Metabolite profiling by LC-MS

1. In-vitro metabolite profiling of by
LC-MS/MS
Tejaswini Kailas Jadhav
MS
(Pharmaceutical Analysis)
NIPER-A

2. KEYWORDS
METABOLITE
PROFILING
IN-VITRO
HUMAN
LIVER
MICROSOMES
HEPATOCYTES
HPLC,
LC-MS/MS

3. 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.

4. DMPK studies is most
important part during
process of drug
discovery,
in-vitro ADMET
screens, thereby
accelerating the drug
discovery process.
DRUG DISCOVERY

6. Approaches in
Metabolite profiling
Metabolic
stability
Reaction
phenotyping
Metabolite
identification
Rate and
nature of
metabolism in
in-vitro
species Reactive
metabolite
formation
Enzyme
induction and
enzyme
inhibition-DDI

9. IN-SILICO
PREDICTION
Meta
Print 2D
METASITE
SYBYL
METEOR
Metabol
Expert

10. IN-
VITRO
S9 FRACTIONS
CYTOSOLIC
FRACTIONS
CELL LINES
LIVER SLICES
HUMAN LIVER
MICROSOMES
HEPATOCYTES
RAT LIVER
MICROSOMES

11. PROTEIN
PRECIPITATION
LIQUID-LIQUID
EXTRACTION
SOLID PHASE
EXTRACTION
EXTRACTION

12. MASS HPLC
LC-
NMR
GC-MS LC-
MS/MS
LC-ICP-
MS FT-IR RAMAN
TOF
Q-TOF
ION TRAP
QqQ
ORBITRAP
ANALYTICAL TECHNIQUES

13. 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

15. •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

17. TEJASWINI KAILAS JADHAV
MS(PHARMACEUTICAL ANALYSIS)
NIPER-A