University Place  Curtis Hedman Prescription Drug Traces Found in Lake Michigan  February 26, 2014 UP988   cc >> Hello, everyone. Thank you for watching Wednesday Nite at the Lab. I'm Cassie Immel from the UW Biotech Center. This evening we have Curtis Hedman from the Wisconsin State Laboratory of Hygiene. He'll be talking to us about some of those pharmaceutical traces found in Lake Michigan. He has a PhD degree in environmental chemistry and technology from UW Madison. He has 25 years of laboratory experience spanning the disciplines of microbiology, inorganic chemistry, and organic chemistry. In his current position, Dr. Hedman collaborates with the university government and industry researchers to develop, validate, and run analytical methods utilizing high performance liquid chromatography mass spectrometry, which are for small molecules and environmental and biological matrices. The main focus of Dr. Hedman's research is identification and quantification of emerging environmental contaminants as well as their metabolites and the breakdown products that they may have. So without further ado, let's welcome Dr. Hedman. 

[APPLAUSE]  >> Well, thank you all for coming tonight, and I really appreciate the invitation to come and speak with you about my research and a real exciting inter-campus collaboration we had with UW Milwaukee Center for Great Lakes Research and a topic that's personally of interest to me because I've been looking at pharmaceutical personal care products and other emerging contaminants for about 12 years now. So when I was looking at the archived webcasts for Wednesday Nite at the Lab, I saw a good talk from Patrick McIntyre last May, and he talked about great stakes in the Great Lakes. And, as you know, the Great Lakes are the largest system of fresh surface water on Earth. And really we have to do what we can to protect this precious resource. And to give you an overall view of how I want to approach this talk, I wanted to of course fill you in on the research project itself, but then I wanted to kind of take a look from different perspectives and how it relates to the topic in general, pharmaceuticals in the environment. And I'll start out with my most comfortable topic, environment chemistry, but I'm going to make an attempt to talk about these other points that relate to that topic. So, first of all, I'll bring you through the project that we did with Rebecca Klaper's lab. Rebecca Klaper is an associate professor at the School of Freshwater Sciences at UW Milwaukee's School of Freshwater Sciences, and she works at the UW Milwaukee Water Institute, which is located right on the shores of Lake Michigan. They have access to a great number of resources for the lake research, including the research vessel Neeskay, which they utilize for this research. Two of her graduate students worked very closely with me on this project, Benjamin Blair and Jordan Crago. 

And the Wisconsin State Laboratory of Hygiene, as you know, is located right down the block from the Biotechnology Center at Henry Mall, that's the clinical division, and the environmental health division, which is off campus at Agriculture Drive. Our laboratory director Charles Brokopp, Dr. Brokopp talked last year about the newborn screening program at the clinical division of State Lab of Hygiene. That program has been using mass spectrometry detection for many years. The environmental health division only started using HPLC mass spec in 2004. And it was soon after that that we brought methods online for these emerging contaminants and pharmaceuticals, and that's when we started talking with Dr. Klaper and colleagues about doing some lakes research. The other main player or stakeholder in this research would be the Milwaukee Metropolitan Sewage District and Veolia Water Milwaukee, which is under a 10-year contract to operate MMSD. They were good enough to allow us access to their facilities and also funding for the monitoring. To the right you see an overhead view of the South Shore Water Reclamation Facility in Milwaukee. So using that research vessel, the Neeskay, the vessel was sent out on voyages from the harbor to the wastewater treatment plant outfall off of Jones Island and then at a location called South Gap. Also, they went down shore to the South Shore Water Reclamation Facility and took samples at sites called SS Outfall and then 1.6 kilometers south and east and 3.2 kilometers south and east of that facility. 

And for sampling, two different techniques were used for the water samples. They utilized what's called a Niskin bottle sampler, and that is a tube that is open on both ends but has caps that left open in spring loaded. As they're lowered down to the desired depth, those caps stay open until a triggered spring loaded mechanism closes both ends. So you get sampling, a grab sample at a specific depth. For sediment sampling, we first tried cores but we found that later on just sending scuba divers down to get grabs of the sediment was a better technique. Back at the lab, I would receive these samples in either one-liter amber glass bottles or mason quart jars for the sediments, and I utilized methods that were recently developed by the United States Environmental Protection Agency. Method 1694 for pharmaceuticals and personal care products and method 1698 that covered the steroids and hormones analyses. I did modify these methods a bit, the steroids method in particular. We utilized the high performance liquid chromatography mass spec for both methods for the analysis. And for the solids and hormones, I utilized a newer solids extraction technique called accelerated solvent extraction. 

