Impurities, Degradants &

Stabiity of API’s

By
Dr. G.K.B. Prasad
Q_Pharma Consulting India
Hyderabad
 Quality of a Drug
TWO MOST IMPORTANT FEATURES OF A
DRUG

 EFFICACY
 SAFETY
 Quality of a Drug

‘SAFETY’ DETERMINED BY :

 TOXICITY PROFILE OF THE DRUG

 TOXICITY OF THE IMPURITIES

Drug must be ‘SAFE’ and ‘EFFECTIVE’ through out the

shelf life.
 ‘Impurities’ Under Focus

 ‘Assay’ ‘Impurities’

 ‘Toxicity Potential’ at low concentrations.

 Definitive prediction of long term toxic

effects of ‘impurities’ difficult.

 Drug substances can degrade during

storage.
 ICH Guidelines on Impurities & Stability
Study

ICH Q1A (R) : Stability testing of new drug substances

ICH Q3A : Impurities in new drug substances
ICH Q3C : Residual Solvents in Drug Substances

Also applies to existing drugs (impurity profile may be different

than the inventor).

‘Pharmacopoeial’ specifications alone may not be adequate.

 Nature of Impurities
Organic:
- Drug related (degradants)
- Process related (related substances)
In-Organic:
Reagents, Catalysts etc.
Inorganic salts
Metal residues
Others (charcoal, Filter aid etc.)
(Generally addressed in Pharmcopoeial specs)

Residual Solvents
Microrganisms
External Contaminants:

- From environment
- Cross contamination
(other materials)
GMP Issues (controlled through GMP).
 In-Organic Impurities

 Generally addressed by Pharmacopoieal

specifications and test methods.

Alkali metal salts: Sulfated ash test

Residue of toxic metal: Heavy metal test
Residue of Metal catalyst: Specific tests (ex; Arsenic)
Atomic absorption
 Organic Impurities
Process related:
Residues of,
- Raw material & Intermediates
- Side products
- Residual solvents

Degradants:
Possible degradation of API due to its exposure
to environment over long period of time.
- Heat, Sun light, Air, Acid/ basic medium
 Process Related Impurities
Ex: Four Stage Process

R1 R2 R3
A B C D Finished product
SM
Side product Side product Side product
S1 S2 S3

Residues of A, B, C, S1, S2, S3, R1, R2 and R3 are

‘Potential Impurities’ in the finished product (D)
 Potential Impurities in Finished
Product

 All potential impurities in drug substance to be

examined.
 Passover of early stage impurities to be
examined.
 Potential impurities from earlier stage could be
excluded, if established that they are absent in
intermediates.
 Suitable method to determine impurities shall
be established ….Generally HPLC, GC etc.
 Potential Impurities
 ‘May’ or ‘May Not’ be actually present in
the Drug Substance.

 Required to establish the suitability of the

test method (Validation)

 Demonstrate that the method can separate

and detect all the potential impurities.
 Identification of Impurities
 Synthesize
 Isolate from ‘Deviation Trails’
 RRT’s based on reported data
 In case not possible to synthesize, document
various efforts made.
 Quantification of Impurities
Chromatographic methods (HPLC / GC):

Based on Impurity Standards

Assuming Response Factor same as that of the

drug substance.
- Impurities with response factor similar or less
than that of drug substance.

- Unknown Impurities
(Caution to be exercised in case of residues with
toxicity at low concentrations, Ex: hormones).
 Impurities in Plant based Drugs

 More Complex
 Mixture of large number of organic compounds
 Identify potential impurities which can pass
over.
 Chromatographic finger printing
 RRT’s in literature data
 Specifications for Impurities
3 ‘Threshhold Levels’:
(New and Existing drug substances)

Reporting Level :Above which, should be reported

Identification Level :Above which, should be identified

Qualification level :Above which, Toxicity data is

required
 Recommended Impurity Limits
Max daily dose < or = 2g >2g

Reporting level 0.05% 0.05%

ID Level 0.1% or 1mg TDI* 0.05%

(which is lower)

Qualfn level 0.15% 0.03%

*TDI – Total daily intake

 Recommended Impurity Limits
Existing drug Substances:

Impurity profile based on mfg process.

Impurities described in pharmacopoieal

monograph

New impurities to be reported based on

threshold limits
 Impurity Limits for Drug Products
 Reporting level:
Max. daily dose Threshold limits
< or = 1 g 0.1%
>1g 0.05%

 Identification level:
<1 mg 1% or 5 μg TDI
1 mg – 10 mg 0.5% or 20 μg TDI
>10 mg – 2 g 0.2% or 2 mg TDI
>2g 0.1%
 Impurity Limits for Drug Products
Qualification level:

Max Daily Dose Threhold Limits

<10 mg 1% or 50 μg
10 mg – 100 mg 0.5% or 200 μg
>100 mg – 2 g 0.2% or 2 mg TDI
>2 g 0.1%
 Stability Study Program For API’s

API Molecules can degrade over a period of time under

following environmental conditions:

 Temperature
 Humidity
 Air (oxidation)
 Ph Conditions (acidic / basic)
 Stability Program for API’s

 Determine Shelf Life of the Drug

 Define Expiry date / Retest
 Degradants within limits through out shelf life

API specifications:
- Actual Limits
- Limits for Batch Release
(More stringent)
 Stability Under Controlled Conditions
Environmental conditions change periodically;
Morning, Afternoon, Evening, Night
Summer, winter, Rainy season etc.

