A THESIS ON

IN VITRO EVALUATION STUDIES OF ECOSPIRIN TABLETS

Dissertation submitted to

Jawaharlal Nehru Technological University, Kakinada, A.P.,

In partial fulfillment of the requirement for the award of the degree of

Bachelor of Pharmacy

By
CH.RAJASRI (17101R0041),
G.BHAVANI(17101R0046),
G.SAI LAKSHMI (17101R0049)
V.SRAVANI (17101R0003)
Under the guidance of
Dr.K.RAJYA LAKSHMI, M.Pharm, Ph.D
ASSOCIATE PROFESSOR

DEPARTMENT OF PHARMACEUTICS

BAPATLA COLLEGE OF PHARMACY

BAPATLA-522101

2017-2021.
 CERTIFICATE

 This is to certify that the dissertation entitled “In-vitro evaluation studies of Ecospirin
tablets’’done for the partial fulfillment for the award of degree of Bachelor of Pharmacy
has been carried out by CH.RAJASRI,G.BHAVANI,G.SAI LAKSHMI,V.SRAVANI
under my supervision andguidance at the Department of Pharmaceutics, Bapatla
College of Pharmacy during the period 2019-2020 to the Jawaharlal Nehru
Technological University, Kakinada.

Dr. T.E.G.K.Murthy, M.Pharm, Ph.D.

Principal &Professor
Department of Pharmaceutics
Bapatla College of Pharmacy,
Bapatla.

Place: Bapatla
Date:
 CERTIFICATE

 This is to certify that the dissertation entitled “In-vitro Evaluation studies of Ecospirin
tablets’’done for the partial fulfillment for the award of degree of Bachelor of Pharmacy
has been carried out by CH.RAJASRI, G.BHAVANI, G.SAI LAKSHMI,V.SRAVANI
under my supervision andguidance at the Department of Pharmaceutics, Bapatla
College of Pharmacy during the period 2019-2020 to the Jawaharlal Nehru
Technological University, Kakinada.

Dr. K.RAJYA LAKSHMI, M. Pharm , Ph.D

.Associate Professor

Department of Pharmaceutics
Bapatla College of Pharmacy, Bapatla.

Place: Bapatla
Date:
DECLARATION

The project embodied in this thesis entitled “In-vitro Evaluation studies of Ecospirin
tablets’’ was carried out in the department of pharmaceutics under the esteemed guidance of
Dr.k.Rajyalakshmi, M.Pharm, Ph.D., Assistant Professor in Bapatla College of Pharmacy,
Bapatla. The extent and source of information derived from the existence literature have been
indicated throughout the project work at appropriate places. The work is original and has not
been submitted in part or full for any diploma or degree of J.N.T.U University or any other
university.

CH.RAJASRI------------------------

G.SAILAKSHMI__________________

G.BHAVANI------------------------

V.SRAVANI------------------------
ACKNOWLEDGEMENT

It is a wonderful opportunity for me to thank God for his blessings to accomplish this task.

At the outset I express my deep sense of gratitude, and sincere thanks to Dr. K..RAJYA
LAKSHMI, M.Pharm, PhD; associate professor of Bapatla College of pharmacy, Bapatla for
her valuable guidance support and advice, encouragement, functional freedom and constant
supervision.
I specially trade my respected gratitude to faculty of pharmaceutics, Bapatla college of
pharmacy, Bapatla for her constant support throughout my academics and splendid guidance
regarding every aspect of my decisions for my future aspirations.

I take this opportunity to express my deep sense of gratitude and sincere thanks to
Mr.RangaRao, Mr.srinivasa Rao, Lab asst. of pharmaceutics department for their help
during experimental work.

I am thankful to Mr.Venkataswamy, Librarian, and Mr. Baji, Library assistant of

Bapatla College of pharmacy, Bapatla for providing the valuable books, journals and
references during this project work.

I am fortunate to have such a lovable parents. I am also grateful to whole

Family members for their affection and lovable support, without which this degree would not
have been possible.

I am specially thankful to all my senior M.pharmcy students of Bapatla College of

pharmacy, Bapatla for providing their help during this project work.

I am indebted to the management “Bapatla Educational Society, Bapatla” for providing me

the necessary infrastructural facilities to carry out the project work.

I take great privilege and pleasure to acknowledge the contributions of many individuals who
have been inspirational and supportive throughout my work undertaken and endowed me with
the most precious knowledge to success in my endeavor. My work bears the imprint of all
those people.

