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PURE ONE PetroXclean

PROVIDES THE NATURAL BIOREMEDIATION SOLUTION


INTRODUCTION

PURE ONE ENVIRONMENTAL PRODUCTS is a manufacture of a family of environmentally safe products and is proud to offer this product for the cleanup of petroleum-based contaminants in soil, ground water and marine environments. PURE ONE ENVIRONMENTAL PRODUCTS offers a unique remediation technology that is unmatched in the environmental market. PetroXclean, has been tested and proven in use in many countries around the world, with outstanding and unequaled results. PetroXclean, a propriety formulation of enzymes and organic nutrients that enhances bioremediation processes, is scientifically sound, environmentally friendly and very competitive in cost-effectiveness.


ENHANCED BIOREMEDIATION: THE PetroXclean ADVANTAGE

PetroXclean is a multifaceted bioremediation enhancing agent which is highly effective, simple to use, cost-effective and completely non-toxic. PetroXclean is a complex non-bacterial organic formulation, designed to enhance bioremediation of a wide range of petroleum-based contaminants in soil and water. It contains no bacteria nor genetically engineered microbes, but rather it stimulates indigenous bacteria to degrade such products.

Tests have now confirmed that the PetroXclean-enhanced bioremediation of petroleum-contaminated soil has resulted in concentrations of petroleum constituents below regulatory cleanup requirements in multiple field projects totaling more than 2,000,000 tons of contaminated soil. the ultimate measure of success has been the rapid disappearance of both target and non-target contaminants without the buildup of toxic by-products following PetroXclean-enhanced bioremediation.

PetroXclean technology, while elegantly simple, is scientifically sound and highly cost-effective. It is environmentally friendly, non-toxic, and has been approved for use by many governmental agencies. all ingredients of PetroXclean are organic in origin and biodegradable.


PetroXclean MECHANISM OF ACTION

Soil and its complex microbial community can accept and degrade to some degree a variety of petroleum-based pollutants into non-toxic forms. There exists in nature microbial forms (i.e., bacteria, yeast, molds, etc.) which have the capability to biodegrade many petroleum-based agents which are noxious to the environment. these microbes have the capability to produce extracellular enzymes which lead to the breakdown (mineralization) of the petroleum compounds, which transforms them into food sources for the microbe’s own carbon dioxide (CO2), water (H20), and biomass (fatty acids which are used for a food source by other microorganisms).

The biodegradation of petroleum-based compounds can be accomplished both in oxygen-rich (aerobic) and in oxygen-poor (anaerobic) environments. The aerobic mineralization of contaminants is much faster and more complete than the anaerobic process. However, both aerobic and anaerobic treatments may be necessary in some cases.

At sites contaminated with petroleum-based compounds, a small percentage of the overall bacterial population is capable of utilizing the petroleum compounds as a food source for energy and growth. This limited bacterial population, combined with a lack of enhanced stimulation, is what limits most normal bioremediation projects. With the PetroXclean stimulation, the petroleum-consuming microbes are favored in the biological chain and increase in number proportional to those without that special capability. PetroXclean acts in a three-stage process to enhance bioremediation. In the first stage, native soil bacterial populations in a petroleum-Rich environment are stimulated with the PetroXclean growth enhancement substrate. At the same time, PetroXcleanS biodegradable digestive enzymes (nature’s catalysts) are provided to break down the petroleum complexes into more digestible fragments. the presence of the petroleum break-down products stimulated more of the native soil bacterial to "switch" and begin to consume petroleum compounds.

In the second stage, the enlarged native bacterial population undergoes natural bioselectivity. those bacteria capable of utilizing petroleum compounds as their natural "food source" are presented with a rich source of digestible petroleum compounds to continue their growth.

