in vitro metabolite profiling
TRANSCRIPT
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In-vitro metabolite profiling of by LC-MS/MS
Tejaswini Kailas Jadhav
MS(Pharmaceutical Analysis)
NIPER-A
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KEYWORDS
METABOLITE PROFILING IN-VITRO HUMAN LIVER
MICROSOMES HEPATOCYTESHPLC,
LC-MS/MS
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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.
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DMPK studies is most important part during
process of drug discovery,
in-vitro ADMET screens, thereby
accelerating the drug discovery process.
DRUG DISCOVERY
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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
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IN-SILICO PREDICTION
Meta Print 2D
METASITE
SYBYL
METEOR
MetabolExpert
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IN-VITRO
S9 FRACTIONS
CYTOSOLIC FRACTIONS
CELL LINES
LIVER SLICESHUMAN LIVER MICROSOMES
HEPATOCYTES
RAT LIVER MICROSOMES
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PROTEIN PRECIPITATION LIQUID-LIQUID
EXTRACTIONSOLID PHASE EXTRACTION
EXTRACTION
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MASS HPLC LC-NMR GC-MS LC-
MS/MSLC-ICP-
MS FT-IR RAMAN
TOF
Q-TOF
ION TRAP
QqQ
ORBITRAP
ANALYTICAL TECHNIQUES
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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
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•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
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TEJASWINI KAILAS JADHAV
MS(PHARMACEUTICAL ANALYSIS)NIPER-A