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EEA/Norway Grants 2009-2014 Programme LV05 NORWEGIAN-LATVIAN RESEARCH COOPERATION ACTIVITY

EEA/Norway Grants 2009-2014 Programme LV05 NORWEGIAN-LATVIAN RESEARCH COOPERATION ACTIVITY. Project Progress Project leader at UiO prof. Elsa LUNDANES Presented by Dr Silvija ABELE 15th of February, 2016 University of Oslo.

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EEA/Norway Grants 2009-2014 Programme LV05 NORWEGIAN-LATVIAN RESEARCH COOPERATION ACTIVITY

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  1. EEA/Norway Grants 2009-2014 Programme LV05NORWEGIAN-LATVIANRESEARCH COOPERATION ACTIVITY Project Progress Project leader at UiO prof. Elsa LUNDANES Presented by Dr Silvija ABELE 15th of February, 2016 University of Oslo

  2. EEA and Norway Grants 2009-2014 Research and ScholarshipProgramme LV05 Research Scholarship EEZ/NFI/S/2015/003 Educational collaboration between Norway and Latvia for establishment of an effective risk assessment and control of pharmaceutical residues 5971.00 EUR 01.08.2015 - 31.12.2015 • NFI/R/2014/010 • Establishing of the scientific capacity for the management of pharmaceutical products residues in the environment of Latvia and Norway • 218432.00 EUR • 01.06.2015 - 30.04.2017

  3. EEA/Norway Grants 2009-2014 Programme LV05NORWEGIAN-LATVIAN RESEARCH COOPERATION ACTIVITY Establishing of the Scientific Capacity for the Management of Pharmaceutical Product Residues in the Environment of Latvia and Norway NFI/R/2014/010

  4. NO-LV RESEARCH COOPERATION ACTIVITYResearch Project NFI/R/2014/010 Overall objective of the Project is 1) to enhance the growth of research-based knowledge 2) through the scientific cooperation between Norway and Latvia, 3) developing a scientific capacity for the administration of pharmaceutical residues in the Latvian and Norwegian environment

  5. ProjectBackground and Justification • Continuous increasing consumption of pharmaceutical products (PP) • PP residues are introduced into the aquatic environment from: • municipal treatment plants • hospital effluents • livestock farming • households • PP presence in the environment -a serious threat to environment and human health • antibiotics  resistant bacteria strains • hormones  endocrine disrupting effects

  6. Publications onOccurenceofPharmaceuticals in theEnvironment Journal of Hazardous Materials 175 (2010) 45–95

  7. Project Concept • The knowledge about PP, their short and long-term environmental impacts, possible degradation mechanisms and management strategies is essential • Advanced high resolution liquid chromatography - mass spectrometric method will be developed to obtain information about the PP contamination status

  8. The Aim of the Study • To develop and validate an ultrasensitive and selective analytical method for combined quantitative analysis ofpharmaceuticalsin the environmental water samples • Ultra-High Performance Liquid Chromatography coupled to a hybrid quadrupole Orbitrapmass spectrometer (UHPLC-q-Orbitrap) will be used together with on-line sample preparation (SPE) • Results obtained in Riga and Oslo will be compared • 2-3 Peer-reviewed publications on analytical performance and application advantages of the elaborated method

  9. Pharmaceuticals Studied

  10. Stages ofthe Research Project Method developmentusingindividual standard compounds Sample collection Sample preparation Method optimization Method validation

  11. UV-HPLC Caffeine standard RT 15 min • Reverse phase chromatography • Gradient 2% B to 95% B in 20 min • Buff A = water with 0.1% FA • Buff B = 95% ACN / 5 % water 0.1 % FA • Buff A flow rate 50 µL/min • Concentration 0.01 mg/mL • Injection 50 µL • UV-Detection 254 nm

  12. UV-HPLC Ketoprofen Analgesic/ Anti-inflammatory Ketoprofen • Reverse phase chromatography • Gradient 2% B to 95% B in 20 min • Buff A = water with 0.1% FA • Buff B = 95% ACN / 5 % water 0.1 % FA • Buff A flow rate 50 µL/min • Concentration 0.01 mg/mL • Injection 50 µL • UV-Detection 280 nm

