Chemical quality control lecture 3

Compendial methods for QC of bulk drugs & drug products

Drug Compendia use various methods in order to identify, check the purity and to
quantitate drugs
• Physico-chemical methods used in quality control laboratories.
• Physical criteria
• Stability criteria
A-Gravimetric methods

It is used in the compendium either through:

A) Precipitation:
Thiamine HCl (BP) is precipitated with silicotungestic acid solution, filtered,
washed, dried, and weighed.

B) Solvent Extraction:
Butabarbiturate sodium (BP), acidification of aqueous solution with HCl
where butabarbituric acid precipitates out and is extracted with ether.
B-Titrimetric methods
Provide standard pharmaceutical methods for the assay of unformulated drugs
and excipients and some formulated drugs. e.g., those that lack strong
chromophore.
Used for standardization of raw materials and intermediates used in drug
synthesis.
Certain specialist titration, such as Karl Fisher titration, used to estimate water
content & are widely used in pharmaceutical analysis.
Advantages:
• They give higher degree of precision and accuracy than instrumental
methods of analysis. Precision ± 0.1% is being achievable.
• They are generally robust.
• Can be automated.
• Cheap to perform and don’t require specialized apparatus.
• They are not dependent on the calibration of an instrument.
Limitations:

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• Non-selective
• Time consuming if not automated.
• Needs greater operator skills than instrumental methods.
• Require large amount of sample and reagent.
• Reactions should be rapid and complete.
✓ Titrimetric methods include.
Acid-base, Precipitimetric (Argentometric), EDTA and Redox titration
C-Electrochemical methods
Potentiometry
• The main application is the determination of pH.
(To control the quality specifications of raw materials as well as many Dosage forms
especially injections).
• Very common in determination of e.p. of titrimetric methods. (Indirect
application of Potentiometry)

D-Spectroscopic methods
1-UV-VIS Spectroscopy in QC of drugs
Identification tests:
Most drug compendia use absorption spectra in UV-VIS region to identify
most drugs unless the dug is saturated and will not have any absorbance in the
region 200-800 nm.
Check Purity of the drug:
Some drug compendia use absorbance ratio as purity index of some
pharmaceuticals. As BP for cyanocobalamin and amphotericin B.
Limit test for impurities.
UV/VIS spectroscopy is used to quantitate concomitant compounds or
degradation products.
e,g. determination of hepatotoxic 5-HMF in dextrose injection.

For single component:

o BP gives the value of specific absorbance, A( 1 %, 1 cm) at the
corresponding ƛmax of the drug.
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o Single point procedure used by USP involves measurement of sample

solution and of standard solution of USPRS.
o Both solutions are prepared in the same manner Ctest = A test* Cstd/
Astd

For binary mixture:

Verdot’s method where two simultaneous equations are formulated and used
in calculations.

2- Colorimetry in QC of drugs
Used when the sample is colored and if colorless, we add chromogen to give
colored product.

General requirement of the colored product

• Should be intensely colored
• Unaffected over a reasonable pH
• Stable with time
• Quantitative color formation
• The formed colored products obey beer's law over a reasonable
concentration range.

General requirements of an ideal chromogen

✓ Colorless or easily separated.
✓ Selective
✓ Known color formation mechanism
✓ Rapid color development
✓ Give single product of specified ƛmaX
E.g. Determination of Amines:
a) Diazotization/ coupling with Bratton-Marshall reagent
b) Schiff base formation; with aldehydes : as Anisaldehyde and Vanillin

3-Spectrofluorimetry
o If the drug is naturally fluorescent → direct fluorimetry
o However, if the drug is non – fluorescent → indirect fluorimetry.
Two approaches. can be followed either via
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o Treatment with suitable reagent to give a fluorescent product; treatment

of thiamine HCI with alk. fericyanide,
o Derivatization with fluorogen, so that a fluorophore is added to the drug
and thus will become fluorescent; Fluroescamines react with primary
amine and amino acids to give strongly fluorescent derivatives.

Advantages of spectrofluorimetry

• High selectivity
• High sensitivity
Disadvantages of spectrofluorimetry
• No permanent calibration curve; should be done at each run.
• The linearity is limited.
5-IR spectroscopy
• The application of IR in QC is mainly qualitative to check the identity and
purity of drugs. Most drug compendia use it.
• The best solvents used are CCl4 or CHCl3 or CS2. IR spectra are generally
obtained as KBr or KCl discs.
• The fingerprint region of the spectrum (2000-400 cm-1) is very useful in
the identification of drugs.
• Formulations are usually, extracted with a specified solvent and it is
stipulated that adequate spectra will be obtained only if excipients in the
formulation are adequately removed.
• In addition to its major use in checking the identity and purity of drugs it
can be used to solve some pharmaceutical problems as: Quantitative
determination of cyclophosphamide with a fixed amount of ferric
thiocyanate as an internal standard (USP)·

IR Spectrophotometry
• IR spectrophotometry is primarily used for identification of raw materials
and products.
• The Infrared region is divided into near, mid and far-infrared.
• Near-IR refers to the part of the infrared spectrum that is closest to visible
light and far-IR refers to the part that is closer to the microwave region.

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• Mid-IR is the region between these two. For chemical analysis, we are
interested in mid IR region (4000 cm-1 to 650 cm-1).

• IR spectra can roughly be divided into two regions:

a) Functional groups (1200–3600 cm-1)
b) Fingerprint region (600–1200 cm-1)
• Advanced IR spectrophotometers are supplied with a library of IR spectra
that are stored on a computer.
• IR spectra of unknown compounds can thus be compared electronically
with reference spectra.

