Session: Emerging Topics in Analytical Toxicology, Forensics, and Doping Control I

Session Chair: Prof. Dr. Hans Maurer
Certified by GTFCh with 2 Credit Points (Forensic Toxicologists GTFCh, Clinical Toxicologists GTFCh, Forensic Chemists GTFCh, and Forensic-Clinical Chemists GTFCh)
Englisch

Role of IRMS in Life Sciences with Focus on Forensic Sciences

Andreas Rossmann, isolab GmbH
Stable isotope analyses of bio elements (H, C, N, O, S) in food and food components have been used for food authenticity and origin control since the late 1970th. In 1990, the EU introduced official methods for stable isotope analyses for wine (1). Since 2001, stable isotope analyses on human tissues are used for provenancing of unknown individuals (2). The isotope composition of the environment is transferred into human bodies through diet, thus the premise of “you are what you eat isotopically”. Regional specific stable isotope ratios of the elements from food and drink are integrated into body tissues of living organisms. Examination of the isotope composition of different body tissues (teeth, bones, hair, nail) provide geographical information from childhood until death. Some tissues, such as teeth, are formed during childhood whereas tissues such as hair or nail are continuously growing until death. Stable isotope values of the bio-elements (δ2H, δ13C, δ15N, δ18O and δ34S) in body tissues are linked to the composition of an individual’s diet and the origin of food and drink. Basically, δ13C values show the content of C4 or C3 plants or marine versus terrestrial sources, δ15N values indicate the amount of animal protein (meat, dairy products, fish), and δ34S values give a hint to any marine influences (sea products, sea-spray effect, sediments of marine origin) in a person’s diet. δ2H and δ18O values predominantly contain information about the climatic conditions at the whereabouts of individuals, as the isotopic composition of regional meteoric water is strongly linked to annual air temperature. Further information can be obtained from stable isotope data of geo elements as strontium or lead. Human provenancing based on isotopic analysis contributes in providing geographical origin information from different time-windows in an individual’s life and aids the police with the investigation leading to the identification of the victim in some cases. In collaboration with the Institute of Legal Medicine, LMU Munich, we further develop isotope methods for human provenancing, ensuring capacity and continuity for this type of forensic intelligence investigations. The application of the isotope methods on human remains of unidentified decedents will be explained through case studies: 1) a provenance study of a deceased found inside a suitcase in Berlin, Germany; 2) a provenance study of an unknown woman found dead in Burgenland, Austria, in 1993.
Englisch

Role of IRMS in Life Sciences with Focus on Forensic Sciences

Andreas Rossmann, isolab GmbH
Stable isotope analyses of bio elements (H, C, N, O, S) in food and food components have been used for food authenticity and origin control since the late 1970th. In 1990, the EU introduced official methods for stable isotope analyses for wine (1). Since 2001, stable isotope analyses on human tissues are used for provenancing of unknown individuals (2). The isotope composition of the environment is transferred into human bodies through diet, thus the premise of “you are what you eat isotopically”. Regional specific stable isotope ratios of the elements from food and drink are integrated into body tissues of living organisms. Examination of the isotope composition of different body tissues (teeth, bones, hair, nail) provide geographical information from childhood until death. Some tissues, such as teeth, are formed during childhood whereas tissues such as hair or nail are continuously growing until death. Stable isotope values of the bio-elements (δ2H, δ13C, δ15N, δ18O and δ34S) in body tissues are linked to the composition of an individual’s diet and the origin of food and drink. Basically, δ13C values show the content of C4 or C3 plants or marine versus terrestrial sources, δ15N values indicate the amount of animal protein (meat, dairy products, fish), and δ34S values give a hint to any marine influences (sea products, sea-spray effect, sediments of marine origin) in a person’s diet. δ2H and δ18O values predominantly contain information about the climatic conditions at the whereabouts of individuals, as the isotopic composition of regional meteoric water is strongly linked to annual air temperature. Further information can be obtained from stable isotope data of geo elements as strontium or lead. Human provenancing based on isotopic analysis contributes in providing geographical origin information from different time-windows in an individual’s life and aids the police with the investigation leading to the identification of the victim in some cases. In collaboration with the Institute of Legal Medicine, LMU Munich, we further develop isotope methods for human provenancing, ensuring capacity and continuity for this type of forensic intelligence investigations. The application of the isotope methods on human remains of unidentified decedents will be explained through case studies: 1) a provenance study of a deceased found inside a suitcase in Berlin, Germany; 2) a provenance study of an unknown woman found dead in Burgenland, Austria, in 1993.
Englisch