CH.RAJASRI,

G.BHAVANI

G.SAI LAKSHMI,

V.SRAVANI
DEDICATED TO

MY FAMILY
 ABBREVATIONS

ABBREVATIONS
GIT Gastro Intestinal Tract

Hr Hour

Min Minutes
RPM Rotations Per Minute

N Normality

X Mean

S.D Standard Deviation

K Dissolution Rate Constant

DE Extent of Dissolution

DF Degree of Freedom

MS Mean Sum of Squares

SS Sum of Squares

F Similarity factor

P Probability

IP Indian Pharmacopoeia

Fig Figure

Tab Table
INDEX

INDEX
S.NO TITLE PAGE NO.

1. AIM, OBJECTIVE AND SCOPE OF WORK

2. INTRODUCTION

3. LITERATURE REVIEW

4. METHODOLOGY

5. RESULTS

6. DISCUSSION AND CONCLUSION

 Chapter – 1

AIM, OBJECTIVE

Chapter – 1
AIM, OBJECTIVE AND SCOPE
1.1 AIM:-

To carry out the In-vitro evaluation studies of ECOSPIRIN tablets

1.2 OBJECTIVE:-

• To conduct the INVITRO evaluation study of selected anti-platelet that is

Ecospirin drug

• 1.3 SCOPE :

• Ecospirin tablets are widely used to treat and prevent heart attacks ,strokes

and heart related chest pain.

• It helps to prevent the formation of blood clots in blood vessels.

• It is a very widely used medicine for heart problems.

• In view of its wide usage and commercial availability ,in our study we aimed

to carry out the invitro evaluation studies of Ecospirin tablets.

 Chapter 2

INTRODUCTION

Chapter 2

INTRODUCTION
2.1 TABLETS(1)
A tablet is a pharmaceutical dosage form comprising a mixture of active
substances and excipients, usually in powder form, pressed or compacted from
a powder into a solid dose. The excipients can include glidants (flow aids),
diluents, binders or granulating agents and lubricants to ensure efficient
tableting; disintegrants to promote tablet break-up in the digestive tract;
sweeteners or flavours to enhance taste; and pigments to make the tablets
visually attractive. A polymer coating is often applied to enhance the tablet's
appearance or to make the tablet smoother and easier to swallow and to
control the release rate of the active ingredient, to make it more resistant to
the environment (extending its shelf life). “Caplets” are those tablets which are
in the shape of capsules. Medicinal tablets and capsules are often called pills.

Tablet coating :

Coating is a process by which an essentially dry, outer layer of coating material

is applied to the surface of a dosage form in order to confer specific benefits
that broadly ranges from facilitating product identification to modifying drug
release from the dosage form. After making a good tablet, one must often coat
it. Coating may be applied to multiple range of oral solid dosage form, including
tablets, capsules, multiparticulates and drug crystals. When coating
composition is applied to a batch of tablets in a coating pan, the tablet surfaces
become covered with a tacky polymeric film. Before the tablet surface dries,
the applied coating changes from a sticky liquid to tacky semisolid and
eventually to a non-sticky dry surface pans. The entire coating process is
conducted in a series of mechanically operated acorn-shaped coating pans of
galvanized iron stainless steel or copper. The smaller pans are used for
experimental, developmental, and pilot plant operations, the larger pans for
industrial production.

Primary components involved in tablet coating

 Tablet properties
 Coating process
 Coating equipments
 Parameters of the coating process
 Facility and ancillary equipments
 Automation in coating processes

Coating Process Design & Control

In most coating methods, when the tablets are being agitated in a pan, fluid
bed, etc. at that time spraying on tablets by coating solution takes place. As the
solution is being sprayed, a thin film is formed that adheres directly to each
tablet. The coating may either be formed by a single application or may be built
up in layers through the use of multiple spraying cycles.In pharmaceutical
industry, rotating coating pans are often used. Firstly, uncoated tablets are
placed in the pan, which is typically tilted at an angle from the horizontal, and
then the liquid coating solution is introduced into the pan while the tablets are
tumbling. By passing air over the surface of the tumbling tablets, the liquid
portion of the coating solution is then evaporated. In comparison, a fluid bed
coater operates by passing air through a bed of tablets at a velocity sufficient to
support and separate the tablets as individual units. Once separation takes
place, then the tablets are sprayed with the coating composition.

The coating process is usually a batch operating task consisting of the

following phases:

 Identification of batch and Recipe selection (film or sugar coating)

 Loading/Dispensing (accurate dosing of all required raw materials)

 Warming

 Spraying (Both application and rolling are carried out simultaneously)

 Drying

 Cooling

 Unloading

Coating equipment

A modern tablet coating system combines several components:

 A coating pan

 A spraying system

 An air handling unit

 A dust collector

Advantages of tablet coating

  Tablet coatings must not make tablets stick together during the coating
process, must follow the fine contours of embossed characters or logos on
tablets and must be stable and strong enough to survive the handling of
the tablet.
 Printing on tablets can also be done by coatings, if required. Coatings are
necessary for tablets giving a smoother finish, makes large tablets easier to
swallow and also to mask the unpleasant taste.

