Assignment No.

1 Pharmaceutical Quality

The University of Lahore

Department of Pharmacy

Pharmaceutical Quality

Assignment 1

Topic Monographs of lipid and blood sugar modifying

agents
and Cough suppressant

Section 7B (Pharm -D)

Submitted by
Raheel Asghar

Reg. No.:

70116221

Submitted to

Dr. Maryam Shabbir

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 Assignment No. 1 Pharmaceutical Quality

Submission date 18-11-2023

MONOGRAPH OF LIPID AND SUGAR MODIFYING AGENTS

1-Rosuvastatin

Reference: USP 43, page no. 3950

Classification:
It’s belongs to statins family.

Statins:
Statins also classify as lipophilic or hydrophilic.

Lipophilic statins Hydrophilic statins

Simvastatin Pravastatin
lovastatin Fluvastatin
Atorvastatin Rosuvastatin

Generic name:

➢ Rosuvastatin

Brand Name Synonyms:

➢ Crestor
➢ Ezallor
➢ Ezallor Sprinkle

Structure of Rosuvastatin:

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 Assignment No. 1 Pharmaceutical Quality

Identification:
• A. The UV absorption spectra of the rosuvastatin peak of the Sample solution exhibit maxima and
minima at the same wavelengths as those of the corresponding peak of the Standard solution, as
obtained in the Assay.
• B. The retention time of the major peak of the Sample solution corresponds to that of the Standard
solution, as obtained in the Assay.

Assay
• Procedure
Protect all solutions containing rosuvastatin from light.
Solution A: 1% trifluoroacetic acid in water Mobile phase: Acetonitrile, Solution A, and water
(37:1:62)
Diluent: Acetonitrile and water (25:75)
Standard stock solution: 1 mg/mL of USP Rosuvastatin Calcium RS prepared as follows. To a
suitable amount of USP Rosuvastatin Calcium RS in a suitable volumetric flask add water equal to
about 50% of the flask volume.Vigorously mix or sonicate the flask to dissolve the material.Add
acetonitrile equal to about 25% of the total volume and then dilute with water to volume.
Standard solution: 25 µg/mL of USP Rosuvastatin Calcium RS in Diluent from the Standard stock
solution Sample solution: Nominally 25 µg/mL of rosuvastatin prepared as follows. Transfer a
suitable number of Tablets,NLT 5 Tablets for 80-mg Tablet strength and NLT 10Tablets for all other
Tablet strengths, into a suitable extraction flask. Add water and vigorously mix to disintegrate the
Tablets. Add acetonitrile and mix vigorously. Add more water to obtain a 25:75 composition of
acetonitrile and water. Pass the solution through a suitable filter. Dilute the filtrate with Diluent, if
necessary, to the desired concentration.
Chromatographic system
(See Chromatography á621ñ, System Suitability.)Mode: LC
Detector: UV 242 nm. For Identification A, use a diodear ray detector in the range of 200–440
nm.Column: 3.2-mm × 25-cm; 5-µm packing L1. [NOTE—A suitable guard column may be used.]

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 Assignment No. 1 Pharmaceutical Quality

Column temperature: 40°Flow rate: 0.75 mL/min Injection volume: 10 µL

Run time: NLT 1.3 times the retention time of rosuvastatin
System suitability
Sample: Standard solution
Suitability requirements
Tailing factor: NMT 1.8
Relative standard deviation: NMT 2.0%

Analysis
Samples: Standard solution and Sample solution Calculate the percentage of the labeled amount of
rosuvastatin (C22H28FN3O6S) in the portion of Tablets taken:
Result = (rU/rS) × (CS/CU) × [M × (Mr1/Mr2)] × 100rU = peak response of rosuvastatin from the Sample
solution S = peak response of rosuvastatin from the Standard solution
CS= concentration of USP Rosuvastatin Calcium RS in the Standard solution (µg/mL)
CU= nominal concentration of rosuvastatin in the Sample solution (µg/mL)
M = number of moles of rosuvastatin per mole of rosuvastatin calcium, 2
Mr1 = molecular weight of rosuvastatin, 481.54Mr2 = molecular weight of rosuvastatin calcium,
1001.14
Acceptance criteria: 90%–110

