Download anhydride which can be used for quantitative estimation of

“DEVELOPMENT OF NEW ANALYTICAL METHODS FOR THE
ESTIMATION OF CILNIDIPINE IN BULK DRUG AND
PHARMACEUTICAL FORMULATIONS”
M. PHARM DISSERTATION PROTOCOL
SUBMITTED TO
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
BANGALORE, KARNATAKA
By
SRAVANTHI GUDAVARTHI
Under the guidance of
Mr. PRAKASH S.SARSAMBI M.Pharm
DEPARTMENT OF PHARMACEUTICAL ANALYSIS
H.K.E.S’s COLLEGE OF PHARMACY,
SEDAM ROAD, GULBARGA – 585105
2010-2011
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA,
BANGALORE
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1.
Name of the Candidate and
SRAVANTHI GUDAVARTHI
Address (in block letters)
D/O G.JANARDHAN RAO, FLAT NO:307, SUN RISE
PARK APPARTMENTS, CZECH COLONY, STREET
NO:3,SANATHNAGAR,HYDERABAD-500016,
ANDHRA PRADESH.
2.
Name of the Institution
H.K.E.S’s COLLEGE OF PHARMACY, Sedam Road,
GULBARGA-585105, KARNATAKA.
3.
Course of Study and Subject
M. Pharm
(PHARMACEUTICAL ANALYSIS)
4.
Date of Admission to Course
5.
Title of the Research topic
18-06-2010
“DEVELOPMENT OF NEW ANALYTICAL
METHODS FOR THE ESTIMATION OF
CILNIDIPINE IN BULK DRUG AND
PHARMACEUTICAL FORMULATIONS”
6.
Brief Resume of Intended Work
6.1 Need for study
Cilnidipine is a new calcium channel blocker of the dihydropyridine type. It is used
as antihypertensive. It is a dual blocker of L-type voltage gated calcium channels in
vascular smooth muscle and N-type calcium channels in sympathetic nerve terminals that
supply blood vessels, which makes it an efficient antihypertensive drug. It is not official
in any pharmacopeia. And no spectrophotometric methods have been reported. Therefore
Investigations of some new instrumental methods are in need for the quantitative
estimation of cilnidipine in bulk drug and its pharmaceutical dosage forms with high
sensitivity, accuracy, precision and economical too.
6.2 Review of Literature
Cilnidipine1-2
is
chemically
1,4-Dihydro-2,6-dimethyl-4-(3-nitrophenyl)-3,5-
pyridinedicarboxylic acid 2-methoxyethyl (2E)-3-phenyl-2-propenyl ester. The Molecular
formula for Cilnidipine is C27H28N2O7. Molecular weight is 492.5.Its melting point is
115.5-116.6oc. It is used in the treatment of hypertension. It is soluble in chloroform,
acetone, slightly soluble in methanol, ethanol and water. Literature Survey reveals that
few physico chemical methods are reported for Cilnidipine in plasma. i.e., HPTLC
determination of Cilnidipine in pharmaceutical formulation3, LC-MS in plasma4, HPLC
with tandem Mass spectroscopic detection in plasma5-6. The structural formula for
Cilnidipine is as below
H3C
H
N
CH3
O
O
H3CO
O
O
NO2
Cilnidipine
6.3 Objectives of the Study:
The
analytical important function groups of Cilnidipine has not been completely
exploited for designing sensitive, precise, accurate and flexible spectrophotometric
methods for the determination of Cilnidipine in bulk drug and pharmaceutical
formulations. Hence, with the above mentioned fact the following instrumental methods
are planned to develop.
Since the drug Cilnidipine is sufficiently soluble in chloroform, acetone and slightly
soluble in methanol, ethanol and water a number of UV spectrophotometric methods can
be developed for its quantitative estimation in bulk drug and pharmaceutical dosage
forms.
(i). Since Cilnidipine is having the aromatic nitro group which can be reduced to primary
aromatic amino group with Zinc in HCl.
