DELHI PHARMACEUTICAL SCIENCES & RESEARCH UNIVERSITY

(Govt of NCT Delhi)

Sector 3, M.B. Road, Pushp Vihar, New Delhi-110017
2023-24

PRACTICE SCHOOL REPORT

Submitted By: XXXX XX

B. Pharm 7th Semester
XX/BPH/DIPSAR/2020
 Table of contents

SUBJECT S. NO. TOPIC AIM

PHARMACEUTICS 1 Preparation and To prepare and evaluate
evaluation of calcium microbeads from sodium alginate
alginate microbeads and calcium chloride

PHARMACOLOGY 3 Evaluation of To evaluate the pharmacokinetic

pharmacokinetic parameters of the given drug
parameters
(bioavailability,
volume of
distribution,
clearance) using
literature review
4 Evaluation of To determine IC50 and LC50 for
toxicology parameters the given drug
(IC50, LC50) using
literature review
PHARMACEUTICAL 5 Synthesis of para-dyes To synthesise para red dye from
CHEMISTRY acetanilide

6 Recrystallisation of To purify the given potash alum

potash alum sample by recrystallisation

PHARMACOGNOSY 7 Soxhlet Extraction of To extract sennosides from Senna

& Sennosides using Soxhlet Extraction
PHYTOCHEMISTRY

8 Extraction of alkaloids To extract quinine and quinidine

from Cinchona from Cinchona bark
Subject: PHARMACEUTICS

EXPERIMENT 1
Aim:
To prepare and evaluate calcium alginate microbeads from sodium alginate and calcium chloride.

Requirements:
Sodium Alginate 90 cps (1% w/v solution in water at 25 °C), Calcium chloride dihydrate, magnetic
stirrer, magnetic stirrer bead, 22-gauge needle, oven

Principle:

Objectives:

Procedure:
1. Preparation of Solutions
1. Sodium Alginates solution (2.5% w/v) was prepared by mixing 1.25g of solute in
50ml water by heating and stirring.
2. Calcium chloride solution (2% w/v) was prepared by mixing 2g of solute in 100 ml
water by stirring.
2. Transfer of solutions
1. Using a 22-gauge needle, sodium alginate was introduced drop by drop into the
calcium chloride solution at 400 rpm.
2. The solution was allowed to stir for 1 hr.
3. Beads Removal
1. Beads were filtered out.
4. Drying
1. The beads were dried in the oven overnight.
5. Evaluation
1. Flow properties were evaluated for the given beads.
Observations:

Bulk Volume Bulk Weight Bulk Density Tapped Volume Tapped Weight Tapped Density

Hausner’s Ratio =

Carr’s Index =

Learning Outcomes:

References:
Mandal, Sanchita & Kumar, S. & Krishnamoorthy, Balakrishnam & Basu, Sanat. (2010).
Development and evaluation of calcium alginate beads prepared by sequential and simultaneous
methods. Brazilian Journal of Pharmaceutical Sciences. 46. 785-793. 10.1590/S1984-
82502010000400021.
EXPERIMENT 2
Aim:

Requirements:

Principle:

Objectives:

Procedure:

Observations:

Learning Outcomes:

References:
Subject: PHARMACOLOGY

EXPERIMENT 3
Aim:
To determine the pharmacokinetic properties of the given drug using literature review.

Requirements:
Internet connection.

Principle:
1. Bioavailability:
Bioavailability refers to the extent a substance or drug becomes completely available to its intended
biological destination(s). More accurately, bioavailability is a measure of the rate and fraction of the
initial dose of a drug that successfully reaches either; the site of action or the bodily fluid domain
from which the drug's intended targets have unimpeded access.

2. Volume of Distribution:
The volume of distribution (Vd) is a pharmacokinetic parameter representing an individual drug’s
propensity to either remain in the plasma or redistribute to other tissue compartments. V d is a
proportionality constant that relates the total amount of drug in the body to the plasma concentration
of the drug at a given time. The following equation can represent Vd:
Volume of Distribution (L) = Amount of drug in the body (mg) / Plasma concentration of drug
(mg/L)
3. Clearance Rate
Drug clearance is defined as the volume of plasma cleared of a drug over a specified time. Thus, the
unit of measurement for drug clearance is volume/time. Another equation can calculate clearance.
Clearance is equal to the rate at which a drug is removed from plasma(mg/min) divided by the
concentration of that drug in the plasma (mg/mL). The total ability of the body to clear a drug from
the plasma is renal clearance plus hepatic clearance plus clearance from all other tissues.