Disadvantages of tablet coating

 Limitations of sugar coating such as relatively high cost, long coating

time and high bulk have led to the use of other coating materials.
 However the process of coating is tedious and time-consuming and it
requires the expertise of highly skilled technician

2.2 ENTERIC COATED TABLETS(1)

 An enteric coating is a barrier that controls the location of oral

medication in the digestive system where it is absorbed. The word
 “enteric” indicates small intestine; therefore enteric coatings prevent
release of medication before it reaches the small intestine.
 The enteric coated polymers remain unionise at low pH, and therefore
remain insoluble. But as the pH increases in the GIT, the acidic functional
groups are capable of ionisation, and the polymer swells or becomes
soluble in the intestinal fluid.
 Materials used for enteric coatings include CAP, CAT, PVAP and HPMCP,
fatty acids, waxes, shellac, plastics and plant fibers.
 There are four reasons for putting such a coating on a tablet or capsule
ingredient: Protection of active pharmaceutical ingredients, from the
acidic environment of the stomach (e.g. enzymes and certain
antibiotics).
 To prevent gastric distress or nausea from a drug due to irritation (e.g.
sodium salicylate). For the delivery of drugs that are optimally absorbed
in the small intestine to their primary absorption site in their most
concentrated form.
 To provide a delayed-release component for repeat action. Required
for minimizing first pass metabolism of drugs. The choice of the
polymer and the thickness of the coated layer are critical to control the
pH solubility profile of the enteric coated dosage form. The most
common drugs which cause stomach ulcers like aspirin, diclofenac and
naproxen are frequently available with enteric coatings.
 Omeprazole, which is a drug which stops the stomach from producing
acid, is itself broken down in acid and therefore the drug generally has an
 enteric coating around it either as a granule in the capsules or as a
granule in the dispersible form.
 Sulfasalazine is used either for the treatment of Crohn's disease which is
inflammation of the intestines or for the treatment of arthritis. When
used for Crohn's disease where it is needed in the intestines to work, it is
given with an enteric coating whereas for arthritis it is very often given
without an enteric coating so that it can be absorbed more quickly.
 A review on recent advances of enteric coating 7 ERY-TAB is an
antibacterial product containing erythromycin base in an especially
enteric-coated tablet to protect it from the inactivating effects of gastric
acidity and to permit efficient absorption of the antibiotic in the small
intestine. ERY-TAB (erythromycin delayed-release tablets) are available
for oral administration in three dosage strengths, each white oval tablet
containing 250 mg, 333 mg, or 500 mg of erythromycin as the free base.
Other commercially available tablets are enteric coated aspirin. E.g.
Micropirin® 75mg EC tablets and enteric coated peppermint oil. E.g.
Colpermin®
 2.1 Ideal properties of enteric coating material
 Resistance to gastric fluids.
 Susceptible/permeable to intestinal fluid.
 Compatibility with most coating solution components and the
drug substrate.
 Formation of continuous film
 Nontoxic, cheap and ease of application
 Ability to be readily printed
Polymers used for enteric coating

Different polymer used for enteric coating

POLYMERS DISSOLUTION PH
Shellac (esters of aleurtic acid) 7.0
Cellulose acetate phthalate (CAP) 6.2
Poly(methacrylic acid-co-methyl 5.5-7.0
methacrylate)
Cellulose acetate trimellitate (CAT) 5.0
Poly(vinyl acetate phthalate) (PVAP) 5.0
Hydroxypropyl methylcellulose 4.5-5.5
phthalate (HPMCP)

New materials used for tablet coating

 Zein

 Aqua-Zein®, which is an aqueous zein formulation containing no alcohol.

 Amylose starch and starch derivatives

 Dextrins

Drug profile:
2.3 Aspirin(4) ;

Name : Aspirin

Brand name /proprietary name : Ecospirin

Chemical structure :

Molecular formula : C H O   
9 8 4

Weight :180.1574

Chemical name :2-acetyloxybenzoic acid

PROPERTIES:
Appeareance : White , Crystalline powder

Melting point :275 °F

PKa : 3.5

Solubility :soluble in water, chloroform,less soluble in anhydrous

ether.

MECHANISM OF ACTION(4):

 Acetylsalicylic acid disrupts the production of prostaglandins throughout

the body by targeting cyclooxygenase-1 (COX-1) and cyclooxygenase-2

(COX-2). Prostaglandins are potent, irritating substances that have been

shown to cause headaches and pain upon injection into humans.