H
N
H3C
CH3
H3C
O
O
CH3
O
O
H3CO
H3CO
Zn/Hcl
O
H
N
O
O
NO2
O
NH2
CILNIDIPINE
(I) REDUCED CILNIDIPINE
I = R – NH2
Since, reduced cilnidipine (I) is having aromatic amino group it can be condensed with
various
aromatic
aldehydes
like
p-dimethylaminobenzaldehyde
(PDAB)
(II),
p-
dimethylamino cinnamaldehyde (PDACA) (III), vanillin to get colored Sciff’s base and can
be used for quantitative estimation of drug by visible spectrophotometry.
R NH2
R'
+ OHC
(I)
Reduced Cilnidipine
R N
HC
R'
(II)
Colored chromogen or schiff’s base
CH3
R - NH2
+
OHC - HC = HC
N
CH3
PDACA
(I)
(III)
CH3
R - N = HC - HC = HC
N
CH3
Schiff's Base
ii). Reduced Cilnidipine can be diazotised with nitrous acid (HNO2) and coupled with
chromogenic agents like N-(1-naphthyl) ethelenediamine dihydrochloride (BMR) to get
colored chromogens and drug can be estimated quantitatively by colorimetry.
R-NH2
NaNO2 / HCl
R-N+
NCl- +
NHCH2CH2NH2 . 2HCl
Reduced
Cilnidipine
Bratton-Marshall reagent
[N-(1-naphthyl) ethelenediamine
dihydrochloride]
R - N=N
NHCH2CH2NH2
Colored azo compound
Along with BMR reagent we can also use other coupling agents like -naphthol,
diphenylamine, N-phenyl aniline, methyl aniline to develop number of new analytical
method.
iii). The amino group in reduced Cilnidipine undergoes oxidative coupling reaction with 3methyl-2-benzothiazolinone hydrazone (MBTH) in presence of Ferric Chloride or
Cerric ammonium sulphate or Ferrous ammonium sulphate and forms the colored
chromogen by which the drug can be estimated quantitatively by colorimetry.
S
N
+
R-NH2
Reduced
Cilnidipine
N NH2
CH3
MBTH
FeCl3
S
+
N
CH3
Colored chromogen
N
N-R-NH2
iv). Reduced Cilnidipine forms colored complexes with 1,10-Phenanthroline and 2,2′bipyridine in presence of Fe (III) which can be utilized for quantitative estimation of
Cilnidipine in pharmaceutical formulations.
R - NH2
+
Fe3+
Fe2+
10
-
O-
na
o
Ph
sp
ho
ro
li
ric
ac
id
e
d
in aci
r id ic
py or
Bi ph
2'- os
2 , - Ph
O
1,
e
Ph
h
nt
ne
N
N
N
N
Fe2+
N
N
N
N
N
N
Fe2+
N
N
Colored chromogen
Colored chromogen
v). Phosphomolybdo tungstic acid well known as Folin-ciocalteu(FC reagent) reagent in
alkaline pH form colored chromogens with reduced Cilnidipine due to presence of
aromatic amino group by redox reaction which can be used for quantitative estimation
of Cilnidipine in bulk drug and its pharmaceutical dosage forms.
vi). Reduced Cilnidipine forms colored internal salt with Citric acid in presence of acetic
anhydride which can be used for quantitative estimation of Cilnidipine in bulk drug and
Pharmaceutical dosage forms.
vii). Reduced Cilnidipine reacts with Ninhydrin reagent in N,N-dimethylformamide medium
(DMF) producing colored product (Ruhemenn’s purple) which can be used for
quantitative estimation of Cilnidipine in bulk drug and its pharmaceutical dosage
forms.
O
OH
C
C
O
OH
C
R-NH2
+
2
C
Reduced
CILNIDIPINE
C
C
OH
O
N
C
C
O
O
Ninhydrin
Ruhemenn's purple
+
H3C
H3CO
O
H
N
CH3
O
O
O
NH2
Viii). Reduced Cilnidipine reacts with p-benzoquinone (PBQ) to form colored product which
can be used quantitatively for estimation of Cilnidipine in bulk and pure forms.
