Laura Bush is the Editor in Chief for BioPharm International

Laura Bush

In 2009, a group of researchers conducted a study using inductively coupled plasma–mass spectrometry (ICP-MS) to determine if heavy metal contaminants were present in pet foods (1,2). The team conducted a follow-up study in 2019, using updated cryogenic and microwave technologies for sample preparation before ICP-MS analysis. We spoke to the study’s lead author, Patricia Atkins about these investigations.

What prompted the original study (1,2) and the update?


In my role at Spex CertiPrep I am tasked with creating new and interesting research projects, I often draw from my life experiences and interests for topics. In 2009, I was a grieving pet owner. I felt that I had fed my cats the best foods, but one had died from cancer. I wondered if the food was as good as purported. This was just a year or two after the melamine scandal in pet food. I had the ability to do some testing, so I did.

Since that original study, 10 years had passed and the U.S. Food Safety Modernization Act (FMSA) had been enacted. It seemed time to revisit the study.

In the new study, we focused only on dry food, as opposed to wet and dry food of the first study. We primarily did that to unify our preparation and analysis. Our dry foods were purchased or donated by employees and ground using cryogenic milling. Each pet food was sampled three times, and run in triplicate with cleaning runs and blanks between samples. All replicates were examined by ICP-MS, and if one replicate appeared contaminated, it was either rerun or eliminated.

Did you see any significant changes since the first study? Have you changed your sample preparation or analysis method over time?

Overall, we found that the levels of heavy metals had not significantly changed in 10 years. Both dog and cat foods had higher levels of arsenic and nickel than what had been found in 2009. Arsenic in 2009 had a maximum of 290 ppb, while in 2019 we saw it up to 690 ppb. Nickel in 2009 had a maximum of 3200 ppb, but in 2019 we found up to 5900 ppb. The levels of lead overall in the 2019 samples had decreased compared to 2009 from a maximum of 5.9 ppm to 0.5 ppm. In our first study, we had surprisingly found uranium up to 0.9 ppm in our dog food samples; in this study, we have found cat and dog food samples containing up to 1.7 ppm of uranium.

Our sample preparation and analysis methods over the decade have change some with new advances in microwave and ICP-MS technologies. Our instruments are more sensitive, and see smaller amounts with the ability to filter out interferences, which in turn increases accuracy.

The first study attracted attention. Did you get any pushback from manufacturers, or any response from any governmental bodies? Do you expect any additional response from this study?

Yes, the first study did receive a lot of attention, both good and bad. I spoke with many pet food manufacturers at conferences, and while they would not publicly comment, many personally agreed with our findings. We also had the Royal Military College of Canada request our pet food samples to try analysis with a new type of instrumentation. We shared our samples, and, overall, their results corresponded with ours. Some of the governmental bodies, including the FDA, did have some feedback. I take these responses and any that may come from our current study as helpful criticism to create better studies in the future. Some requests from this current study was to bring back the wet food studies and include new frozen or refrigerated pet foods.

Have you had any regulatory group or pet food supplier criticize your work on a scientific basis?

There were a few points of contention from the FDA in our 2009 study, based upon how we chose to prepare samples and compare them to feeding amounts and regulatory limits. The criticisms focused around calculating for dry weight of wet food samples, and the amount we standardized to feed pets and using human limits for pets. So, in this study, we focused only on dry food, and used pet food manufacturer feeding guide- lines, but still kept the human reference dosage models.

(1) P. Atkins, L. Ernyei, W. Driscoll, and R. Thomas, 

(2) P. Atkins, L. Ernyei, W. Driscoll, R. Obenauf, and R. Thomas, 

See the full interview at: https://www.spectroscopyonline.com/view/assessing-heavy-metals-pet-food-icp-ms-update.

 is a Senior Application Scientist at Spex CertiPrep’s Certified Reference Material’s division. ●

Articles

A 2009 study found many heavy metals in pet food. Has anything changed?

An Update on Assessing Heavy Metals in Pet Food with ICP-MS

We are pleased to present here part II of our celebration of 

 35th year of publishing. For this special coverage, we asked leading spectroscopists to share their perspectives on a range of spectroscopic techniques and applications. Their articles, split between the June and July issues, highlight important developments, look at opportunities, and consider what challenges must be overcome for future advances. We hope that these contributions-26 in total over the two issues-give you a good sense of the current state of analytical spectroscopy, and what the future might hold. 