Prostaglandins increase the sensitivity of pain receptors and substances

such as histamine and bradykinin. Through the disruption of the

production and prevention of release of prostaglandins in inflammation,

this drug may stop their action at pain receptors , preventing symptoms

of pain. Acetylsalicylic acid is considered an antipyretic agent because of

its ability to interfere with the production of brain prostaglandin

E1. Prostaglandin E1 is known to be an extremely powerful fever-

inducing agent. **Effects on platelet aggregation** The inhibition

of  platelet aggregation by ASA occurs because of its interference

with thromboxane A2 in platelets, caused by COX-1

inhibition. Thromboxane A2 is an important lipid responsible for platelet

aggregation, which can lead to clot formation and future risk of heart

attack or stroke. 

PHARMACOKINETICS(4) :

 tmax = 3 to 4 hours .

 Mean Elimination half life is 48hours .

 When ingested orally, acetylsalicylic acid is rapidly absorbed in both

the stomach and proximal small intestine.

 Aspirin is a lipophilic drug.0

 The Peak plasma salicylate concentrations occur between 1-2 hours

post-administration.

 The drug crosses the placenta

 The rate of salicylate is often variable, ranging from 10% to 85% in the

urine, and heavily depends on urinary pH.

ADVERSE EFFECTS (2)

 Cardiac disorders

 Congenital , familial and genetic disorders

 Ear and labyrinth disorders

 Endocrine disorders

 Eye disorders

 Gastrointestinal disorders

 General disorders and administration site conditions

 Hepatobiliary disorders

 Blood and lymphatic system disorders

 Immune system disorders

 Infections and infestations

 Injury, poisoning and procedural complications

 Investigation

 Metabolism and nutrition disorders

 Nervous system disorders

 Pregnancy, puerperium and perinatal conditions

 Product issues

 Psychiatric disorders

 Renal and urinary disorders

 Reproductive system and breast disorders

 Respiratory , thoracic and mediastinal disorders

 Skin and subcutaneous tissue disorders

 Social circumstances

 Surgical and medical procedures

 Vascular disorders

CONTRAINDICATIONS (2):

 Ischaemic heart disease, hypertension, epilepsy, hepatic or renal

impairment and pregnancy are the contraindications. Patients should be

cautioned not to drive. Sumatriptan and ergotamine should not be

administered within 24 hours of each other. Interaction with 5-HT

reuptake inhibitors, MAO inhibitors and lithium has been reported

responding to the first dose should not be given the second dose. It is the

only triptan available for parenteral use; 6 mg s.c. may be given to

patients who cannot take the drug orally or in whom the pain develops

very rapidly. After injection, it acts in 10–20 min and is more consistently
 effective. Alternatively, for rapid action and in patients who vomit out the

oral tablet, 25 mg nasal spray can be used. It may be repeated once after 2

hours. A bitter taste may be felt after the nasal spray.

 Aspirin displaces warfarin, naproxen, sulfonylureas, phenytoin and

methotrexate from binding sites on plasma proteins: toxicity of these

drugs may occur. Its antiplatelet action increases the risk of bleeding in

patients on oral anticoagulants. 2. Aspirin at analgesic doses inhibits

tubular secretion of uric acid and antagonizes uricosuric action of

probenecid. Tubular secretion of methotrexate is also interfered. 3.

Aspirin blunt diuretic action of furosemide and thiazides and reduces K+

conserving action of spironolactone. Competition between canrenone

(active metabolite of spironolactone) and asprin reduces protein bound

iodine levels by displacement of thyroxine; but hypothyroidism does not

occurs

DOSAGE AND ADMINISTRATION :

 ADULTS-in low doses (50-325) mg per day produces anti platelet effect

which lasts for 8-10 days .

 Aspirin in high doses (2-3) grams per day which inhibits both PGI2 and

thromboxane synthesis.
  Anti rheumatic fever doses are 75-100 mg per kg per day.

 Rheumatoid arthritis aspirin a dose of 3-5 grams per day effective.

 Analgesic 300-600 mg during 6-8 hours.

DRUG INTERACTIONS (1)

1. Aspirin displaces warfarin, naproxen, sulfonylureas, phenytoin and

methotrexate from binding sites on plasma proteins: toxicity of these drugs

may occur. Its antiplatelet action increases the risk of bleeding in patients

on oral anticoagulants.

2. Aspirin at analgesic doses inhibits tubular secretion of uric acid and

antagonizes uricosuric action of probenecid. Tubular secretion of

methotrexate is also interfered.

3. Aspirin blunts diuretic action of furosemide and thiazides and reduces K+

conserving action of spironolactone. Competition between canrenone

(active metabolite of spironolactone) and aspirin for active transport in

proximal tubules has been demonstrated.

