Under TSCA, the US Environmental Protection Agency (EPA) is authorized to screen chemicals, require reporting or testing on chemicals, ban manufacturing or importing of chemicals, and track new chemicals. However, in the 35 years since TSCA was passed, EPA has required testing on less than 300 substances. EPA has only attempted to ban nine chemicals, and was unsuccessful in its attempt to ban one of them (asbestos). For this and numerous other reasons, TSCA has undergone significant criticism in recent years, making clear the need for TSCA reform. 

Among the primary criticisms of TSCA are the complex and burdensome requirements that EPA must meet to require testing of chemicals. There is also little transparency regarding how EPA determines and prioritizes chemicals of concern, assesses the safety of these chemicals, and decides when risk assessment and/or management is needed. In addition, confidential business information (CBI) claims limit the public’s access to substantial amounts of risk-related information.

In response to criticisms of TSCA, EPA published principles of TSCA reform in 2009. The principles emphasized the need for sound science, risk-based criteria for reviewing chemicals, manufacturers to supply safety information, consideration of sensitive subpopulations in assessments, consideration of cost and substitutes in assessments, prioritization, incorporation of green chemistry, and sustained funding for EPA to conduct assessments. Also in 2009, EPA released an Enhanced Chemical Management Program. EPA then expanded on this effort in the Existing Chemicals Program, which was published in 2012. The 2012 Program described a two-step prioritization process for identifying chemicals for expedited review. From 1000 candidates, 83 chemicals were selected. As of May 2013, 30 have been listed for assessment and five draft assessments have been published.

In April 2013, the US Government Accountability Office (GAO) published a report in which they evaluated EPA’s efforts to strengthen chemicals management. Their findings were mixed. On a positive note, EPA has increased efforts to obtain toxicological and exposure data. However, it may take years to obtain the data using the vehicles in place, and it is not clear that this review process will result ultimately in risk reduction. GAO also found that EPA has missed some opportunities to obtain data submitted by companies to foreign governments. For example, EPA could take further action to report toxicity and exposure data submitted to the European Chemicals Agency under the REACH regulation. 

Also in April 2013, the Safe Chemicals Act was reintroduced in identical form to an act with the same name that was introduced (but not passed) in 2011. Among other things, the Safe Chemicals Act would shift the burden of safety to chemical companies and give EPA full authority to require data beyond a minimum data set. The Act has been co-sponsored by Democrats and Republicans in the Senate.

CPS risk assessor, Elizabeth Dederick, PhD, will present on TSCA and TSCA reform at a workshop in Germany hosted by Knoell. Details on the workshop are available here: http://www.knoell.com/en/event/workshop-asia-north-america-2013.

One important topic covered in this meeting included implementation issues with U.S. OSHA HazCom 2012 and in particular, the training requirements employers must provide by December 1, 2013. Topics such as important subjects to address, how to develop a curriculum, and how to ensure that all employees are properly trained to understand the new workplace labeling system, the new safety data sheets (SDS), and how employees can obtain and use the appropriate hazard information were discussed. Maureen Ruskin, the Office Director of the Office of Chemical Hazards within OSHA, spoke about how the new labels and SDSs must be consistent and in compliance with the international harmonization system known as the Globally Harmonized System of Classification and Labelling of Chemicals (GHS). New labels and SDSs are required to be provided by suppliers by June 1, 2015 and employers have until June 1, 2016 to make adjustments to their workplace programs.

With the increasing global adoption of GHS, SDSs are becoming more streamlined. However, various countries have unique minimum requirements and additional national and/or regional requirements. If a company wants to market a product in a country where GHS has been adopted, the company’s labels and SDSs must be in compliance with that country’s specific GHS requirements. Speakers covered the information required in each of the 16 sections of the SDS for the EU, US, Japan, Korea, and Brazil.

Other topics discussed include the dis-harmonization within the transportation sector of the US and UN Model regulations, the Safer Consumer Products (SCP) regulations proposed by the California Department of Toxic Substances Control (DTSC), Mexico and Brazil regulatory updates, current amendments to Taiwan’s Chemical Substances Control Act (TCSCA), and NFPA combustible dust standard updates. The SCHC meeting was a great opportunity for Critical Path Services, LLC to learn about new and developing issues related to hazard communication that may be of interest to existing and potential clients.

