Unit-I

Impurities in pharmaceutical
substances
Objective: Identification and quantification
of impurities in drug compounds
Dr. Saleemulla Khan
What is an Impurity?
• An impurity, as defined by the ICH (The International Conference on
Harmonisation of Technical Requirements for Registration of
Pharmaceuticals for Human Use) guidelines is “any component
of the medicinal product which is not the chemical
entity defined as the active substance or an excipient
in the product”
Classification of Impurities
• USP Classification According to united state pharmacopoeia
impurities classify as
• Impurities in official articles
• Ordinary impurities
• Organic volatile impurities
• ICH Classification
• Organic impurities
• Inorganic impurities
• Residual solvents
• Organic impurities may arise from starting materials, by products,
synthetic intermediates and degradation products.
• Organic impurities sources are complex; Drugs prepared by multi-step
synthesis results in various impurities, their number and the variety of
their structures are almost unlimited.
• Several other factors such as;
• Purity of the starting material,
• Method of isolation, Intermediates
• Purification, Byproducts,
• Conditions of storage, Degradation product, etc.
Inorganic impurities may be derived from the manufacturing process
and are normally known and identified as reagents, ligands, inorganic
salts, heavy metals, catalysts, filter aids and charcoal etc.

Residual solvents are the impurities introduced with solvents

 Sources of Impurities
Sources of organic impurities
Impurities originating from drug substance synthetic processes
• Most of the drug substances (low molecular weight) are chemically synthesized.
• Chemical entities that carry forward to the product include,
• A. raw materials,
• B. intermediates,
• C. solvents,
• D. chemical reagents,
• E. catalysts,
• F. by-products,
• G. impurities present in the starting materials,
• Also those chemical entities formed from starting material impurities (particularly those
involved in the last steps of the synthesis).
• These impurities are usually referred to as process impurities
Starting materials and intermediates: Most Common
• Starting materials and intermediates are the chemical building blocks
used to construct the final form of a drug substance.
• Unreacted starting materials and intermediates, particularly those
involved in the last steps of the synthesis, can appear in the final
product as impurities
• For example, in the synthesis of tipranavir drug substance, aniline is
the intermediate in the last step of the synthesis.
• Due to the similarity between the structures of aniline and the final
product, it is difficult to totally eliminate it in the subsequent
purification step.
• Another example: In paracetamol bulk, there is a limit test for p-
aminophenol, which could be a starting material for some one
manufacturer
Reagents, ligands and catalysts
• These chemicals are less commonly found in APIs; however, in some cases
they may pose a problem as impurities.
• Chemical reagents, ligands, and catalysts used in the synthesis of a drug
substance can be carried over to the final products as trace level
impurities.
• For example, carbonic acid chloromethyl tetrahydro-pyran-4-yl ester
(CCMTHP), which is used as an alkylating agent in the synthesis of a ß
lactam drug substance, was observed in the final product as an impurity.
• Many chemical reactions are promoted by metal based catalysts.
• In some cases, reagents or catalysts may react with intermediates or final
products to form by-products.
• Example: Pyridine, a catalyst used in the course of synthesis of
mazipredone, reacts with an intermediate to form a pyridinium impurity.
By-products of the synthesis
• All chemical reactions are not 100% selective; the side-reactions are
common during the synthesis of drug substances.
• By-products from the side reactions are among the most common
process impurities in drugs.
• By-products can be formed through a variety of side reactions, such
as incomplete reaction, over reaction, isomerisation, dimerisation,
rearrangement, or unwanted reactions of starting materials or
intermediates with chemical reagents or catalysts.
Degradation products
Impurities can also be formed by degradation of the end product
during manufacturing of bulk drugs.
However, degradation products resulting from storage or formulation
to different dosage forms or aging are common impurities in the
medicines.
The deg-radation of penicillins and cephalosporins is a well-known
example of degradation products.
Enantiomeric impurities
• The single enantiomeric form of a chiral drug is now considered as an
im-proved chemical entity that may offer a better pharmacological
profile and an increased therapeu-tic index with a more favorable
adverse reaction profile.
• However, the pharmacokinetic profile of levofloxacin (SIsomeric form)
and ofloxacin (R- isomeric form) are comparable, suggesting the lack
of advantages of single isomer in this regard.
• The prominent single isomer drugs, which are being marketed,
include levofloxacin (S-ofloxacin), levalbuterol (R-albuterol) and
esomeprazole (S-omeprazole)
 Inorganic impurities
Reagents, ligands, and catalysts:
• The chances of having these impurities are rare: however, in some processes,
these could create a problem unless the manufacturers take proper care
during production
Heavy metals
• The main sources of heavy metals are the water used in the processes and
the reactors (if stainless steel reactors are used), where acidifi-cation or acid
hydrolysis takes place.
• These impuri- ties of heavy metals can easily be avoided using demineralized
water and glass-lined reactors.
• Other materials (eg, filter aids, charcoal)
• The filters or filtering aids such as centrifuge bags are routinely used in the
bulk drugs manufacturing plants, and, in many cases, activated carbon is also
used.
Solvent residues
• Residual solvents are organic volatile chemicals used during the manufacturing
process or generated during the production.
• It is very difficult to remove these solvents completely by the work-up process;
however, efforts should be taken to the extent pos-sible to meet the safety data.
• Some solvents that are known to cause toxicity should be avoided in the
production of bulk drugs.
• Depending on the possi-ble risk to human health, residual solvents are di-vided
into 3 classes
• Solvents such as benzene (Class I, 2 ppm limit) and carbon tetrachloride (Class I, 4
ppm limit) are to be avoided
• On the other hand, the most commonly used solvents such as methylene chloride
(600 ppm), methanol (3000 ppm), pyridine (200 ppm), toluene (890 ppm), N,N-
dimethylformamide (880 ppm), and acetonitrile (410 ppm) are of Class II
• Class III solvents (acetic acid, acetone, isopropyl alcohol, butanol, ethanol, and
ethyl acetate) have permitted daily exposures of 50 mg or less per day.
• In this regard, ICH guidelines for limits should be strictly followed
Impurity forms during formulation
• Method related
• A known impurity, 1-(2,6-diclorophenyl)indolin-2-one is formed in the
production of a parenteral dosage form of diclofenac sodium if it is
terminally sterilized by autoclave
• It was the condition of the autoclave method (ie, 123 + 2°C) that
enforced the intramolecular cyclic reaction of diclofenac so-dium
forming the indolinone derivative and sodium hydroxide.
Environmental related
• Exposures to adverse temperatures - There are many APIs that are
labile to heat or tropical tem-peratures. For example, vitamins as drug
substances are very heat –sensitive
• Light-especially UV light - Several studies have reported that
ergometrine as well as methyl er-gometrine injection is unstable
under tropical con-ditions such as light and heat, and a very low level
of active ingredient was found in many field sam-ples
• Humidity- For hygroscopic products, humidity is considered
detrimental to both bulk powder and formulated solid dosage forms.
Aspirin and ranitidine are classical examples.
Pharmacopoeial status
• The quality of a chemically active substance with respect to organic
impurities is controlled by a set of tests within a pharmacopoeial
monograph
• Individual monographs are periodically updated to keep place with
scientific progress and regulatory developments.
• Major pharmacopoeias will continue publishing new or revised
relevant monographs and general chapters
• Two general chapters (466 & 1086) of the US Pharmacopoeia (USP)
deal with organic impurity testing.
• Concepts and definitions are clearly described although different
terminology from that of ICH is used.
Indian Pharmacopoeia
• Until now, one of three types of tests in bulk pharmaceutical
chemicals is ordered