4. Aspirin reduces protein bound iodine levels by displacement of thyroxine;

but hypothyroidism does not occur

 TRADE NAME : Trade names of some of the marketed Brand Name(s):
Zorprin, Bayer Buffered Aspirin, Durlaza, Asatab, Adprin-B, Alka-Seltzer Extra Strength
with Aspirin, Alka-Seltzer with Aspirin, Arthritis Pain Formula, Ascriptin, Ascriptin
Maximum Strength, ASA, Bayer Children's Aspirin, Bayer Women's Low Dose, Bayer Low
Adult Strength, Bayer Advanced Aspirin, Bayer Extra Strength, Bayer Extra Strength
Plus, Bufferin, Bufferin Extra Strength, Ecotrin, Ecotrin Maximum Strength, Empirin,
Extended Release Bayer 8-Hour Caplets, Extra Strength Bayer Plus Caplets, Genuine
Bayer Aspirin, Halfprin, Maximum Bayer Aspirin, St. Joseph Adult Chewable Aspirin, St.
Joseph Regular Strength, acetylsalicylic acid

STORAGE AND STABILITY : Stored in well closed container at room

temperature (20°C -25°C ).

REFERENCES :

1.Singh Deep Hussan : A review on recent advances of enteric

coating.

2.K.D.Tripati ,Essential of medical pharmacology , 7th Edition ,

Jaypee Brothers medical publishers pg.,no.58-571 .

3.Government of India , Indian pharmacopeia , sixth Edition 2010 ,

The Indian pharmacopeia commission ,volume-2

4.pubchem.ncbi.nlm.gov:aspirin.
 Chapter – 3
LITERATURE REVIEW
Chapter – 3
3.1 LITERATURE REVIEW:
1. Mohamad Mroueh. et.al (2003) Aspicot® is an enteric-coated aspirin
that is being extensively used in the Middle East, including
Lebanon where this drug is manufactured, without any clinical in
vivo implication showing or confirming its bioequivalence. For this
reason, this investigation was carried out to evaluate the in vitro
dissolution as well as the bioavailability and pharmacokinetic
properties of 2 tablet oral dosage forms of enteric-coated aspirin,
Aspirin protect® and Aspicot®, in a single dose of 200 mg among
healthy volunteers.
2. Samira karim.et.al(2016) The study aims at the design of a sustained
release dosage form for potential use of aspirin which is currently
used as the cardiotonic, analgesic, antipyretic and anti-
inflammatory agent to investigate the effect of polymers on the
release pattern from tablets. Ethyl cellulose and hydroxypropyl
methyl cellulose-K15 MCR polymers had been used in the
formulation of sustained release tablets which were prepared by
direct compression method. Drug release study was evaluated for
8 hrs in 50 mM phosphate buffer, pH 6.8 at 50 rpm. Standard
physicochemical tests were performed for all the formulations.
3. Singh deep hussan.et.al(2012) Enteric coated tablets are solid unit
dosage forms which are designed to bypass the stomach and release the
drug in small intestine and are meant for oral administration. The word
 “enteric” indicates small intestine; therefore enteric coatings prevent
release of medication before it reaches the small intestine. Most enteric
coatings work by presenting a coated surface that is stable at the highly
acidic pH found in the stomach, but breaks down rapidly at a less acidic
(relatively more basic) pH. Materials used for enteric coatings include
CAP, CAT, PVAP and HPMCP, fatty acids, waxes, shellac, plastics and plant
fibers. The present review describes enteric coating, their ideal
properties, benefits and limitation, various polymers used, their chemical
structure, criteria for drug selection and mechanism, methods of
manufacturing and evaluation of enteric coated tablets. Recently, these
have attracted the interest of many formulators due to their advantages
over the conventional drug delivery systems as they prolong the dosing
intervals and also increase patient compliance. The study provides an
overview of the recent advances that have taken place in this arena.
4. Kunal Kanani.et.al(2015) Aspirin has been used therapeutically for over
100 years. As the originator and an important marketer of aspirin-
containing products, Bayer’s clinical trial database contains numerous
reports of the pharmacokinetics of various aspirin formulations. These
include evaluations of plain tablets, effervescent tablets, granules,
chewable tablets, and fast-release tablets. This publication seeks to
expand upon the available pharmacokinetic information concerning
aspirin formulations. In the pre-systemic circulation, acetylsalicylic acid
(ASA) is rapidly converted into its main active metabolite, salicylic acid
(SA). Therefore, both substances are measured in plasma and reported in
the results. The 500 mg strength of each formulation was chosen for
 analysis as this is the most commonly used for analgesia. A total of 22
studies were included in the analysis. All formulations of 500 mg aspirin
result in comparable plasma exposure to ASA and SA as evidenced by
AUC. Tablets and dry granules provide a consistently lower Cmax
compared to effervescent, granules in suspension and fast release
tablets. Effervescent tablets, fast release tablets, and granules in
suspension provide a consistently lower median Tmax compared to dry
granules and tablets for both ASA and SA. This report reinforces the
importance of formulation differences and their impact on
pharmacokinetic parameters.
5. Jennifer B. Dressman.et.al(2012) A biowaiver monograph for
acetylsalicylic acid (ASA) is presented. Literature and experimental data
indicate that ASA is a highly soluble and highly permeable drug, leading to
assignment of this active pharmaceutical ingredient (API) to Class I of the
Biopharmaceutics Classification System (BCS). Limited bioequivalence (BE)
studies reported in the literature indicate that products that have been tested
are bioequivalent. Most of the excipients used in products with a marketing
authorization in Europe are not considered to have an impact on
gastrointestinal motility or permeability. Furthermore, ASA has a wide
therapeutic index. Thus, the risks to the patient that might occur if a non-
bioequivalent product were to be incorrectly deemed bioequivalent according
to the biowaiver procedure appear to be minimal. As a result, the BCS-based
biowaiver procedure can be recommended for approval of new formulations of
solid oral dosage forms containing ASA as the only API, including both
multisource and reformulated products, under the following conditions: (1)
excipients are chosen from those used in ASA products already registered in
International Conference on Harmonization and associated countries and (2)
the dissolution profiles of the test and the comparator products comply with
the BE guidance.
6. Deepak L. Bhatt.et.al(2020) Dyspeptic symptoms are common with aspirin
and clinicians frequently recommend that it be taken with food to reduce these
side effects. However, food can interfere with absorption, especially with
enteric-coated aspirin formulations. We evaluated whether food interferes
with the bioavailability of a new, pharmaceutical lipid-aspirin complex (PL-ASA)
liquid-flled capsule formulation. In this randomized, open label, crossover
study, 20 healthy volunteers fasted for≥10 h and then randomized as either
“fasted”, receiving 650 mg of PL-ASA, or as “fed”, with a standard high-fat meal
and 650 mg of PL-ASA 30 min later. After a washout of 7 days, participants
crossed over to the other arm. The primary outcome was comparison of PK
parameters of the stable aspirin metabolite salicylic acid (SA) between fasted
and fed states. Mean age of participants was 36.8 years and 55% were male.
The ratios for the fed to fasted states of the primary SA PK parameters of
AUC0−t and AUC 0−∞ were 88.7% and 88.8% respectively, with
90% confidence intervals between 80 and 125%, which is consistent with FDA
bioequivalence guidance. Mean peak SA concentration was about 22% lower
and occurred about 1.5 h later in the fed state. Food had a modest effect on
peak SA levels and the time required to reach them after PL-ASA
administration, but did not impact the extent of exposure (AUC) compared with
intake in a fasted state. These data demonstrate that PL-ASA may be co-
administered with food without signifcant impact on aspirin bioavailability.
 REFERENCES
1.Mohamad Mroueh A Comparitive Single Dose Bioequivalance Study of
Two Enteric Coated Aspirin Brands Among Healthy Volounteers(2003).
2.Karim Formulation and In vitro Evaluation of Aspirin Sustained Relaese
Tablets Using Hydrophyllic Polymer(2016).
3.Singh Deep Hussan A review on recent advances of enteric coating(2012).
4.Kunal Kanani Influence of differing Anaslgesic Formulations of Aspirin
on Pharmacokinetic Parameters(2015).
5.Jennifer B. Dressman Biowaiver Monograph for Immediate -Release Solid
Oral Dosage Forms: Acetylsalicylic Acid(2012).
6.Deepak L. Bhatt Pharmacokinetic/pharmacodynamic assesment of a novel
pharmaceutical lipid-aspirin complex: results of a randomized
crossover ,bioequivalence study.
 CHAPTER -4
METHODOLOGY
Chapter - 4
METHODOLOGY:-

