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Analytical Department and Chemical Synthesis

Staff
Staff
• Claudia Eser
   laboratory technician
• Monika Seiler
   laboratory technician
Portrait
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 metabolomics projects in the Virtual Liver Network (a major national initiative on systems biology of the liver, funded by the German Federal Ministry for Education and Research) intracellular and extracellular concentrations of endogenous metabolites in hepatocytes and liver tissue 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
 

 
 
 
Mass spectrometers

 

 

  • GC-MS
  • GC-MSD 5975C inert XL, Agilent

    LC-MS(-MS)
  • Single Quadrupol 6120, Agilent
  • Triple Quadrupol 6460, Agilent
  • Triple Quadrupol 6495B, Agilent
  • Q-TOF iFunnel 6550 with UHPLC, Agilent
  • Ion Trap HCT plus, Bruker

HPLC Systems

  • 2 gradient HPLC systems (Agilent 1100/1200) with UV-Diode array and fluorescence detector, fraction collector
  • Devices for sample preparation

Chemical synthesis

  • Microwave reactor
  • Preparative flash chromatography system

 

Established quantitative methods for endogenous metabolites, mainly for cells and tissue samples

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 (all 20 proteinogenic amino acids, ornithine, urea); co-metabolites for detoxification (UDP-glucuronic acid); cholesterol biosynthesis and metabolites (mevalonate, desmosterol, lathosterol, cholesterol, 4ß-hydroxycholesterol); bile acids (glycocholate, taurocholate, glycochenodeoxycholate, muricholate); nucleotides (energy charge, ATP, ADP, AMP); nucleosides (deoxycytidine, methyldeoxycytidine for determination of total DNA methylation); nucleobases (uracil, thymine and metabolites); lipids (about 25 individual triglycerides, present in liver, phosphatidylcholines, sphingomyelins, fatty acids, phosphatidylethanolamines semiquantitative, phosphatidylglycerols semiquantitative, phosphatidylserines semiquantitative); steroid hormones (cortisol, 6ß-hydroxycortisol, estradiol, estrone, estrone sulfate, DHEA, DHEAS, testosterone, androstenedione, progesterone, epitestosterone)

Established quantitative methods for drugs and drug metabolites (plasma, urine, cells, tissue, in vitro experiments)

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; doxorubicin (doxorubicinol); fexofenadine; ifosfamide; lansoprazole; letrozole (carbinol metabolite, carbinol glucuronide); MAP kinase inhibitors; mefloquine (mefloquine enantiomers, carboxymefloquine); 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); rabeprazol; statins (atorvastatin, lovastatin, simvastatin, pravastatin, rosuvastatin and metabolites); tamoxifen (endoxifen and further 30 metabolites); tenatoprazol; thiopurine drugs (6-thioguanine in plasma, all nucleotide metabolites in RBC); tolbutamide (hydroxytolbutamide); tyrosine kinase inhibitors (dasatinib, imatinib, N-desmethylimatinib, nilotinib); verapamil (all major metabolites)