The third stage of PetroXclean enhancement is one in which PetroXclean is applied at regular intervals to continually stimulate the selective growth of petroleum-consuming bacteria and to provide surplus enzymes to break down the petroleum compounds for bacterial digestion. By adding the PetroXclean enhancers during this stage, the natural Gaussian decline in bacteria populations is curtailed until all of the available petroleum complex has been reduced. Once the petroleum compounds have been totally scavenged, the petroleum-consuming bacteria rapidly decline to their natural population in the soil matrix. The following describes in more detail how PetroXclean interacts in petroleum-affected soil and/or water by catalyzing the natural biochemical reactions:


MICROBIAL GROWTH

While microbes are capable of utilizing petroleum hydrocarbons as an energy source, they preferentially utilize simple sugars (glucose) for rapid growth and energy. Other growth factors required by some microbes include amino acids, purines and pyrimidines. PetroXclean provides a rich source of these simple sugars, amino acids, and the other related growth factors. Once the glucose energy source is exhausted, those microbes capable of utilizing petroleum for such growth are naturally favored and their absolute, as well as relative, numbers increase dramatically.

The microbial growth phase is characterized by an initial lag phase while the DNA and enzyme phase develops. The extracellular enzymes necessary to break down organic molecules for microbial growth are readily available in PetroXclean, shortening the lag phase. As soon as the basic requirements for enzyme and DNA production are met, the soil microbes enter a phase where the exponential growth develops. During this rapid growth phase, microbes are particularly sensitive to adverse conditions, and thus respond well with the presence of PetroXclean. By continually adding fresh nutrients, microbial growth is pulsed applications, as discussed further below, and monitoring of oxygen, water saturation and microbial activity is so important.


ENZYME INDUCTION

PetroXclean provides a rich mixture of extracellular enzymes capable of initiating and catalyzing the breakdown of a wide variety of petroleum compounds. Extracellular enzymes, which initiate the metabolic (oxidative) process for biodegradation of petroleum compounds are produced by certain microbes. In the first step of such oxidation, these extracellular enzymes instantaneously break off two carbon units from the saturated hydrocarbon chains, which are typical of most petroleum compounds. The transformed petroleum molecule is then released from the enzyme, which is then freed to react with other petroleum molecules are transferred into the microbe for its own metabolic process. The microbe "reads" from whence the food source originated and alters its own enzyme production to attack the identified food source (in this case the petroleum molecules) directly. This is called "enzyme induction". Such enzyme induction in the enlargedsoil bacterial community results in an exponential increase in petroleum bacterial growth.


CO-METABOLISM AND SECONDARY SUBSTRATE ACTIVITY

PetroXclean further enhances the cleanup process through co-metabolism and secondary substrate activity. Co-metabolic breakdown occurs as a result of a non-target compound being in the presence of the high energy metabolic field created with PetroXclean. elaborated during the metabolism of petroleum compounds are capable of altering non-target organic compounds and transforming them into a more readily oxidizable and non-toxic form which can then be metabolized by the soil microbial consortium. As the target contaminant decreases below a critical concentration, microbial populations decrease and the reaction slows or stops completely. This explains why most bioremediation soil venting operations cease to be effective as the petroleum compound concentrations decrease. By maintaining a rich microbial population with PetroXclean’S carbon and food sources, even small residuals of petroleum compounds continue to be metabolized to very low concentrations. This secondary substrate enhancement by the use of PetroXclean allows for a cost-effective treatment of large volumes of petroleum-affected soil with low hydrocarbon concentrations.


MOBILIZATION OF PETROLEUM COMPOUNDS FOR BETTER MICROBIAL ACTION

PetroXclean contains organic biodegradable bioemulsifiers (surfactants) to break down the adsorption of petroleum onto soil particles. This allows the hydrocarbons to move more freely through the soil pores where the less mobile microbial community lives. the bioemulsifiers break down macroscopic clumps of petroleum into smaller units with increase the surface area for microbial effect. The surfactant action also helps the nutrients in PetroXclean migrate through the soil matrix more easily.


OPERATIONAL OVERVIEW

There are multiple factors which determine the speed and completion of the breakdown of petroleum compounds in both soil and water. The acceleration of the naturally occurring microorganic metabolic and enzyme production process is accomplished by way of delivering to the naturally occurring bacteria present in the soil an optimal environment and food source. The various components necessary to this process include:

The environmental factors must be continually monitored throughout the treatment process in order to create the optimal breakdown process. Addressing all of the above variables provides a sound basis for a successful project. PURE ONE ENVIRONMENTAL PRODUCTS provides technical guidance for specific sites undergoing consideration of PetroXclean - enhanced bioremediation treatment.