The new standard covers over 43 million workers who produce or handle hazardous chemicals in more than five million workplaces across the country. The modification is expected to prevent over 500 workplace injuries and illnesses and 43 fatalities annually. Once implemented, the new rule will:

• Enhance worker comprehension of hazards, especially for low and limited-literacy workers;
• Reduce confusion in the workplace;
• Facilitate safety training;
• Result in safer handling and use of chemicals;
• Provide workers quicker and more efficient access to information on the safety data sheets;
• Result in cost savings to American businesses of more than $475 million in productivity  improvements, fewer safety data sheet and label updates and simpler new hazard communication training; and
• Reduce trade barriers by harmonizing with systems around the world.

The revised standard requires a common approach to hazard determination and communication that results in better quality and consistent hazard classification across the workplace, making it easier for workers to understand the properties of the materials they handle. HazCom 2012 requires that all manufacturers, importers, and distributors revise product labels and SDSs by June 2015. Employers are also required to train workers by December 1, 2013 on the new labels elements and safety data sheets format to facilitate recognition and understanding.

Product stewardship experts and toxicologists at CPS are currently helping companies implement the new HazCom 2012 standard.

Herbicide resistance, notably resistance to glyphosate, known by the trade name, Round-up®, was the first commercially-viable trait introduced. The benefit to the farmer is the ability to adopt a no-till system, which saves costs and reduces loss of topsoil. The next major trait that has been widely adopted is resistance to lepidopteran insects through the use of the CryIA gene from Bacillus thuringiensis, hence the name, BT. BT cotton is resistant to the cotton bollworm (Helicoverpa zea), which gives farmers the advantage of not having to use expensive and potentially environmentally-damaging pesticides. The same gene in corn gives resistance to European corn borer (Ostrinia nubilalis).

Before a GM crop can be planted commercially in the US, it must first past inspection by the Animal Health and Plant Inspection Service, or APHIS. APHIS is primarily concerned with whether an introduced crop, by biotechnology or importation, will become a pest. APHIS lists approximately 140 crops that have passed regulation status. Industries hold the majority of the permits, but also publically funded entities such as USDA-ARS and University of Florida hold permits on GM crops. Testing of a new crop can take 5 to 10 years. Crops are evaluated for stability of phenotype, environmental effects, and vulnerability to pests and disease.

Any new agricultural food crop, whether bred through biotechnology or traditional means, must pass FDA regulation, which evaluates the characteristics of the food. According to guidance issued by the FDA (see FDA.gov, statement of policy – Foods derived from New Plant Varieties), known toxicants, nutrients, and allergenicity of the crop must be screened. This applies to foods destined for both animal and human consumption. Although the testing is carried out by the developer, the FDA’s Biotechnology Evaluation Team evaluates safety and compliance with the law. The FDA lists a total of 95 consultations on GM crops.

Finally, a plant that expresses a pesticide, such as the CryIA gene in BT corn, must pass EPA’s regulation of pesticidal substances (Federal Register Volume 66, Number 139, 2001). These biopesticides undergo the same rigorous inspection as any other pesticide. Toxicology and digestibility tests were conducted on the protein product of the CryIA gene when it was first introduced in corn, and the endotoxin was found to be non-toxic for mammalian consumption.

Critical Path Services, LLC (CPS), recently presented a poster on a method for testing allergen proteins in soybean. CPS’s GLP compliant laboratory partners with a number of crop protection companies to test their genetically modified foods for alterations in known metabolites and proteins.

*Business casual attire, defined as a collared shirt and pressed slacks for men and comparable attire for women (per the dress code of The Union League).

Contact sales@criticalpathservices.com for information on how CPS can assist you with your chromatography needs.

