in vitro metabolite profiling
TRANSCRIPT
In-vitro metabolite profiling of by LC-MS/MS
Tejaswini Kailas Jadhav
MS(Pharmaceutical Analysis)
NIPER-A
KEYWORDS
METABOLITE PROFILING IN-VITRO HUMAN LIVER
MICROSOMES HEPATOCYTESHPLC,
LC-MS/MS
METABOLITE PROFILING
Metabolites are the intermediates and products of metabolism.The measurement and interpretation of the endogenous metabolite profile from a biological sample (typically urine, serum, or biological tissue extract) have provided many opportunities to investigate drug and metabolite that are present in body.
DMPK studies is most important part during
process of drug discovery,
in-vitro ADMET screens, thereby
accelerating the drug discovery process.
DRUG DISCOVERY
Approaches in Metabolite profiling
Metabolic stability
Reaction phenotyping
Metabolite identificatio
n
Rate and nature of
metabolism in in-vitro species Reactive
metabolite formation
Enzyme induction and
enzyme inhibition-DDI
IN-SILICO PREDICTION
Meta Print 2D
METASITE
SYBYL
METEOR
MetabolExpert
IN-VITRO
S9 FRACTIONS
CYTOSOLIC FRACTIONS
CELL LINES
LIVER SLICESHUMAN LIVER MICROSOMES
HEPATOCYTES
RAT LIVER MICROSOMES
PROTEIN PRECIPITATION LIQUID-LIQUID
EXTRACTIONSOLID PHASE EXTRACTION
EXTRACTION
MASS HPLC LC-NMR GC-MS LC-
MS/MSLC-ICP-
MS FT-IR RAMAN
TOF
Q-TOF
ION TRAP
QqQ
ORBITRAP
ANALYTICAL TECHNIQUES
LC-MS/MS
Mass spectrometry plays a key role in drug metabolite identification, It has improved accuracy and stability, along with new data processing techniques, has improved the quality and productivity of metabolite identification processes.
LC-MS/MS
•Incubation of test drug sample with hepatocytes/HLM will lead to formation of Phase 1 (Oxidation / Reduction / hydrolysis) and Phase 2 conjugated metabolites.
•Which will be identified by validated LC-MS/MS method.
•Metabolism (percentage of test compound metabolized after a certain time) and qualitative determination of metabolite pattern (minor, intermediate, and major abundance of each metabolite)
EXPECTED OUTCOMES
TEJASWINI KAILAS JADHAV
MS(PHARMACEUTICAL ANALYSIS)NIPER-A