Climatic Zones:
Zone I - Temperate (ICH region)
Zone II - Sub tropical (ICH region)
Zone III - Hot & Dry
Zone IV - Hot & Humid (Tropical)
 Mean Kinetic Temperature
Mkt (TK) is the single calculated temperature which simulates
the kinetic effect of time temperature distribution.
∆h / R
Tk = ____________________________________________
-ln{e-∆H/RT1H + e-∆H/RT1L +….+e-∆H/RTnH + e-∆H/RTnL}
2n
Where,

Tk = mean kinetic temperature

∆H = heat of activation (83.144 kJ/mole-1)
R = Universal gas constant
TiH = high temperature in °K during the 1st week
T1L = Low temperature in °K during nth week
TnH = High temperature in °K during nth week
TnL = Low temperaturer in °K during nth week
n = total number of weeks (i,e 52)
T = Absolute temperature in °K
°K = °C + 273.2
 Stability Study Conditions
Mean Kinetic Temperature (MKT):

Climatic Zone MKT Derived Temp

(°C) (°C)
I 20 21
II 22 25
III 27.9 30
IV 27.4 30

Ref: WHO Tech Report Series no 790

 Stability Study Conditions

Long Term Accelerated

ICH region (Zone II & II)

Temp: 25 ± 2 °c } 40 ± 2 °c }
RH : 60 ± 5% RH } 60 ± 5% }

INDIA (Zone III)

Temp: 30 ± 2 °c } 40 ± 2 °c }
RH: 65 ± 2 °c } 75 ± 5% }
 Stability Study Conditions
Intermediate Conditions:

Recently revised to,

Temp: 30 ± 2°c
RH: 65 ± 5%

(Same as Long Term Conditions for Zone III & IV)

 Stability Study Conditions

Packaging of Samples:
 Simulate Market dispatch packing:
(Identical material for Primary & Secondary PM, )

 Sample enough for at least two analysis

 Pack separately for each schedule
 Label (Sample name, B.No., Mfg Date, S.No. etc.)
 Stability Indicating Test Methods
Monitoring of Stability Study samples:

 The test methods must be able to detect degradation of the

drug substance
(Physical Description, pH, solubility, TLC etc.)

 Quantification of Degradants (HPLC, GC etc.)

- HPLC: Generally preferred stability indicating method
- The method must have been validated (for impurities)
- Trending of degradents profile
 Forced Degradation Study (Stress
Testing)
 Technique to identify potential degradant.
 Degradation of API molecule depends on its intrinsic
stability (API may or may not form degrade)
 Stress conditions to force the API to degrade.
 Stress conditions – Similar to the ones to which API is
exposed in environment .
- Degradation due to Temperature
- Oxidation due to exposure to Air
- Degradation due to exposure to Sun light / UV light
- Degradation due to Acid / Basic conditions (in Mfg process)
 Forced Degradation Study - Technique

 Treat the drug substance to extreme Heat, pH,

Direct sun light Light/ UV, Chemical oxidation.

 Develop methods to separate degradants

 Isolate degradants & characterize (if required!)

Forced Degradation Study - Technique

 pH: Aq. Hydrochloric Acid

Aq. Sodium Hydroxide

 Temp: Incements by 10 °c above 40 °c

Max temp 105 °c

 Oxidation: Dilute aq. Hydrogen Peroxide

 Forced Degradation Study - Technique

PHOTODEGRADATION (Photo stability):

Light source:

Artificial day light fluorescent lamp (UV +

Visible) equivalent to D65 / D165 standard (ex.
ISO 10977; 1993).
Ex: Xenon or metal halide lamp
 Characterisation of Degradents

 Isolation: Preparative HPLC

Preparative TLC

 Characterisation:

NMR, IR, Mass Spectra, X-ray diffraction

Known references etc.
 Analysis of Stability Samples

 Must conform to all specifications

 Use stability indicating methods
 Monitor degradants
 Report / Identify / Qualify based on threshold
limits
 Evaluate any deviations from set conditions
(Analyse samples on significant deviations)
 Frequency of Analysis:

 LONG TERM:
- 3rd, 6th , 9th , 12th , 18th , 24th , 36th , 48th and 60th
months

 ACCELERATED:
- 3rd and 6th month
- More frequently if known degrade
more readily.
 Stability Data Evaluation

 - Degree of variability of Batches

 - Worst Batch data
 - Statistical Evaluation
 - Commitment to verify ‘Long Term’ Study
 Extrapolation of ‘Retest Date’ Beyond ‘Long
Term’
Accelerated Long Term Extrapolation

1. Not Significant Not Significant 2 X Long term

Max: Long term+12 months

2. Change Change 1.5 X Long term (No stat)

Observed Observed Max: Long term + 6 months

2X Long term (with Stat)

Max: Long term+ 12 months

3. Significant Not significant Max: Long term +3months (no stat)

& Not significant Max: Long term + 6months (with Stat)
at Intermediate)

Significant at --- “ ----- Max : Long Term data ( May be less)

at intermediate
 Degradants of Biological Drugs
 Mostly proteins and peptides; more complex
 Functions related to secondary & tertiary
structure
 More sensitive to Light, Heat, PH, Oxidation,
ionic concentration and shear
 Require precisely defined storage conditions.

Stress Conditions:

 Based on intrinsic stability of the individual

drug.