	One theme that arose naturally in pulling together these articles is the contributions that various spectroscopic techniques can make in biomedical applications. In this issue, Katsumasa Fujita of Osaka University starts this off with a look at biomedical Raman imaging. Frank Vanhaecke of Ghent University, in turn, highlights the power of multicollector inductively coupled plasma–mass spectrometry (MC-ICP-MS) for isotope metallomics. Vincent Motto-Ross of the University of Lyon, with collaborators from medical institutions, illustrates the advance of laser-induced breakdown spectroscopy (LIBS) imaging into clinical diagnostics. Gunda Koellensperger and colleagues at the University of Vienna explain the central role that ICP-MS is playing in addressing the metallome at the single-cell level.

	Nanoscale measurements with ICP-MS is another theme. John Olesik of the Ohio State University assesses progress in the use of single-particle ICP-MS for the analysis of engineered and natural nanoparticles, and the factors that can affect measurement accuracy. Joerg Bettmer and colleagues at the University of Oviedo discuss the challenges of analyzing the smallest of nanoparticles and potential strategies to overcome those difficulties.

	With a look at instrumental developments, Steve Buckley of Ocean Insight, who is also our “Lasers and Optics Interface” columnist, summarizes recent advances in light sources for spectroscopy, discussing where further developments are expected and the broader impact on instruments and applications. Richard Crocombe, in turn, assesses the future of portable spectroscopy.

	Addressing the use of spectroscopy in forensics, Brooke Kammrath of the University of New Haven explains how the combination of spectroscopy and microscopy can advance the analysis of forensically relevant traces. Diana Wright of the FBI discusses efforts to develop more uniform comparison language in forensic science reporting.

	Taking a broader view, Gary Hieftje considers the current and future status of atomic spectroscopy, with an ominously titled article (“Is the End Near?”). In a companion piece, Rick Russo asks what must be done to ensure sufficient funding for the next generation of atomic spectroscopists.

	Burkhard Beckhoff of Physikalisch-Technische Bundesanstalt wraps up our special coverage with a discussion of an application with broad potential impact in our modern lives: the use of X-ray spectrometry to investigate aging mechanisms in the high-energy-density battery materials that will power important devices in the future.

	Anniversaries cause us to reflect on how far we have come, where we stand, and where we are going. The articles in our two anniversary issues do exactly that-highlighting important accomplishments as well as significant challenges that remain. Based on what I have seen so far, I believe that the spectroscopy community will embrace the opportunities of today, to bring even more exciting advances tomorrow.

We are pleased to present here part II of our celebration of Spectroscopy’s 35th year of publishing. For this special coverage, we asked leading spectroscopists to share their perspectives on a range of spectroscopic techniques and applications.

Where Spectroscopy Is Heading

are excited to be celebrating our 35th year of publishing. To honor the occasion, we asked leading spectroscopists to share their perspectives on their areas of focus, whether that be a specific technique or an area of application. Some of these articles assess the significant advances that have been made in recent decades. Many address an important current challenge that spectroscopists are working to overcome. Others look at opportunities for the future. Some do all three.

	In total, we received 26 excellent contributions, and we are grateful to the thought leaders who took the time to share their views with you, our readers. Given the enormous response, we have made what was originally going to be a single anniversary issue into two, this month and next-for double the impact! In each issue, we include articles that address a range of analytical spectroscopy techniques and applications, to give you a broad sense of where the field of analytical spectroscopy stands and where it is heading.

	In this issue, we start with a look at Raman spectroscopy. Zac Schultz of the Ohio State University assesses machine learning for enabling complex bioanalysis with surface-enhanced Raman scattering (SERS). Mun Seok Jeong of Sungkyunkwan University evaluates at the evolution and future of tip-enhanced Raman spectroscopy (TERS) as a quantitative and qualitative analysis technique.

	Robert Hannah looks at the gaps in teaching spectral interpretation of both Raman and infrared (IR) spectra. Heinz Siesler of the University of Duisburg-Essen and Hu Yan of Jiangsu University of Science and Technology assess the reality of handheld near-infrared (NIR) spectrometers, and how that compares to the hype. 

	John Olesik of the Ohio State University considers the strengths and shortcomings of inductively coupled plasma–optical emission spectroscopy (ICP-OES), and in what ways other techniques may (or may not) be able to challenge this technique. Norbert Jakubowski, a former division head at the German Federal Institute for Materials Research and Testing (BAM), considers the promise of ICP-mass spectrometry (ICP-MS) for single-cell and single-protein analysis.

