Pharmaceutical Analysis

(I) Spring 2024

Dosage form analysis
By
Prof. Tariq Almog
REFERNCES
1. Handbook of Pharmaceutical Analysis, edited by Lena
Ohannesian and Anthony J. Streeter, 1st edition 2001.
2. USP & PB.
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 Introduction
Dosage forms are pharmaceutical drug products marketed for use,
consisting of a specific mixture of active ingredients and inactive
components.
Dosage form analysis refers to the process of determining the
composition, quality, and characteristics of pharmaceutical drug products
in the form in which they are marketed for use, such as tablets, capsules,
solutions, suspensions, etc

Common analytical techniques used for dosage form analysis include:

✓ Titrations
✓ Ultraviolet (UV) and Infrared (IR) Spectroscopy
✓ High Performance Liquid Chromatography (HPLC)
✓ Gas Chromatography (GC)
✓ Atomic Absorption (AA) Spectroscopy.
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 Aerosol inhalation
Aerosol is the dosage form that the unit dose contain the
medicament in suspension form that can be delivered by
propellant which can be:
Dichloro-difluro tetramethane-
Dichloro-tetrafluro tetramethane -
Pulse:
➢ Surface active agent
➢ Stabilizing agents
➢ Other adjuvants.

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The Aerosol inhalation device consist from:
➢ Aerosol dispenser which is used to containing the medicament under
pressure.
➢ Metering valve is used for precision of the dose or amount of
medicament.
Examples of aerosol inhalation :
1. Isoprenaline Aerosols
2. Salbutamol Aerosols
3. Sodium chromoglycate Aerosols
4. Benzoin inhalation B.P.
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Isoprenaline Aerosols Inhalation bronchodilator

Assay based on the formation of the complex, when the

phenol group of isoprenaline react with iron salts producing
quinone products having a colour measured at 530 nm using
spectrophotometer. The result obtained is the average of
10 spray.

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 Particle size is an important part of specification for

isoprenaline aerosol inhalation, it should be 2-8 µm.

➢ If the particle too small it will coming out during

breath.

➢ If too big it will not able to penetrate the lung to

give there action in the bronchitis and alveoli.

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 Capsule
➢ It is a shell containing medicament, made from
gelatin, it can be hard or soft according to
content % of glycerol or sorbitol solution.
➢ Capsule usually containing a single dose of active
ingredient.
➢ Capsule can be: hard capsule; soft capsule;
modified release capsules or gastro resistance
capsules.

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 Quality control of capsules
1. Disintegration test: Apply for all types of capsule
except modified release capsules.
2. Dissolution test: Apply for all types of capsule
3. Uniformity of content: should apply for all types of
capsules.
4. Uniformity of weight: Should apply for all types of
capsules.
5. Determination of active ingredient in capsule: Chemical
assay apply for determination PAI in the capsule.
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 5. Determination of active ingredient in capsule
Direct method of analysis: By using assay spectrophotometry or chromatography
as for the assay of ingredient in dry capsules if not possible we use:
➢ Solvent extraction method:
Used when the direct method is unsuitable as the assay of amylobarbiton
sodium capsule and phenytoin capsule in which the drug dissolve in organic
solvents to separate it from excipients and other components present
in the capsule formulation.
➢ Digestion method:
This done by digest the capsule with acid and heat using 0.1N HCl either in
water bath, autoclave or microwave, the resulted is filtered then the assay
method is applied.
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 Name four capsules and mention their method of assay?

Examples method of assay ingredient in capsule

❖ Clofibrate capsules (UV method)
❖ Ampicillin Capsules (HPLC method)
❖ Cephalexin Capsules (Iodimetric method)
❖ Disopyramide Capsules (non-aqueous titration).

