Team:TEC GenetiX CCM/HP/BPlan

iGem Tec CCM/Home

Business plan

1-Project core and benefit

In the 21st century, most of the materials we use are made of plastics. From car chassis to food containers, these materials revolutionized the world; but, are they innocuous? Recent studies demonstrate that they release toxins, such as BPA, when they're exposed to high temperatures (Gonzales 2011, p.62). Among these toxins are PCBs and PBDE, which enter the human body through contaminated air, food or through direct contact with the skin. According to Faroon and Olson and the Syracuse Research Corporation (2000), several studies found PCBs were associated with cancer within the liver, intestines, skin and the gastrointestinal tract. For PBDEs evidence has been found that they affect negatively the levels of T4 in infants, as well as having an effect on neurodevelopment and other endocrine effects (Pohl et al., 2015). These facts create a conflict within our society in terms of health issues and environmental problems. The purpose of our developing project is to detect phthalates, PCB, PBDE and BPA in plastics that come in contact with food or water and issue a certification to enterprises that use the adequate amounts of these chemicals in their products or are free of them. To do this, our first course of action would be to propose a law to the government in which the regulation of these compounds is implemented. The certification mentioned above would consist in detecting the concentration of these substances in the companies’ products, and determining if such products are above or below the possibility of potential risks to human health, for example, the EFSA proposed a Tolerable Daily Intake (TDI for their acronym) of 0.05 milligrams of bisphenol A per kilogram body mass per day. If the levels of these compounds are higher than the established as the maximum (by this regulation, we shall give enterprises viable alternatives to lower their levels of these toxins, or recommendations so as to reduce the amount of these in their plastics and bioremediation opportunities. In case that the law proposal to regulate phthalates and BPA is not approved, we could be an optional certification. Our company would then begin a conscientization campaign regarding these compounds, and if a bottler decided to have our certification, that would increase customer trust towards certified bottling, increasing their sales and showing they are socially responsible.
Our company would therefore benefit companies economically by increasing their sales compared to the competition. It would also improve public health regarding issues related to the compounds, such as ovarian cancer. Another benefit from our business would be increasing society’s interest in what they consume every day, encouraging critical thinking.

2-. Name and logo

3. Mission and vision

Mission: Being the first Mexican company with a certification program regarding the rules and regulations of additives in plastics implemented on the main bottling businesses to ensure the population’s well-being.
Vision: To become a national service in the account of legislative and regulatory proposals of additives in plastics, and certify all the bottlers in the country.

4. Target market

The main scenery in which our company could act is the plastics’ commercial market in Mexico. During the last decade, Mexico’s plastic industry has been growing, and it recently became one of the most attractive areas in the country’s market, especially in the automobile, aerospace and packaging sectors, according to the SPI’s trade association. With a profit margin of approximately 30%, this market is now an open field of investment possibilities for small and medium enterprises from foreign countries, such as the United States and Canada. In 2014, Mexico’s trade surplus regarding plastic imports and exports was worth $11 billion (Missouri’s Department of Economic Development, 2016).
In the packaging sector, Mexico ranked as the second largest consumer of PET bottles in the world, according to the United Nations University (Smith, C. and Medina, M. 2013). A study realised by the same institution states that the national consumption in a year equals a total of about 750,000 tons of PET.