Honey bees are susceptible to a number of parasites and pathogens, as well as management and environmental stressors.  Parasitic mites are the most common and include two species of Varroa mite and three species of tracheal mite that may destroy a hive if not controlled (Shimanuki and Knox, 2000)[2].  Varroa mites were introduced to US honey bees in 1987 and now occur in every US state except Hawaii.  Varroa infestations can be reduced or eliminated with the use of commercial miticides like Apiguard^Ò (thymol gel), ApiLife VAR^Ò (thymol strips), Mite‑Away IIÔ (formic acid) (S. Praise, 2006)[3], or tau‑fluvalinate.  Tracheal mites are microscopic internal parasites.  Tracheal mites can be controlled by using mite‑resistant bees in combination with the use of menthol, vegetable oil, or formic acid (Hunt, 2010)[4].

A parasitic fly known as the Zombee fly (Apocephalus borealis) can attack honey bees and cause them to become disoriented and abandon their hives, a symptom common to CCD.  Honey bees that have been parasitized by the Zombee fly often leave the hive at night and cannot navigate back home.  Documented cases of parasitizaton by the Zombee fly were originally confined to the west coast, but citizen science groups are helping to map the movement of this pest across the US.  The Zombee fly is not the sole cause of CCD.  Although up to 85% of the workers in collapsing hives have been seen to be infested with Zombee fly larvae, it has not been proven conclusively that CCD is caused by parasitization of honey bees by Zombee fly (Kaplan, 2012 and Core, 2012)[5]^,[6].

Bacterial and fungal diseases also injure honey bees and their hives.  Bacterial diseases include American foulbrood and powdery scale caused by Paenibacillus larvae, European foulbrood caused by Melissococcus pluton, septicemia, and spiroplasmosis.  Often the only way to eliminate the bacterial diseases is by destruction of infected colonies and contaminated equipment.  The use of antibiotics has proven effective in some cases.  There have been control failures, however, due to the development of resistance to the antibiotics commonly used to control foulbrood infections in beehives.  Although these sanitation measures and use of antibiotics are effective in the year the infection is detected, recurrence of infection in subsequent years is not unusual (Wilson and Skinner, 2009)[7].  Powdery scale, septicemia, and spiroplasmosis are only rarely encountered.  Septicemia has been treated effectively in the past with the use of erythromycin, but effective treatments of the other diseases have not been developed because they only cause minor economic losses (Alippi et al, 1999)[8].

The fungal diseases chalkbrood (Ascosphaera apis) and stonebrood (Aspergillus spp.) are common in the US but only occasionally cause serious disease.  Since the brood diseases caused by these fungi are usually eliminated naturally without treatment, we have not discussed here interventions to control these fungi.

Microscopic protozoa can affect adult bees and, in combination with other diseases, have dramatic effects on bee and hive health.  There are several species of protozoans that cause dysentery and/or disturbances in nitrogen excretion; these include Nosema species, amoeba, and flagellates (Shamuki and Knox, 2000).  These species do not affect brood but affect the gut and malpighian tubules of adult bees.  Colonies can be disinfected by electron beam radiation, acetic acid fumigation of the comb, or treatment with Fumigilin‑B.  Nosema ceranae and Nosema apis, in combination with viral diseases like Israel acute paralysis virus (IPV) or invertebrate iridescent virus, have been linked to colonies that were susceptible to CCD from 2006 to 2011.

There are approximately a dozen viral diseases found in honey bees including sacbrood virus, deformed wing virus, chronic bee paralysis virus, acute bee paralysis virus, Kashmir bee virus, and Israeli acute paralysis virus.  Hives can be asymptomatic for some of these viruses.  Viruses can become symptomatic or infective within the honey bee host, but it is not clear what factors are involved in converting an asymptomatic infection to a symptomatic infection (Chen et al., 2006)[9]; poor nutrition or stress from transportation of commercial beehives from crop to crop during the growing season have been suggested as causes.  A healthy hive can usually overcome a virus.  A hive that has been weakened by fungal and bacterial infections, overcrowding, and environmental stress may become more susceptible to a virus[10].

Although the joint presence of Nosema species and virus species is a good marker for the likelihood of colony collapse disorder, it is not likely that these are the only joint causative agents.  A relationship between a variety of diseases, pathogens, parasites, and pesticide use has also been implicated in CCD.

