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Chemistry of Chocolate

Chemistry of Chocolate

Swiss… Belgian… Pure… Milk…

Here we are in mid-February again, bombarded by chocolate from all sides in preparation for Valentine’s Day on the 14th. Whether in a solid bar, as a chewy truffle, or as a luxurious drink, chocolate has completely infiltrated our lives. Most people can agree that this confectionary treat is fantastic for any occasion – to be given as a gift, to recover after having a bad day, as well as to celebrate a good one – chocolate is certainly meant to be enjoyed.

Even if you don’t like the taste, the chances are high that someone close to you does. So how can you be certain of its quality?

Components of a chocolate bar

For the sake of this article, let us consider the humble chocolate bar, without any extra additions (not to mention any Golden Tickets). This form can be found worldwide in nearly any grocery store or candy shop, generally designated as white, milk, or dark.

All of this variability comes from the edible seeds in the fruit of the cacao tree, which grows in hot, tropical regions around the equator. They must be fermented and then roasted after cleaning. From this, cocoa mass is produced, which is a starting base for several uses. Cocoa butter and cocoa solids are prepared from the cocoa mass and are utilized in products ranging from foods and beverages to personal care items.

As for chocolate bars, these are generally sweetened and modified from the pure form, which is very bitter. Milk (liquid, condensed, or powdered) is added to many types, but does not necessarily have to be present. Varying the content of the cocoa solids and cocoa butter in chocolate to different degrees results in the classifications of dark to white. While some dark chocolates do not contain any milk, white chocolates do to add to the significant amounts of cocoa fat used to produce them.

In general, dark chocolate contains a high ratio of cocoa solids to cocoa butter and may or may not contain any milk. It may be sweetened or unsweetened. Milk chocolate is a much broader category, containing less cocoa solids but not necessarily a different cocoa butter content compared to dark chocolate, as milk fats are also introduced. Milk chocolate is also sweetened, either with sugar or other substitutes. White chocolate contains no cocoa solids at all, but a blend of cocoa butter and milk, along with sweeteners.

Depending on the country, there are different regulations in place regarding the classification of the type of chocolate. If you are interested, you can find a selection of them here.

What makes your favorite chocolate unique?

Of course, more ingredients are added to chocolate bars to affect a number of things like the aroma, texture/mouthfeel, and certainly to enhance the flavor. The origin of the cacao beans, much like coffee, can impart certain characteristics to the resulting chocolate. The manufacturing process also plays a major part in determining e.g., whether the chocolate has a characteristic snap or has a distinct scent, setting it apart from other brands.

In some cases, vegetable fats are used to replace a portion of the cocoa fats, although this may not legally be considered «chocolate» in some countries. The adjustment of long-standing recipes for certain chocolate brands has sometimes led to customer backlash, as quality is perceived to have changed. Truly, chocolate is inextricably tied to our hearts.

Applications for chocolate quality analysis

Nobody wants to give their Valentine a bad gift, especially out-of-date chocolate from a dubious source. Here, we have prepared some interesting analyses for different chocolate quality parameters in the laboratory.

Sugar analysis via Ion Chromatography (IC)

Most types of chocolate contain sugars or sugar substitutes to sweeten the underlying bitterness. Considering different regulations regarding food labeling and also nutritional content, the accurate reporting of sugars is important for manufacturers and consumers alike.

Sugar analysis in chocolate can be performed with Metrohm IC and Pulsed Amperometric Detection (PAD). An example chromatogram of this analysis is given below.

A small amount of commercially produced sweetened milk chocolate was weighed and dissolved into ultrapure water. After further sample preparation using Metrohm Inline Ultrafiltration, the sample (20 µL) was injected on to the Metrosep Carb 2 – 150/4.0 separation column and separated using alkaline eluent. As shown, both lactose and sucrose elute without overlap in less than 20 minutes.

Learn more about Metrohm Inline Ultrafiltration for difficult sample matrices and safeguard your IC system!

In this example, the sugar content was listed on the label as 47 g per 100 g portion (470 g/kg). Lactose was determined to be 94.6 g/kg, and sucrose was measured at 385.6 g/kg. To learn about what other carbohydrates, sweeteners, and more can be determined in chocolate and other foods with Metrohm IC, download our free brochure about Food Analysis and check out the table on page 25!

Lactose content in lactose-free chocolate

The accurate measurement of lactose in lactose-free products, including chocolate, is of special importance to consumers who are lactose-intolerant and suffer from digestive issues after eating it. Foods which are labelled as lactose-free must adhere to guidelines concerning the actual non-zero lactose content. Foodstuff containing less than 0.1 g lactose per100 g (or 100 mL) is most frequently declared as lactose-free.