Objectives:

Procedure:
Observations:

Learning Outcomes:

References:
1. Price G, Patel DA. Drug Bioavailability. [Updated 2023 Jul 30]. In: StatPearls [Internet].
Treasure Island (FL): StatPearls Publishing; 2023 Jan-. Available from:
https://www.ncbi.nlm.nih.gov/books/NBK557852/
2. Mansoor A, Mahabadi N. Volume of Distribution. [Updated 2023 Jul 24]. In: StatPearls
[Internet]. Treasure Island (FL): StatPearls Publishing; 2023 Jan-. Available from:
https://www.ncbi.nlm.nih.gov/books/NBK545280/
3. Horde GW, Gupta V. Drug Clearance. [Updated 2023 Jun 20]. In: StatPearls [Internet].
Treasure Island (FL): StatPearls Publishing; 2023 Jan-. Available from:
https://www.ncbi.nlm.nih.gov/books/NBK557758/#
EXPERIMENT 4
Aim:

Requirements:

Principle:

Objectives:

Procedure:

Observations:

Learning Outcomes:

References:
Subject: PHARMACEUTICAL CHEMISTRY

EXPERIMENT 5
Aim:
To synthesise para red dye from acetanilide.

Requirements:
Chemicals and materials. 2-Naphthol, p-nitroaniline, sodium nitrite, sodium hydroxide,
trisodium phosphate, HCl, and ethanol.
Glassware and equipment. Three beakers (a 250 mL and two 100 mL), a pipette, a pipette
sucker, a magnetic bar, a hot plate, hardened filter paper, Buchner funnel, an Erlenmeyer
flask (100 mL), a glass rod, a graduated cylinder, a spatula, an ice bath and a water bath etc.

Principle:

Objectives:

Procedure:
1. In a 500 mL beaker, dissolve 1 g (0.025 mol) of sodium hydroxide and 9.5 g (0.025 mol) of
commercial tri-sodium phosphate (Na3PO4.12H2O) in 200 mL of water to create a buffer
solution.
2. Add 1.4 g (0.01 mol) of 2-naphthol into the beaker and stir until the 2-naphthol is completely
dissolved.
3. Chill the solution by placing it in an ice bath for 5 minutes, stirring intermittently.
4. In a 100 mL beaker, add 3 mL of distilled water.
5. Pipette out 3 mL of concentrated hydrochloric acid and add it to the beaker containing water.
6. Dissolve 1.4 g of p-nitro aniline in the acidic solution by gently warming on a hot plate until all
of it is dissolved.
7. Allow the solution to cool down to room temperature and then pour it into a 100 mL
Erlenmeyer flask containing about 10 g of chopped ice.
8. Swirl the mixture to obtain a fine suspension of p-nitro aniline hydrochloride crystals, while
keeping the mixture at 5-10 °C in the ice bath and agitating it vigorously.
9. Dissolve 0.8 g (0.011 moles) of sodium nitrite in 3-4 mL of water and pour it quickly into the
cold solution of p-nitro aniline hydrochloride.
10. Swirl the mixture until most of the hydrochloride has dissolved (2-3 minutes), then let it stand
for a few minutes to complete the diazotization process.
11. Pour the diazonium salt solution into the chilled alkaline solution of 2-naphthol all at once.
12. Stir the mixture vigorously for a few minutes to ensure complete reaction.
 13. Add 5 mL of concentrated hydrochloric acid and raise the temperature to about 30 °C on a hot
plate.
14. Stir the reaction mixture for 30-40 minutes using a magnetic stirrer.
15. Collect the bright red dye by vacuum filtration.
16. Wash the dye with distilled water until the filtrate is essentially free of chloride ion (neutral
pH).
17. Finally, wash the crystals of the azo dye with small portions of ethanol.
18. After drying, weigh the product.

Observations:
Theoretical Yield= 2.93 g
Actual Yield= 11.68g
Percentage Yield= 11.68/2.93∗100=398.6 %
Note: The yield crossed 100%, as the water vapour did not really evaporate completely. (Boiling
point of water is 100 °C).