Assay of Alcofenac capsule

Sample: 20 capsule equivalent to 0.5 g of alclofenac.
Solvent : 100 ml 96% neutralized ethanol.
Titrant: 0.1 M NaOH.
Indicator: ph.ph.
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 Cough Capsules Promethazine Hcl

❖ Promethazine HCl. 36 mg
❖ Noscapine . 12.5 mg
❖ Ephedrine HCl. 7.2 mg

❖ promethazine Hcl
To determine promethazine HCl add to the sample
Palidium chlorate
.
PdCl3 ORANGE COLOUR
+ mesure at 427 nm
Palidium chlorate
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❖ Noscapine

Noscapine
Detected at UV 310 nm

10 ml noscapine has diluted to 100 ml water then measured

spectrophotometry at 310 nm.

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❖ Ephedrine HCl

Ephedrine HCl
Detected at UV 225 nm

Ephedrine HCl, has the same max of Noscapine 310nm,

therefore, it oxidized in alkaline solution (NaIO4 + NaHCO3)
and extracted with sulphuric acid and cyclohexane then
measured spectrophotometry at 225 nm.

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 CREAMS
➢ Creams are defines as “a semisolid dosage form containing one
or more drug substances dissolved or dispersed in a suitable
base”

➢ Semisolid emulsions either O/W or W/O type.

➢ They are used to apply drugs to the skin for protective and
therapeutic purposes.

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 Quality control applied for creams
1.Microbial quality control.
2. Physical quality control.
3. Chemical quality control: for many creams determination of
active ingredient can be done without separation as the basis
does not interfere with the assay process
Ex: cetrimide cream; salicylic acid cream ; sulphur cream
while the others separation should be done prior the assay
❖ Choice of correct solvent is important for assay; as it avoid
a lengthy separation procedure.

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 Cream Solvent
Hydrocortisone Dehydrated ethanol
Nexenone Ethanol
Proflavine Chloroform/alcohol

➢ Extraction and separation applied when the concentration of

medicament is small e.g . Steroids preparation.
➢ Spectrophotometric or titrimetric analysis is applied for
detection the PAI at the end of extraction.

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 ➢ Cetrimide Cream
How you can prepare cetrimide cream for the assay?
What are the assay condition of cerimide?

Sample preparation: Dissolve the cream in, hot water, then add sulfuric
acid, chloroform and few drops of dimethyl yellow as indicator.
Assay condition:
Analyte: Cetrimide
Titrant: Dioctyl sodium sulfosuccinate.
Indicator: dimethyl yellow solution.
End point: The yellow color is obtained.
Each 1ml Dioctyl sodium sulfosuccinate = 0.3364 Cetrimide

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 Eye Drops

Eye drops are sterile dosage forms, consist of

solutions or suspensions of one or more
components intended for use in the conjunctional
sac for therapeutic or diagnostic purpose.
Eye drops are sterile preparation and those that
are aqueous containing a bactericidal and
fungicidal.

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 Quality control of Eye drops
The preparation of eye drops must comply with the following test :
1. Limit of particle size.
2. Sterility.
3. Chemical quality control:
4. Containers.
5. Labeling.

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 Chemical quality control of eye drop
Name three eye drops and mention their method of assay?

Chemical assay depend on the types of PAI in the formulation of eye

drop, the method of analysis can be volumetric method or instrumental
method as spectrophotometer or chromatographic method.

➢ Atropine eye drops: Assay by Gas chromatography.

➢ Chloramphenicol eye drops: Assay by UV spectroscopy at 278 nm

➢ Pilocarpine eye drops: Can be titrated with cetyl pyridinium chloride

using bromophenol blue as indicator. Since 2013, the assay of
Pilocarpine is done by HPLC .
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 Eye ointment
❖Ointments are semisolid dosage forms for external use,
usually consisting of solid or semisolid hydrocarbon
base of melting point close to human body temperature.
❖The base, usually containing antioxidant, stabilizing
agents, and antimicrobial preservatives.
❖ After applying ointment to the eye, it decomposes into
small drops, which stay for a longer time period in
conjunctival sac, thus increasing drug’s bioavailability.
❖Eye ointment must be prepared in sterile conditions.