ENGINEERING MANAGEMENT OF BIOREMEDIATION

Under normal PetroXclean enhanced soil remediation operations, a "wavefront" of PetroXclean and water is moved through the soil in a weekly pulsed fashion under aerobic conditions. Water is critical for metabolic activity (growth and reproduction) of the microbial community as well as hydrolysis. Ideal moisture content is usually at 60% to 80% of field soil holding capacity.

Since the vast majority of petroleum breakdown is performed by bacteria that utilize oxygen in their metabolic process, it is necessary to ensure that oxygen is available insufficient quantities. this is provided in the upper soil layers by mixing PetroXclean with domestic water, oxygenated aquifer water, and/or air-sparged water. In extreme cases, a small dose of hydrogen peroxide can be added to the injection solution.

If the petroleum compounds have penetrated deep into the soil and the conditions are anaerobic, a combination of PetroXclean, oxygenated water and other sources of oxygen can be injected. Movement of PetroXclean, water and oxygen through the deep layers of the vadose zone and beyond depend on sound engineering studies to develop a plan to utilize PetroXclean technology to its fullest potential.

PetroXclean is provided in a concentrated form. One gallon of the concentrate is used to completely bioremediate approximately eight cubic yards of petroleum-affected soil. A 6% solution (one gallon/16 gallons water) of PetroXclean is applied to the affected soil in weekly applications the moisture content of the bioremediation bed needs to be maintained at 60% to 80% field capacity between PetroXclean applications for successful treatment.


EXAMPLE

A property has 1,000 cubic yards of petroleum-affected soil. The soil has Been placed in a lined berm, 18 inches high with a soil depth of 14 inches. Using the ratio of one gallon of PetroXclean per 8 cubic yards, 125 gallons of PetroXclean will be needed. Each weed, 20% of the PetroXclean (25 gallons) is applied in a single dose, diluted first with 400 gallons of water.

For the first application, 25 gallons of PetroXclean is diluted in a mixing tank with 400 gallons of water (6% solution) and then applied to the bioremediation bed slowly enough for percolation through the soil. If the surface is impermeable, roughening with a mechanical or hand rake is conducted first to increase permeability. For the remainder of the first week, the moisture content is maintained by adding water. No additional PetroXclean is used. At the end of the week, the soil may be turned or tiled to augment oxygenation, if warranted. For each subsequent week of treatment, 25 gallons of PetroXclean, diluted with 400 gallons of water, is applied followed with maintenance of the moisture content.

In many cases, the most rapid and cost-efficient method of treatment is to spread the petroleum-affected soil on the surface in single layer 14-inch lifts and treated under optimal aerobic conditions. However, one can move a weekly front of PetroXclean laden water and/or air through a larger ex-situ soil pile. Manifolded, slotted PVC pipe installed in grids throughout larger piles can be used to maintain appropriate application of PetroXclean, water and oxygen.

PetroXclean can be used in-situ by utilizing horizontal, vertical, or tangential venting/injection wells, injection galleries or air-sparging systems. A sound engineering plan must be developed for the unique conditions existing at each site, taking into account the many variables involved in moving the wave of PETRO CLEAN, water and oxygen through the petroleum plume. In some cases, petroleum-affected ground water can be recycled through the bioremediation bed in conjunction with the PetroXclean application, this cleaning the ground water simultaneously. PetroXclean has been used in conjunction with many other remediation technologies for performance enhancement. These applications should be coordinated closely with CII to ensure successful application.


TOXICOLOGY

PetroXclean has been subjected to repeated toxicological testing in mammals and marine life to assess its safety in intended field uses. Under normal operating parameters, PetroXclean is applied at a dosage of 0.0555 mg/l. Marine life studies demonstrated that the 48-hour medial lethal concentration (LC50) determined on brine shrimp was greater than 100 mg/l. the 96-hour LC50 determined on killfish was found to be 5.3 mg/l.

Mammalian studies demonstrated no skin sensitizing reaction in guinea pig testing at a 500 mg/l concentration. No corneal nor iris abnormalities were found in rabbits exposed for 13 days to 100 mg of undiluted product. Testing performed in Sprague-Dawley rats indicated a LD50 of 75 mg/kg body weight of undiluted PetroXclean. Autopsy results, however, indicated that due to the large volume of PetroXclean necessary to produce toxic results, death was due to drowning, thus invalidating the LD50 results. Based on these tests and other not mentioned here, greater than a quart of undiluted PetroXclean ingested by normal adult human at one time could be toxic.