  13. UV-HPLC Coffeine Fluoexetine Diclofenac Analgesic/ Anti-inflammatory Ketoprofen Metoprolol Carbamazepine Anti-hypertensive Losartan Azythromycine

  14. Conditions 0.0001 mg/mL 800 uL injection 5 uL/min flow rate Aim to detect10 pg/mL LC-MS-MS Results Acetaminophen (Paracetamol) Analgesic/ Antipyretic Sulfamethoxazole Antibiotics Ketoprofen Analgesic/ Anti-inflammatory

  15. Experimental Setup Load Particles Pharmaceuticals Salts/solvent To waste From autosampler/Pump 1 Flow: 50 µL/min 0.1 % FA in H2O Injection volume: 100 µL Filter Credit Hanne Røberg-Larsen To column (ID 0.3 mm) SPE From pump 2 Flow: 5 µL/min

  16. Experimental Setup Particles Pharmaceuticals Salts/solvent To waste From autosampler/Pump 1 Flow: 50 µL/min 0.1 % FA in H2O Injection volume: 800 µL Inject Filter Credit Hanne Røberg-Larsen To column (ID 0.3 mm) SPE From pump 2 Flow: 5 µL/min

  17. Conditions 0.0001 mg/mL 800 uL injection 5 uL/min flow rate 30 min gradient LC-MS-MS Results 23 Compounds Retention time, min • Reverse phase chromatography • The Thermo Scientific™ TSQ Quantiva™ Triple Quadrupole Mass Spectrometer

  18. Sample preparation OSLO • On-line SPE (solid phaseextraction) • Small volumes (1 mL) • Automatic • No contamination • Time effective RIGA • Off-line (extract purification onphospholipid columns) • Large volumes (5 L) • Manual • Possible lab contamination • Time consuming Results to be compared

  19. Occurence study of AB by UHPLC-Orbitrap-HRMSRIGA • Occurence studies of 34 multi-class antibiotics were performed in fish samples obtained from 5 inland water farms in Latvia • Presence of two compounds was determined:

  20. Outcome • cooperation between the Latvian and Norwegian scientists on application of PPs analysis methods • benefiting short and long-term management on the distribution of pharmaceuticals in the Latvian and Norwegian environment • determination of their biodegradation processes and the methods reducing their hazardous impact on human health

  21. EEA/Norway Grants 2009-2014 Programme LV05NORWEGIAN-LATVIAN RESEARCH COOPERATION ACTIVITY Educational collaboration between Norway and Latvia for establishment of an effective risk assessment and control of pharmaceutical residues EEZ/NFI/S/2015/003

  22. SynergyAspect. ScholarshipActivity • Visits and/or the internship of several academic staff members and PhD students from the University of Latvia to the University of Oslo • Visit of academic staff members of UiO to UL, and holding several lectures devoted to activities of the current Project to students of the Faculty of Chemistry

  23. Mobility: November/ December 2015

  24. Synergy Aspect. Research Activity • Successful long-term scientific and educational cooperationbetween researchersofLatvian and Norwegian Universities • Acquired knowledge of environmental object analysis • Transfer ofknowledgebetweenresearchgroups • Involvement of PhD students, master and bachelor degree students • Development of training and studymaterials • Joint research publications

  25. People involvedUniversityof Latvia / BIOR • Assoc. professor Vadims Bartkevics • Professor Arturs Viksna • Researcher Dr Silvija Abele • Dr Dzintars Zacs • Dr Ingars Reinholds • Iveta Pugajeva • Olga Muter

  26. People involvedOslo University • Professor Elsa Lundanes • Assoc. professor Steven Ray Wilson • Researcher Dr Silvija Abele • Master student Deniz Demir • Bachelor student Sunniva F. Amundsen

  27. Future Tasks • Sampling of inland water bodies and industrial waste waters in Latvia and Norway • Application of the elaborated analytical methods (UHPLC-Orbitrap-HRMS) for the assessment of PPs in collected inland water and waste water samples • Preparation of peer-reviewed publications (in progress) of analytical performance and application advantages of the elaborated sensitive high resolution MSmethods for the analysis of pharmaceuticals

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