5. Law proposal

In Mexico, there is little control over the great variety of chemical substances that make up our products. The first issue is that there is no formal definition of chemical substance in the Ley General del Equilibrio Ecológico y la Protección del Ambiente (General Law of Ecological Equilibrium and Environment Protection, LGEEPA). This has caused confusion among institutions that look for a classification of chemical substances. This law has not been updated since 2012. The only official organism with a definition for this term is the Sistema Armonizado de Clasificación y Comunicación de Peligros de los Productos Químicos (INECC, 2014).
Enterprises look at reference lists for making their decisions; however, some of these lists contain a variety of mistakes that make them inaccurate. For example, there are substances mentioned more than once, unofficial or colloquial terms are used indistinctively, and generic names are used when trying to mention a specific chemical. It is important for organizations to have a uniform list of chemical substances, their toxicology and important characteristics, in order to make decisions based on the consumers’ health, ecological impact and possible hazards that may come handling these substances. While it is of utmost importance for enterprises and consumers alike to be informed of the risks the substances they come in contact every day may pose to their health and the environment, creating or modifying norms, laws or agreements is often a slow process (SEMARNAT, 2007).
The Registro de Emisiones y Transferencias de Contaminantes (Emissions and Contaminant Transference Registry, RETC) is a list updated yearly with information on the contaminant emissions of enterprises in Mexico. This Registry is open to public access, and there is also a list of enterprises that do not fit the required levels of contaminants. However, the list of regulated substances is consists of only 104, as of 2013 (SEMARNAT, 2013). According to Chemical Abstract Services (CAS), there were around 343,000 regulated substances in the United States, Europe and China (INECC, 2014).
Substances are classified as dangerous if they fit any of the categories specified in the Apéndice 19 del Anexo 22 de Reglas de Carácter General en Materia de Comercio Exterior. These are explosives; compressed, refrigerated or liquefied gasses; flammable liquids; flammable solids; organic oxides or peroxides; acute toxins, or infectious agents; radioactive; corrosives; and others (Reglas Generales de Comercio Exterior, 2016). However, BPA, phthalates such as the benzyl butyl phthalate, PCBs and PDBEs are not being taken into account, despite sharing the characteristics of acute toxins (INECC, 2014). To classify as an acute toxin, the substance must be proved or, in compliance with the precautionary principle, strongly suspected to cause death, grave injuries or negative effects on human health upon inhalation, ingestion or contact with the skin. In the case of Europe, the European Food Safety Authority (EFSA) established in 2006 a tolerable daily intake of 0,05mg of BPA/kg of corporal weight/day, however, an uncertainty factor of 100 was applied to the TDI. In the case of the United States, the U.S. Food and Drug Administration (USFDA) in 2008 emitted a report in which the tolerable daily intake is 5 mg of BPA/kg of corporal weight/day and in Japan, the TDI is 0,05mg of BPA/kg of corporal weight/day, according to the Food Safety Commission of Japan (FSC) the maximum quantity of BPA in containers mustn't get over the 2,5 ppm (WHO, 2010).
In the case of Europe, the European Food Safety Authority (EFSA) established in 2006 a tolerable daily intake of 0,05mg of BPA/kg of corporal weight/day, however, an uncertainty factor of 100 was applied to the TDI, in the case of the United States, the U.S. Food and Drug Administration (USFDA) in 2008 emitted a report in which the tolerable daily intake is 5 mg of BPA/kg of corporal weight/day and in Japan, the TDI is 0,05mg of BPA/kg of corporal weight/day, according to the Food Safety Commission of Japan (FSC) the maximum quantity of BPA in containers mustn't get over the 2,5 ppm.
We have consulted different studies regarding BPA and the phthalate family, and we have found convincing evidence that both of these need a regulation since they fit the criteria for class six of dangerous substances, which means that they pose certain degree of health hazards and, according to the precautionary principle, they would have to be regulated; our investigation and bibliography is available in this same document. Therefore, both BPA and phthalates have to be regulated by some organism if they continue to be used in plastics, both national and foreign. In order to be able to do this, a clear maximum has to be established for the use of these substances by the relevant authorities, and a faculty must make sure the norms are respected. We could offer the service of quantifying phthalates and BPA in different products that come in contact with food and water and certify them as safe according to this established limit.
Article number 71 of the Mexican Constitution explains that in order for a law proposal to be discussed by the Mexican Congress, it is necessary to have at least 0.13 % of unique signatures from voting citizens. This means that we require around 130 thousand signatures in order to send the law proposal to congress.The law proposal requires certain characteristics, which are: applicable legal fundamentals, the date when the consensus is going to be done, brief description of the topic, the date when the proposal was done.

