
z-0
r- 0
t-
r-
-
-
ju l
ju l
fe v
ju n
ju n
ja n
ma
ma
se
ou
ab
ma
no
ag
de

28. Brazil Pilot Test Data
CVOCs – Anaerobic Degradation
MW-G203
100.000 Carbon Tetrachloride
Total CT
(Daughter Products)
Tetracloroethene
10.000
Total PCE
(Daughter Products)
umol/L
Expon. (Carbon
1.000 Tetrachloride)
Expon. (Total CT
(Daughter Products))
Expon. (Tetracloroethene)
0.100
Expon. (Total PCE
(Daughter Products))
Expon. (Total VOC Mass)
0.010
-05
5
5
6
- 05
- 06
- 06
6
5
6
5
5
5
- 06
t- 0
r- 0
i -0
v -0
i -0
r -0
t- 0
z -0
o-0
jul
jun
jan
jun
fev
ma
ma
ou
se
ab
ma
no
de
ag

29. Brazil Pilot Test Data
CVOCs – Anaerobic Degradation
MW-G208
100.000
Carbon Tetrachloride
Total CT
(Daughter Products)
Tetracloroethene
10.000
Total PCE
(Daughter Products)
Expon. (Carbon
umol/L
Tetrachloride)
1.000 Expon. (Total CT
(Daughter Products))
Expon. (Total PCE
(Daughter Products))
Expon. (Tetracloroethene)
0.100
Expon. (Total VOC Mass)
0.010
-05
5
5
6
- 05
- 06
- 06
6
5
6
5
5
5
- 06
t- 0
r- 0
i -0
v -0
i -0
r -0
t- 0
z -0
o-0
jul
jun
jan
jun
fev
ma
ma
ou
se
ab
ma
no
de
ag

30. Butane Biostimulation of CVOCs
Brasil – Full Scale

31. Butane Biostimulation of CVOCs
Brasil – Full Scale

32. Sunoco Gasoline Station in Canton, MA (Closed)
Time Period Start-up +11 Months
Parameters
(ug/L) MW-3 MW-3
Methyl tert-butyl ether (MBTE) 370 52
Benzene 850 <5.0
Toluene 830 <5.0
Ethylbenzene 380 <5.0
Xylenes 2,000 <5.0
Naphthalene 110 <10
C5-C8 Aliphatic Hydrocarbons 1,900 <50
C9-C12 Aliphatic Hydrocarbons 660 <50
C9-C10 Aromatic Hydrocarbons 1,700 <50
Site closed after one year of post-remedial monitoring

33. Former Union 76 Gasoline Station in Tallahassee,
Florida
Time Period Start-up +6 Months
Parameters
(ug/L) MW-7 MW-7
Methyl tert-butyl ether (MBTE) 38 2.4
Benzene 1,900 1.6
Toluene 1,400 0.60
Ethylbenzene 900 0.51
Xylenes 4,000 1.20
Napthalene 49 0.097
System start-up date 03/4/2007

34. Sunoco Gasoline Station in Waltham, MA (Closed)
Time Period Start-up + 4 months
Parameters
VPH (ug/L) MW-4 MW-4
Methyl tert-butyl ether (MBTE) 31,800 2,330
Benzene 2,010 403
Toluene 87.2 <5.0
Ethylbenzene 867 76
Xylenes 853.8 18
Naphthalene 229 <20
C5-C8 Aliphatic Hydrocarbons <2,000 557
C9-C12 Aliphatic Hydrocarbons <750 906
C9-C10 Aromatic Hydrocarbons 5,770 1,130
Site closed after six months of post-remedial monitoring

35. Butane Biostimulation Summary
• Simple, rapid, effective for a wide variety of
environmental pollutants
• Degradation of CVOCs simultaneous
• Cost-effective and readily adaptable to large or
small clean-ups
• No soil excavation or disposal necessary (very
minimal surface disruption)
• Subsurface bacterial populations return to
background levels after process is
stopped

36. Aquatane
• A site-specific enhancement of
aerobic microorganisms
• Liquid delivery system
• Low molecular weight hydrocarbon
substrate
• Hydrogen peroxide oxygen source
• Nutrients may be added
• May degrade free product
– Likely through re-solubilization

37. Aquatane
Treats many types of contaminants
– Petroleum products
(aromatics and PAHs)
– Chlorinated solvents
– Others
Effective in soil and ground water

38. Aquatane
• Can be mixed on site
• Simple infrastructure
• Low cost
• Approved in Indiana, Tennessee,
Texas and Pennsylvania

39. Aquatane
Degradation of Chlorinated Solvents in Ground Water

40. Aquatane

41. Aquatane

42. Aquatane Injection
REMOTE SITE: SURFACE
BIOSTIMULATION INJECTION
DOWN-HOLE INJECTION FOR
WELL WITH LIMITED SURFACE
ACCESS

43. Aquatane
Bio-stimulant
Addition to Injection
Wells

44. Aquatane
Dripper System
- Gravity Flow
- Needle valves
allow large range
of flow control

45. Aquatane Biostimulation Summary
• Effective for a wide variety of environmental
contaminants
• Degradation of CVOCs simultaneous
• Continuous injection of biostimulant optimal
• Free product may be degraded
• Simple infrastructure
• Low cost

46. Global BioSciences, Inc.
Questions?
Additional technical information available on our website at:
http://www.globalbiosciences.com