	Jerry Workman and Howard Mark-who share our anniversary, given that they have been writing the “Chemometrics in Spectroscopy” column for our publication for 35 years-address an important remaining challenge in chemometrics. Juergen Popp and colleagues at the Leibniz Institute of Photonic Technology consider how molecular spectroscopy techniques can be combined with artificial intelligence methods to make them more effective as process analytical technology (PAT).

	Yukihiro Ozaki of Kwansei Gakuin University discusses the strengths of far-ultraviolet spectroscopy, and describes a few applications where those features can provide great benefits. José Manuel Costa-Fernandez of the University of Oviedo describes the advantages of using nanoparticle labels with ICP-MS for the analysis of clinical biomarkers. Dmitry Kurouski of Texas A&M University shows how Raman spectroscopy is aiding agriculture, and Igor Lednev of the State University of New York at Albany explains how Raman spectroscopy can help determine the time of a crime.

	Duncan Graham of the University of Strathclyde wraps up our special coverage this month by addressing a very timely question: Can Raman spectroscopy aid in the fight against the current coronavirus pandemic?

	These articles highlight the important contributions that analytical spectroscopy techniques have made, and are continuing to make, in the broader field of analytical chemistry and in a wide range of scientific disciplines and industries. They also provide help focus our attention on remaining gaps and significant opportunities.

	Please tune in next month for the second half of our special anniversary coverage. There, another set of experts will address questions in these and other techniques and applications.

In celebration of Spectroscopy’s 35th Anniversary, leading spectroscopists discuss important issues and challenges in analytical spectroscopy.

Celebrating 35 Years of Spectroscopy

As the use of nanoparticles in a wide range of applications continues to increase, the need for techniques to analyze these particles also grows. Javier Laserna of the University of Málaga, in Spain, has developed an approach to measure individual nanoparticles in air using optical trapping (OT) combined with laser-induced breakdown spectroscopy (LIBS). Laserna received the 2018 Lester W. Strock award for this development as well as his broader research on analytical methods and instrumentation using LIBS. He recently spoke to us about this work.

What led you to investigate LIBS for nanoparticles?

	Well, to start with, nanoparticles, as everybody knows, are of extreme interest because of the growing number of applications for these objects in many areas of technology. There are only a few analytical techniques for measuring the composition of nanoparticles, and with most of those techniques, it is only possible to analyze nanoparticles in bulk or when they are attached to a surface or suspended in solution. The ability to analyze individual particles is almost nonexistent; only a few techniques are available, and none of them provide the complete analytical composition of the nanoparticle, as far as I know.

	LIBS is well known for its ability to provide panoramic spectra-from the ultraviolet (UV) to the infrared (IR)-in a single shot, in bulk materials. But in the case of nanoparticles, we wanted to analyze these extremely light and extremely low-mass particles in a single shot.

	So, we tried it and to our surprise, it worked rather well. The signal-to-noise ratio we obtained was very reasonable considering the very low mass of the nanoparticles.

You developed a method for elemental identification of nanoparticles using LIBS that involves optical trapping. How are the optical trapping and LIBS measurement carried out?

	One of the most difficult tasks in this work is to isolate an individual nanoparticle for analysis. This is by no means an easy task. For this purpose we used an optical trap. The concept behind optical trapping technology is rather simple. A continuous-wave (CW) laser beam, propagating upwards, is focused on sample nanoparticles in air or in solution (Figure 1a). The radiation pressure created in this nanoparticle medium is able to counteract the gravity forces that are acting on the particles. So, in principle, this optical field is able to trap the particles. However, the trapping is not always easy, particularly when the particles are very small and when the trapping is performed in air. The air currents in the laboratory, for example, tend to disrupt the particles and they fall out of the trap.

	So, we created a confined nanoparticle medium by using a cuvette that is commonly used for spectrophotometry. That way, it is possible to keep the particles inside the trap and free from the air currents. But before you can get to the optical trapping, you have to create an aerosol of the particles you want to analyze. We did that using optical catapulting.

	This is done by putting a powder containing nanoparticles on top of a microscope slide. Then we shoot a laser pulse into the nanopowder from under the slide. The shockwave created from the laser pulse beneath the slide creates the nanoparticle aerosol. The aerosol then interacts with the CW laser, which is sent colinearly with the catapulting beam, propagating upwards through the microscope slide. This CW laser is tightly focused by a microscope objective, which is a 20x laser objective with a numerical aperture of 0.4. All of this is done inside the cuvette to keep the nanoparticles from blowing away (Figure 1b).