2.REPAGLINIDE
Reference USP 43: PAGE :3856

Classification:
It’s belongs to antidiabetic class
➢ Six classes of oral antidiabetic drugs.
➢ Biguanides
➢ Sulfonylureas
➢ Meglitinides
➢ Thiazolidinediones
➢ Dipeptidyl peptidase IV inhibitors
➢ α-glucosidase inhibitors

Generic name:
➢ Repaglinide

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 Assignment No. 1 Pharmaceutical Quality

Structure of Repaglinide:

Identification:
• A. thin-layer chromatographic identification test
á201ñ
Sample solution: To a quantity of powdered Tablets equivalent to 10 mg of repaglinide, add 10 mL of a
mix tureof methanol and methylene chloride (1:1), shake for15 min, and centrifuge.
Developing solvent system: Toluene, methylene chloride and methanol (2:2:1)
Acceptance criteria: Meet the requirements
• B. The retention time of the major peak of the Sample solution corresponds to that of the Standard
solution, as obtained in the Assay.
• C. The UV spectrum of the major peak of the Sample solution corresponds to that of the Standard
solution, as obtained in the Assay.

ASSAY
• Procedure

Buffer pH 4.0: 2 g/L of monobasic ammonium phosphate solution. Adjust with phosphoric acid to a pH
of 4.0Buffer pH 2.5: 2 g/L of monobasic ammonium phosphate solution. Adjust with phosphoric acid
to a pH of 2.5Mobile phase: Methanol and Buffer pH 2.5 (7:3) Diluent: Methanol and Buffer pH 4.0
(7:3)
Standard solution 1: 800 µg/mL of USP Repaglinide RS in methanol
Standard solution 2: 80 µg/mL of USP Repaglinide RS prepared by diluting 5.0 mL of Standard solution
1 with Diluent to 50.0 mL
System suitability stock solution: 80 µg/mL of USPR epaglinide Related Compound A RS in methanol
System suitability solution: 80 µg/mL of USP
Repaglinide RS and 1.6 µg/mL of USP Repaglinide Related Compound A RS prepared as follows.
Transfer 1.0 mL of System suitability stock solution to a 50-mL volumetric flask add 5.0 mL of Standard
solution 1, and dilute with Diluentto volume.

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 Assignment No. 1 Pharmaceutical Quality

Sample solution: Transfer 8 whole Tablets to a suitable volumetric flask, and dissolve in and dilute with
Diluent to volume to obtain a solution containing 80 µg/mL Stir for20 min, and filter or centrifuge a
portion of the solution.
Chromatographic system
(See Chromatography á621ñ, System Suitability.) Mode: LC
Detector: UV 245 nm or diode array. [NOTE—Use diode array detector to perform Identification test
C.] Column: 4.0-mm × 6-cm; 5-µm packing L1
Column temperature: 40°Flow rate: 1 mL/min Injection volume: 20 µL
System suitability
Samples: Standard solution 2 and System suitability solution
[NOTE—The typical relative retention times for repaglinide related compound A and repaglinide are
about 0.4 and 1.0, respectively.]
Suitability requirements
Resolution: NLT 7.0 between repaglinide related compound A and repaglinide, System suitability
solution Tailing factor: 0.8–2.0 for the repaglinide peak, System suitability solution
Relative standard deviation: NMT 2.0% for replicate injections, Standard solution 2

Analysis
Samples: Standard solution 2 and Sample solution Calculate the percentage of the labeled amount of
repaglinide (C27H36N2O4) in the portion of Tablets taken:
Result = (rU/rS) × (CS/CU) × 100rU = peak response from the Sample solution S = peak response from
Standard solution 2
CS= concentration of USP Repaglinide RS in Standard solution 2 (µg/mL)
CU= nominal concentration of repaglinide in the Sample solution (µg/mL)
Acceptance criteria: 95.0%–105.0%
Composition and temperature of the mobile phase
Ionic strength and pH of the aqueous component, of the mobile phase
Flow rate, column dimensions, column temperature, and pressure
Stationary phase characteristics including, type of chromatographic support (particle-based or
monolithic), particle or pore size, and porosity, and specific surface area
Reverse phase and other surface modification of the stationary phases, the extent of chemical
modification (as expressed by end-capping, carbon loading, etc.)