 This graphic was produced by Ben Blair, and I think it does a really good job of describing what we saw and where. And here we have the different sampling sites as depicted in the earlier graphic, but here we can see by color the different compounds that were detected. And each square represents 25 nanograms per liter, or part per trillion. So, certainly, at the wastewater treatment plant outfalls we're seeing the greatest concentrations, and then things taper off as you get further away from the wastewater treatment plant outfalls. A couple things that were striking were the amount of one compound in particular, the Metformin, and that was present in quite high concentrations in microgram per liter actually at the outfall. And it's a compound that wasn't studied as much up until our current study because it requires a different type of extraction. We actually have to run a second extraction at basic pH to be able to capture that compound. So it might have been something that was missed in earlier studies because it's a little more difficult to extract. Other compounds that are a little bit higher concentration were the cotinine, listed in red. Caffeine was certainly present. 

And the next couple of slides go over the pharmaceutical personal care products detected and their uses. You'll see some themes, many of them are hormones. Interestingly, the hormones that were detected were mainly the androgenic hormones and also progesterone. So we did not see the estrogen, estriol, estrone that we would expect to see along with those others. And certain over-the-counter prescription meds, such as acetaminophen, codeine found in cold medications, metabolites of caffeine and nicotine. The Metformin, interestingly, is an antidiabetic drug. So I don't know if that makes a comment on where we are with society if the number of people are taking Metformin and it's ending up in the harbor at those concentrations. Again, some other antibiotics, macrolides, sulfonamides, Triclosan, the antimicrobial, and some of the soft soaps and other personal care products. And the result of our study also show that these concentrations were fairly consistent over several different sampling events. We went out six different times over the course of one year, and with the exception of the June 2010 outing, the sample concentrations in nanogram per liter group together pretty closely. 

And that brings up the point that these pharmaceutical personal care products, even though they might breakdown in the environment over time you get a certain pseudo steady state concentration because they're constantly being input into the water from the wastewater treatment facility. Looking at the sediment samples, we had again a wide variety of compounds that were detected, and again the larger concentrations were in the sediments immediately after the wastewater treatment plant facilities at the outfalls. But we also had some significant concentrations of Azithromycin, Diphenhydramine, Metformin at the south gap location, which was a couple of kilometers south of the outfall. Interesting to note that the compounds in the red boxes were not detected in the water samples, and that could be due to a couple of different effects. Some of the compounds' chemistries and functional groups allow them to significantly bind to the sediments and be pulled out right away. And it could be that if they bind to the sediments and are somewhat present in the water they might be more susceptible to microbial or UV breakdown, but they're protected once they're absorbed to the solids. So after looking at the project as a whole, I wanted to go in a little more detail about the environmental chemistry behind these pharmaceutical and personal care products and the environment. Pharmaceutical personal care products could fall into several broad categories that you've probably seen in the literature. They've been called emerging contaminants, compounds of emerging concern, anthropogenic waste indicators. A certain component of PPCPs are also endocrine disrupting compounds, and the wastewater treatment plant engineers like to group these in with microconstituents. 

The study of hormones and pharmaceutical personal care products in United States surface waters really took off around the year 2000 with Dana Kolpin and his colleagues at United States Geological Survey. They looked at these compounds in United States streams all across the country, including our area of study close to Lake Michigan. And their group looked at an even larger array of compounds, including antibiotics, prescription drugs, nonprescription drugs, the hormones, and household industrial compounds. And as a summary that they published in 2002 in Environment Science and Technology, the percent of samples with detection were quite high, and it was shocking to me that antibiotics were detected in 80% of the drinking water samples that were studied. Shifting back to the Milwaukee harbor, these were pharmaceutical personal care product samples that I analyzed for Dr. Klaper's lab before the study that was published. We did these with grab samples that were taken in the fall of 2007. And that was before the US EPA methods came out. We were using methods developed by Shane Snyder's lab when he was with the Southern Nevada Water Authority. And looking at wastewater treatment plant influent certainly had a large array of compounds and high concentrations. And what stands out again is caffeine, which is taken out once you get the effluent. 

If we remove the caffeine, it's a little bit easier to see how the concentrations relate to one another. You see Triclosan being present in the effluent, downstream, and the harbor and less so for the other compounds Carbamazepine, Fluoxetine, Paraxanthine, Carisoprodol. Looks like the Bisphenol A was completely removed in the wastewater treatment plant process. Another group of compounds, the antibiotics, showed a very high amount of Tetracycline in the influent, but that was very successfully taken out in the treatment process. A small amount found in the effluent, but by the time we got to the downstream samples it was nonexistent. Taking the Tetracycline out of that influent bar graph, we get a better view of the other compounds. Ofloxacin, Norfloxacin, and Trimethoprim, and Azithromycin are predominant but also taper off pretty quickly once you get to the harbor. Shifting again, looking at the hormones. There were the hormones that we saw in the current study. We did also detect the 17 beta estradiol, but only in the influent. 