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 How you can assay the following eye ointment.[Chloramphenicol,
Oxyphenbutazone, and Sulphacetamide].
❖Chloramphenicol eye ointment:
Assay: Suspend a quantity eq. to 10 mg of CPL in petroleum spirit
(boiling range of 40-60°C) and extract with water.
Measure absorbance at 278 nm (UV).

❖Oxyphenbutazone eye ointment:

Assay: By extraction from the ointment
base. Measure UV at 254 nm.

❖Sulphacetamide eye ointment:

Assay: After extraction from the ointment base.
Amperometric titration using sodium nitrite to determine
the content of SCD.
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 Ear drops
➢ Ear drops are liquid preparation, suspension,
emulsion or solution of drug(s) in, diluted alcohol,
glycerin or polyethylene glycol intended for
instillation into the ear.
➢ Aqueous solutions are not suitable because the
secretions in ear are mainly fatty and aqueous
solutions do not easily mix with them.
➢ The ingredients of ear drops are different
depending on the purpose.

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 Chemical Assay of ear drops
How you can make a Chemical Assay for Sodium
Bicarbonate ear drop?
➢ Sodium Bicarbonate ear drop.
Analyte: Sodium Bicarbonate .
Solvent: Water.
Titrant: 0.1 M standard hydrochloric acid.
Indicator: Methyl orange solution. End point: Red-
orange color is obtained.
Titration reaction: acid –base
1 ml 0.1 M HCl = 8.401 mg of NaHCO3
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 How you can make a Chemical Assay for Boric Acid
Ear Drop?

➢ Boric Acid Ear Drops

Analyte: Boric Acid.
Solvent: Water containing mannitol.
Titrant: 1M Standard sodium hydroxide,
Indicator: ph.ph solution.
End point: A pink color is obtained.
Titration reaction: acid –base
1ml 1 M NaOH = 61.8 mg H3 BO3
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 Suppositories
They are solid preparations, usually administered in a single dose
for local or systematic action.

The medication is incorporated in a suitable base that liquefies at

the body temperature and releases the medicament.

Quality Control in Suppositories

1. Appearance: 2. Uniformity of weight:

3. Disintegration test: 4. Chemical Assay

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 How you can make a Chemical Assay for
Aminophylline suppositories (USP-29)?
Aminophylline is a compound that consists of theophylline and
ethylenediamine in a 2:1 ratio.
❖ Aminophylline suppositories (USP-29):
Assay for ethylinediamine,
30 min 500mg 150 ml
Sample preparation: Worm and reflex the sample with equal
volume of alcohol and ether.

Analyte: Ethylinediamine

Titrant: 0.1N HCl

End point: detected by modified calomel electrode.

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 Tablets

Tablets may be defined as solid

pharmaceutical dosage forms containing
drug substances with or without suitable
diluents and prepared either by
compression or moulding methods.
This dosage form is intended to be
administered through oral route

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 QC of tablets
In addition to the assay for active ingredient content the
following test is apply:
Uniformity of weight;
Disintegration, and where appropriate,
Uniformity of content and
Dissolution.

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 How you can assay Co-trimoxazole tablets?

❖ Co-trimoxazole tablets: (sulfamethoxazole + trimethoprim )

Preparation the sample: separate the two components by
dissolve tablet in NaOH solution then extract trimethoprim
with chloroform.
Assay of Sulfamethoxazol: (titrmetric)
Analyte: Sulfamethoxazol
Titrant: 0.1 N sodium nitrite
End point detection: By amperometric.
Assay of Trimethoprim: (spectrophotometric)
Trimethoprim is determined by Uv. at λmax 271 nm.
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 Ointments
➢ A highly viscous or semisolid preparation usually consist of single
phase base in which solid or liquid medicinal substances dispersed
in it. Ointments intended for external application.
➢ Ointments are divided into:
✓ Hydrophobic ointments,
✓ Hydrophilic ointments.
➢ Main test: uniformity of content
Examples: to relieve itching and irritation
Beclomethazone oint., Calamine oint., Salicylic acid ointment.