PROJECT RESULTS

Results of selected bench tests, pilot-scale tests and actual field project implementation are appended. Please note that the proprietary PetroXclean formulation has been tested historically under several different product names, including Bactozyme, FryeZyme and PetroXclean.


PetroXclean TEST PROTOCOL

I. PetroXclean Test Protocol Methodology

The test soil contaminated with petroleum product should first be characterized and then divided into two parts, one for control and one for testing. when in the full-scale mode, it is still advised to reserve a small control area so relative degradation observation can be correlated. In the laboratory, however, each soil test volume should be placed in open-topped test pans to depth of six inches. The calculated volume of PetroXclean mixed solution should be added to the test bed and an equivalent amount of water should be added to the control bed. Then both test beds should be brought to approximately 20 percent moisture by adding water. This procedure should be repeated four additional times ( for a total application of five doses) on a weekly basis. On a daily, both matrix. Moisture levels should be maintained at 20 to 25 percent at all times, or an amount to maintain moist conditions without producing leachate or standing water. In actual field conditions, the same protocol should be observed, but the turning of the soil may be conducted on a less frequent basis, depending on the soil characteristics.

A bacterial count is recommended prior to commencement of testing to check for soil sterility. Additionally, the bacteria in the soil should have been exposed to petroleum hydrocarbons, allowing some forms to already be acclimated. This shortens the lag time for exponential bacterial growth once the PetroXclean has been added. If the soil is devoid of bacteria, please contact PURE ONE ENVIRONMENT PRODUCTS for recommendations.

II. PetroXclean Test Protocol Product Utilization

One gallon of PetroXclean concentrate will treat eight cubic yards of petroleum-affected soil.

One part of PetroXclean concentrate (5.88%) should be pre-mixed with 16 parts water (94.12%).

For each eight cubic yards of soil to be treated, the one gallon of PetroXclean concentrate (mixed with water) should be applied on-fifth at a time for five weekly doses.

To treat one cu. yd (0.756 cu. m) of soil, use a total (over five weeks) of 2.125 gallons (8.043 liters) of PetroXclean mix.

  1. Total PetroXclean = 0.125 gallons (473 ml) = 2 cups
  2. Total Water = 2.000 gallons (7.5701)
  3. For each weekly doses: 6.8 cups (1.609) mix:
    PetroXclean = 0.400 cups (95 ml)
    Water = 6.400 cups (1.514 l)
  4. For each gallon of PetroXclean/water mixture, use 0.94 cup (223 ml) PETRO- CLEAN concentrate.

MAJOR GENERA OF OIL-DEGRADING BACTERIA AND FUNGI

BACTERIA
  • Achromobacter
  • Acinetobacher
  • Actinomyces
  • Aeromonas
  • Alcaligenes
  • Arthrobacter
  • Bacillus
  • Beneckea
  • Brevebacterium
  • Coryneforms
  • Erwinia
  • Flavobacterium
  • Klebsiella
  • Lactobacillus
  • Leucothrix
  • Moraxella
  • Nocardia
  • Peptococcus
  • Pseudomonas
  • Sarcina
  • Spherotilus
  • Spirillum
  • Streptomyces
  • Vibrio
  • Xanthomyces
FUNGI
  • Allescheria
  • Asperglilus
  • Aurebasidium
  • Botrytis
  • Candida
  • Cephalosporium
  • Cladosporium
  • Cumminghamella
  • Debaromyces
  • Fusarium
  • Gonytrichum
  • Hansenula
  • Helminthosporium
  • Mucor
  • Oldiodendrum
  • Paecylomyces
  • Phialophora
  • Penicillium
  • Rhodosporidium
  • Rhodotorula
  • Sccharomyces
  • Saccharomycopisis
  • Scopulariopsis
  • Sporobolomyces
  • Torulopsis
  • Trichoderma
  • Trichosporon


olinda01.jpg (35530 bytes) Oil Well on Property

Heavy asphaltic crude oil has been pumped from the southern California oilfields since the 1890’s. Over the years, this oil has spilled out onto the surrounding ground and weathered, resembling road tar. To develop these properties, the weathered crude oil must be cleaned up, but conventional methods of cleanup are generally too expensive for most developers. Bioremediation, enhanced with PetroXclean, provides a fast and cost-effective solution for property developers for cleaning up these old oilfield sites.