	The aerosol created in the cell is highly dispersed, so we have to wait for the nanoparticle to deposit by gravity. Usually, after 3 to 5 minutes, the particle is trapped; the exact time depends on factors such as particle size, particle geometry, and the optical properties of the particle, but most particles are trapped within about 5 minutes.

	In the optical trap it is possible to capture a cluster of particles rather than a single particle. So, we repeat measurements several times until the signal has a low range S/N. This measurement response indicates that the particles creating this signal are individual particles.

	We can modify the trapping beam to make the particles go up or down in the trap by changing the power of the trapping laser. We can manipulate the particles so that particles that are too large fall outside the trap. Also, we sometimes observe strings of particles in the trap. We manipulate the laser system so that we end up with only one particle in the trap.

	The next thing that is important is to have the particle in a precise position in space so that the nanoparticle can be effectively interacted with by the LIBS excitation beam, which is shot perpendicular to the particle trapping beam. This analyzing laser is a moderate-power laser (200 millijoules per pulse), with excitation wavelength set at 1064 nm. We send a single shot of this laser at the nanoparticle that is trapped. It is important that the nanoparticle targeted is precisely located where laser is going to hit. That way, we can measure the spectrum of the particle. If the nanoparticle is not in that precise spot (at 0,0,0 coordinates), the S/N is nil and we don’t see any signal from the analyte. The only thing we see in that case is the spectrum of air.

	Then what we observe in the spectra is the mixture of the spectra of the nanoparticle plus the spectrum of air. So how do we get rid of the air?

	There are several strategies we can use. The first is to delay integration of the signal by several microseconds. In this way, many of the ionic spectral lines of nitrogen and oxygen in particular are reduced, virtually to the point of disappearing, and so then we have only the weak ionic lines of air-nitrogen of oxygen-and the spectral lines belonging to the nanoparticle. Some atomic lines of oxygen and nitrogen are still present but they can be resolved from the analyte lines by our spectrometer.

	The second strategy is to focus the analytical beam a little ahead of the position of the particle.

	The most intense lines of air are a little bit off axis of where we are observing the signal. The plasma of air and the nanoparticle are both observed simultaneously. We then, through data analysis, discriminate between the air and particle signals.

	This is a complex process, but the important thing is that we are able to discriminate the plasma of air from the plasma of the particle.

Your method enabled detection of nanoparticles smaller than 100 nm-in other words, in the attogram (10

 gram) regime. What challenges did you have to overcome to achieve detection of particles that small?

	At first, we were a bit surprised that this technology, LIBS, could detect such tiny particles. We had heard many times that one of the barriers to wider adoption of LIBS is low sensitivity. We have been working with LIBS for many years, so we knew that what is meant by low sensitivity is low sensitivity when analyzing trace elements in a matrix. For example, in solid materials, such as when we are analyzing an alloy with one element at trace level, LIBS can rarely achieve a limit of detection (LOD) below the low ppm level.

	However, the ability to analyze signals from monoelemental materials, like pure copper particles, is much improved and lower LODs than ppm can be achieved.

	So, we started investigating this. We started with microparticles in the 2-micrometer size and achieved a reasonable S/N. Then we went down to 100-nm particles and still achieved a very nice signal-not only with metal nanoparticles like nickel particles but also with silica nanoparticles. We analyzed smaller and smaller particles and still obtained an acceptable S/N. So, we learned that LIBS is able to detect extremely low masses when particles are single-phase or single-component particles.

	Then, we discovered that we are not limited to analyzing mono-elemental particles. We were able to analyze multi-elemental particles, such as ferrite particles composed of from three to four elements, and we were able to analyze all the components at same time.

	So, we kept investigating ever smaller particles, and we were able to analyze 25-nm-sized particles; this is close to the LOD. Ultimately, at this size of nanoparticle, we achieved an LOD on the order of 57 attograms of mass, which is a spectacular tiny size and mass for LIBS and for comparable technologies.

What novel measurements have you been able to achieve with this approach so far?

	For the moment we are still doing research work on this approach. We are still working to understand the many processes involved and how to optimize the many variables that affect the system and trying to understand how the signals can be captured in the most efficient manner possible. The solid angle that we use for signal collection in LIBS is very low in comparison with the 4π emitting geometry of a plasma.