Determination of lactose in chocolate is possible with IC. Here is an chromatographic overlay of a dissolved chocolate sample with lactose spikes which was analyzed via Metrohm IC using the flexiPAD detection mode.

Milk chocolate, labelled lactose-free measured via Metrohm IC (0.57 ± 0.06 mg/100 g lactose, n = 6).

The sample contained 0.6 mg lactose per 100 g, with measurement of the lactose peak occurring at 13.2 minutes. The black line is the unspiked lactose-free chocolate sample, red and blue are spiked samples of increasing concentration. To prepare the samples, approximately 2.5–5 g chocolate was dissolved in heated ultrapure water, using Carrez reagents to remove excess proteins and fats from the sample matrix. Afterward, centrifugation of the samples was performed, followed by the direct injection of the supernatant (10 µL) into the IC system. Measurement was performed with the Metrosep Carb 2 – 250/4.0 separation column and an alkaline eluent.

Interested in lactose determinations with ion chromatography? Download our free Application Notes on the Metrohm website!

Water determination with Karl Fischer Titration

The amount of water in foods, including chocolate, can affect their shelf life and stability, as well as contribute to other physical and chemical factors. Aside from this, during the processing stage, the amount of water present affects the flow characteristics of the chocolate mass.

AOAC Official Method 977.10 lists Karl Fischer titration as the accepted analysis method for moisture in cacao products.

The determination of moisture in different chocolate products is exhibited in the following downloadable poster. As an example, several samples (n = 10) of dark chocolate (45% cocoa content) were analyzed for their moisture content with Metrohm Karl Fischer titration.  Results were found to be 0.96% water with a relative standard deviation (RSD) of 2.73%. More information about this analysis can be found in our poster about automated water determination in chocolate, or in chapter 11.6 of our comprehensive Monograph about Karl Fischer titration.

Oxidation stability with the Rancimat test

Oxidation stability is an important quality criterion of chocolate as it provides information about the long-term stability of the product. Cocoa contains various flavonoids that act as antioxidants. Although the flavonoid content may vary amongst chocolate type, in general, the greater the content of cocoa solids in the chocolate, the greater its antioxidant effect.

The 892 Professional Rancimat from Metrohm determines the oxidation stability of fat-containing foods and cosmetics. The Rancimat method accelerates the aging process of the sample and measures the induction time or oxidation stability index (OSI).

Chocolate cannot be measured directly with the classical Rancimat method, as no evaluable induction time is obtained. There are many reasons for this: e.g., the fat content is too low. Traditionally, extraction of the fat from the chocolate is necessary, but not always.

Learn more about the Rancimat method on our website, and download our free Application Note about the oxidation stability of chocolate. In this Application Note, the oxidation stability of white, milk and dark chocolate is determined without extraction.

Cadmium in chocolate by Voltammetric analysis

The toxic element cadmium (Cd) can be found in elevated concentrations with high bioavailability in some soils. Under such conditions, cacao trees can accumulate cadmium in the beans. Chocolate produced from the affected beans will contain elevated cadmium levels.

Typical limit values for Cd in chocolate in the European Union are between 100 µg/kg and 800 µg/kg (EU Commission Regulation 1881/2006) depending on the cocoa content of the chocolate. Anodic stripping voltammetry (ASV) can be used to accurately determine trace quantities of cadmium in chocolate down to approximately 10 µg/kg. The method is simple to perform, specific, and free of interferences.

Chocolate samples are first mineralized by dry ashing in a furnace at 450 °C for several hours. The remaining ash is then dissolved in an acidified matrix. The cadmium determination is then carried out on the 884 Professional VA instrument from Metrohm. To learn more about how to perform the analysis, download our free Application Note.

Happy Valentine’s Day from us all at Metrohm!

Post written by Dr. Alyson Lanciki, Scientific Editor at Metrohm International Headquarters, Herisau, Switzerland.

«Analyze This»: 2020 in review

«Analyze This»: 2020 in review

I wanted to end 2020 by thanking all of you for making «Analyze This» – the Metrohm blog for chemists such a success! For our 60th blog post, I’d like to look back and focus on the wealth of interesting topics we have published this year. There is truly something for everyone: it doesn’t matter whether your lab focuses on titration or spectroscopic techniques, or analyzes water samples or illicit substances – we’ve got you covered! If you’re looking to answer your most burning chemical analysis questions, we have FAQs and other series full of advice from the experts. Or if you’re just in the mood to learn something new in a few minutes, there are several posts about the chemical world to discover.