• 1. A chromatographic purity test coupled with a non- specific assay

• 2. A chromatographic purity- indicating method that also serves as an
assay
• 3. A specific limit test for known impurities, a procedure that requires
reference standards for these impurities
ICH guidelines:
• According to ICH guidelines, each impurity must be investigated with
respect to both chemistry and safety aspects.
• The former include identification (structural characterization),
reporting and quantitation using suitable analytical procedures, while
the latter include a process of acquiring and evaluating data
concerning the biological safety of an impurity (qualification).
• Individually listed impurities, limited with specific acceptance criteria,
are referred to as specified and they can be either identified or
unidentified.
General Method for impurity detection
• 1. Isolation and characterization
• 2. Column chromatography
• 3. Gas chromatography
• 4. Flash chromatography
• 5. TLC
• 6. GC
• 7. HPLC
• 8. HPTLC
• 9. Capillary electrophoresis (CE)
Following methods are commonly used for the
isolation, they are
• i. Extraction
• ii. Column chromatography
• iii. Preparative separation
• iv. Extraction:
• v. Liquid –solid extraction
• vi. Liquid-liquid extraction
Inorganic Impurities
• The number of inorganic impurities and residual solvents are limited.
• These are easily identified and their physiological effects and toxicity
are well known.
• For this reason the limits set by the pharmacopoeias and the ICH
guidelines can guarantee that the harmful effects of these impurities
do not contribute to the toxicity or the side effects of the drug
substances.
Limit Tests