It looks like the wastewater treatment plant process did a really good job of removing the hormones. Now shifting gears to look at the study of pharmaceutical personal care products from environmental toxicology standpoint. I bring up the interesting point that when you're looking at nanogram per liter quantities as very, very tiny. One nanogram per liter is equivalent to one drop in about 20 Olympic sized swimming pools. So when you're looking at concentrations that are that low it's orders of magnitude lower than a therapeutic dose of a prescription drug. Having said that, when you look at the human health relevance of the results of the compounds in Lake Michigan, I'd refer you to a study that was performed by Shane Snyder and colleagues, actually a colleague of mine at State Lab of Hygiene Dr. Jocelyn Hemming performed some of the endocrine disruption analyses for this study. And they looked at, again, drinking water and wastewater treatment plant facility water from facilities across the United States. And they also like to add food and drink samples, such as milks, yogurts, etc. And the conclusion of the study was that the human health relevance, the indicators that were detected in the concentrations that were present in drinking water were not of human health relevance. In fact, they found that some of the food and drink products, the milks and some of the packaged products, had more of the xenoestrogens, and some of those things. 

 Again, looking at the Great Lakes ecosystem from a macro scale, the fish that you would eat, again it would not be likely impacted by the concentrations that we detected in the water. But looking a little bit more closely at the lower lifeforms in the food web, this is where we start to have interest in the toxicological significance below this line and we're starting to look at the microinvertebrates, the phytoplankton, and things that the larger fish are going to eventually be eating for their diet. And we want to make sure that these concentrations aren't affecting these lower lifeforms because if you affect one part of the food chain, you could impact all the way up into the higher parts of the food web. And a study that many of you are probably familiar with, Karen Kidd and colleagues dosed an entire test lake in northwest Ontario at only five parts per trillion with alpha-ethinyl estradiol contraceptive hormone. And over the course of a couple of years it caused the collapse of the fathead minnow population in that lake. So even though these are extremely small concentrations, depending on the compound they could have a great impact on the smaller lifeforms and benthic organisms. And for the current study analysis by Ben Blair, they looked at what they call a toxicity risk portion for these lower lifeforms in the food web and the benthic organisms. And depending on the compound, with a risk quotient of greater than one showing a high risk, compounds such as caffeine, paraxanthine. Some of them that were present in higher concentrations throughout the spatial part of the study you can see that the risk quotients are well above one for those compounds. So their conclusion is that at the concentrations seen, there could be some toxicological significance for the lower lifeforms and benthic organisms in Lake Michigan, and it warrants the further study of these compounds in Lake Michigan. 

Now moving to what this means from an environmental engineering standpoint. Looking at Milwaukee in the early 1900s, this was a great picture that was actually on the Milwaukee Metropolitan Sewage District web page. In the late 1800s, tunnels were made to channel the wastewater into the rivers and into the lake. And they kept track of the wild rice growing close to those outflows and figured if the wild rice was growing, everything was okay. So we've come a long way in several decades. The wastewater treatment plant process in general has a process where large particulates are screened out. Then the water is sent through clarification chambers where oils tend to go to the surface and get skimmed out and other particulates will settle down. Then the water goes to a secondary treatment that has microbiological component that helps to breakdown some of these organic compounds. And then if the organics are still present, they go into a disinfection stage. And depending on the wastewater treatment facility, this disinfection can be in many different forms, UV, ozone, carbon sequestration, activated carbon. 

And then at the end they're making sure that that water meets permit limits as established by the Clean Water Act. And I really have to appreciate MMSD and Veolia for supporting this research, and not only looking at the occurrence of the PPCPs but also working with Dr. Klaper's group on looking at the wastewater treatment plant processes and how we might be able to improve the capture of these organics as they're passing through. This was another recent publication from this group that I collaborated with them on. And we were looking at a US EPA model called the -- system looking at microbial degradation of compounds and looking at the presence of these compounds and how they would model with this program. And overall we found a pretty good relationship with a slope of 0.77 for the different compounds that we tested. So we could predict pretty closely how the removal efficiency of these different compounds, with the exceptions of a couple of compounds that have high KOW constants. You can see Triclosan and Triclocarban tend to swerve a lot to the solids, and that didn't predict well for them. We also have another open project where we're looking at another aspect of biological degradation of these pharmaceutical compounds. Now moving into what this means from an environmental policy standpoint. I'm going to take a stab at it even though it's not my area of expertise. 