olinda02.jpg (33081 bytes) Tar Seeps

In addition to spillage of heavy oil from oil well pumping, southern California has many natural tar seeps, causing similar cleanup problems for property developers. The tar seep material, heavily weathered with dirt and plant debris, is commonly known as Brea. This material is very difficult to clean up, short of digging it up and hauling to a disposal facility. PetroXclean enhanced bioremediation has been proven to clean up Brea material to concentrations readily acceptable to all regulatory agencies

olinda03.jpg (52949 bytes) Well Abandonment

As a part of developing old oilfields, the oil wells must be properly abandoned. As these wells are abandoned, much contaminated soil is usually revealed as the old pumps and casings are removed. Note the crude oil on the ground around the casing that is being removed. Bioremediating this contaminated soil on-site with PetroXclean avoids the costs of hauling this soil to an off-site disposal facility, and then bringing in clean soil to replace it.

olinda04.jpg (30188 bytes) PetroXclean in Buckets

PetroXclean concentrate is being pre-mixed in 5-gallon buckets for application in bioremediating heavy crude oil-affected soils in a pilot test. Note how easily PetroXclean mixes with water in the buckets.

olinda05.jpg (20843 bytes) Bioremediation Bed

Soil contaminated with weathered heavy asphaltic crude oil has been prepared into a bioremediation bed. Note the dark color of the soil from the crude oil, prior to application of PetroXclean.

olinda06.jpg (29340 bytes) Discing Bioremediation Bed

After the bioremediation bed is prepared and during bioremediation with PetroXclean, the bed is disced regularly to provide air to the soil. This allows for the bacteria and fungi to aerobically digest the heavy oil, once they have been activated by the enzymes in PetroXclean.

olinda07.jpg (12529 bytes) Water Truck

Water is added routinely to the bioremediation bed using a water truck. Once a week, PetroXclean is added to the water in the water truck at a ratio of 1 part PetroXclean to 16 parts water.

olinda08.jpg (17643 bytes) Water Truck

The PetroXclean application is repeated weekly. The rest of the time, the water truck keeps the bed damp (60% - 80% of holding capacity) in order for the microbes to completely digest the oil efficiently.

olinda09.jpg (17848 bytes) Sampling by Consultants

Routinely the project consultants take soil samples to check the progress of the soil bioremediation. In this particular project, the Total Petroleum Hydrocarbon (TPH) concentrations at the beginning averaged more than 70,000 ppm, with most of the hydrocarbon chains more than C28.

olinda10.jpg (30759 bytes) Bioremediation Bed

After 9 weeks of bioremediation using PetroXclean, the heavy crude oil-contaminated soil was checked by scrapping an area of the bed. Note that the soil has no signs of crude oil in it and the soil is easily turned, unlike the sticky tar-like soil that was present of the bioremediation project using PetroXclean.

olinda11.jpg (29627 bytes) Bioremediation Bed 9 Weeks Later.

For this particular project in southern California, the heavy crude oil-contaminated soil was bioremediated in less than 9 weeks using PetroXclean. Note the color change in the soil. The TPH concentrations were reduced from 70,000 ppm to less than 500 ppm, with no hydrocarbon molecules left with less than a C28 chain (no light or leachable hydrocarbons). This site was declared "clean" by California regulatory officials, so the soil was not only cleaned up with PetroXclean, but the developer was able to use the soil for construction of residential housing pads.

olinda12.jpg (22910 bytes) Bioremediated Soil Being Used for Fill

After 12 weeks, the soil was removed from this southern California site with scrappers and used for fill for luxury home building pads. With the use of PetroXclean, the residential developer not only cleaned up the property in record time, but no imported soil was needed to complete this hillside development, saving the developer millions of dollars in construction costs

PetroXclean (HCZyme) Bioremediation Article
From Soil & Groundwater Cleanup - Annual Bioremediation Issue

PetroXclean MSDS


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Last modified: April 20, 2006