Ignorance is not bliss for “inert” pesticide ingredients

One of the complicated parts of assessing the hazards and risks of glyphosate is that the product that everyone uses (for example, Round-Up) is not just glyphosate. The active ingredient is glyphosate, but the final formulation sold in stores is a combination of glyphosate and other “inert” ingredients.

[Note: I’m going to stubbornly use quotation marks around the words “inert” throughout this article, to emphasize my point that this is not an accurate characterization. “Inert” implies inactive, which is not true. Read on for more.]

These “inert” ingredients are subject to essentially no testing, disclosure, and regulatory requirements, even though they almost always make up a larger percentage of the final product than active ingredients. And, evidence indicates that combinations of the “inert” and active ingredients can actually be more toxic than the pure active compound (for example, see here, here, and here).

A new publication by Mesnage et al. in Food and Chemical Toxicology reviews the problems with the status quo and the implications for health effects research. Given the relevance of this topic to my previous blog posts on glyphosate (see here and here) and pesticides in general, I’ll summarize some of the authors’ key points below.

But first, some terminology: what is the difference between active and “inert” pesticide ingredients?

Under the U.S. Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA), an active ingredient is one that is intended to be toxic to the target species. For example, glyphosate, the active ingredient used in glyphosate-based herbicides (GBHs), blocks an essential enzyme pathway in plants. All other ingredients in a pesticide product, often added to improve effectiveness, are classified as “inert.”

Abounding uncertainty about “inerts”

Contrary to what their name suggests, however, “inert” ingredients may not actually be inert. In fact, the U.S. Environmental Protection Agency (EPA) explicitly states that this designation “does not mean non-toxic.”

But, it’s challenging to get extensive and accurate information about these chemicals because:

  • Neither the “inert” ingredients nor the final formulations (the combination of active + “inert” ingredients) are subject to most standard regulatory toxicity tests, such as evaluation of cancer and reproductive effects. As a result, pesticide approvals are based on the unrealistic scenario of exposure to the active ingredient alone.
  • Companies can routinely claim the identity and concentration of these “inert” ingredients as confidential business information (CBI). That is why you’ll often see labels like the extremely vague one below. As a result, it’s difficult – actually, essentially impossible – for scientists to independently evaluate possible toxicity. We are kept blind to these final products.
Image: Beyond Pesticides
  • Because we don’t know the identity of these “inert” ingredients, there are essentially no monitoring data on environmental or human exposure.

So, in summary, we don’t know how toxic the “inert” ingredients or final formulations are; the identity of these “inert” ingredients is kept secret from the public; and we aren’t monitoring any of these chemicals for their presence in our bodies or the environment.

All of this makes it challenging for the EPA to conduct accurate pesticide risk assessments, which require information on both hazard (ie: toxicity) and exposure.

Constant change 

An added barrier is that companies often change their formulations over time and across regions. Apparently, there are usually between 100-150 different formulations of GBHs on the market at any given time, and over 2000 different varieties have been registered in Europe since 2012.

How are we supposed to evaluate the health effects of such a moving target? Robust epidemiological studies require precise definitions of exposure (referred to as the “consistency” principle) to prove causality. In essence, the exposure under investigation should be defined very specifically, such that it is not possible for variations in versions of the exposure to have different effects, which could muddy the overall conclusion of the study.

(As a concrete example, think about investigating the impact of “exercise” on health. Exercise is very broad, so it wouldn’t be helpful or informative to evaluate the effect of general “exercise,” which could span everything from a 30-minute walk once per month to a 2-hour run each day. The effects of these different types of exercise could have very different impacts on health. So, a better study question would be focused on a more specific type of exercise.)

For pesticide epidemiology, all of these changing formulations make it very challenging to draw conclusions on health effects across time and space. It’s quite likely that one study based in multiple locations could be evaluating the effects of different products at the same time. A study looking at one region over a period of several years also faces the same problem. As the authors of the recent publication stated, “formulated GBHs with the same product name from different countries may not be the same mixture of chemicals, nor the same as the brand-name product bought previously, or in the future.”

This is one possible reason for differing conclusions about hazard, and it makes reproducibility nearly impossible.

Overcoming INERTia 

The authors put forth a few suggestions to improve this murky situation. Some can be acted on by researchers now, such as including detailed product information (ex: trade name, dates of manufacture, product ID number) in the methods sections of their papers, to facilitate reproducibility and comparison across studies.