We love to hear back from you as well. Leaving comments on your favorite blog posts or contacting us through social media are great ways to voice your opinion—we at Metrohm are here for you!

Finally, I wish you and your families a safe, restful holiday season. «Analyze This» will return on January 11, 2021, so subscribe if you haven’t already done so, and bookmark this page for an overview of all of our articles grouped by topic!

Stay healthy, and stay curious.

Best wishes,

Dr. Alyson Lanciki, Scientific Editor, Metrohm AG

Quickly jump directly to any section by clicking a topic:

Customer Stories

We are curious by nature, and enjoy hearing about the variety of projects where our products are being used! For some examples of interesting situations where Metrohm analytical equipment is utilized, read on.

From underwater archaeological research to orbiting Earth on the International Space Station, Metrohm is there! We assist on all types of projects, like brewing top quality beers and even growing antibiotic-free shrimp – right here in Switzerland.

Interested in being featured? Contact your local Metrohm dealer for details!


Metrohm is the global market leader in analytical instruments for titration. Who else is better then to advise you in this area? Our experts are eager to share their knowledge with you, and show this with the abundance of topics they have contributed this year to our blog.

For more in-depth information about obtaining the most accurate pH measurements, take a look at our FAQ about pH calibration or read about avoiding the most common mistakes in pH measurement. You may pick up a few tips!

Choose the best electrode for your needs and keep it in top condition with our best practices, and then learn how to standardize titrant properly. Better understand what to consider during back-titration, check out thermometric titration and its advantages and applications, or read about the most common challenges and how to overcome them when carrying out complexometric titrations

If you are interested in improving your conductivity measurements, measuring dissolved oxygen, or the determination of oxidation in edible fats and oils, check out these blog posts and download our free Application Notes and White Papers!

Finally, this article about comprehensive water analysis with a combination of titration and ion chromatography explains the many benefits for laboratories with large sample loads. The history behind the TitrIC analysis system used for these studies can be found in a separate blog post.

Karl Fischer Titration

Metrohm and Karl Fischer titration: a long history of success. Looking back on more than half a century of experience in KFT, Metrohm has shaped what coulometric and volumetric water analysis are today.

Aside from the other titration blog posts, our experts have also written a 2-part series including 20 of the most frequently asked questions for KFT arranged into three categories: instrument preparation and handling, titration troubleshooting, and the oven technique. Our article about how to properly standardize Karl Fischer titrant will take you step by step through the process to obtain correct results.

For more specific questions, read about the oven method for sample preparation, or which is the best technique to choose when measuring moisture in certain situations: Karl Fischer titration, near-infrared spectroscopy, or both?

Ion Chromatography (IC)

Ion chromatography has been a part of the Metrohm portfolio since the late 1980s. From routine IC analysis to research and development, and from stand-alone analyzers to fully automated systems, Metrohm has provided IC solutions for all situations. If you’re curious about the backstory of R&D, check out the ongoing series about the history of IC at Metrohm.

Metrohm IC user sitting at a laboratory bench.

Common questions for users are answered in blog posts about IC column tips and tricks and Metrohm inline ultrafiltration. Clear calculations showing how to increase productivity and profitability in environmental analysis with IC perfectly complement our article about comprehensive water analysis using IC and titration together for faster sample throughput.

On the topic of foods and beverages, you can find out how to determine total sulfite faster and easier than ever, measure herbicides in drinking water, or even learn how Metrohm IC is used in Switzerland to grow shrimp!

Near-Infrared Spectroscopy (NIRS)

Metrohm NIRS analyzers for the lab and for process analysis enable you to perform routine analysis quickly and with confidence – without requiring sample preparation or additional reagents and yielding results in less than a minute. Combining visible (Vis) and near-infrared (NIR) spectroscopy, these analyzers are capable of performing qualitative analysis of various materials and quantitative analysis of a number of physical and chemical parameters in one run.

Our experts have written all about the benefits of NIR spectroscopy in a 4-part series, which includes an explanation of the advantages of NIRS over conventional wet chemical analysis methods, differences between NIR and IR spectroscopy, how to implement NIRS in your laboratory workflow, and examples of how pre-calibrations make implementation even quicker.

A comparison between NIRS and the Karl Fischer titration method for moisture analysis is made in a dedicated article.

A 2-part FAQ about NIRS has also been written in a collaboration between our laboratory and process analysis colleagues, covering all kinds of questions related to both worlds.

Raman Spectroscopy

This latest addition to the Metrohm family expands the Metrohm portfolio to include novel, portable instruments for materials identification and verification. We offer both Metrohm Raman as well as B&W Tek products to cover a variety of needs and requirements.