H3C
H3CO
H
N
CH3
O
O
O
+
O
O
O
PBQ Reagent
NH2
Reduced Cilnidipine
H
N
H3C
H3CO
CH3
O
O
O
O
N
O
( Purple color)
ix). Reversed phase HPLC method can also be developed using (shimadzu HPLC class VP
series 6.01 ) with two LC-10 AT VP pumps variable wave length programmable
UV/Visible detector for quantitative estimation of Cilnidipine in bulk drug and
pharmaceutical dosage forms with high accuracy, sensitivity, precision too.
7.
Material and Methods
In the present investigation of new Instrumental methods for quantitative estimation of
Cilnidipine, we are in need and using Shimadzu 1700 double beam UV/visible
spectrophotometer, HPLC (Shimadzu class VP series 6.01), chromatographic
instruments and volumetric glass apparatus. Drug sample will be provided by J.B
Chemicals & pharmaceuticals Ltd, Daman– 396210.
7.1 Source of Data
1. Internet, Library
2. Gulbarga University, Gulbarga
3. I.I.Sc. Library, Bangalore
4. I.I.C.T. Library, Hyderabad
5. R.G.U.H.S. Library, Bangalore
7.2 Methods of Collection of Data (including sampling procedure, if any)
Data collected from
1. Internet, H.K.E.S. College of Pharmacy, Gulbarga.
2. Analytical abstract and chemical abstract Gulbarga University, I.I.C.T. & I.I.SC.
Libraries.
3. Journals like – Indian J. Pharmaceutical Sciences, Indian Drug, Indian J. Analytical
Chemistry, Pharma review and Asian .J.Chemistry.
4. e-Journals
5. Drug sample of Cilnidipine will be collected from J.B. Chemicals & pharmaceuticals
Ltd, Daman-396210.
7.3 Does study require any investigation or interventions to be conducted on patients or
other humans or animals? If so please describe briefly
-No-
7.4 Has ethical clearance been obtained from your institution in case of 7.3
-Not Applicable-
8.
References
1. O’Neil M.J, editor, The Merck Index An Encyclopedia of Chemicals, Drugs and
Biologicals, Merck & Co. Inc. 14th edition, 2006: 379.
2. Sweetman SC, editor, Martindale The Complete Drug Reference, Pharmaceutical
Press, London (U.K), 35th edition,2007:1118.
3. Karmalkar HS, Vaidya VV, Gomes NA, Chooukekar MP and Kekare MB.
Determination of cilnidipine from pharmaceutical formulation by high performance
thin layer chromatographic method. Analytical chemistry 2008; 7(8).
4. Hatada K, Kimura M, Ono L and Ozaki M. Determination of a new calcium
antagonist and its main metabolite in plasma by thermospray liquid chromatography
– mass spectroscopy. J Chromaogr. 1992; 583: 116-121.
5. Xianhua zhang. Determination of Cilnidipine, a new calcium antagonist, in human plasma
using high performance liquid chromatography with tandem mass spectroscopic detection.
Analytical chemical acta. 2007; 142-146.
6. Lee HW, Seo JH, Lee HS, Jeong SY and Lee KT. Development of a liquid
chromatography/ negative ion electrospray tandem mass spectrometry for the
determination of cilnidipine in human plasma and its application to bioequivalence
study. J Chromatogr B Analyt Technol Biomed Life Sci. 2008; 862: 246-51.
9.
Signature of Candidate
10.
Remarks of the Guide
11.
Name & Designation of (in block
letters)
11.1
Guide
The work undertaken will be novel & Industrial
oriented. The work will be new research
findings. All facilities are available in our
college to undertake the work.
Mr. PRAKASH S.SARSAMBI, M.Pharm
ASSISTANT PROFESSOR
DEPT. OF PHARMACEUTICAL ANALYSIS,
H.K.E.S’s COLLEGE OF PHARMACY,
GULBARGA.
11.2
Signature
11.3
Head of the Department
Dr. S.M.MALIPATIL
M.Pharm., Ph.D
PROFESSOR
DEPT. OF PHARMACEUTICAL ANALYSIS,
H.K.E.S’s COLLEGE OF PHARMACY,
GULBARGA.
12
11.4
Signature
12.1
Remarks of the Chairman &
Principal
12.2
Signature