Download development and validation of analytical methods for the

“DEVELOPMENT AND VALIDATION OF ANALYTICAL METHODS
FOR THE ESTIMATION OF PREDNISOLONE TABLET”
SYNOPSIS FOR
M. PHARM DISSERTATION
SUBMITTED TO
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
BENGALURU, KARNATAKA
BY
SAHANAPRIYA.C.N
M. PHARM Part-I
DEPARTMENT OF PHARMACEUTICAL ANALYSIS
PES COLLEGE OF PHARMACY
BENGALURU-560050
(2011-12)
UNDER THE GUIDANCE OF
Mr. VENKATESH PRASAD S.P
Asst. Professor
DEPARTMENT OF PHARMACEUTICAL ANALYSIS
PES COLLEGE OF PHARMACY
BENGALURU-560 050,KARNATAKA
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
BENGALURU, KARNATAKA.
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1.
Name of the candidate
and address:
SAHANAPRIYA.C.N
Department of Pharmaceutical Analysis,
PES College of Pharmacy,
50 Feet Road, Hanumanthanagar,
BSK I-Stage,
Bengaluru-560050.
Permanent Address:
D/o NAGARAJU.C.K
Chandagalu.
Dudda hobli,
Mandya taluk and Mandya district,
Karnataka.
2.
Name of the institution:
3.
Course of
subject:
4.
Date of the admission:
5.
Title of the topic:
study
and
PES College of Pharmacy
50 Feet Road, Hanumanthanagar,
BSK I- Stage,
Bengaluru-560050
MASTER OF PHARMACY IN
PHARMACEUTICAL ANALYSIS
29/7/2011
“DEVELOPMENT AND VALIDATION OF ANALYTICAL
METHODS FOR THE ESTIMATION OF
PREDNISOLONE TABLET”
2
6.
BRIEF RESUME OF THE INTENDED WORK:6.1 Need for the study:
Prednisolone is a corticosteroid drug with predominant glucocorticoid and low mineralocorticoid
activity, making it useful for the treatment of a wide range of inflammatory and auto-immune
conditions such as asthma, uveitis, pyoderma gangrenosum, rheumatoid arthritis, ulcerative
colitis, temporal arteritis and Crohn's disease, Bell's palsy, multiple sclerosis, cluster headaches,
vasculitis, acute lymphoblastic leukemia and autoimmune hepatitis, systemic lupus erythematous,
and dermatomyositis. It is also used for treatment of sarcoidosis, though the mechanism is
unknown.
Prednisolone is the active metabolite of prednisone, which is also used as a drug. Prednisolone
can also be used as an immunosuppressive drug for organ transplants and in cases of adrenal
insufficiency (Addison's).
Corticosteroids inhibit the inflammatory response to a variety of inciting agents and, it is
presumed, delay or slow healing. They inhibit the edema, fibrin deposition, capillary dilation,
leukocyte migration, capillary proliferation, fibroblast proliferation, deposition of collagen, and
scar formation with inflammation.
Glucocorticoids are most commonly employed for their systemic anti-inflammatory and
immunosuppressive properties.
Drug Profile :
Prednisolone
O
HO
HO
H
H
O
3
H
OH
Synonym : 11,17-dihydroxy-17-(2-hydroxyacetyl)-10,13-dimethyl-6,7,8,9,10,11,12,13,14,15,16,
17- dodecahydrocyclopenta [a]phenanthren-3-one.
Formula : C21H28O5
Molecular Mass : 360.444 g/mol
Mechanism of action:
Prednisolone irreversibly binds with glucocorticoid receptors (GR) alpha and beta for which they
have a high affinity. AlphaGR and BetaGR are found in virtually all tissues with variable
numbers between 3000 and 10000 per cell, depending on the tissue involved. Prednisolone can
activate and influence biochemical behaviour of most cells. The steroid/receptor complexes
dimerise and interact with cellular DNA in the nucleus, binding to steroid-response elements and
modifying gene transcription. They both induce synthesis of some genes and, therefore, some
proteins, and inhibit synthesis of others.
Not all metabolic actions on genes are known. Most mediator proteins are enzymes, e.g., cAMPdependent kinase.(1)
Anti-inflammatory and immunosuppressive actions:
• Inhibition of gene transcription for COX-2, cytokines, cell adhesion molecules, and inducible
NO synthetase.
• Blockage of Vit D3-mediated induction of osteocalcin gene in osteoblasts
• Modification of collagenase gene transcription
• Increase synthesis annexin-1, important in negative feedback on hypothalamus and anterior
pituitary gland.
• Anti-inflammatory action, it is presumed.
Several analytical techniques like spectroscopy(2,4), HPLC(5,10) and HPTLC(11) methods have been
reported for assay of Prednisolone in combination with other drugs. There are no reported
methods for the estimation of prednisolone alone in the formulations containing only
prednisolone , hence there is a need to develop analytical methods.
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6.2 REVIEW OF THE LITERATURE:
have reported Development and validation of
Analytical method for Estimation of Prednisolone in bulk and tablets using uv-visible
spectroscopy.