Role of IRMS in Life Sciences with Focus on Forensic Sciences

Andreas Rossmann, isolab GmbH
Stable isotope analyses of bio elements (H, C, N, O, S) in food and food components have been used for food authenticity and origin control since the late 1970th. In 1990, the EU introduced official methods for stable isotope analyses for wine (1). Since 2001, stable isotope analyses on human tissues are used for provenancing of unknown individuals (2). The isotope composition of the environment is transferred into human bodies through diet, thus the premise of “you are what you eat isotopically”. Regional specific stable isotope ratios of the elements from food and drink are integrated into body tissues of living organisms. Examination of the isotope composition of different body tissues (teeth, bones, hair, nail) provide geographical information from childhood until death. Some tissues, such as teeth, are formed during childhood whereas tissues such as hair or nail are continuously growing until death. Stable isotope values of the bio-elements (δ2H, δ13C, δ15N, δ18O and δ34S) in body tissues are linked to the composition of an individual’s diet and the origin of food and drink. Basically, δ13C values show the content of C4 or C3 plants or marine versus terrestrial sources, δ15N values indicate the amount of animal protein (meat, dairy products, fish), and δ34S values give a hint to any marine influences (sea products, sea-spray effect, sediments of marine origin) in a person’s diet. δ2H and δ18O values predominantly contain information about the climatic conditions at the whereabouts of individuals, as the isotopic composition of regional meteoric water is strongly linked to annual air temperature. Further information can be obtained from stable isotope data of geo elements as strontium or lead. Human provenancing based on isotopic analysis contributes in providing geographical origin information from different time-windows in an individual’s life and aids the police with the investigation leading to the identification of the victim in some cases. In collaboration with the Institute of Legal Medicine, LMU Munich, we further develop isotope methods for human provenancing, ensuring capacity and continuity for this type of forensic intelligence investigations. The application of the isotope methods on human remains of unidentified decedents will be explained through case studies: 1) a provenance study of a deceased found inside a suitcase in Berlin, Germany; 2) a provenance study of an unknown woman found dead in Burgenland, Austria, in 1993.
Englisch

Profiling of New Psychoactive Substances by IRMS

Michael Pütz, Bundeskriminalamt
Drug profiling is a useful tool for the comparative characterization of the physical and chemical properties of illicit drugs, used by police authorities to establish links between samples of different seizures, to obtain information on trafficking routes and to gather background information on the origin of samples. The chemical profiling based on chromatographic signatures of synthesis-related impurities is far more demanding for new psychoactive substances (NPS), specifically synthetic cannabinoid receptor agonists (SRCA´s) compared to classic drugs. Main reasons are the significantly higher chemical purity of SRCA drug substances (often > 98 % (w/w)) and the nonapplicability of acid/base liquid-liquid extraction for API removal. Consequently, for the profiling of SRCA´s in Spice-products, a novel impurity-profiling workflow with the combination of flash-chromatography (F-LC), UHPLC-MSn and multivariate data analysis of signatures with optional complementary and orthogonal stable isotope ratio (SIR) analysis of isolated pure SRCA´s via EA-IRMS or GC-IRMS was developed [1]. In a proof-of-concept study this profiling concept was successfully applied to a range of Spice products containing the SCRA 5F-PB-22, being highly prevalent on the German NPS market between 2013 and 2015, allowing assignment of different products to one manufacturer based on very similar δ13C, δ15N and δ2H isotope ratio data [2]. In a larger field study, one of the most prevalent and hazardous SRCA to date, MDMBCHMICA, was assessed via the F-LC/UHPLC-MSn impurity profiling workflow combined with IRMS analysis (δ13C, δ15N) in 61 pure samples, 141 samples of Spice-products of various brands from test purchases in ca. 40 online-shops as well as 136 Spiceproducts from police seizures [3]. The observed low variance for δ13C and δ15N data for all pure samples and Spice-product extracts of MDMB-CHMICA in relation to the variance of the SIR data for indole precursor samples from different chemical manufacturers, indicated that all available samples have been produced from precursor material with similar SIR. PCA models of the SIR and impurity signatures for the herbal blends resulted in cluster formations of multiple samples, presumably representing individual synthesis batches of MDMB-CHMICA from one source. Additionally, Spiceproducts with similar date of purchase exhibited a tendency for cluster formation [3].
Englisch