Other proposals will need to wait until there is political will for policy change. Most important is the need for full public disclosure of pesticide product composition. (By the way, back in 1997, the American Medical Association urged Congress to “support all efforts to list both active and inert ingredients on pesticide container labels.”) The authors also suggest monitoring of food and feed for concentrations of the “inert” ingredients (that is, if we can get access to information about their identities!), so we can understand patterns of exposure.

Additionally, it is essential to revise the pesticide approval processes to include full testing of “inert” ingredients as well as the final formulated products. We urgently need a regulatory system that accounts for these real-world exposures.

It’s high time for transparency on these formulations and their effects on human health and the environment.

Ancient philosophy, modern toxicology

The whole is greater than the sum of its parts.”
– Aristotle (384 BC-322 BC)


Aristotle was talking about metaphysics and the emergent theory, but his insight corresponds to an important emerging theory in environmental health: combinations of different chemicals acting together in our bodies can produce larger (or different) effects than would be seen if each chemical were acting independently. In technical terms, this is called “synergism.”

Why does this matter? Through the course of our daily lives, we are all exposed to hundreds of different types of chemicals. Most laboratory toxicity studies, however, only assess the effects of a single compound in a carefully controlled environment. Consequently, the (very limited) data that we have on chemical hazard do not actually reflect real-world exposure situations (ie: co-exposures to mixtures of chemicals). Researchers are beginning to address this deficiency, though, and initial results suggest that Aristotle’s ancient wisdom is eerily relevant to modern-day toxicology.

A recent study published in the journal Toxicological Sciences examined the interaction between polycyclic aromatic hydrocarbons (PAHs) and arsenic. PAHs are organic pollutants that are produced during combustion processes (including from tobacco). Many PAHs, such as benzo[a]pyrene, can cause DNA damage and are known or suspected to cause cancer. Arsenic is a naturally occurring element that can exist in different chemical forms. The inorganic form As+3 can interfere with DNA repair and is linked to skin diseases and cancer. Human exposure to As+3 often occurs through ingestion of contaminated drinking water or rice-based products. Many people around the world are exposed to both PAHs and inorganic arsenic simultaneously, but little is known about how these two chemicals — one that causes DNA damage, and one that interferes with DNA repair – act together in the body.

For this work, researchers examined the effects of As+3 and three specific PAHs (benzo[a]pyrene and two metabolites, BP-Diol and BPDE) separately and together in mouse thymus cells (precursors to T-cells). Because T-cells serve a critical function in the immune system, chemical damage could lead to immune dysfunction.

After chemical treatment, the researchers measured the amount of DNA damage and DNA repair inhibition. At specific combinations of doses (one with As+3 and BP-Diol, and one with As+3 and BPDE), they saw a larger effect from treatment with two chemicals simultaneously than what would have been predicted from treatment with the same chemicals individually.

Next, they measured cell death (specifically, apoptosis) and found that while individual exposures to As+3 and BP-Diol did not increase death, exposure to the compounds together caused a synergistic increase in the percentage of dead cells. One possible explanation for this result is that at low levels of separate exposure, the body can adapt to prevent damage. But perhaps with the two chemicals together, the system is overwhelmed and cannot compensate.

Overall, based on these and other related results in this study, the researchers hypothesized that the As+3 increases the toxicity of certain PAHs through its ability to inhibit DNA repair pathways. As I noted above, PAHs alone can cause DNA damage. With the addition of As+3, which interferes with DNA repair during normal cell cycle replication, cell damage is even greater.

Previous work had documented the existence of similar interactions between PAHs and arsenic, but those studies had used high doses that were not representative of potential human exposures. This study, by contrast, investigated the effects of low-level exposures that are more similar to what we might encounter in the environment.

One important caveat of this work is that the researchers conducted the experiment in isolated mouse thymus cells. In vitro systems (or “test tube experiments”) are increasingly common in toxicology, as the field aims to find alternatives to whole animal testing. However, there are obvious limitations to these models. Not only are mouse cells different from human cells, but these mouse thymus cells are separated from the rest of their system and may not represent how a fully functional organism responds and/or adapts to a toxicant exposure. As follow-up, researchers should test this chemical combination in a relevant animal model to see whether similar results are obtained.