Abd El-Maboud I. Mohamed 1, Hesham Salem 2 and Eman Maher 2 have reported
Spectrophotometric determination of binary mixtures of prednisolone with some
antibiotics.

Mohit Rohitas, Abhinav Agrawal, Ashish Jain K, Narendra Lariya K, Anil Kharya K and
Govind Agrawal P. have reported Development of Simultaneous Spectrophotometric
method of Mesalazine and Prednisolone in dosage form.

Ashok R, Prakash PP, and Tamil Selvan R
have reported Spectrophotometric Determination of
Corticosteroids and Its Application in Pharmaceutical Formulation.
Singh DK and
Rohan Verma

Berend Oosterhuis,Inke Ten Berge JM,Peter Schellekens TA,Richard Koopmans P,Chris
Van Boxtel J have reported Prednisolone Concentration-Effect Relations in Humans and
the Influence of Plasma Hydrocortisone.

Alam MDA, Alka Ahuja, Sanjula Baboota, Gidwani SK, Ali J have reported Formulation
and evaluation of pharmaceutically equivalent parenteral depot suspension of methyl
prednisolone acetate.

Bhawani SA1, O Sulaiman1, R Hashim1 and MN Mohamad Ibrahim have reported
Thin-Layer Chromatographic Analysis of Steroids: A Review.

Mary Cheng H, Willium Huang Y,Allen Lipsey L have reported Simultaneous LiquidChromatographic Determination of Prednisone and Prednisolone in Plasma.

Oneeb Majid, Fatemeh Akhlaghi, Terry Lee, David W. Holt, and Andrew Trull have
reported Simultaneous Determination of Plasma Prednisolone, Prednisone, and Cortisol
Levels by High-Performance Liquid Chromatography .

Mallesh kurakula1, Abdul Bari Mohd1. , Samhuidrom2, Padmanabha Rao A2, Prakash V
Diwan have reported Estimation of Prednisolone in Proliposomal formulation using RP
HPLC method.

Al-Habet SM, Rogers HJ. have reported Two chromatographic methods for the
determination of corticosteroids in human biological fluids: pharmacokinetic applications.
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6.3 OBJECTIVES OF THE STUDY:In the proposed work attempts shall be made to:
 To develop new stability indicating analytical method for the estimation of
Prednisolone by chromatographic methods.

Validate the proposed method in accordance with BP, USP and ICH guidelines for
the intended analytical application.

Apply the proposed methods for analysis of Prednisolone in their dosage form and
in raw materials.
7.
Materials and methods:7.1 Source of data:

Literature survey was done at P.E.S. College of Pharmacy using Internet facilities
(RGUHS HELINET) and at Library.
 Strides Arco lab private limited, Bengaluru.
 Reference from library at RGUHS, Bengaluru.
 Indian Institute of Sciences, Bengaluru.
7.2 Method of collection of data:
Analytical method development on Prednisolone shall be carried out in collabration
with M/s STRIDES ARCO LABS PVT LIMITED (Analytical service department). The
analytical methods will be developed by using instruments like UV visible spectrometer,
HPLC, and HPTLC. The data so obtained is treated statistically to determine the compliance
of the experimental result as per ICH and Pharmacopeial guidelines and for the routine use of
the developed analytical methods in industry.
JOURNALS:
1. Indian Journal of Pharmaceutical Sciences.
2. Journal of Chromatography A
3. Journal of Chromatography B.
4. Asian Journal of Pharmaceutical and Clinical Research.
5. International Journal of Pharma Research and Development.
6. Journal of American Society for Mass Spectrometry.
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7. Journal of Pharmaceutical and Biomedical Analysis.
8. Journal of Medical Microbiology.
7.3. DOES THE STUDY REQUIRE ANY INVESTIGATION OR INTERVENTIONS TO BE
CONDUCTED ON PATIENTS OR OTHER HUMANS OR ANIMALS?
-: NO :-
7.4. HAS ETHICAL CLEARENCE BEEN OBTAINED FROM YOUR INSTITUTION
IN CASE OF 7.3?
-: NOT APPLICABLE :REFERENCES
1.
Introduction of Prednisolone. http://enwikipediaorg/wiki/prednisolone.
2.
ASHOK R, Prakash PP, Tamil SR. Development and Validation of Analytical Method for
Estimation of Prednisolone in Bulk and Tablets using UV-Visible Spectroscopy. Int J Pharm
Pharm Sci,. [Research]2011;3:184-6.
3.
El-Maboud A, Mohamed I, Hesham S, Eman M. Spectrophotometric determination of
binary mixtures of prednisolone with some antibiotics. Thai J. Pharm. Sci. 2006 30:1-24.
4.
Mohit R, Abhinav A, Ashish KJ, Narendra KL, Anil KK, Govind PA. Development of
Simultaneous Spectrophotometric Method of Mesalazine and Prednisolone in same dosage form
Int J Appl Pharm , [Research]. 2010;2:8-11.
5.
Singh DK, Rohan V. Spectrophotometric Determination of Corticosteroids and Its
Application in Pharmaceutical Formulation. Iranian Journal of Pharmacology & Therapeutics.
2008;7:61-5.
6.
Berend O, Ineke JMTB, Peter TAS, Richard PK, Chris JVB. Prednisolone Concentration-
Effect Relations in Humans and the Influence of Plasma Hydrocortisone. J. Pharmacol. Exp.
Therapeut. 1986;239:919-26.
7.
Alam MDA, Alka A, Sanjula B, K. GS, Ali J. Formulation and evaluation of
pharmaceutically equivalent parenteral depot suspension of methyl prednisolone acetate. Indian J
pharm sci, [Research]. 2009;71:30-4.
7
8.
Bhawani SA, Sulaiman O, Hashim R, Mohamad MN, Ibrahim. Thin-Layer
Chromatographic Analysis of Steroids: A Review. Trop J Pharm Res, [Review]. 2010;9:30113.
9.
Mary HC, William VH, Allen IL. Simultaneous Liquid-Chromatographic Determination
of Prednisone and Prednisolone in Plasma. Journal of Clinical Chemistry. 1988;34-9.
10.
Oneeb M, Fatemeh A, Terry L, David WH, Andrew T. Simultaneous Determination of
Plasma Prednisolone, Prednisone, and Cortisol Levels by High-Performance Liquid
Chromatography. Journal of Therapeutic Drug Monitoring. 2001;23:163–8
11.
Mallesh k, Abdul BM, Samhuidrom., Padmanabha R, Prakash VD. Estimation of
Prednisolone in Proliposomal formulation using RP HPLC method. International Journal of
Research in Pharmaceutical and Biomedical Sciences. [Research]. 2011;2:1663-9.
12.
Habet AS, Rogers H. Two chromatographic methods for the determination of
corticosteroids in human biological fluids: pharmacokinetic applications. J Pharm Sci.
1989;78(8):660-6.
Websites:
www.rguhs.ac.in
www.pubmed.com
www.medline.com
www.chemindustry.com
www.google.com
www.sciencedirect.com
www.ingentaconnect.com
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9.
Signature of the candidate
(Sahanapriya.C.N)
10. Remarks of the guide
Mr. VENKATESH PRASAD S.P
Asst.Professor
Department of Pharmaceutical Analysis,
P.E.S. College of Pharmacy,
Hanumanthanagar,
Bengaluru-560050
11. Name and Designation of
11.1 Guide
11.2 Signature
Dr. Nagaraj
Professor & Head
P.E.S College of Pharmacy,
Hanumanthanagar,
Bengaluru-560050
11.3 Head of the department
11.4 Signature
Prof. Dr. S. Mohan
Principal & Director,
P.E.S College of Pharmacy,
Hanumanthanagar,
Bengaluru-560050
12. Remarks of the Chairman and
Principal
12.1 Signature
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