Here you can find out some of the history of Raman spectroscopy including the origin story behind Mira, the handheld Raman instrument from Metrohm Raman. For a real-world situation involving methamphetamine identification by law enforcement and first responders, read about Mira DS in action – detecting drugs safely in the field.

Mira - handheld Raman keeping you safe in hazardous situations.

Are you looking for an easier way to detect food fraud? Our article about Misa describes its detection capabilities and provides several free Application Notes for download.

Process Analytics

We cater to both: the laboratory and the production floor. The techniques and methods for laboratory analysis are also available for automated in-process analysis with the Metrohm Process Analytics brand of industrial process analyzers.

Learn about how Metrohm became pioneers in the process world—developing the world’s first online wet chemistry process analyzer, and find out how Metrohm’s modular IC expertise has been used to push the limits in the industrial process optimization.

Additionally, a 2-part FAQ has been written about near-infrared spectroscopy by both laboratory and process analysis experts, which is helpful when starting out or even if you’re an advanced user.

Finally, we offer a 3-part series about the advantages of process analytical technology (PAT) covering the topics of process automation advantages, digital networking of production plants, and error and risk minimization in process analysis.

Voltammetry (VA)

Voltammetry is an electrochemical method for the determination of trace and ultratrace concentrations of heavy metals and other electrochemically active substances. Both benchtop and portable options are available with a variety of electrodes to choose from, allowing analysis in any situation.

A 5-part series about solid-state electrodes covers a range of new sensors suitable for the determination of «heavy metals» using voltammetric methods. This series offers information and example applications for the Bi drop electrode, scTrace Gold electrode (as well as a modified version), screen-printed electrodes, and the glassy carbon rotating disc electrode.

Come underwater with Metrohm and Hublot in our blog post as they try to find the missing pieces of the ancient Antikythera Mechanism in Greece with voltammetry.

If you’d like to learn about the combination of voltammetry with ion chromatography and the expanded application capabilities, take a look at our article about combined analysis techniques.

Electrochemistry (EC)

Electrochemistry plays an important role in groundbreaking technologies such as battery research, fuel cells, and photovoltaics. Metrohm’s electrochemistry portfolio covers everything from potentiostats/galvanostats to accessories and software.

Our two subsidiaries specializing in electrochemistry, Metrohm Autolab (Utrecht, Netherlands) and Metrohm DropSens (Asturias, Spain) develop and produce a comprehensive portfolio of electrochemistry equipment.

This year, the COVID-19 pandemic has been at the top of the news, and with it came the discussion of testing – how reliable or accurate was the data? In our blog post about virus detection with screen-printed electrodes, we explain the differences between different testing methods and their drawbacks, the many benefits of electrochemical testing methods, and provide a free informative White Paper for interested laboratories involved in this research.

Our electrochemistry instruments have also gone to the International Space Station as part of a research project to more efficiently recycle water on board spacecraft for long-term missions.

The History of…

Stories inspire people, illuminating the origins of theories, concepts, and technologies that we may have become to take for granted. Metrohm aims to inspire chemists—young and old—to be the best and never stop learning. Here, you can find our blog posts that tell the stories behind the scenes, including the Metrohm founder Bertold Suhner.

Bertold Suhner, founder of Metrohm.

For more history behind the research and development behind Metrohm products, take a look at our series about the history of IC at Metrohm, or read about how Mira became mobile. If you are more interested in process analysis, then check out the story about the world’s first process analyzer, built by Metrohm Process Analytics.

Need something lighter? Then the 4-part history of chemistry series may be just what you’re looking for.

Specialty Topics

Some articles do not fit neatly into the same groups as the rest, but are nonetheless filled with informative content! Here you can find an overview of Metrohm’s free webinars, grouped by measurement technique.

If you work in a regulated industry such as pharmaceutical manufacturing or food and beverage production, don’t miss our introduction to Analytical Instrument Qualification and what it can mean for consumer safety!


Finally, if you are more interested in reading articles related to the industry you work in, here are some compilations of our blog posts in various areas including pharmaceutical, illicit substances, food and beverages, and of course water analysis. More applications and information can be found on our website.

Food and beverages
All of these products can be measured for total sulfite content.

Oxidation stability is an estimate of how quickly a fat or oil will become rancid. It is a standard parameter of quality control in the production of oils and fats in the food industry or for the incoming goods inspection in processing facilities. To learn more about how to determine if your edible oils are rancid, read our blog post.

Determining total sulfite in foods and beverages has never been faster or easier than with our IC method. Read on about how to perform this notoriously frustrating analysis and get more details in our free LC/GC The Column article available for download within.