6. Market incentive

In order to get the number of signatures needed for the law to be discussed in Congress, we need to have an educational campaign in order to inform the customers about the health hazards that these toxins cause. Giving conferences in schools is an option that we have already implemented, and the idea is to inform the children and their parents about the repercussions in health that these toxins have.
Even if the law is rejected, we will have a large community supporting us and achieving the second way of reaching bottlers, that is through social pressure. By creating environmental awareness through recycling campaigns and sharing information through social media, consumers will be able to decide in a more socially responsible way; they will start rejecting products from bottling enterprises that do not have the certificate. This will lead to a decrease in sales and will encourage enterprises to get it.
With the law proposal, however these companies would then need to have the certificate to keep functioning without getting fined. Another strategy that has been implemented already is a campaign to clean a populated part of the city in order for the people to make questions and get informed about this problem. This has to be followed by a campaign on social media that will help spread the information given as well as videos of interviews of people who asked about the campaign and the project. The certification program will be innovative in Mexico which shall increase its reputation by the use of marketing strategies on both the plastic producers and the plastic consumers. Therefore, taking advantage of its Unique Selling Proposition, Toximon will be the brand leader on certifying the plastic industry, even though other certification companies enter the market, Toximon will hold a patent of its work, but will also consider working in joint venture for a specific project.

7. Product’s cost and price - Financial management

Considering the materials and equipment required to produce the bacterial batches to perform the certification analysis our team decided to make a budget taking into consideration all the aspects involved in the transformation process. The prices are displayed below: Note: All reactants that do not show their laboratory name were retrieved from Sigma-Aldrich Corporation.
Initial investment: 48330.37 USD
Bacterial batches generated: 5L per transformation (Number of bacteria per batch given by the equation n=IB(2)^t/1800)
Bacteria used in each analysis: 5 repetitions of 1ml OD 0.6
Price of certification: 1000.00 USD
1. NEB® PCR Cloning Kit (20 reactions) 275.00 USD

2. BioBrick® Assembly Kit (50 reactions) 261.00 USD

3. QIAprep Spin Miniprep Kit (50) 96.27 USD
4. Antibiotics:


Phthalates are a group of esters that have been used for over 40 years in the plastic industry. These have a variety of applications and are mostly used as plasticizers, in PVC coating and food packages (Fromme et al., 2002). These compounds do not bound chemically to plastics, and therefore humans and other lifeforms may be exposed either by inhalation, dermal exposure, or ingestion. (Zota, Calafat, Woodruff, 2014).
Importance in health
The endocrine disruptors are substances which have the potential to block the hormonal response of the endocrine system, affecting the development, reproduction or behavior of people, some examples are PVC, DDT, Pentachlorobenzene (PCB) phthalates and PBA. Some examples of the effects in health of this substances are the abnormalities in the genitals and cancer (Cosoi, 2006).
Phthalates act as endocrine disruptors. They are some of the most abundant endocrine disruptors found indoors. Studies from 2003 showed that phthalates were found in the air of 120 homes in a concentration of 50-1500 ng∙m-3 (Rudel, Camann, Spengler, Korn, & Brody, 2003). However, their effects vary depending on the dose, the moment of exposure and the type of phthalate. For example, it has been shown that In utero exposure in rats to the BBzP, DnBP, and DEHP phthalates causes reproductive tract malformation if it occurs during the sexual differentiation period. Nevertheless, other phthalates did not show any significant change in the rat’s structure (Zota, Calafat, Woodruff, 2014). Exposure during early development is suspected to have repercussions in adulthood, some of which include hormone responsiveness and decreased semen quality. (Meeker, Sathyanarayana, Swan, 2009). Besides, certain studies developed in the European Union determined that phthalate exposure has a 40 - 69% probability of causing 53,900 cases of obesity and 20,500 new-onset cases of diabetes, all in older women (Legler et al. 2015). Some phthalates show weak estrogenic and mitogenic activity. This was tested on yeast cells, as well as human breast cancer cells for the mitogenic behavior. Thus, they are expected to behave similarly in humans. These phthalates, in a decreasing order of activity, butyl benzyl phthalate (BBP), dibutyl phthalate (DBP), diisobutyl phthalate (DIBP), diethyl phthalate (DEP), diisononyl phthalate (DINP). (Harris, Henttu, Parker, Sumpter, 1997).
Moreover, phthalates are often used in medical devices, causing patients undergoing dialysis, fetuses, and neonatus to be more likely exposed to these compounds. Studies carried out on animals show that there are negative effects on the reproductive system before they reach the levels calculated in clinical exposure. Thus, the possibility of phthalates in medical devices causing long term effects must not be ruled out (Tickner, et al., 2001). Phthalates are the most widely used plasticizers worldwide. In North America, 70% of plasticizers used belong to this family. Due to their widespread use, phthalates have come to the attention of consumers, and so have their health concerns. Since 1999, phthalate demand in Europe has decreased, followed by a ban on some phthalates in 2005 (Chemsystems, 2008).
The effects on human health are not so documented as those of phthalates, and therefore, it is not possible to ensure that these will be less harmful. Like phthalates, the alternatives do not bond to plastics chemically, and thus there is a slight chance that these will leak into the products contained inside them.
Acetyl Triethyl Citrate
Health concerns: Ocular exposure showed moderate negative reaction. However, the reported effects subsided after 48 hours at most (Johnson, 2002). There is apparently no risk of dermal absorption, and the average concentration in air vapor for people working with these materials is 8.37-50.2 mg∙m-3. There is also no evidence that these cause any kind of chromosomal aberration or genetic mutation of any sort. (Stuer-Lauridsen, et al., 2001)
Environmental concerns: It is relatively soluble, 0.0005 g/l, although this number was obtained at unknown temperatures (Stuer-Lauridsen, et al., 2001) Economic concerns: It is estimated that a ton of this material costs 2,120 USD.
DINCH (Di-isononylcyclohexane-1,2-dicarboxylate):
Its commercial name is Hexamoll. This was developed to plasticise materials that come in contact with food and children's toys. Now, DINCH has the approval of the Japanese, German and European for food contact (Chemsystems, 2008). It was first used specifically to replace DEHP and DINP as plasticisers in PVC (Jobwerx, 2006).
Percentage of these substances in a bottle of water, or in a series of them
In some Saudi Arabian brands of plastic bottles, at 4°C, the concentrations of phthalates found were inside the 1.194-21.128 µg/l range. (Al-Saleh, Shinwari, & Alsabbaheen, 2011). In Hungary, water bottles from three different brands were examined to determine the quantity of phthalates that leaked into drinking water, both carbonated and non-carbonated. The phthalates found were the following: diisobutyl phthalate .
Repercussions to the human body’s health
DEHP can affect multiple organ systems such as : kidneys, lungs and heart and mainly reproductive tract, since testicular toxicity is a critical health effect. The effects DEHP has in said organs, is further explained in the following table.

