Design, synthesis and application of carbazole macrocycles in anion sensors

Our supramolecular analytical chemistry team has published an article (Beilstein J. Org. Chem. 2020, 16, 1901-1914) about developing carboxylate sensor prototypes that use macrocyclic carbazole receptors as ionophores.

Usually, reported research concludes with the demonstration of binding abilities of receptors and only suggests possible applications at a theoretical level. This work stands out from most publications by incorporating the entire workflow from receptor design to the characterisation of working sensor prototypes.

By involving this additional step, the team demonstrated the shortcoming of evaluating binding with just the receptor and analyte in solution. The predictions of selectivity can change considerably when measuring binding in an actual sensor membrane. This does not mean that such binding measurements would be obsolete, as the issue of binding anions selectively remains relevant in supramolecular chemistry.

The work was published in collaboration with the group of prof. Johan Bobacka at Åbo Akademi. The open access article was published in the Beilstein Journal of Chemistry thematic issue “Molecular Recognition” (in memory of Prof. Carsten Schmuck).

Symmetric Potentiometric Cells for the Measurement of Unified pH Values

Recently the UnipHied consortium published a new article – Symmetric Potentiometric Cells for the Measurement of Unified pH Values. Symmetry 2020, 12, 1150, where we compared the performance of different cells and instruments used to measure acidities on the unified pH scale (pHabs). The advantage of the unified pH scale is that it enables expressing the acidities of any systems – non-aqueous, solutions, colloidal systems, etc – via pHabsH2O values, which are directly comparable to the conventional pH values of the aqueous pH scale.

This is the first time unified acidities have been measured outside of our group. Nine partners from Europe – national metrology institutes and universities – have set up systems to measure pHabs and validated them with standard aqueous buffers.

The results show that the method is robust and does not depend on the cell nor the instrument (pH meter, potentiostat, or electrometer). The inherent symmetry of the cell design helps to reduce the experimental workload and improve the accuracy of the obtained results.

The next step is to measure unified acidities of non-aqueous systems, which could be used by routine laboratories to calibrate their pH electrodes.

The UnipHied project is funded from the EMPIR programme (project 17FUN09) co-financed by the Participating States and from the European Union’s Horizon 2020 research and innovation programme.

The secrets of two mummies have been unravelled

PLOS ONE has recently published a multidisciplinary paper on two Egyptian child mummies lead and participated by the members of our group. The team of 19 experts unravelled the secrets of two Graeco-Roman child mummies by applying most recent analytical techniques from archaeology, forensic sciences, analytical chemistry, medicine, entomology, and genetics. This is one of the most extensive multidisciplinary study of ancient mummies and the paper is free to download here.

New non-invasive approach to analyze textile fibers

Another new method development in our group has been assembled into a publication. Article by Pilleriin Peets (on the picture), Karl Kaupmees,  Signe Vahur, and Ivo Leito in the journal of Heritage Science introduces non-invasive approach to identify all kinds of different textile fibers using reflectance-FT-IR spectroscopy and chemometry. In this work 61 single-component textile pieces were analyzed, 4000 spectra were recorded and all in all 16 different textile fiber types were characterized. As reflectance-FT-IR spectroscopy is not widely used to analyze samples with an uneven surface, in our best knowledge, this article provides the largest collection of r-FT-IR spectra from textile fibers that is recorded and published. All the r-FT-IR spectra from different fiber classes are now available for other scientists, conservators, museum workers and industries to use!

PCA classification graph of different textile fiber classes. Peets, P., Kaupmees, K., Vahur, S., Leito, I. Reflectance FT-IR spectroscopy as a viable option for textile fiber identification. Herit Sci  7, 93 (2019)

New publication – retention studies in C18 column using novel fluorinated eluent additives

It is essential to understand the interaction between the analyte, mobile phase and the column chemistry for the best possible separation of molecules. Of all three, the mobile phase in HPLC is the easiest to modify and thus, usually contains different eluent additives or buffers. While using mass spectrometry undoubtedly increases the variety of analytes possible to detect, it also introduces limiting factors such as the requirement of the volatility of the eluent. Most common additives in LC-MS systems are formic and acetic acid, their ammonium salts as well as ammonium bicarbonate. In our recent paper, we have proposed novel – fluorinated, eluent additives (hexafluoroisopropanol, hexafluoro-tert-butyl alcohol, trifluoroethanol as well as nonafluoro-tert-butyl alcohol and perfluoropinacol).  Their influence was shown on rather simple exemplary molecules, which are widely spread over different logP values, containing protonated and deprotonated acids and bases. All novel fluorinated eluent additives demonstrated a strong influence on basic polar analytes in basic medium – they drastically increased retention. A decrease in retention was observed for acidic analytes when these novel eluent additives were used.

Moreover, current research displays a comprehensive overview of retention mechanisms for nonafluoro-tert-butyl alcohol and is the first time ever when perfluoropinacol has been introduced as eluent additive for reversed phase chromatography. Additionally, the influence on MS signal was studied when fluoroalcohols were used as eluent additives. This is also the first time when the absolute pH (pHabs) scale was used for expressing the mobile phase pH.

Current fundamental research forms a basis for a better understanding of the influence of fluoroalcohols as eluent additives and will help in the assay development in a wide range of applications.

This work is part of a larger endeavor – to promote a wider usage the unified pH scale (pHabs) by the research and technology communities, which is currently in progress via the UnipHied.

The UnipHied project is funded from the EMPIR programme (project 17FUN09) co-financed by the Participating States and from the European Union’s Horizon 2020 research and innovation programme.



New publication – Instrumental techniques in the analysis of natural red textile dyes

A wide variety of different dyes, complex composition of natural dye sources and low concentrations in samples make the identification of textile dyes challenging. In our cultural heritage group, work has been done for years to overcome some of these problems. Using seven different red dye sources and five instrumental approaches, a method for the analysis of textile dyes has now been developed in our lab to study different archaeological findings, museum artifacts, and other textile pieces. More about the results can be found in the article published in the Journal of Cultural Heritage by P. Peets, S. Vahur, A. Kruve, T. Haljasorg, K. Herodes, T. Pagano and I. Leito.

The usefulness of this developed method has been proved by analyzing several case-study samples from Estonian National Museum, KANUT, and private collections. Thanks to the multiple instrument combination and especially the usage of high-resolution mass-spectrometry, it was also possible to identify synthetic organic dyes without the use of any standard substances.

EACH is featured in Analytical and Bioanalytical Chemistry!

The educational section of the leading analytical chemistry journal Analytical and Bioanalytical Chemistry, the ABCs of Education and Professional Development in Analytical Science has published a paper about the EACH programme: EACH (Excellence in Analytical Chemistry), an Erasmus Mundus Joint Programme: progress and success

The EACH programme is an Erasmus Mundus Joint Master Degree programme coordinated by our group. It has become one of the leading master’s degree programmes focusing on analytical chemistry within its short existence. The programme is run in cooperation by four top European universities: in addition to the coordinating university, the University of Tartu (first-year university), also Uppsala University (Sweden), Åbo Akademi University (Finland) and University Claude Bernard Lyon 1 (France) are involved as second-year and specialisation institutions.

The paper gives a comprehensive overview about the EACH programme, including a detailed overview of what EACH is, what kind of topics are taught during the first study-year at the University of Tartu, what is the nature of the different study-tracks (specialisation during second study-year either in Sweden, Finland or France), what extra activities, such as internship and winter school the EACH programme offers. The paper also gives an explanation on why it is necessary to have such a programme like EACH, and how to be a part of the EACH experience.

One section in the paper is dedicated to the impact of the EACH programme – with the four years that the programme has accepted students, it has reached a lot of countries and the employability rate of the graduates is almost 90%.

This Autumn, the fifth intake of EACH students is starting their studies at the University of Tartu, expanding the blue areas on the map presented above even more.

For more information on the EACH programme, please see the programme’s webpage and also the newly published paper.

New publication – Comparison of derivatization methods for the quantitative gas chromatographic analysis of oils

Often paints contain oils that bind together the components (pigments, fillers, etc) of the paint. Analysis of these oils requires derivatization, but which derivatization method is the best for a quantitative approach?

The work on this topic began with the need of our cultural heritage group to have a routine method for the determination of fatty acid composition of oils with gas chromatography (GC). The analysis of oils with GC requires derivatization and for this a wide variety of procedures have been applied. It turned out there are only few comparisons of derivatization methods in the literature and all are quite limited by scope. For that reason, it was decided to explore this important topic comprehensively and, importantly, in the terms of absolute (not relative) quantification.

A member of our group – PhD student Eliise Tammekivi – implemented four common and well-known derivatization procedures to perform the absolute quantification of fatty acids with GC-MS and GC-FID. The four compared derivatization methods were: (1) methylation with m-(-trifluoromethyl)phenyltrimethylammonium hydroxide (TMTFTH), (2) two-step derivatization with sodium ethoxide (NaOEt) and N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA), (3) two-step derivatization with KOH and BSTFA and (4) acid-catalyzed methylation (ACM).

This study has now been completed and the results have been published in the journal Analytical Methods by E. Tammekivi, S. Vahur, O. Kekišev, I. D. van der Werf, L. Toom, K. Herodes and I. Leito. In the publication, a comprehensive and wide-scale quantitative comparison of the four derivatization methods is presented. The results demonstrate that methylation with TMTFTH is the least work-intensive and most accurate derivatization method – both in terms of reproducibility and derivatization efficiency (yield).  For further information see: Analytical Methods, 2019, 11, 3514 – 3522

Russian roulette with Ecstasy

PhD Fellow Max Hecht and Dr Karin Kipper with collaborators looked into the variability in content and dissolution profiles of MDMA tablets collected in the UK and published the results recently in Drug Testing and Analysis.

Ecstasy comes in all shapes, sizes and colours. 3,4-methylenedioxy-met(h)amf(/ph)etamine (MDMA) is the chemical name for a recreational drug taken for its empathogenic “high”, that alters the sensation of energy, empathy and pleasure. MDMA is mainly consumed in the form of tablets.

The trends in dose

To identify the risks associated with MDMA consumption, researchers from the UK and the University of Tartu analysed the drug dose of 412 tablets and, for the first time, studied the dissolution aspects of another 242 tablets collected in the UK in the time period between 2001-2018. Like in other European countries, the MDMA content in tablets is steadily increasing in the last decade, with some recent tablets containing unprecedented high doses.

The variability in dissolution

Dissolution testing is commonplace in the quality control and batch production of tablets in pharmaceutical laboratories and for bioequivalence testing. For MDMA, different dissolution rates were observed, ranging from fully dissolved in 30 min – the fast group, to up to 2 h until the whole content of the drug is completely released (slow group). No significant differences in a dose per tablet could be found between slow and fast releasing groups. As well as there was no association of colour, logo or shape with the dose or dissolution group. 

In conclusion, this means that, while it always is a gamble to take Ecstasy, the ever-present danger of overdosing is also on the rise. You are welcome to read our freshly published article for more details.

ESI outcompetes other ion sources in LC-MS trace analysis

Choosing the best possible ion source is a very important step in liquid chromatography-mass spectrometry (LC/MS) method development. In a recent paper,  ESI outcompetes other ion sources in LC-MS trace analysis Anal. Bioanal. Chem. 2019 Asko Laaniste from the UT Analytical chemistry group carried out a large scale survey of different LC/MS ion sources (and their different working modes) as applied to pesticide analysis. He compared electrospray ionisation (ESI) source, thermally focused/heated electrospray (HESI), atmospheric pressure photoionisation (APPI) source with and without dopant, and multimode source in ESI mode, atmospheric pressure chemical ionisation (APCI) mode, and combined mode using both ESI and APCI, i.e. altogether seven different ionisation modes for the analysis of 40 pesticides (see list below).

The lowest limits of detection (LoDs) were obtained by ESI and HESI. The widest linear ranges were observed with the conventional ESI source without heated nebulizer gas. ESI source was significantly less affected by matrix effect than the HESI source. APPI ranked second (after ESI) by not being influenced by matrix effect; therefore, it would be a good alternative to ESI if low LoDs are not required.

It was somewhat unexpected that ESI outperformed HESI. This may be caused by the instability of the additional heated gas (sheath gas) in HESI that is used in addition to the nebulising gas.

In conclusion, as a broad generalisation, Asko’s results show that although several new ion sources have been introduced during the last decade, the conventional ESI performs at least equally to these new ion sources in the case of polar to medium polarity pesticides. ESI with thermal focusing (HESI) performed more or less equally with ESI in terms of LoD. At the same time, HESI had significantly poorer intermediate precision of matrix effect values for most compounds. Thus, for trace analysis, ESI is still the ion source of choice.

(Full list of pesticides: pymetrozine, thiamethoxam, methiocarb-sulfoxide, chloridazon, imidacloprid, acetamiprid, methiocarb-sulfone, thiacloprid, imazalil, thiophanate-methyl, metribuzin, pyrimethanil, fenpropimorph, spiroxamine, propoxur, triasulfuron, bupirimate, paclobutrazol, methiocarb, azoxystrobin, epoxiconazole, myclobutanil, fenhexamid, fluquinconazole, flusilazole, mepanipyrim, bitertanol, propiconazole, triazophos, methoxychlor, ditalimfos, tebufenozide, benalaxyl, pyrazophos, buprofezin, indoxocarb, trifloxustrobin, quinoxyfen, pirimiphos-ethyl, hexythiazox)


(Image top left: Asko Laaniste preparing solutions; Image right: Comparison of accuracy of LC-MS analysis using different ion sources)