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 Assignment No. 1 Pharmaceutical Quality

TECHNIQUE USED IN ASSAYS AND IDENTIFICATION FROM

USP:
System suitability:
Reference: USP 43, Volume 4, Page number 6865(621)
[NOTE—This section only covers liquid chromatography and gas chromatography]
The various components of the equipment employed must be qualified and be capable of achieving the
performance required to conduct the test or assay. The system suitability tests represent an integral part
of the analytical procedure and are used to ensure adequate performance of the chromatographic system.
Column plate number, retention factor (mass distribution ratio), system repeatability, signal-to-noise,
symmetry factor, and resolution/peak-to-valley ratio are the parameters that may be employed in
assessing the performance of the chromatographic system. When it is indicated in the individual
monograph, in case of complex chromatographic profiles (e.g., for biotechnological and biological
products) visual comparison of the profiles can be used as a system suitability test. Factors that may
affect the chromatographic behavior include:
Retention times and relative retentions may be provided in monographs for information purposes only,
unless otherwise stated in the monograph. There are no acceptance criteria applied to relative
retentions.
Compliance with the system suitability criteria is required throughout the chromatographic procedure.
No sample analysis is acceptable unless the suitability of the system has been demonstrated.
The following requirements are to be fulfilled, in addition to any other system suitability criteria stated
in the monograph. When specific requirements are stated in the monograph, they supersede the
requirements mentioned in this chapter.

Liquid Chromatography:
Reference: USP 43, Volume 4, Page number 6856(621) The term "liquid
chromatography" (LC), as used in the compendia, is synonymous with high-pressure liquid
chromatography and high-performance liquid chromatography. LC is a separation technique based on a
solid stationary phase and a liquid mobile phase.
Stationary phase:
Separations are achieved by partition, adsorption, or ion-exchange processes,
depending on the type of stationary phase used. The most commonly used stationary phases are modified
silica or polymeric beads. The beads are modified by the addition of long-chain hydrocarbons. The
specific type of packing needed to complete an analysis is indicated by the “L” designation in the
individual monograph (see also the section Chromatographic Columns). The size of the beads is often
described in the monograph as well. Changes in the packing type and size are covered in the System
Suitability section of this chapter.
Chromatographic column:
The term "column" includes stainless steel, lined stainless steel, and
polymeric columns, packed with a stationary phase. The length and inner diameter of the column affects
the separation, and therefore typical column dimensions are included in the individual monograph.
Changes to column dimensions are discussed in the System Suitability section of this chapter.
Compendial monographs do not include the name of appropriate columns; this omission avoids the
appearance of endorsement of a vendor’s product and natural changes in the marketplace. See the section
Chromatographic Columns for more information. In LC procedures, a guard column may be used with

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 Assignment No. 1 Pharmaceutical Quality

the following requirements, unless otherwise is indicated in the individual monograph: (a) the length of
the guard column must be NMT 15% of the length of the analytical column, (b) the inner diameter must
be the same or smaller than that of the analytical column, and (c) the packing material should be the
same as the analytical column (e.g., silica) and contain the same bonded phase (e.g., C18). In any case,
all system suitability requirements specified in the official procedure must be met with the guard column
installed.
Mobile phase:
The mobile phase is a solvent or a mixture of solvents, as defined in the individual
monograph.
Apparatus:
A liquid chromatograph consists of a reservoir containing the mobile phase, a pump to
force the mobile phase through the system at high pressure, an injector to introduce the sample into the
mobile phase, a chromatographic column, a detector, and a data collection device.
Gradient elution:
The technique of continuously changing the solvent composition during the
chromatographic run is called gradient elution or solvent programming. The gradient elution.
proportional composition of the mobile phase at the stated time.
Procedure:
Equilibrate the column and detector with mobile phase at the specified flow rate until a constant signal
is received.
Inject a sample through the injector, or use an autosampler.
Begin the gradient program.
Record the chromatogram.
Analyze as directed in the monograph.