Measuring the true sodium content in foodstuff directly and inexpensively is possible using thermometric titration, which is discussed in more detail here. To find out the best way to determine moisture content in foods, our experts have written a blog post about the differences between Karl Fischer titration and near-infrared spectroscopy methods.

To determine if foods, beverages, spices, and more are adulterated, you no longer have to wait for the lab. With Misa, it is possible to measure a variety of illicit substances in complex matrices within minutes, even on the go.

All of these products can be measured for total sulfite content.

Making high quality products is a subject we are passionate about. This article discusses improving beer brewing practices and focuses on the tailor-made system built for Feldschlösschen, Switzerland’s largest brewer.

Pharmaceutical / healthcare

Like the food sector, pharmaceutical manufacturing is a very tightly regulated industry. Consumer health is on the line if quality drops.

Ensuring that the analytical instruments used in the production processes are professionally qualified is a must, especially when auditors come knocking. Find out more about this step in our blog post about Analytical Instrument Qualification (AIQ).

Moisture content in the excipients, active ingredients, and in the final product is imperative to measure. This can be accomplished with different analytical methods, which we compare and contrast for you here.

The topic of virus detection has been on the minds of everyone this year. In this blog post, we discuss virus detection based on screen-printed electrodes, which are a more cost-effective and customizable option compared to other conventional techniques.

Water analysis

Water is our business. From trace analysis up to high concentration determinations, Metrohm has you covered with a variety of analytical measurement techniques and methods developed by the experts.

Learn how to increase productivity and profitability in environmental analysis laboratories with IC with a real life example and cost calculations, or read about how one of our customers in Switzerland uses automated Metrohm IC to monitor the water quality in shrimp breeding pools.

If heavy metal analysis is what you are interested in, then you may find our 5-part series about trace analysis with solid-state electrodes very handy.

Unwanted substances may find their way into our water supply through agricultural practices. Find out an easier way to determine herbicides in drinking water here!

Water is arguably one of the most important ingredients in the brewing process. Determination of major anions and cations along with other parameters such as alkalinity are described in our blog post celebrating International Beer Day.

All of these products can be measured for total sulfite content.
Illicit / harmful substances

When you are unsure if your expensive spices are real or just a colored powder, if your dairy products have been adulterated with melamine, or fruits and vegetables were sprayed with illegal pesticides, it’s time to test for food fraud. Read our blog post about simple, fast determination of illicit substances in foods and beverages for more information.

Detection of drugs, explosives, and other illegal substances can be performed safely by law enforcement officers and first responders without the need for a lab or chemicals with Mira DS. Here you can read about a real life training to identify a methamphetamine laboratory.

Drinking water regulations are put in place by authorities out of concern for our health. Herbicides are important to measure in our drinking water as they have been found to be carcinogenic in many instances.

Post written by Dr. Alyson Lanciki, Scientific Editor at Metrohm International Headquarters, Herisau, Switzerland.

Electrochemistry in orbit

Electrochemistry in orbit

For over twenty years now, there has been continuous human occupation off our planet.

The International Space Station (ISS), launched in 1998, is a modular satellite in low orbit around the Earth, which is visible even with the naked eye.

Since November 2, 2000, the ISS has had a constantly revolving crew from a variety of nations, working on projects to further push the boundaries of our knowledge. Aside from their important scientific duties, these astronauts must live their daily lives like us – exercising, relaxing, cleaning, and sleeping – albeit in microgravity.

The International Space Station celebrated 20 years of constant habitation in November, 2020.

In October, an Antares rocket carrying a Cygnus resupply ship was launched by NASA at Johnson Space Center. This cargo ship carried an experimental system on board used to study the oxidation of ammonia under microgravity conditions to convert urine into water on the ISS.

Improving this waste management system has far-reaching repercussions for longer exploratory missions where the weight of the payload must be optimized with the amount of water needed (which is heavy) to sustain life during the trip. Given the limited resources aboard a spaceship, the recovery of water from all processes is of great importance.

Future missions which may benefit from this study include trips to the moon (Artemis) and eventually to Mars (Orion).

This system uses Metrohm DropSens screen-printed electrodes (SPEs). The novel nanomaterial coating of the electrodes was developed by researchers at the University of Alicante in Spain in collaboration with the University of Puerto Rico. In this article, we would like to introduce the people behind the project and elaborate on the research they are doing in space with Metrohm products.

Meet the researchers

Dr. José Solla Gullón (Ph.D. 2003, Chemistry)

Dr. José Solla Gullón in his laboratory at the University of Alicante, with Metrohm DropSens and Metrohm Autolab products on the bench.