The collapse of honey bee populations in the US and the phenomenon of CCD were dramatic for beekeepers, scientists, the agricultural community, and the general public.  The factors associated with CCD are complex, but the impact is simple.  Honey bee colonies are dying.  It is therefore important to understand the cause of bee declines and identify ways to sustain sufficient bee populations to support the pollination services that honey bees provide.

CPS would like to thank Tom Priester for the above contribution.

[1]   “Honey Bees Colony Collapse Disorder (CCD),” United States Department of Agriculture, last modified October 01, 2012, http://www.ars.usda.gov/is/br/ccd.

[2]   Shimanuki, H. and D.A. Knox.  “Diagnosis of Honey Bee Diseases.”  Agricultural Handbook No. 690 (2000).  United States Department of Agriculture. Agricultural Research Service.

[3]   Praise, S.  “Recommendations for the Control of Honey bee Tracheal Mites.”  Vermont Agency of Agriculture (2006).

[4]   Hunt, G.  “Parasitic Mites of Honey Bees.”  Purdue University Extension (2010).  E‑201‑W. http://extension.entm.purdue.edu/publications/E-201.pdf

[5]   Kaplan, J.K.  “Colony Collapse Disorder: an Incomplete Puzzle,” United States Department of Agriculture, last modified July 24, 2012, www.nps.ars.usda.gov.

[6]  Core, A., C. Runckel, J. Ivers, C. Quock, T. Siapn, S. DeNault, B. Brow, J. DeRisi, C.D. Smith, and J. Hafernik.  “A New Threat to Honey Bees, the Parasitic Phorid Fly Apocephalus borealis.”  PLoS ONE 7, no. 1 (2012):  e29639. doi:  10.1371/journal.pone.0029639.

[7]   Wilson, M. and J. Skinner.  “European Foulbrood:  A Bacterial Infection Affecting Honey Bee Brood.”  Washington State University Cooperative Extension (2010).  http://www.extension.org/pages/23693/european-foulbrood:-a-bacterial-disease-affecting-honey-bee-brood

[8]   Alippi, A.M., G.N. Albo, D. Leniz, I. Rivera, M.L. Zanelli, and A.E. Roca.  “Comparative study of tylosin, erythromycin and oxytetracycline to control American foulbrood of honey bees.”  Journal of Apicultural Research 38, no. 3–4 (1999):  149–158.

[9]   Chen, Y.P., J.S. Petis, A. Collins, and F. Feldlaufer.  “Prevelance and Transmission of Honey bee Viruses.” 2006.  Applied and Environmental Microbiology 72, no. 1 (2006):  606-611.

[10]  “Honey Bee Diseases,” Diapharm LTD, accessed November 2012, http://www.diafarm.org/index.php/en/honey-bee-diseases/77.

These experiments were designed to create a robust method for protein quantitation within various sample matrixes. The developed gel/label-free method was applied to quantitate levels of Kunitz Trypsin inhibitor (KTI), a prominent soy allergen, in soybeans (Glycine max). Reproducible protein extraction and isolation techniques coupled with the selectivity of LC-MS/MS analysis allowed for accurate determination of endogenous KTI concentration.

The method is applicable to all sample matrix types (plasma, plant tissue, etc.). Utilizing small proteome reactors coupled with highly selective LC-MS/MS analysis, this technique allows detection and quantitation of virtually any protein (intercellular, membrane, etc.) Additionally, by examining the mass of constituent peptides, post-translational modifications can be illuminated. With such high selectivity, this robust method eliminates the need for sample pre-fractionation, such as two-dimensional gel electrophoresis or strong anion exchange. Isotopically labeled standards may be used to further increase quantitative capabilities; however, this method greatly reduces the need for such analytical standards.

