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