I am currently a Distinguished Researcher at the Institute of Electrochemistry of the University of Alicante, Spain. My research mainly focuses on the synthesis, characterization and electrochemical properties of different types of nanoparticles with well-defined size, composition, shape, and surface structure. My overall publication record includes about 175 publications (h-index 53). I have also given more than 250 contributions in international and national meetings.

Ms. Camila Morales Navas

Camila Morales Navas holding the Nanoracks 2U, where the electrochemical equipment is kept inside.

I am a senior graduate student in the Department of Chemistry at University of Puerto Rico (UPR). I am working on a research project in collaboration with NASA, titled «Elucidating the Ammonia Electrochemical Oxidation Mechanism via Electrochemical Techniques at the ISS», or «Ammonia Electrooxidation Lab at the ISS (AELISS)» for short. The purpose of this project is to improve the water processing system and to identify new technology for long-term missions in space.

The project is attributed to NASA-ESPCoR, University of Puerto Rico, University of Alicante, NuVant Systems, and Nanoracks, with support from Metrohm DropSens.

Read more about the project on the NASA website:

The AELISS project

For a brief overview by Camila and her graduate advisor, have a look at the video provided below by NASA:

Here, you can see the Metrohm DropSens instruments used for this study: the screen-printed carbon electrode (SPCE8X110) and its corresponding flow-cell (FLWCL8X1C).

The Metrohm DropSens 8X110 carbon SPE (left) and the FLWCL8X1C flow-cell (right).
Instrumentation setup for the AELISS project which was launched to the ISS in October, 2020.

How did the AELISS project begin?

About five years ago, the groups from the University of Alicante and the University of Puerto Rico (UPR) began working together on microgravity experiments which led them to collaborate again for this project, which now resides on the ISS.

The electrochemical oxidation of ammonia using platinum as a catalyst is a well-established reaction, first published almost two decades ago by José’s group. The ammonia is extremely sensitive to the surface structure of platinum. However, this is well-known on earth. How does this reaction process behave in a microgravity environment? The groups sought to determine this by performing experiments in the US using a special airplane which mimics weightlessness for brief periods by flying in a parabolic motion.

SPE modification process: droplets of platinum nanoparticle ink provided by the University of Alicante deposited on the carbon SPEs. Platinum acts as a catalyst for the oxidation reaction. Click image to enlarge.

At first, this was purely for research, but later Camila’s group in Puerto Rico thought more about its potential use in space. Urea from urine is converted to ammonia, which then goes through the electrochemical oxidation process, resulting in N2 gas, water, and energy. Perhaps it was possible to use this technology to improve the onboard water recovery and recycling system in the ISS and other spaceships?

Because the UPR group often writes research proposals that are funded by NASA, they are quite knowledgeable in this area regarding the project requirements, as well as what materials are allowed on board a mission. The UPR group has been working in conjunction with NASA for about 20 years.

Unassembled equipment: plastic protector frame (grey), Metrohm DropSens FLWCL8X1C electrochemical flow-cells with 8X110 carbon SPEs (blue/white), and Nanoracks 2U (green). Click image to enlarge.

Combining the expertise in ammonia oxidation research from José’s lab in Spain with the knowledge of Camila’s group in Puerto Rico about NASA’s engineering and safety requirements made the construction and realization of the complex AELISS project possible. However, launching something to the ISS isn’t without its issues…

Has the COVID-19 pandemic had a significant effect on the research? 

Camila Morales Navas assembling the AELISS equipment in the UPR laboratory. Click image to enlarge.

Aside from the usual problems and delays that can pop up during collaborative research projects, the introduction of a global pandemic at the last stages did not help the situation. The COVID-19 pandemic affected the timeline of the AELISS project, especially when it came to traveling and working within the extremely regulated environment of NASA. Additionally, Puerto Rico had already dealt with several large earthquakes and hurricanes in this period.

Keeping each other on track became difficult at times, particularly when Camila had to bring the entire setup back home to finish the engineering. In June, she was able to return to the laboratory and complete the project. However, the stressful part was not yet over because there was still a flight to NASA in the US, and with that the ever present threat of COVID-19 infection during travel.

One positive test result would mean a denial of entry – there can be no chance of infecting the ISS crew.

Ultimately, everything went to plan before and during the launch, and the instrumentation was sent to the International Space Station in October along with other precious cargo for the astronauts. Now that this part of the puzzle is finished, the rest of the work begins…

How will AELISS differ from similar experiments on Earth?