Profiling of New Psychoactive Substances by IRMS

Michael Pütz, Bundeskriminalamt
Drug profiling is a useful tool for the comparative characterization of the physical and chemical properties of illicit drugs, used by police authorities to establish links between samples of different seizures, to obtain information on trafficking routes and to gather background information on the origin of samples. The chemical profiling based on chromatographic signatures of synthesis-related impurities is far more demanding for new psychoactive substances (NPS), specifically synthetic cannabinoid receptor agonists (SRCA´s) compared to classic drugs. Main reasons are the significantly higher chemical purity of SRCA drug substances (often > 98 % (w/w)) and the nonapplicability of acid/base liquid-liquid extraction for API removal. Consequently, for the profiling of SRCA´s in Spice-products, a novel impurity-profiling workflow with the combination of flash-chromatography (F-LC), UHPLC-MSn and multivariate data analysis of signatures with optional complementary and orthogonal stable isotope ratio (SIR) analysis of isolated pure SRCA´s via EA-IRMS or GC-IRMS was developed [1]. In a proof-of-concept study this profiling concept was successfully applied to a range of Spice products containing the SCRA 5F-PB-22, being highly prevalent on the German NPS market between 2013 and 2015, allowing assignment of different products to one manufacturer based on very similar δ13C, δ15N and δ2H isotope ratio data [2]. In a larger field study, one of the most prevalent and hazardous SRCA to date, MDMBCHMICA, was assessed via the F-LC/UHPLC-MSn impurity profiling workflow combined with IRMS analysis (δ13C, δ15N) in 61 pure samples, 141 samples of Spice-products of various brands from test purchases in ca. 40 online-shops as well as 136 Spiceproducts from police seizures [3]. The observed low variance for δ13C and δ15N data for all pure samples and Spice-product extracts of MDMB-CHMICA in relation to the variance of the SIR data for indole precursor samples from different chemical manufacturers, indicated that all available samples have been produced from precursor material with similar SIR. PCA models of the SIR and impurity signatures for the herbal blends resulted in cluster formations of multiple samples, presumably representing individual synthesis batches of MDMB-CHMICA from one source. Additionally, Spiceproducts with similar date of purchase exhibited a tendency for cluster formation [3].
Englisch

Profiling of New Psychoactive Substances by IRMS

Michael Pütz, Bundeskriminalamt
Drug profiling is a useful tool for the comparative characterization of the physical and chemical properties of illicit drugs, used by police authorities to establish links between samples of different seizures, to obtain information on trafficking routes and to gather background information on the origin of samples. The chemical profiling based on chromatographic signatures of synthesis-related impurities is far more demanding for new psychoactive substances (NPS), specifically synthetic cannabinoid receptor agonists (SRCA´s) compared to classic drugs. Main reasons are the significantly higher chemical purity of SRCA drug substances (often > 98 % (w/w)) and the nonapplicability of acid/base liquid-liquid extraction for API removal. Consequently, for the profiling of SRCA´s in Spice-products, a novel impurity-profiling workflow with the combination of flash-chromatography (F-LC), UHPLC-MSn and multivariate data analysis of signatures with optional complementary and orthogonal stable isotope ratio (SIR) analysis of isolated pure SRCA´s via EA-IRMS or GC-IRMS was developed [1]. In a proof-of-concept study this profiling concept was successfully applied to a range of Spice products containing the SCRA 5F-PB-22, being highly prevalent on the German NPS market between 2013 and 2015, allowing assignment of different products to one manufacturer based on very similar δ13C, δ15N and δ2H isotope ratio data [2]. In a larger field study, one of the most prevalent and hazardous SRCA to date, MDMBCHMICA, was assessed via the F-LC/UHPLC-MSn impurity profiling workflow combined with IRMS analysis (δ13C, δ15N) in 61 pure samples, 141 samples of Spice-products of various brands from test purchases in ca. 40 online-shops as well as 136 Spiceproducts from police seizures [3]. The observed low variance for δ13C and δ15N data for all pure samples and Spice-product extracts of MDMB-CHMICA in relation to the variance of the SIR data for indole precursor samples from different chemical manufacturers, indicated that all available samples have been produced from precursor material with similar SIR. PCA models of the SIR and impurity signatures for the herbal blends resulted in cluster formations of multiple samples, presumably representing individual synthesis batches of MDMB-CHMICA from one source. Additionally, Spiceproducts with similar date of purchase exhibited a tendency for cluster formation [3].
Englisch

