API analysis using Analytical instrumentation

1. SIGNIFICANCE OF ANALYTICAL
INSTRUMENTATION IN API DEVELOPMENT
AND MANUFACTURING
DR. V. CHANDRASEKHARAN

2. THE PHARMACEUTICAL INDUSTRY
• ONE OF THE MOST REGULATED
• HIGHLY RESEARCH AND INNOVATION DRIVEN
• 25 MAJOR THERAPEUTIC SEGMENTS
• OVER 1800 DIFFERENT STRUCTRUAL TYPES
• 44 NEW MOLECULES IN 2014 INTRODUCED
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3. THE PHARMACEUTICAL INDUSTRY HAS THREE
SEGMENTS.
• DRUG DISCOVERY AND DEVELOPMENT
• API MANUFACTURE
• FINISHED PHARMACEUTICAL PRODUCT DEVELOPMET AND
MANUFACTURE
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4. DRUG DISCOVERY ASPECTS
 Antihypertensives:
 50’s Direct Vasodilators Eg: Hydralazine
 70’s Beta blockers Eg: Atenalol
 70’s Ca channel blockers Eg: Nifedipine
 80’s ACE inhibitors Eg: Captopril
 90’s Angiotensin II receptor antagonist Eg:
Losartan
 2010 Renin Inhibtors Eg: Aliskerin
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5. ACTIVE PHARMACEUTICL INGREDIENT
 It is defined as a substance or compound used in a finished
pharmaceutical product intended to furnish desired
pharamacological and or therapeutic activity
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6. STRUCTRUAL TYPES
 An Active Pharmaceutical ingredient can be from a simple
structural type to a very complex type.
 It covers a wide variety of classes like aliphatics, aromatics,
heterocyclics, carbohydrates, peptides, nucleotides ,
organometallics etc
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7. DIMETHYL FUMARATE USED FOR
MULTIPLE SCLEROSIS
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8.  AZITHROMYCIN-MACROLIDE ANITIBIOTIC
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9. API SYNTHESIS
 It can be totally synthetic. Eg. Ibuprofen
 It can be semisynthetic ( starting material from natural sources,
fermentation product etc) Eg. Paclitaxol, Amoxicillin
 It can be biotechnology based. Eg. Insulin, vaccines
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10.  Route of synthesis should lead to the correct structure,
stereochemistry , crystalline form and size.
 Should be well controlled and validated.
 Should produce the API which meets acceptable standards of
quality attributes and other criteria.
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11. Cl Cl
NO2
OH
NO2
OH
NO2 NHOH
OH
NH2
OH
OH
NHCOCH3
OH
SYNTHESIS ROUTES TO ACETAMINOPHEN
Ammonium
Acetate
Acetic acid
Acetaminophen
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Synthetic route of Acetaminophen (Paracetamol)

12. Key starting material quality
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13. ISOMERS OF BUTYL BENZENE
n-Butyl benzene iso-Butyl benzene sec-Butyl benzene t-Butyl benzene
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15. ANALYTICAL ASPECTS DURING REACTION
SEQUENCING
 PARAMETER OPTIMISING AND DEFINING CRITICAL
PROCESS PARAMETERS.
 DEFINING AND OPTIMISING REACTION MONITORING eg.
TLC, GC, HPLC, UPLC etc
 STRUCTURAL CONFIRMATION OF INTERMEDIATES, SIDE
PRODUCTS FORMED. eg. IR, UV, HPLC, NMR, MASS ETC.
 STABILITY ASPECTS OF INTERMEDIATES
 DEFINING QUALITY ASEPCTS OF INTERMEDIATES.
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16. AZITHROMYCIN SYNTHESIS
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17. FINAL ASPECTS OF API
 STRUCTURAL CONFIRMATION (IR, UV, NMR, MASS, HPLC
ETC)
 IMPURITY PROFILE INCLUDING QUANTIFICATION,
STRUCTURAL ASPECTS.
 RESIDUAL SOLVENTS (GC-MS)
 GENOTOXIC IMPURITIES
 INORGANIC INCLUDING METAL OTHER IMPURITIES.
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18. ANALYTICAL ASPECTS OF CHIRAL API’S
 R isomer is sedative and S isomer is teratogenic.
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19. OH
NH
OH
N
H
N
N
S
H
CH3
O O
N
CH3
OMe
OMe
S,S-ETHAMBUTOL (R,R-ETHAMBUTOL CAUSES BLINDNESS)
ESOMEPRAZOLE
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20. Ibuprofen enantiomer separation by chiral HPLC
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21. SOLID STATE PHENOMENA
 PARTICLE SIZE
 POLYMORPHISM
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22.  Particle size can vary from 0.001 um to 1000 um and this will
have significant effect on final drug product performance
particularly in the solid dosage forms like dissolution,
bioavailability etc.
 The most common methods are either by Sieve analysis or by
Laser diffraction method. A number of other techniques are
also available
 A recent advancement in this area is Image analysis-Raman
Spectroscopy which gives the following parameters-Particle
shape, particle size and Chemical identifcation in one shot.
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23. POLYMORPHISM
Polymorphs are different crystalline forms of the
Same pure substance in which molecules have
Different crystal lattice formation and or
Different molecular conformations.
It has been shown different polymorphs often
Have different physico chemical properties,
Solubility, dissolution rates, bioavailability etc.
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24.  Piroxicam and its salts exist in two crystalline forms, one needle
shape and the other cubic form.
 Form one melts at 196-198 oC where as Form 2 melts at 199-
201 oC.
 The IR spectra clearly differentiates the two forms in the amide
(N-H) and the C=O of amide and ester.
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25. IR spectra of Piroxicam Polymorphs
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PIROXICAM
N
O
N
S
OH
NH
CH3
O O

26. X-ray diffraction pattern of Piroxicam polymorphs 2
(needle ) and 1 (cubic)
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27.  So far three polymorphic forms of paracetamol have been
reported.
 Form 1 monoclinic which is commercially available and more
stable.
 Form 2 orthorhombic, difficult to prepare, metastable, but has the
advantage that sold dosage form can be made by direct
compression without the aid of binders.
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28. DSC-Raman spectra of Paracetamol polymorphs
PARACETAMOL EXISTS IN THE FOLLOWING FORMS:
1. THERMODYNAMICALLY STABLE MONOCLINIC FORM. POOR COMPRESSION PROPERTIES.
2. ORTHORHOMBIC METASTABLE FORM BUT DIRECTLY COMPRESSIBLE
3. A NEW FORM
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29. OTHER EXAMPLES
 Ranitidine , Piroxicam, Clopidogrel, Famotidine, Ritonavir.
 A number of instrumental analytical techniques are used for the
characterisation of different polymorphs, like IR, Raman, X-ray
diffraction, DSC, solid state NMR, AFM (atomic force
microscope) etc.
 Raman can measure low freqency vibrations better than IR.
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30. Ibuprofen degradation studies by HPLC
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31. CONCLUSION
 With stringent regulatory-purity-impurity profile emanating day by
day as new , high potent, complex structrual types API being
discovered and developed, Analytical instrumentation has taken
the pivotal role in all facets of API development and
manufacture.
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32. Thank You