The final goal of this research is to determine how gravity affects the oxidation of ammonia, and also to test out different catalysts for the reaction in microgravity. While several other parameters can be adjusted in the lab such as pH, nanoparticle shape, and more – gravity is a universal constraint we cannot avoid. On Earth, we are only able to mimic the effects of microgravity for a few seconds with freefall. The previous collaboration between the groups in this project also involved performing experiments on special flights that allowed weightless conditions for less than 15 seconds at a time. This is certainly not enough time to draw long-term conclusions, and hence the push to launch the project into orbit. Only then can a true comparison be made, and conclusions drawn about the effects of gravity and the future applicability of this technology.

Dr. José Solla Gullón shown in his lab at the University of Alicante depositing Pt nanocubes on the 8X110 substrates which are used in the FLWCL8X1C electrochemical cell. Click image to enlarge.

One of the major concerns regarding this project is to achieve the most efficient conversion of waste urine into usable water for long-term space missions. Here, water recycling is a critical point. Also, it is important to note that the product of the oxidation of ammonia is nitrogen gas, but the behavior of gases is not the same on Earth as in space. Understanding how the N2 bubbles behave in the absence of gravity is a critical step to study.

Camila’s doctoral research project aims to answer these questions and more, using the realistic conditions of space rather than short periods of weightlessness in flight. So how did the researchers come to use Metrohm products?

There’s Something About Metrohm

So, why choose Metrohm over other providers? I asked José and Camila just what it was that drew them to our products.

«In my case, I have been working with Metrohm DropSens for many, many years. We have a very good collaboration, not only in the in the case of the nanomaterials, but also in the electrochemical cells, and the use of the screen-printed electrodes for electroanalysis. So, we have a very long history together

Dr. José Solla Gullón

Distinguished Researcher at the Institute of Electrochemistry, University of Alicante

Additionally, José mentioned that it was the fact that the electrochemical cells from Metrohm DropSens were very small, perfectly fitting into their conceptual system, which was another critical point. In fact, only cosmetic changes were needed to the products to be used in this project – all of the used materials were already approved for use by NASA..

For Camila, this was her first time using these products, and she found their out-of-the-box usage incredibly helpful.

«This was my first experience since José suggested it. And I trust them because they’re the people that really know about this subject

Camila Morales Navas

Senior graduate student in the Department of Chemistry, University of Puerto Rico

In the past, José has asked Metrohm DropSens several times to custom design SPEs for his research needs, and has always found them responsive and agreeable.

«I know that I can send an email and in two hours, I will have some response. This is wonderful for me. They are always open to new solutions

Dr. José Solla Gullón

Distinguished Researcher at the Institute of Electrochemistry, University of Alicante

We wish the very best to the research groups behind the ambitious AELISS project at the University of Puerto Rico and the University of Alicante. We at Metrohm are proud that our products can contribute to space exploration.

Post written by Dr. Alyson Lanciki, Scientific Editor at Metrohm International Headquarters, Herisau, Switzerland.

Special thanks go to Dr. José Solla Gullón and Ms. Camila Morales Navas for their important research and taking time to contribute to this article.

How to determine if your edible oils are rancid

How to determine if your edible oils are rancid

Rancidity is the process through which oils and fats become partially or completely oxidized after exposure to moisture, air, or even light. Though not always that obvious, foods can go rancid long before they become old. For oils, whose antioxidant properties are highly valued, such as for olive oil, this is especially problematic. A simple (and free) test for rancidity of oils can be performed at home using your own analytical instruments: your senses of smell and taste.

  1.  Pour a few milliliters of the oil into a shallow bowl or cup, and breathe in the scent.
  2.  If the smell is slightly sweet (like adhesive paste), or gives off a fermented odor, then the oil is probably rancid.
  3.  A taste test should be performed to be sure, since some oils may have a naturally sweet scent.
  4.  Ensure the oil sample is at room temperature, then sip a small amount into your mouth without swallowing. Similar to tasting wine, slurp air across the oil in your mouth, then exhale to determine if the oil has flavor.
  5.  If the oil has no flavor, it is most likely rancid. Do not consume it!

Once food has turned rancid, there is no way to go back and fix it. So, if you find out by means of the sensory test that the oil is rancid, it is already too late. For those of us who would rather skip this step to avoid having rancid food in our mouths, the possibility to accurately predict the future oxidation behavior of edible oils would be great. In fact, this is exactly what the Rancimat from Metrohm can do if you follow our tips and tricks in this article.

Rancimat to the rescue!

With the 892 Rancimat and the 893 Biodiesel Rancimat, Metrohm offers two instruments for the simple and reliable determination of the oxidation stability of natural fats and oils and of biodiesel, respectively. The method, also known as the Rancimat method or Rancimat test, is the same in both cases. It is based on a simple principle of reaction kinetics, according to which the rate of a chemical reaction (here the oxidation of fatty acids) can be accelerated by increasing the temperature.