Nevertheless, this study provides important evidence of synergistic effects from low-level exposures to two common environmental contaminants. And these data may be just the tip of the iceberg. What other potential interactions exists between the thousands of other chemicals that we are exposed to over the course of our lives? The challenge with synergistic interactions is that they cannot always be predicted from testing individual chemicals. (I’ve written about this previously, specifically with regard to cancer processes.) It is daunting to think about testing all of the potential combinations that may exist, since our public health agencies are struggling to generate even basic toxicity data on all of these chemicals individually.

I wish we could consult Aristotle on this problem.

One strategy to start to address this challenge could be to prioritize testing combinations of chemicals that – like the pair chosen in the study described here – are most common across the population. Existing biomonitoring efforts, such as the U.S. National Health and Nutrition Examination Study (NHANES), could guide the selection of appropriate mixtures. Testing these highly relevant chemical combinations could provide valuable information that could be immediately translated into risk calculations or regulatory standards.

As Plato, another great ancient thinker said, “the beginning is the most important part of the work.” So, while it is definitely overwhelming to think about tackling the question of chemical combinations, it is crucial that we take first steps to make a start.

Cancer Risk Assessment: Are We Missing the Forest for the Trees?

In recent years, national and international environmental public health organizations (including the US Environmental Protection Agency and the World Health Organization) have begun to use the adverse outcome pathway (AOP) and/or mode of action (MOA) as unifying frameworks for chemical testing and risk assessment. While the details of these frameworks vary, their underlying ideas are similar: researchers link specific molecular changes caused by environmental chemicals with adverse outcomes at the organism level (ie: disease), and then risk assessment is conducted based on the premise that preventing the early molecular disruption will prevent the development of the end-stage adverse event.

While there are practical advantages and real logic to this mechanism-based approach, a new review article published in Carcinogenesis suggests that this strategy may be overly simplistic and could potentially hinder our ability to adequately identify chemicals that contribute to the development of cancer.

This international team of cancer biologists and environmental health scientists organized their discussion around the “Hallmarks of Cancer,” a list of acquired characteristics that commonly occur in cancer (for example: continued growth, resistance to cell death, and tissue invasion). For each key characteristic, they identified typical target sites for disruption as well as environmental chemicals that have been shown to act on those targets. The researchers focused their discussion solely on chemicals that were not already categorized as human carcinogens by the International Agency for Research on Cancer (IARC), and they took careful note of effects observed at low doses. In addition, they specifically mapped connections between different pathways to highlight cases in which alterations leading to a given cancer hallmark could also lead to another.

Their lengthy review provides an important overview of the procarcinogenic effects of numerous common chemicals, but perhaps the most significant conclusion of this work is to emphasize the pitfalls in the status quo for risk assessment. By focusing on categorizing single chemicals as ‘carcinogens,’ we neglect to acknowledge that combinations of chemicals that individually do not meet criteria to be categorized as ‘carcinogenic’ may act in synergistic ways to promote the development of cancer. Even recent efforts to evaluate the effects of chemical mixtures may be inadequate, as they mostly focus on chemicals with common cellular pathways or targets. What about the numerous compounds, as identified in this review, that act on disparate pathways and organs to contribute to a similar disease process in the body?

To address these problems, the authors propose several key principles for an improved framework for cumulative risk assessment, including consideration of the synergistic activity of:

  • chemicals that act via different pathways
  • chemicals that act on different target tissues
  • non-carcinogens that act at low doses to contribute to pro-carcinogenic processes
  • chemicals that are not structurally similar

Carcinogenesis, like many disease processes, is complicated, and identifying the numerous pathways and organs involved is – and will continue to be – an enormous scientific challenge. Slow progress can be made, nevertheless, with a shift towards testing real-world combinations of chemicals and by using the ‘Hallmarks of Cancer’ to guide relevant and appropriate research. New technologies, such as high throughput screening, computational modeling and systems biology-based analysis, can aid in this process. However, the authors stress that traditional in vivo testing still holds an important place in cancer-related research – at least until there is appropriate validation of these emerging tools.

This publication highlights that our current chemical testing and risk assessment system is overly narrow and negates the complexity with which chemicals can interact in the body. We must broaden our approach to acknowledge that distinct chemicals can act in distinct ways at distinct sites – even at low doses – to contribute synergistically to a specific disease process. Reframing our perspective is daunting, and it will emphasize our limited knowledge about the mixtures of chemicals that we are exposed to everyday. But, if we can look up to see the forest, we may begin to make our way towards safer territory.