Competing methodology, typically immunoassays like ELISA, are often less expensive, faster, and can analyze multiple samples simultaneously. However, the antibodies manufactured to bind the protein of interest are not always as selective as desired. Non-specific binding results in a high frequency of false positives, especially in disease diagnosis such as HIV. Additional issues arise when these antibodies cannot bind their target protein due to a change in physical structure, such as denaturation, a common result of extraction procedures using strong organic solvents. Such primary antibodies do not bind, or bind inconsistently, to many proteins in their denatured or non-denatured form, adding to the immunoassay’s loss of sensitivity.

All quantitative strategies have their own strengths and weaknesses, so method selection should be based on individual research needs. The desired results, however, are always the same—accurate and reproducible determination of target protein identification and concentration. The method developed here provides a strategy for precise quantitation of multiple analytes from any biological sample via LC‑MS/MS.

Investigations continue in the CPS laboratory to assess the applicability of the method to multiplexed analysis.

Specifically, these guidance documents address key scientific and regulatory issues relating to an abbreviated development of biosimilars.  “Biosimilars: Questions and Answers Regarding Implementation of the Biologics Price Competition and Innovation Act of 2009” focuses on the questions that may arise early in the application process, such as how to request meetings with the FDA and acceptable differences between biosimilars and their references.  “Scientific Considerations in Demonstrating Biosimilarity to a Reference Product” outlines considerations in demonstrating biosimilarity between a therapeutic protein and reference product.  The FDA advocates both a stepwise and totality of the evidence approach in this guidance to assemble data in support of biosimilarity.  Lastly, “Quality Considerations in Demonstrating Biosimilarity to a Reference Protein Product” provides guidelines for a number of factors which can influence biosimilarity, including expression systems, manufacturing processes, physiochemical properties, functional activities, and impurities.

The FDA received comments until April 16 and has announced a public hearing on May 11 to obtain input for the finalization of these guidances.

Contact sales@criticalpathservices.com for information on how CPS can assist you with your submission.

In its fact sheet, available online, OSHA lists the following major changes to the HCS:

• Hazard classification: Chemical manufacturers and importers are required to determine the hazards of the chemicals they produce or import. Hazard classification under the new, updated HCS provides specific criteria to address health and physical hazards, as well as classification of chemical mixtures;
• Labels: Chemical manufacturers and importers must provide a label that includes a signal word, pictogram, hazard statement, and precautionary statement for each hazard class and category;
• Safety Data Sheets (SDSs): The new format requires 16 specific sections, ensuring consistency in presentation of important protection information; and
• Information and training: To facilitate understanding of the new system, the new standard requires that workers be trained by December 1, 2013 on the new label elements and SDS format, in addition to the current training requirements.

Modifications to the HCS include revised criteria for classification of chemical hazards; revised labeling provisions that include requirements for use of standardized signal words, pictograms, hazard statements, and precautionary statements; a specified format for SDSs; related revisions to definitions of terms used in the HCS, and requirements for employee training on labels and SDSs. The final rule also modifies provisions of other standards, including standards for flammable and combustible liquids, process safety management, and most substance-specific health standards to ensure consistency with the modified HCS requirements. OSHA states that the results of these modifications will be improved safety, facilitation of global harmonization of standards, and production of hundreds of millions of dollars in annual savings.

This long awaited Final Rule on the Hazard Communication Standard (HCS) is now available at http://www.osha.gov/dsg/hazcom/ghs-final-rule.html.

We are poised to assist you in this transition, from reviewing your chemical inventory, authoring your new MSDSs, training your employees, or managing the implementation. Call to schedule a Hazard Communication Standard audit and begin the implementation.

As proposed, UCMR 3 would require public water systems (PWSs) to monitor for 28 chemicals and two viruses (see link for additional information). All PWSs serving more than 10,000 people, and a representative sample of 800 PWSs serving 10,000 or fewer people, would be required to conduct Assessment Monitoring for 28 "List 1" chemicals during a continuous 12-month period from January 2013 through December 2015. In addition, a targeted group of 800 PWSs serving 1,000 or fewer people would be required to conduct Pre-Screen Testing for two "List 3" viruses during a 12-month period from January 2013 though December 2015.

For more information about the UCMR-3 program go to: http://water.epa.gov/lawsregs/rulesregs/sdwa/ucmr/ucmr3/basicinformation.cfm