	We are trying to improve the capture of the photons emitted by the plasma. Also, we are trying to avoid one of the most inefficient steps in spectroscopy. When analyzing emission from a light source with a traditional monochromator or spectrograph, a considerable amount of photons from the source are lost at the entrance slit. This is the very nature of these devices; it is what they are designed to do.

	We are trying to avoid this loss of photon signal from traditional monochromator-based measurements by applying a hyperspectral approach to collect the signal from the plasma. By using an appropriate set of filters that collects all the signals from the analyte of interest we expect to capture all the photons and analyze them all.

	This research is ongoing, and we still don’t have definitive results. But if this works, we will have much better LODs at a much lower attogram sample mass level.

How could this technique actually be used in a practical application?

	To think about this, we are reviewing the sampling efficiency of the approach. As I said before, we can capture one particle about every 3 minutes on average. So, if we need to analyze hundreds of thousands of particles, this rate would be prohibitive, and we could not use this approach as a screening technique for large numbers of particles.

	So, what we believe is the main application of our technology is the analysis conditions where we have only very tiny amounts of sample, such as functionalized nanoparticles that are synthesized in very small amounts for biology research. These samples are hard to analyze by other technologies. Most technologies need relatively large amounts of sample. So, when the amount of sample available for analysis is on the order of micrograms, our technology provides a solution.

	At the same time, we also intend to increase our sample throughput by analyzing the nanoparticles in a flow. This is another approach that has nothing to do with the optical trapping or catapulting we have been discussing. In this approach, we would create a sample suspension and create an aerosol of microdroplets and a device to dry the sample, so that we can analyze them on the fly, with a kind of time-of-flight (TOF) analyzer using LIBS. We are only starting this work but are hopeful that this will provide much better sample throughput for analyzing single nanoparticles.

	For now, this is a laboratory technique, but we think this can evolve into a commercial product in the future.

	Remember that the need for this technology is growing because there are more and more nanoparticles being used in many types of technologies. That means the need to characterize the chemical properties-not only the optical or magnetic properties-of nanoparticles is increasing. Those chemical properties are very important in many applications. And currently, very few technologies are available to analyze the chemical composition of nanoparticles, so we see opportunity here. So, we plan to protect our work through a patent, and we are preparing a patent application now.

		P. Purohit, F.J. Fortes, and J.J. Laserna, 

Javier Laserna is a professor of analytical chemistry at the University of Málaga, in Málaga, Spain. His current research interests include the use of lasers in chemical analysis, laser-induced plasma spectroscopy; time-of-flight mass spectrometry; secondary ionization mass spectrometry; surface analysis using laser ablation with optical and ion detection, and laser remote chemical analysis. In the 1990s, he succeeded in demonstrating large-scale optics standoff laser induced breakdown spectroscopy for analysis of distant objects. Since then, this technique has been used for the analysis of explosives and in space exploration. Currently, he is a co-investigator of the SuperCam instrument for the NASA Mars 2020 mission. Laserna has participated in several research projects funded by the European Commission’s Framework Programme as both a consortium member and coordinator. He has also been the principal investigator of more than 25 research projects funded by the Spanish central government and the regional goverment of Andalucía. He has published over 300 scientific papers and six books or book chapters and is a co-inventor of seven patents held by the University of Málaga. Laserna was awarded the Spanish Royal Society of Chemistry National Award for Research in Analytical Chemistry in 2009 and the Spanish Society for Applied Spectroscopy National Award for his research career in Applied Spectroscopy in 2010. He is also the 2018 recipient of the Lester W. Strock Award from the Society for Applied Spectroscopy.

As the use of nanoparticles in a wide range of applications continues to increase, the need for techniques to analyze these particles also increases. Javier Laserna of the University of Málaga, in Spain, has developed an approach to measure individual nanoparticles in air using optical trapping (OT) combined with laser-induced breakdown spectroscopy (LIBS). Laserna received the 2018 Lester W. Strock award for this development as well as his broader research on analytical methods and instrumentation using LIBS. He recently spoke to us about this work. 

Laura Bush

Articles

An Update on Assessing Heavy Metals in Pet Food with ICP-MS

Where Spectroscopy Is Heading

Celebrating 35 Years of Spectroscopy

Detection and Identification of Individual Nanoparticles Using Laser-Induced Breakdown Spectroscopy (LIBS)