Chemical Analysis and Synthesis
Team
Thomas Mürdter, PhD, chemst, deputy head
Georg Heinkele, chemical engineer
Claudia Eser, laboratory technician
Robert Karandi, laboratory technician
Frank Schönberger, laboratory technician
Sonja Seefried, laboratory technician, safety inspector
Monika Seiler, laboratory technician
Main focus of the department is quantitative, mass spectrometric analysis. Analytical methods for quantification have been developed for more than 100 different drugs and metabolites, being used primarily for pharmacokinetic studies and for in vitro investigations of enzyme kinetics and drug transport.
As part of a metabolomics project in the HepatoSys competence network (a funding initiative of the German Federal Ministry of Education and Research on systems biology of hepatocytes) intracellular and extracellular concentrations of endogenous metabolites in hepatocytes are determined. Liver-specific intermediates of glucose metabolism (glycolysis, gluconeogenesis, TCA cycle, pentose phosphate pathway), lipid metabolism (ketone bodies, bile acids, intermediates in cholesterol synthesis) and amino acid metabolism are quantitatively determined.
The main task of the synthesis section is the chemical synthesis of otherwise not available substances (mostly drugs and their metabolites and stable isotope-labeled compounds). These substances are needed in virtually every area: In the analysis as reference substances and internal standards, for conducting in vitro studies on metabolism and transport of drugs and stable isotope-labeled drugs for clinical trials.
Equipment
• GC-MS (GC-MSD 5975C inert XL, Agilent)
• LC-MS (LC-MSD, Agilent)
• LC-MS-MS (Triple Quadrupol G6460A, Agilent)
• LC-MS-MS (HCT plus, Bruker)
• MALDI-TOF-(TOF)-MS (Ultraflex, Bruker)
HPLC-systems
• Gradient HPLC (Agilent 1200 with UV-dode array and fluorescence dtection)
• Several isocratic HPLCs with UV or fluorescence detection
Devices for sample preparation
| Glycolytic metabolites | glucose, pyruvate, lactate, glucose 6-P, glucose 1-P, fructose 6-P, fructose 1,6-P2, 3-phosphoglycerate, phosphoenolpyruvate |
| Pentose phosphate pathway | 6-phosphogluconate, ribulose 5-P, ribose 5-P |
| TCA cycle | malate, fumarate, citrate, isocitrate, aconitate, 2-oxoglutarate |
| Amino acids and urea | 20 proteinogenic amino acids, urea |
| Co-metabolites for detoxification | UDP-glucuronic acid |
| Cholesterol biosynthesis and metabolites | mevalonate, desmosterol, lathosterol, cholesterol, 4β-hydroxycholesterol |
| Bile acids | glycocholate, taurocholate, glycochenodeoxycholate |
| Nucleotides (energy charge) | ATP, ADP, AMP |
| Nucleosides | deoxycytidine, methyldeoxycytidine |
| Nucleobases | uracil, thymine and metabolites |
| Lipids |
about 25 individual triglycerides, present in liver phosphatidylcholines, sphingomyelins fatty acids phosphatidylethanolamines (semiquantitative) phosphatidylglycerols (semiquantitative) phosphatidylserines (semiquantitative) |
| Drug | Metabolites |
| 5-ASA | Acetyl-5-ASA |
| 5-fluorouracil | |
| amodiaquine | desethyl amodiaquine |
| antipyrin | all major metabolites |
| budesonide | |
| bupropion | hydroxybupropion |
| caffeine | paraxanthine |
| clomiphene | 8 metabolites |
| cyclophosphamide | |
| dextromethorphan | dextrorphan, hydroxymorphinan, methoxymorphinan, glucuronides |
| digoxin | |
| fexofenadine | |
| ifosfamide | |
| lansoprazole | |
| MAP kinase inhibitors | |
| mephenytoin | nirvanol, 4-hydroxymephenytoin metformin |
| metoprolol | α-hydroxymetoprolol |
| midazolam | 1'-hydroxymidazolam, 4-hydroxymidazolam |
| nabumetone | 5 metabolites |
| omeprazole | 5-hydroxyomeprazole, omeprazole sulfone |
| opiates | codeine, morphine, dihydrocodeine and desmethyl and glucuronide metabolites |
| phenacetine | acetaminophen |
| piritramide | |
| pridinol | hydroxylated metabolites |
| propafenone | 5-hydroxypropafenone, propafenone glucuronide, N-desalkylpropafenone |
| statins | atorvastatin, lovastatin, simvastatin, pravastatin, rosuvastatin and metabolites |
| tamoxifen | 30 metabolites |
| thiopurine drugs | 6-thioguanine in plasma, all nucleotide metabolites in RBC |
| tolbutamide | hydroxytolbutamide |
| tyrosine kinase inhibitors | |
| verapamil | all major metabolites |