There was another Wednesday Nite at the Lab session last year that talked about great stakes in the Great Lakes, and it was a colleague from UW Limnology. He talked about the Great Lakes Restoration Initiative. And I was happy to see that last month the second round of funding came through for this big project in the tune of $300 million. It's a great collaboration between multiple federal agencies, USDA, EPA, USGS all working together. And as part of the first round of GLRI studies, the United States Geological Survey National Water Quality Lab, the same group that did the analysis for Dana Kolpin's reconnaissance study, looked at different areas of concern on the Great Lakes, including sampling location in the Milwaukee harbor near Jones Island wastewater treatment plant. And they did detect a number of these pharmaceutical personal care products and other industrial chemicals, and the range, I just put the range, was 32 to 135 nanogram per liter. And just a sampling of the compounds that they detected. Atrazine and Bisphenol-A, diethyl phthalate, DEET which is the active ingredient in mosquito repellent. Again Triclosan and some of the other compounds that were detected by our study. The point I'd like to make here is that the methods that we're using are looking at unique targets. 

We have to know what we're looking for, optimize the compounds so we can detect it at such low levels. So I'm looking at several dozen, and USGS lab is looking at a couple of hundred. Well, according to the general accounting office, we have more than seven million recognized chemicals and approximately 80,000 in common use in industry. So you start to think about how can we even hope to track what's being put into the environment and determine what in that mixture might be causing toxicity at a low level. And to make a complicated situation worse is that it's tough for the US EPA to look at that huge amounts of compounds and then determine which ones should we really focus on with the resources that we have. And I think a good way that they're trying to accomplish this is with the contaminant candidate list. Now we're looking, every six years they look at a new set of contaminants that they're going to hold water treatment facilities to monitor. And they're currently looking at nominations for the CCL 4 list. On the last round, I thought it would be interesting to show how they go through that process of weeding out candidates and getting to a final value. First they look at the whole universe of compounds, and they came down to about 7500 potential chemical and microbial contaminants. 

Then they prioritize down to about 600 using the contaminant's potential to occur in public water systems and the potential for public health concern. Finally, they weed it down to 116 contaminants based on detailed evaluation of occurrence and health effects and expert judgment from toxicologists. In that first step, between universe and the PPCL, a lot is based on the structure activity relationships. So they're looking at toxicity of known compounds and looking at compounds with similar structure. But a new exciting program, again it's a multi-federal agency collaboration between EPA, the NIH, and FDA, is called the Tox21 program which is looking at a much wider variety of compounds and using high throughput robotic screening with cellular based assays to try to determine the toxicology on a cellular level. So we're looking at cellular receptor biology such as the endocrine disruption, but we're also looking at here the gene arrays and how genes might be upregulated or downregulated based on the exposure to toxicants or combinations of toxicants. So that's an exciting program that I think will inform not only EPA for their contaminants but FDA for food safety testing.   Now looking at environmental sustainability, I look at this from a consumer standpoint. And I recently came across a documentary that was made by the state of Michigan on the pharmaceutical and emerging contaminants in water and found this half hour program that was shown on Michigan Public Television. It's also available on YouTube if you want to search for it. 

But there are a number of different pharmaceutical take back initiatives. And the DEA is looking at a national drug take back initiative, and they work with local municipalities to create these take back events. And they also recently were able to place drug drop-off boxes by police stations and municipal buildings. And I was just picking Stoughton because that's my home community, and I've actually been to the drug disposal event which is held at the local fire station there. And I was happy to see the drug drop-off box outside of our police station. And just today I saw an email stating that soon the DEA is going to issue a note, a ruling that pharmacies, such as CVS, Walgreens, if they're willing to participate, will be able to have these drug drop-off boxes by their facilities as well. So I hope this has been a real interesting way to look at not only the little project that we did but pharmaceuticals and the environment as a whole and how they relate to different aspects and looking at different perspectives. But for the project itself, I wanted to acknowledge Benjamin Blair and Jordan Crago from the Klaper Lab and colleagues that helped me with the sample preparation: Archie Degnan, Steven Quinlan, Nicole Nicksic, and Yajaira Pluess. And I'm missing a lot of people from the UW Milwaukee side, the people who man the Neeskay, the divers that went down for the sediments. Again, the project was funded by the Milwaukee Metropolitan Sewage District, but also Ben Blair had a research assistantship from the National Oceanic and Atmospheric Administration. 

 And with that, I thank you for your attention, and I left my email. If any of you have questions, feel free to contact me at that, and thank you for your attention. [APPLAUSE]