4.1 Glass ware used to perform project(11)

Volumetric flask - 10,25,100,250,500,1000ml

Measuring cylinder - 10,100,1000ml

Beaker - 100,500,1000ml

Pipette - 1ml and 10ml

Test tubes - 10ml

Funnel

Glass rod

Spatula

4.2 Chemicals used(12)

Sodium hydroxide -4.48gm -

Potassium dihydrogen phosphate - 34 gm

0.1 N HCl - 42.5 ml

4.3Tablets brands used for Aspirin :

TABLE -4.3

Brand strength Strength Mfg.date Exp.date Batch. No

1 ECOSPIRIN-325 Aspirin-325mg 02-2020 01-2022 04007647

Quality control parameters

Table :- 4.4

Parameters Equipments Model number

High Precision Balance CITIZEN – CX265N

Weight

CAMPBELL ELECTRONICS
Roche Friabilator Model
Friability THERMONIK

ZINEO SCIENTIFIC
Monsanto Hardness Tester
Hardness INSTRUMENTS

Tablet Disintegration Test CAMPBELL ELECTRONICS

Disintegration Machine THERMONIK

Dissolution Dissolution apparatus AARKEY LABTRONIX INDIA

4.5 Construction of Calibration Curve for ECOSPIRINTABLETS :

The calibration curve was constructed in PH6.8phosphate buffer100mg of aspirin was
accurately weighed, transferred into 100 ml volumetric flask and add 10ml of P H phosphate
buffer to dissolve then the final volume was adjusted by use of the pH 6.8 phosphate buffer ,
which gives a stock solution 1 mg/ ml. Further dilutions were made with 6.8 phosphate buffer
to obtain 2 to 10 µg/ml concentrations of Atenolol and the absorbance was measured at
265nm respectively (refer table 5.1and fig.1).