Learning Outcomes:

References:
1. Zollinger, H., Color chemistry: syntheses, properties, and applications of organic dyes and
pigments. John Wiley & Sons: 2003.
2. Al-Rubaie, L.; Mhessn, R. J., Synthesis and characterization of azo dye para red and new
derivatives. Journal of Chemistry 2012, 9 (1), 465-470.
3. Stolz, A., Basic and applied aspects in the microbial degradation of azo dyes. Applied
microbiology and biotechnology 2001, 56 (1-2), 69-80.
4. Almashal, F.; Jabar, A. M.; Dhumad, A. M., Synthesis, characterization and DFT computational
studies of new heterocyclic azo compounds. European Journal of Chemistry 2018, 9 (2), 84-88.
5. Brown, D.; Laboureur, P., The aerobic biodegradability of primary aromatic amines. Chemosphere
1983, 12 (3), 405-414.
EXPERIMENT 6
Aim:
To purify the given potash alum sample by recrystallisation.

Requirements:

Principle:
The principle behind the crystallisation is that the amount of solute that can be dissolved by a solvent
increase with temperature.
In crystallisation, the impure substance is dissolved in a suitable solvent to reach its nearly saturated
solution at a temperature higher than the room temperature. At this high temperature, the solute has
very high solubility in that solvent, so a much smaller quantity of hot solvent is needed for dissolving
the solute than the solvent at room temperature. When the solution is cooled, the pure substance is
crystallised. The solution left behind is called mother liquor. All the impurities are left behind in the
mother liquor. The purification method depends on the differences in solubility between the
compound and the impurity.

Objectives:

Procedure:
1. Solvent and solute selection
Choosing an appropriate solvent is the important process of crystallisation, as crystallisation works
only when a proper solvent is used. It is important to choose a solvent that will not dissolve the
substance at room temperature. But as the temperature of the solvent increases, the solubility of the
solute also increases. At the same time, the impurities that are present must either be soluble in the
solvent at room temperature or must be insoluble in the solvent at a high temperature. If the solvent
is not hot when the dissolution is carried out, too much solvent will be used, leading to diminished
yield.
2. Dissolving the solute in the solvent
Add a small portion of the solvent to the beaker containing impure sample and boiling chips while
the sample is heating. Stir the contents gently. Add enough solvent to dissolve the solute to get a
saturated solution at the boiling point of the solvent. If too much solvent is used, the recovery of the
substance will be decreased.
3. Filtration of the hot solution
If the hot solution contains insoluble impurities, it can be removed by the process of filtration. For
this process, place a filter paper cone in a funnel and wet the filter paper with a spray of water to fix
it inside the funnel properly. Place the funnel on a funnel stand and put a China dish below the
funnel.
Note: The stem of the funnel should touch the wall of the China dish to avoid the solution splashing
out.
4. Crystallisation of the filtrate
To concentrate the filtrate, heat the China dish containing filtrate gently with constant stirring. This is
done to ensure uniform evaporation and to prevent formation of a solid crust. When the volume of
the solution is reduced to one half, dip one end of a glass rod in the concentrated solution and cool
the drop by blowing on it. The formation of a thin crust indicates that the crystallisation point has
been obtained.
5. Cooling the concentrated solution
Once it is determined that the solution is saturated with the compound, it is allowed to cool slowly at
room temperature.
In order to cool the concentrated solution, pour the solution into a crystallising dish and keep it
undisturbed. As the solution cools, crystals separate. Once the sample is cooled to room temperature,
place it in an ice bath or in cold water to complete the crystallisation.
If the crystallisation does not start immediately, add a seed crystal or scratch inside the vessel
containing the concentrated solution with a glass rod.
6. Separation and drying of crystals
The crystals formed are separated by either decanting the mother liquor or by the process of
filtration. Wash the crystals with cold water or alcohol. The crystals can be dried by pressing them
gently between sheets of filter paper. They can also be dried by spreading them on a porous plate or
by placing the crystals in a vacuum desiccator. The crystals have a definite geometry, and therefore a
definite shape.

Observations:

Learning Outcomes:

References:
https://www.olabs.edu.in/?sub=73&brch=7&sim=110&cnt=1
Subject: PHARMACOGNOSY & PHYTOCHEMISTRY

EXPERIMENT 7
Aim:
To extract sennosides from Senna using Soxhlet Extraction.