Power of IRMS in Doping Control

Corinne Buisson, afld - Département des analyses / French Antidoping laboratory
Among the list of prohibited substances by the World Anti-Doping Agency, anabolic androgenic steroids remain the most reported substances in doping controls. If the presence of exogenous steroids can be easily achieved by conventional mass spectrometry methods, it is more challenging to determine whether endogenous steroids such as testosterone have been misused or not. Indeed, testosterone, when administered exogenously will lead to the same structural substance in biological fluids. The most powerful tool allowing unequivocal proof of its administration is the Isotope Ratio Mass Spectrometry (IRMS) which is able to establish the isotopic signature of a target compound and thus provide a proof of exogenous administration. Nowadays, IRMS is routinely applied by all anti-doping laboratories to investigate suspicious steroid profiles determined from each athlete. However, if testosterone remained the most analyzed substance by IRMS, since the last decade, more and more applications on other substances have been developed such as the analysis of 19-norandrosterone, boldenone, AICAR or glucocorticoids for example. In addition to clearly determining the endogenous or exogenous origin of naturally occurring prohibited substances, IRMS can also be used as an investigation tool. Indeed, thanks to the IRMS results obtained on testosterone and its metabolites, it is for example possible to differentiate a local administration from an oral or intra-muscular administration. It is also possible to identify the kind of drug administered when some drugs present a specific isotopic signature. IRMS can also be a helpful tool for a better understanding the metabolism or finding metabolites.
Englisch

Power of IRMS in Doping Control

Corinne Buisson, afld - Département des analyses / French Antidoping laboratory
Among the list of prohibited substances by the World Anti-Doping Agency, anabolic androgenic steroids remain the most reported substances in doping controls. If the presence of exogenous steroids can be easily achieved by conventional mass spectrometry methods, it is more challenging to determine whether endogenous steroids such as testosterone have been misused or not. Indeed, testosterone, when administered exogenously will lead to the same structural substance in biological fluids. The most powerful tool allowing unequivocal proof of its administration is the Isotope Ratio Mass Spectrometry (IRMS) which is able to establish the isotopic signature of a target compound and thus provide a proof of exogenous administration. Nowadays, IRMS is routinely applied by all anti-doping laboratories to investigate suspicious steroid profiles determined from each athlete. However, if testosterone remained the most analyzed substance by IRMS, since the last decade, more and more applications on other substances have been developed such as the analysis of 19-norandrosterone, boldenone, AICAR or glucocorticoids for example. In addition to clearly determining the endogenous or exogenous origin of naturally occurring prohibited substances, IRMS can also be used as an investigation tool. Indeed, thanks to the IRMS results obtained on testosterone and its metabolites, it is for example possible to differentiate a local administration from an oral or intra-muscular administration. It is also possible to identify the kind of drug administered when some drugs present a specific isotopic signature. IRMS can also be a helpful tool for a better understanding the metabolism or finding metabolites.
Englisch

Power of IRMS in Doping Control

Corinne Buisson, afld - Département des analyses / French Antidoping laboratory
Among the list of prohibited substances by the World Anti-Doping Agency, anabolic androgenic steroids remain the most reported substances in doping controls. If the presence of exogenous steroids can be easily achieved by conventional mass spectrometry methods, it is more challenging to determine whether endogenous steroids such as testosterone have been misused or not. Indeed, testosterone, when administered exogenously will lead to the same structural substance in biological fluids. The most powerful tool allowing unequivocal proof of its administration is the Isotope Ratio Mass Spectrometry (IRMS) which is able to establish the isotopic signature of a target compound and thus provide a proof of exogenous administration. Nowadays, IRMS is routinely applied by all anti-doping laboratories to investigate suspicious steroid profiles determined from each athlete. However, if testosterone remained the most analyzed substance by IRMS, since the last decade, more and more applications on other substances have been developed such as the analysis of 19-norandrosterone, boldenone, AICAR or glucocorticoids for example. In addition to clearly determining the endogenous or exogenous origin of naturally occurring prohibited substances, IRMS can also be used as an investigation tool. Indeed, thanks to the IRMS results obtained on testosterone and its metabolites, it is for example possible to differentiate a local administration from an oral or intra-muscular administration. It is also possible to identify the kind of drug administered when some drugs present a specific isotopic signature. IRMS can also be a helpful tool for a better understanding the metabolism or finding metabolites.