Construction of calibration curve for ASPIRIN :

The calibration curve was constructed in PH0.1N HCl buffers. 100mg of aspirin was
accurately weighed, transferred into 100 ml volumetric flask and add 10ml of 0.1N HCl
buffer to dissolve then the final volume was adjusted by use of the 0.1 N HCl buffer , which
gives a stock solution 1 mg/ ml.
Further dilutions were made with 0.1N HCL buffer to obtain 2 to 10 µg/ml concentrations of
Aspirin the absorbance was measured at 265nm respectively (refer table 5.1 and fig.2).

4.6 IN-VITRO EVALUATION OF MARKETED DRUG PRODUCTS USING

QUALITY CONTROL TESTS (15)
PARAMETERS :
* Weight variation
* Hardness test
* Friability
* Tablet disintegration
* Tablet dissolution

4.6.1 Weight variation(15.1):-

To study weight variation, 20 tablets of each formulation were weighed using
an electronic balance and the test was performed according to the official
method.(refer table 5.2).

Formula:-

% Deviation =

Average Weight Of Tablet % Deviation

<80 10%

80-250 7.5%

>250 5%
Reference :- INDIAN PHARMACOPOIEA 2010

4.6.2 Hardness(15.3 , 16) :-

For each formulation, the hardness of 6 tablets was determined using

calibrated Monsanto hardness tester. The tablet was held along its oblong axis
in between the two jaws of the tester. At this point, reading should be zero
kg/cm2 . Then constant force was applied by rotating the knob until the tablet
fractured. The value at this point was noted in kg/cm2 .(refer table 5.3).
 fig :- 4.6.2

Reference :- INDIAN PHRAMACOPOEIA 2010

4.6.3Disintegration :-

If the tablet has a soluble external coating immerse the basket in a water at
room temperature for 5 minutes. suspend the assembly in the beaker
containing 0.1Mhydrochloric acid and operate without the discs for 120
minutes, unless otherwise stated in the individual monograph. remove thre
assembly from the liquid .No tablet shows signs of cracks that would allow
the escape of the contents of disintegration apart from fragments of
coating .replace the liquid in the beaker with mixed phosphate buffer ph
6.8,add a disc to each to and operate the apparatus for a further 16 minutes.
remove the assembly from the liquid. the tablets pass the test if all the 6 have
disintegrated.(refer table 5.4).
Reference :- INDIAN PHRAMACOPOEIA 2010

4.6.4 Dissolution :- In-vitro Dissolution Parameters for ecospirin of strength 325mg

ASPIRIN(16) :

In-vitro drug release study for the prepared coated tablets was conducted for a period of 90 min
using a six-station USP type II (paddle) apparatus at 37 ºC and 75 rpm speed. Tablets were kept in 0.1
N HCl for 2 h and later dissolution media replaced with phosphate buffer pH 6.8. Dissolution study
was carried out for 90 min in phosphate buffer pH 6.8. Sampling was done after 10, 20, 30, 45, 60
and 90 min interval; samples of 10 ml were withdrawn from dissolution medium and replaced with
fresh medium to maintain the volume constant.The sample solution was analyzed at 265 nm for
estiation of Aspirin by a UV-spectrophotometer. The amounts of drug present in the samples were
calculated with the help of appropriate calibration curve.(refer table 5.5)
characteristics ASPIRIN ASPIRIN
Buffer 0.1N HCL PH 6.8 Phosphate buffer

Volume 900ml 900ml

rpm 75 rpm 75rpm

Dissolution Apparatus (USP) TYPE-2 (paddle) TYPE -2(Paddle)

ƛmax 265 nm 265nm

Time of samples withdrawn 10min 10min

Total no. samples Withdrawn 7 7

Total time 2hrs 1hr

REFERENCE:-

11. Indian pharmacopoeia Edition 2010

12. Indian pharmacopoeia Edition 2010

13. Indian pharmacopoeia Edition 2010

14. Indian pharmacopoeia Edition 2010

15. Indian pharmacopoeia Edition 2010

16. Shradha S.Twari : Formulation and evaluation of enteric coated asprin tablet by using
bioepoxy resin as coating material.
Chapter - 5

RESULTS
RESULTS & DISCUSSION

Table 5.1 Calibration curve for aspirin using pH phosphate buffer:

Absorbance of Aspirin at 265 nm

Conc.
Trail 1 Trail 2 Trail 3 Mean ± SD

0 0 0 0 0

20 0.065 0.070 0.060 0.195

40 0.118 0.120 0.110 0.245

60 0.192 0.195 0.190 0.360

80 0.250 0.255 0.240 0.512

0.3
100 0.310 15 0.305 0.555
fig:1 construction of calibration curve of ASPIRIN using pH 6.8 phosphate buffer

table 5.1 calibration curve for aspirin using 0.1 N Hcl :

Absorbance of Aspirin at 265nm

Conc.
Trail 1 Trail 2 Trail 3 Mean ± SD

0 0 0 0 0

2 0.105
0.095 0.105 0.100

4 0.199 0.200 0.179 0.165

6 0.262 0.265 0.248 0.207

8 0.403 0.405 0.343 0.279

fig 2:consruction of calibartion curve for aspirin using 0.1 N Hcl

Table 5.2 weight variation test results observed from the marketed formulations OF
ASPIRIN
 S.NO ECOSPIRIN
1 0.400
2 0.415
3 0.400
4 0.405
5 0.410
6 0.400
7 0.405
8 0.415
9 0.410
10 0.415
11 0.415
12 0.420
13 0.400
14 0.410
15 0.400
16 0.405
17 0.415
18 0.405
19 0.400
20 0.410
Avg wt 0.4077

The percent deviation in weight variation of 20 tablets was found to be -1.88%.

Table 5.3 Hardness test results observed from the marked formulations of ASPIRIN

2nd 3rd 4th Mean±SD

Brands 1st Tablet tablet tablet tablet

5.2 4.9 4.97±

ECOSPIRIN 5 4.8

TABLE 5.4 Disintegration test results observed from the marked

formulations of ASPIRIN in

Tablet Dis integration time

1 10
2 12
3 15
4 10
5 15
6 10

ASPIRIN :

BRAND : ECOSPIRIN

Table:5.5 In-vitro dissolution data table for ASPIRIN

 Time
%Drug
(min
released
)
10 45.2

20 69.13

30 85.10

40 87.23

50 105.2

Table 5.6First order data for Aspirin for brand ecospirin

Time
Log % drug unreleased
(min)

10 1.738

20 1.489

30 1.173

40 1.106
 Parameters Trail 1
Table:5.7

Slope 0.0316

K(min-1) 0.072

t50 (min) 9.625

t90(min) 1.458
fig:3 First order plots of ASPIRIN obtained from ECOSPIRIN
CHAPTER 6

DISSCUSSION&CONCLUSION
CHAPTER 6

DISSCUSSION

DISCUSSION OF RESULTS:
Calibration curve was constructed using 0.1N HCL and phosphate buffer 6.8
from the results given in table 5.1 and fig 1 we found that good correlation
was observed between concentration and absorbance in both buffers.
INVITRO EVALUATION STUDIES FOR ECOSPIRIN:
1) AVERAGE WEWIGHT OF TABLETS:
The weight of 20 tablets were noted in table 5.2 and average weight calculated
and was found to be 0.4077 gm.
The percent deviation in weight variation of 20 tablets was found to be within
pharmacopeial limits.
2) HARDNESS TEST
The hardness test was conducted for 4 tablets randomly. Hardness of tablet was
found to be 4.97±
3) DISINTEGRATIOIN STUDIES:
Disintegration test was conducted in two buffers namely 0.1 N HCL and 6.8 ph
buffer the results were given in table 5.4
The result indicated the tablet was not disintegrated in 0.1N HCL and the tablet
was disintegrated within 10 minutes in 6.8 phosphate buffer.
DISSOLUTION STUDIES:
Dissolution studies were conducted in 0.1N HCL for period of 2 hrs later on the
buffer was replaced with 6.8 phosphate buffer. the results were given in table no
5.5 and fig 3
The results indicated that maximum drug release was observed with in a study
period of 50 minutes.
so the dissolution was found to be within the pharmacopoeia limits.
From the dissolution data obtained data the results were subjected to calculate
the release rate constant K1(min-1) ,t50% and t90% .
 Graph was constructed figure no3 using log (% un released) vs time .
 From the slope of this graph , K1 value was calculated.
 Further K1 (min-1) value was used to calculate t50% and t90% .
 The value K obtained was 0.072 from the table 5.7
 t50% was found to be 9.625 min
 t90% was found to be 1.458 min
CHAPTER-7
CONCLUSION :

 Aspirin is the drug for treatment of rheumatic fever.

 It is widely used drug for prevention of blood clots.
 In our work, Ecospirin tablets were procured from market and the tablets
were subjected to evaluation.
 All the results were found to be within pharmacopoeial limits.