SPECTROSCOPIC IDENTIFICATION TESTS:

Reference: USP 43, Volume 4, Page number 6593(197)
Introduction and scope:
This chapter provides several tests and procedures that are used to confirm the chemical identity of a
specific material in its respective monograph. When one of the tests listed in this chapter is referenced
in a monograph, it must be used to confirm the identity of the material. Alternative identification
procedures may be used, provided they are demonstrated to be equivalent to or better than the specified
procedure and if they meet the requirements specified in the Equivalent/Alternative Tests section.
Spectroscopic tests contribute meaningfully toward the identification of many official articles. The test
procedures that follow are applicable to substances that absorb, transmit, reflect, or scatter
electromagnetic radiation in the near-infrared (NIR), infrared (IR), visible or near-infrared (Raman),
ultraviolet (UV), visible (Vis), or X-ray ranges (see Mid-Infrared Spectroscopy (854), Near-Infrared
Spectroscopy (856), Ultraviolet-Visible Spectroscopy (857), (ERR 1-Sep-2021) Raman Spectroscopy
(858), and X-Ray Powder Diffraction 941). The NIR, IR, and Raman spectra, or X-ray diffraction pattern
of a substance, compared with the spectrum or diffraction pattern obtained with the corresponding USP
Reference Standard, provides perhaps the most conclusive evidence of the identity of the substance that

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 Assignment No. 1 Pharmaceutical Quality

can be realized from any single test. The UV or Vis absorption spectrum of a substance, on the other
hand, does not exhibit a high degree of specificity in most cases. To provide unambiguous confirmation
of the identity of a substance, it may be necessary to execute two (or more) identity tests, as specified in
a large proportion of compendial monographs.

Ultraviolet-visible spectroscopy:
Reference: USP 43, Volume 4, Page number 6595(197U)
The reference (197U) in a monograph signifies that a sample solution and a Standard solution are
examined spectroscopically, in 1-cm cells, over the spectral range from 200 to 400 nm, unless otherwise
specified in the individual monograph (see 857). Dissolve a portion of the substance under examination
in the designated medium to obtain a sample solution having the concentration specified in the
monograph. Record and compare the spectra obtained for the sample solution and the Standard solution.
Review or calculate the absorptivities and/or absorbance ratios and compare the results, and where
appropriate, compare to criteria specified in an individual monograph.
The comparison must establish that the UV spectrum of the preparation of the sample exhibits absorption
maxima and minima only at the same wavelengths as those of the appropriately prepared corresponding
USP Reference Standard, and that the absorptivities and/or absorbance ratios are within the specified
limits. If there are differences in the spectra, and the sample spectrum was compared with a previously
obtained and electronically stored spectrum of the USP Reference Standard, the comparison must be
concomitantly repeated with a freshly prepared USP Reference Standard.
Unless otherwise specified in the monograph, absorbances indicated for the calculations of the
absorptivities and/or absorbance ratios are those measured at the maximum absorbance wavelength
(within ±2 nm) specified in the individual monograph. Where the absorbance is to be measured at about
the specified wavelength other than that of maximum absorbance, the abbreviations for minimum (min)
and shoulder (sh) are used, respectively, in an absorption spectrum.
The reference (197U-LC) in a monograph signifies that, when a diode array detector is used in tandem
with an LC procedure test in the monograph, the spectra of the major chromatographic peak(s) of the
sample solution and Standard solution are examined spectroscopically.
The requirements are met if the UV spectra of the sample solution and of the Standard solution exhibit
maxima and minima at the same wavelengths, and, if applicable, the absorptivities and/or absorbance
ratios are within specified limits.

MONOGRAPH OF COUGH SUPPRESSANTS AND OTIC

PREPARATION
Reference BP 2023:PAGE : 134,135

1 Ambroxol hydrochloride:

Classification:
➢ Ambroxol hydrochloride is an aromatic amine
➢ Classification of aromatic amines

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 Assignment No. 1 Pharmaceutical Quality

Aromatic amines are further classified into aryl amine and aryl alkyl amines
Synonyms
• 4-(((2-Amino-3,5-dibromophenyl)methyl)amino)cyclohexanol.
• Abrohexal.
• AM, Bisolvon.
• Ambril.
• Ambro Puren.
• Ambro-Puren.
• Ambrobeta.
• Ambrofur.

Generic Name:
➢ Ambroxol hydrochloride

Structure of Ambroxol hydrochloride:

2 Ammonium Bicarbonate

Reference BP 2023:PAGE : 161,162

Generic name:
Ammonium Bicarbonate

Structure of Ammonium Bicarbonate:

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 Assignment No. 1 Pharmaceutical Quality

Identification:
A. It gives the reaction of carbonates and bicarbonates.
B. Dissolve 50 mg in 2 mL of water R. The solution gives the reaction of ammonium salts
Tests.

Solution S
Dissolve 14.0 g in 100 mL of distilled water R. Boil to remove the ammonia, allow to cool and dilute to
100.0 mL with distilled water R

Chlorides:
Maximum 70 ppm.
Dilute 5 mL of solution S to 15 mL with water R.

Sulfates:
Maximum 70 ppm, determined on solution S.
Iron
Maximum 40 ppm.
Dilute 1.8 mL of solution S to 10 mL with water R.

ASSAY:
Dissolve cautiously 0.500 g in 50 mL of carbon dioxide-free water R. Titrate with 1 M hydrochloric
acid, determining the end-point potentiometrically (2.2.20). Read the volume added at the 2nd point of
inflection, or at the point of inflection if only 1 point is detected.
1 mL of 1 M hydrochloric acid is equivalent to 79.1 mg of NH4HCO3.
Structure.

TECHNIQUE USED IN ASSAYS AND IDENTIFICATION FROM

BP:
Thin-layer Chromatography:

Reference: BP 2023, V-A232 (2.2.27)

Thin-layer chromatography is a separation technique in which a stationary phase consisting of an
appropriate material is spread in a uniform thin layer on a support (plate) of glass. metal or plastic.

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 Assignment No. 1 Pharmaceutical Quality

Solutions of analytes are deposited on the plate prior to development. The separation is based on
adsorption, partition, ion-exchange or on combinations of these mechanisms and is carried out by
migration (development) of solutes (solutions of analytes) in a solvent of a suitable mixture of solvents
(mobile phase) through the thin-layer (stationary phase).

APPARATUS:
1. Plates: The chromatography is carried out using pre-coated plates as described under Reagents
(417). The particle size of the silica gel is indicated after the name of the reagent in the tests where
it is used.
2. Pre-treatment of the plates: It may be necessary to wash the plates prior to separation. This can be
done by migration if an appropriate solvent. The plates may also be impregnated by procedures such
as development, immersion or spraying. At the time of use, the plates may be activated, if necessary,
by heating in an oven at 120 °C for 20 min.
3. Chromatographic tank: With a flat bottom or twin trough, of inert, transparent material, of a size
suitable for the plates used and provided with a tightly fitting lid. For horizontal development the
tank is provided with a trough for the mobile phase and it additionally contains a device for directing
the mobile phase to the stationary phase.
4. Micropipette, micro syringes, Or other application devices suitable for the proper application of
the solutions
5. Florescence detection device: To measure direct fluorescence or the inhibition of fluorescence.
6. Visualization devices and reagents: Suitable devices are used for derivatization to transfer to the
plate reagents by spraying, immersion or exposure to vapor and, where applicable, to facilitate
heating for visualization of separated components.
7. Documentation: a device may be used to provide documentation of the visualized chromatogram,
for example a photograph or a computer file.

Method:
1 Sample application
2 Vertical development
3 Horizontal development
4 Visual evaluation
5 Identification
6 Verification of separating power for identification
7 Related substances test
8 Verification of the separating power
9 Verification of the detecting power
10 Quantitative measurement

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 Assignment No. 1 Pharmaceutical Quality

B-Infrared Spectrophotometry or Absorption Spectrophotometry:

Reference: BP 2023, V-A184 (2.2.24)
PRINCIPLE:
➢ Infrared absorption spectrophotometry (also known as infrared (IR) spectroscopy) is based on
the interaction of infrared radiation with matter. As a result of interaction between a molecule and IR
radiation, absorption of frequencies specific to that molecule can occur, and some intermolecular and
intramolecular vibrations can be excited to higher vibrational levels. This results in an infrared
absorption spectrum with characteristic bands that correspond to the functional groups of the molecule.
➢ The infrared wavelength region can be further divided into 3 subregions, namely near-infrared,
mid-infrared and far- infrared. These subregions have wavelength ranges that are generally accepted by
convention to be 0.8-2.5 um, 2.5-25 μm and 25-1000 um respectively. However, in IR spectroscopy,
wavenumber is more commonly used than wavelength, and can be calculated using the following
equation:

V=1/ λ.104
➢ where v is the wavenumber in reciprocal centimeters (cm¹) and λ is the wavelength in
micrometers. Thus 12 500-4000 cm is near-infrared, 4000-400 cm is mid- infrared and 400-10 cm is far-
infrared.
➢ This chapter concerns only spectroscopy in the mid-infrared region, i.e., 4000-400 cm (2.5-25
um), which hereafter is referred to as infrared for simplicity. This region is where the fundamental
Assignment No. 1 Pharmaceutical Quality
19

➢ below 1500 cm is known as the 'fingerprint region', a very complex and informative part of the
spectrum which characterizes the molecule being investigated.
➢ The mid-infrared region is flanked by the near-infrared region, where overtones and combinations
of fundamental vibrations, mainly C-H, N-H and O-H functional groups, are detected (2.2.40) and
the far-infrared region, where absorption bands associated with crystal lattice modes, hydrogen
bonds, angle deformation vibrations of heavy atoms and molecular rotations are observed.

APPLICATIONS:
➢ As the absorption bands in IR spectra are characteristic of the constituent functional groups of a
compound, IR spectroscopy is widely used to identify substances and provide information on their
structure. It can also be used for quantitative applications, which requires establishing a mathematical
relationship between the intensity of the radiation absorbed by the sample and the concentration of the
investigated component in the sample.
➢ IR spectroscopy is widely used in the pharmaceutical field for chemical and physical analysis in
the laboratory, and has a wide variety of applications during the manufacturing process as outlined
below. IR spectroscopy thereby enables the application of Process Analytical Technology (PAT) as part
of an advanced control strategy.

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 Assignment No. 1 Pharmaceutical Quality

Chemical analysis:
➢ identification of active substances, excipients, dosage forms, manufacturing intermediates,
chemicals and packaging materials;
➢ quality assessment of active substances, excipients, dosage forms, manufacturing intermediates
and packaging materials, including batch-to-batch spectral comparison and supplier change assessment;
➢ quantification of active substances in a sample matrix, determination of water and solvent
content;
➢ quantification of impurities, e.g., in gases, inorganic materials;
➢ reaction monitoring, e.g., chemical synthesis.

Physical analysis:
➢ determination of solid-state properties such as polymorphism.

Limitations:
Notable limitations to the use of IR spectroscopy include the following:
➢ it may be necessary to use additional techniques to unambiguously identify a substance;

➢ pure enantiomers of a substance cannot be discriminated;

➢ it may not be a suitable method for trace analysis;
➢ sample preparation conditions (e.g., pressure, solvent) may
➢ change the crystalline form of a substance that exhibits polymorphism;
➢ for heterogeneous samples, the limited sampling volume may be problematic.

MEASUREMENT MODES:
1. Transmission mode
2. Attenuated total reflection mode

EQUIPMENT:
The most commonly used IR spectrometers are Fourier- transform (FT-IR) spectrometers which typically consist
of:
➢ A suitable polychromatic light source, e.g., a conducting ceramic rod;
➢ An interferometer;
➢ A sample presentation accessory, e.g., a sample holder;
➢ A detector;
➢ Appropriate software for controlling the spectrometer, and for spectral evaluation and data processing.

Other spectrometers based on alternative principles may also be used if the requirements described under Control
of equipment performance are fulfilled.IR spectrometers can also be used in association with a microscope for

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Assignment No. 1 Pharmaceutical Quality

the study of a small part of the sample or for chemical imaging.IR spectroscopy can be coupled to other analytical
techniques such as thermal analysis or chromatography.

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 Assignment No. 1 Pharmaceutical Quality

References:
Reference: USP 43, page no. 3950

Reference USP 43: PAGE :3856

Reference: USP 43, Volume 4, Page number 6865(621)

Reference: USP 43, Volume 4, Page number 6593(197)

Reference: USP 43, Volume 4, Page number 6595(197U)

Reference BP 2023:PAGE : 134,135

Reference BP 2023:PAGE : 161,162

Reference: BP 2023, V-A184 (2.2.24)

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