NIR Spectrophotometry
➢ The spectra have only a few significant peaks, but they are information-
rich due to the number of overlapping absorption bands.
➢ Direct comparison of the spectra is not appropriate.
➢ Suitable validated mathematical treatment of the data is required.
➢ Advanced software is usually used for multivariate calibrations
(chemometrics) to extract the desired information.

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FT-IR Sampling
Liquids
Place a small drop of the compound on one of the NaCl plates, then
place the second plate on top.
Solids
a) In solution (Liquids).
b) Nujol mulls:
Sample are placed onto the face of a NaCl plate, a small drop of mineral
oil is added and the second plate is placed on top.
c) KBr pellets/disks
Mix sample powder with KBr & press into disk.
IR Modes
a) Transmission/Absorption
b) Attenuated total reflection.
• ATR is a sampling technique used in conjunction with infrared spectroscopy
which enables samples to be examined directly in the solid or liquid state
without further preparation.
• Internal reflection material such as diamond, germanium, zinc selenide or
other suitable material having a high refractive index, and which do not
absorb infrared radiation.

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c) Transflectance mode
NIR can be carried out in transmission, reflectance, ATR & in transflectance mode.

6-NMR spectroscopy
• NMR spectrometry was adopted by· BP and USP to solve pharmaceutical
problems as quantitation of gentamycin concomitant components.
• NMR. spectrometry was also adopted by USP to check the purity and
quantitate amyl nitrile.
• NMR spectroscopy is used in pharmaceutical research to aid elucidating the
chemical structure of new medicines derived from natural sources or via
synthesis.

E-Chromatographic methods
➢ Chromatography is defined as a procedure by which solutes are separated
by a dynamic, differential migration process in a system consisting of two
or more phases, one of which moves continuously in a given direction and
in which the individual substances exhibit different mobilities by reason
of differences in adsorption, partition, solubility, vapor pressure,
molecular size, or ionic charge density.
➢ specific requirements for chromatographic procedures for drug substances
and dosage forms, including adsorbent and developing solvents, are given
in the individual monographs.
TLC
The method is relatively quick to perform and is cost effective because it
requires only little equipment and reagents, so TLC is used for second
identification. It is not used as the only identification test but must be
combined with other methods.
HPLC
HPLC is mainly used to test for related substances, it has also been
introduced as an identification parameter and for Assay.
Photodiode Array Detector

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➢ The Photo Diode Array detector (PDA) or Diode Array Detector (DAD)
delivers the entire spectrum of light available from the source lamp to the
flow cell. The light passing through the cell is diffracted into a spectrum
that is projected on a linear array of photodiodes.
➢ In this manner it is possible to record the entire absorbance spectrum of
analytes as they pass by the flow cell.

II- Physical criteria

• Disintegration / dissolution rates in tablets and capsules
• Weight variation
• Net content or declared content of tablets, capsules, and injections in
solid DF.
Certain physical tests could also be applied in determining the quality of
a product by visual inspection only.

General requirements for description of the product should include:

Color, consistency, clarity, stability, contamination with foreign matter or
fungal growth defects such as chipping of tablets, cracking, moisture
absorption, discoloration of the coating, decomposition, mottled
appearance· and various other characteristic defects which could be
checked by visual inspection.

Identification
▪ IR Spectrophotometry (most important technique) for final
dosage form
▪ UV-Visible Spectrophotometry (final dosage form)
▪ TLC (final dosage form)
▪ Melting Point
▪ Polarimetry
▪ HPLC (final dosage form)
▪ Chloride (AgNO3) and Sulfate (BaSO4) Identification

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Assay
• Titration
• HPLC
• UV-Visible Spectrophotometry

Tests
Test Impurities

Particles and colored

Appearance of solution
impurities

Absorbance UV-absorbing impurities

Acidity/alkalinity Acid and alkaline impurities

Specific optical rotation Optically active impurities

Loss on drying Volatile substances

Residual solvents Organic solvents

Heavy metals Heavy metals

Sulfated ash Inorganic material

Water Water

Related substances Related organic impurities

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Ash Value
The ash value usually represents the inorganic residue present in official
herbal drugs and pharmaceutical substances.

Total Ash
It is the amount of residual substances not volatilized from a sample when
the sample is ignited.
Acid-Insoluble Ash
• Residue in total ash is treated with HCl.
• A better test to detect soil in the drug than does the total ash.

Water-Soluble Ash
• Residue in total ash is treated with water.
• It is useful in detecting samples extracted with water.

Sulphated Ash
• It measures the amount of residual substance not volatilized from a
sample when the sample is ignited in the presence of sulfuric acid.
• It is usually used for determination of inorganic impurities in an organic
substance (herbal & synthetic).

Tests for Final Pharmaceutical Products

Related Substances
• It is intended to check that the content of impurities structurally related
to API is below their limit.
• Test for related substances can be performed as a part of the release
control for a production batch.
• It can be performed for stability testing during pharmaceutical
development.
• The test is normally performed by HPLC.

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Uniformity of Content
• To check that the amount of active pharmaceutical ingredient does
not vary too much from dose unit to dose unit.
• This test is typically used for:
a) Final pharmaceutical products containing 2 mg or less of API
per dosage unit.
b) Or where the dose-to-dose variation is critical.

• In the uniformity of content test:

1) Collect 10 individual dosage units randomly (10 tablets).
2) Analyze each dosage unit individually (determine the amount of
API in each unit). Their results should be within a certain
specified limit.
➢ The quantitative method used for Uniformity of content is often the
same method as used for the assay, the only difference being that
the assay gives an average value for the content.

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