The 892 Rancimat (L) and the 893 Biodiesel Rancimat (R) from Metrohm (click to enlarge image).

How does it work?

During the determination, a stream of air is passed through the sample at a constant temperature (e.g. 110 °C according to standard EN 14214 for biodiesel). Any oxidation products that develop are transferred by the air stream to a measuring vessel, where they are detected by the change in conductivity of an absorption solution. In addition to the temperature (both the accuracy and stability of which are guaranteed by the Rancimat system), the preparation of the measurement and the condition of the accessories also influence the quality and reproducibility of the results. In this blog post, we have compiled some practical experience in using the Rancimat to help you.

Oxidation stability: practical tips and tricks from the experts

Remove foreign particles from the reaction vessel

Foreign particles in the reaction vessel can catalyze reactions in the sample, leading to measurement results which are not reproducible. Remove foreign objects such as packaging remains from the reaction vessels using a strong gas stream (preferably nitrogen).

Weigh sample with a plastic spatula

Weigh the sample directly into the reaction vessel. Make sure that the maximum filling height does not exceed 3.5 cm. An error of ±10% in the sample weight has no influence on the final result.

Metal spatulas should not be used for weighing, as the metal ions could catalytically accelerate oxidation.

Reaction vessel lid

The green reaction vessel lid (see following image, article number: 6.2753.100) must seal the reaction vessel tightly. If this is no longer possible, the lid must be replaced. Leaky reaction vessel lids lead to incorrect and non-reproducible measurement results!

Tip: to make it easier to seal or to remove the lid, a fine film of silicone oil can be applied with a finger to the upper outer edge of the reaction vessel, to a height of about 1 cm.

Position and stability of the air tube

The stable, vertical positioning of the air tube (article number: 6.2418.100 or 6.2418.130) in the reaction vessel increases the reproducibility of the measurement results.

The air tube should protrude straight down into the vessel as illustrated in the following graphic representation (click image to enlarge).

Absorbent solution in the measuring vessel

Deionized water is used as the absorption solution with the Rancimat. Prior to beginning the analysis, the electrical conductivity of the water in the measuring vessel should not exceed 5 µS/cm.

If this value is higher, check the filters of the water system, and also ensure that there are no other sources of contamination.

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Positioning of the cannula for air supply

The PTFE (polytetrafluoroethylene) cannula for the air supply into the absorption solution (article number: 6.1819.080) must be aligned properly so that no air passes over the electrodes of the conductivity measuring cell, as shown in the graphic (click image to enlarge).

Air bubbles at the electrodes lead to noisy measurement curves that are difficult to evaluate.

Is it time to start the determination yet?

First, the temperature of the heating block (which is defined in the method) must be reached and stabilize before the reaction vessel is inserted into the instrument.

The sample identification data is then entered in the  StabNet software by the operator.

After connecting all of the tubing for the air supply, the reaction vessel can be inserted into the heating block. The sample measurement begins immediately after pressing the button on the Rancimat.

Cleaning: important for reproducible results

To obtain reliable analysis results, cleaning all accessories is of the utmost importance.

Both the reaction vessel and the inlet tube are disposable items. You can dispose of these materials immediately after cooling down. The rest of the accessories can be cleaned with a laboratory dishwasher (or equivalent) at maximum temperature and maximum drying time.

If you use glass or polycarbonate materials for the measuring vessels, you can of course also clean them in the same manner. The same applies to the measuring vessel lid with integrated conductivity electrode, the transparent silicone tubing, or the black Iso-Versinic tube, as well as the reaction vessel lid.

Tip: the silicone or Iso-Versinic tubing should be washed in a vertical position inside of the dishwasher to ensure it is thoroughly cleaned inside.

After washing, the transfer tubes and the reaction vessel lids should be heated at 80 °C for at least two hours in a drying cabinet, since the materials of these accessories absorb reaction products. This step further reduces the possibility of carryover to the next measurement which leads to unstable measurement results.


Depending on the use of the Rancimat, a regular visual control of the air filter on the back of the instrument is recommended. A clogged filter will lead to fluctuating air flows. The molecular sieve may also need a regular change depending on the instrument usage.

I hope that these tips have given you some helpful suggestions which will save you a little time and troubleshooting when using the Rancimat for determination of the oxidation stability of edible oils and other products. Good luck with your determinations!

Want to learn more?

Rancimat test: Quality control of natural fats and oils in cosmetics

Post written by Simon Lüthi, PM Titration (Meters & Measuring Instruments) at Metrohm International Headquarters, Herisau, Switzerland.