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 What is the titrimetric technique and the condition assay for Salicylic
acid ointment (BP2013)

❖ Salicylic acid ointment (BP2013)

Assay is done by titrimetric technique and the condition for
the titration is as a following
Analyte: Salicylic acid
Solvent: Ethanol (96%), and ether .
Titrant: 0.1M sodium hydroxide
Indicator: Phenol red solution.
Each mL of 0.1 M sodium hydroxide is equivalent to 13.81 mg of
C7H6O3.
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 Injectable preparations
➢ Injectable preparations are sterile products containing one
or more components, to be intended for administration by
injection, infusion or implantation into the body.
➢ Sterile drug product, which is presented in the form of:
solution, suspension, emulsion, or reconstituted lyophilized
powder.
➢ These preparations can be divided into three forms:
✓ Injections,
✓ Intravenous infusions, and
✓ Powder for injections.

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 Quality Control of injectable preparations
In addition to the assay for active ingredient content the following
test is apply:

1. Sterility test.
2. Test for pyrogen.
3. Test for bacterial endotoxin.
4. Leakage test
5. Particulate matter testing.
6. Chemical test: including pH, viscosity,conductivity .
7. Clarity of solution.
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 What are the assay technique can applied for Alprenolol
injection and Cyanocobalamin injection?

❖Alprenolol injection: (UV- Spectrophotometric)

Assay: Take volume eq. to 5 mg of Alprenolol HCl add 5 ml of 0.1
N HCl and sufficient water to produce 50 ml, and measure at 271
nm. (UV).

❖Cyanocobalamin Injection: vitamin B12

Dissolve 25 mg sample in 1000 ml water and measure the

absorbance at UV 361 nm.
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 What are the tests technique can applied for Dextrose
intravenous infusion according to BP?
❖Dextrose intravenous infusion:
Identification Tests: for Dextrose, and presence of Sodium and Chlorides.

Content Determination: Both Dextrose and Sodium Chloride content must be

between 95.0% and 105.0%.
(3.5 and 6.5),
Physical test: As optical rotation, pH Value, Clarity and Color.

Sterility, Endotoxin and Pyrogen Tests:

Particulate Matter:

Absorbance for Related Substances: should not exceed 0.25 of 5-

Hydroxymethylfurfural.

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 Elixirs

Elixirs are clear, liquid, flavored hydroalcoholic

preparation intended for oral use. They contain one or
more medicaments, pleasantly flavored, usually attractive
color containing high proportion of alcohol or sucrose or
suitable polyhydric alcohols together with suitable
additives (including antimicrobial agents)

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 What are the assay techniques can applied for Pediatric
paracetamol elixir
1.Preparation of Sample: Dissolve the pediatric paracetamol elixir in a suitable
volume of water. The concentration should be within the range that allows accurate
measurement at 257 nm.

2.Titration with NaOH: Add NaOH to the solution to achieve the desired pH level.
NaOH added to stop the ionization of paracetamol since can affect its absorbance
properties.

3.Dilution: After titration, dilute the solution further with water if necessary to
ensure that the absorbance falls within the linear range of the spectrophotometer.

4.Filtration: Filter the solution to remove any particulate matter that could interfere
with the absorbance measurement.

5.Measurement: Measure the absorbance of the filtered solution at 257 nm using a

UV-Vis spectrophotometer. Ensure that the instrument is calibrated correctly
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 Syrups
Syrups are concentrated aqueous solutions of Sucrose, other

sugars or sweetening agents, to which small quantities of

suitable polyhydric alcohol may be added for retard

crystallization or to increase solubility. As examples of syrup

are BENYLIN® Children's Dry Cough and Sore Throat Syrup

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As outlined in the British Pharmacopoeia. For syrup the following test
may include:-
1. Identification Tests: To confirm the presence of active ingredients.
2. Assay: Quantitative analysis of the active ingredients to ensure they
meet specified limits.
3. Purity Tests: To check for the presence of impurities or contaminants.
4. Microbial Testing: To ensure the product is free from harmful
microorganisms.
5. Stability Testing: To assess the product's shelf life and storage
conditions.
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