Requirements:
Apparatus: Porcelain dish, Conical flask, Beaker, Measuring cylinder, Funnel, Stirrer, Glass plate.
Reagents: Methanol / Ethanol, Silica gel, Acetic acid, Senna leaves, Benzene, HNO 3, Calcium
chloride, Potassium hydroxide, Sodium sulphate.

Principle:
It is obtained from the dried leaflets of Cassia angustifolia, Cassia acutifolia belonging to family
Leguminosae (Caesalpiniaceae). Sennosides are the anthraquinone glycosides. Senna glycoside, also
known as sennoside, is a medication used to treat constipation and empty the large intestine before
surgery. The medication is taken by mouth or via the rectum. It typically begins working in minutes
when given by rectum and within twelve hours when given by mouth. It is a weaker laxative than
castor oil. Molecular formula is C42H38020.

Objectives:

Procedure:
 The weighted quantity of dried powdered Senna leaves was extracted with benzene for 30 min
on an electric shaker, filtered in vacuum and benzene distilled off.
 The left-over marc was dried at room temperature and extracted with 70% methanol for three
hrs. and filtered under vacuum.
 The marc was re-extracted with 50 ml of 70% methanol for 15 min, filtered and the methanolic
extracts were combined.
 The methanolic extract was concentrated to 1/4th volume, acidified to pH 3.2 by adding HNO3
with constant stirring.
 It was set aside for 15 min. at 5°C, filtered.
 1 g of anhydrous calcium chloride in 10 ml of denatured spirit was added with constant stirring.
 The pH of the solution was adjusted to 8 by addition of potassium hydroxide (KOH) and set
aside for 15 min., filter and collect the precipitate obtained as calcium sennoside dried and
weighed.
 The percentage yield was also calculated.
Identification by chemical tests:
Borntrager’s Test:
To 3 ml sample, dilute sulphuric acid (H2SO4) was added, boiled and filtered. To the cold filtrate,
equal volume of benzene was added and shaken. The organic solvent was separated and ammonia
was added. It was observed that the ammoniacal layer turned pink confirming the presence of
anthraquinone glycoside presence.
Modified Borntrager's Test:
To the 5 ml sample, 5 ml ferric chloride and 5 ml dilute hydrochloric acid was added. It was then
heated for 5 min in a boiling water bath. cooled and benzene was added. It was shaken well,
separated and an equal volume of dilute ammonia was added. It was observed that the ammoniacal
layer turned pink confirming the presence of anthraquinone glycoside.
Identification by TLC:
• Sennoside is identified by thin layer chromatography.
• Stationary phase: Silica gel GF
• Solvent System: n-propanol: ethyl acetate: water: glacial acetic acid (40:40:29:1)
• Spraying reagent: HNO3 - KOH reagent.
• After drying the TLC plate, Sennosides, is detected at Rf value.

Observations:
1. The percentage yield of total sennosides from senna leaves was found to be ____
2. The Rf value of sennosides from senna leaves was found to be ____

Learning Outcomes:

References:
1. Practical Pharmacognosy by K. R. Khandelwal
2. Practical Pharmacognosy by S. Joshi and V. Aeri
EXPERIMENT 8
Aim:
To extract quinine and quinidine from Cinchona bark.

Requirements:

Principle:

Objectives:

Procedure:
1. Moisten powdered cinchona (50gm) with ammonia water and allow it to stand for an hour,
then hot water is added. To the mixture, after cooling, milk of lime is added and the whole
evaporated to dryness.
2. Dry at room temperature or below 60°C.
3. Pack the material in a Soxhlet apparatus and extract with toluene for 6 hours.
4. Extract the toluene extract with dilute sulfuric acid under stirring conditions.
5. Separate the acidic aqueous liquid, neutralise the acidulated layer and allow standing when
neutral sulphates of the alkaloid (quinine, cinchonine, cinchonidine) are crystallised out.
6. Dissolve the crude quinine sulphate in water, decolorized with charcoal and recrystallized
until the cinchonidine and cinchonine are reduced to the required percentage.
7. Weigh and determine its melting point (177°C).
8. Report.

Observations:

Learning Outcomes:

References: