E!10050 New stable Isotope Labelled Certified Reference Materials for Hydrocarbon Analysis

WP1: Project Management The last project meeting was held at Chiron on 13. Dec. 2018 WP2: Exploitation and Dissemination GEUS published project results in Organic Geochemistry 129 (2019) 14-23. A new manuscript for triaromatic oleanane is planned also for Organic Geochemistry. WP4: Synthesis of labelled compounds Synthesis of 13C-labelled PAHs and PBDEs is ongoing, the work will be continued after this project is ended. Synthesis of 13C-labelled cholestane has been done. The synthesis has been optimized and the product purity of 98-99% was achieved. WP5: Isolation and Purification of native biomarkers from geological samples for 12C/13C measurements Five triaromatic triterpanes were obtained very pure from Serbian and Colombian coal. The structure of triaromatic oleanane, triaromatic ursane and triaromatic lupane was confirmed using NMR. The fourth compound appears to be a triaromatic lupane stereoisomer. Determination of the extract structure is underway. The last compound is a ring-E demethylated oleanane/ursane. Extract of immature Chinese coal obtained. Isolation of diaromatic triterpanes (some still unidentified) took place early 2019. One compound isolated is almost pure, some minor compounds need more purification. NMR analysis of isolated compounds is planned. WP6: Product certification and implementation of the CRMs in analytical procedures Ongoing with 13C-labelled PAHs.

Biomarkers such as hopanes and steranes are unique products for Chiron, PAHs, alkyl PAHs and PBDEs are among the best sold products in Chiron?s product ranges. With the outcome of 13C-labelled reference materials and isotope ratio (13C/12C) reference materials developed in the project, Chiron has now a stronger and more competitive marked position for supply CRMs for forensic environmental analysis and petroleum exploration.

The 13C-CRM project aims at seizing a major market opportunity in developing unique isotope-labelled Certified Reference Materials (CRMs) for a more accurate analysis of environmental and petroleum compounds using mass spectrometry techniques. 13C labelled CRMs will be produced, certified, and commerciallized as reference stardards for hydrocarbon analysis, in quantitative GC-MS (gas chromatography - mass spectrometry analysis ) and GC-IRMS (gas chromatography-isotope ratio mass spectrometry). The lack of widely available CRMs has hindered the practice when analysing organic materials with GC-MS and GC-IRMS. The existing CRMs are usually based on structural analogues of the analytes or deuterium labelled versions. However, it has been shown that due to ion suppression effects, the quantitaive analytical results with deuterated standards may vary. Using of 13C labelled standards always yields the true value. In addition, 13C labelled standards can overcome the problems of deuterated standards of stability, recovery, instrument variations and ion suppression. In this 13C-CRM project, CRMs will be developed based on the combination of chemical synthesis and biosynthesis (fermentation) methods, which is not only unique in itself as far as the authors are aware, but essential for developing an attractive spectrum of new petroleum analytical standards. An integrated approach for continuous validation of results and advancements with cross-checking procedures between different techniques will be pursued as research methodology. Different development approaches will be applied for the different target products, e.g. 13C-labelled alkylated PAHs, 13C-labelled hopanoids and sterols.