Bisphenol A is a chemical compound used in plastics to provide them with certain characteristics. It is part of the big group known as plasticizers, just like phthalates. This means it can leak into food or beverages contained inside plastics made up of this compound. The risk increases when the plastic is stressed, either by pressure or temperature. (Nordqvist, 2014).
Bisphenol A is a chemical compound that is usually employed in the fabrication of flame retardants and different resins. It is also found in can coating, additives in different kinds of plastics and even dental fillings. Since, like phthalates, these are not chemically bound to the plastics, it is possible for them to be released either during the manufacturing process or from the final product (Fromme et al., 2002).
Health effects of BPA
The effects of BPA, as well as those of phthalates, vary greatly depending on the dose and time of exposure to the compound. If a fetus is exposed to it during organogenesis, which is the development of organs, it may lead to negative consequences later on in life. It has been shown to cause organizational changes in some organs of the body, as well as altering brain structure and chemistry, even going as far so as to affect behaviour in animals (vom Saal et al., 2007). BPA has a certain degree of estrogenic activity, according to a study carried out in breast cancer cells. This is what led the world to pay special attention to BPA and their potential effects on human health, especially regarding its similarities with estrogen. (Vogel, 2009).
A study from 2005 showed a relationship between BPA levels and frequent miscarriages. Women that had gone through 3 to 11 miscarriages had their BPA levels compared with those of a control group of women with healthy pregnancies and deliveries. There was a proportional relationship between BPA levels and the chance of miscarriage in the results. While more research needs to be done, there is evidence to be alert of these compound’s effect on human health, especially regarding pregnant women ( Sugiura-Ogasawara et al., 2005).
Male genitalia is also affected by prenatal exposure to BPA. Sons aged 0-17 had their anogenital distance measured. The groups were divided in two: those whose parents had been occupationally exposed to BPA and those who hadn’t. Boys whose mother had been exposed to BPA showed a shorter anogenital distance than those whose parents were BPA free. The results depended on the dose: the more BPA consumed by the parents, the shorter the distance was. This is strong evidence in favour of regulating BPA and its contact with pregnant women and newborns (Miao et al., 2011).

References and bibliography:

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