‘‘NEW ANALYTICAL METHODS AND THEIR VALIDATION FOR
THE ESTIMATION OF VORICONAZOLE AND LOPERAMIDE IN
BULK AND MARKETED FORMULATIONS’’
MASTER OF PHARMACY DISSERTATION PROTOCOL
SUBMITTED TO THE
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES KARNATAKA,
BANGALORE
BY
MAKVANA KEVIN KUMAR R.
Under The Guidance of
Dr. E.V.S. Subrahmanyam M.PHARM. Ph.D.
DEPARTMENT OF QUALITY ASSURANCE,
SRINIVAS COLLEGE OF PHARMACY, MANGALORE – 574143
2010 – 2012
1
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
BANGALORE, KARNATAKA
ANNEXURE - II
REGISTRATION OF SUBJECT FOR DISSERTATION
NAME OF THE
1.0
CANDIDATE
MAKVANA KEVIN KUMAR R.
DEPARTMENT OF Q.A.,
SRINIVAS COLLEGE OF PHARMACY,
ADDRESS
VALACHIL,POST PARENGIPITTE,
MANGALORE -574143
2.0
NAME OF THE
INSTITUTION
3.0
COURSE OF STUDY &
SUBJECT
4.0
DATE OF ADMISSION
5.0
TITLE OF THE TOPIC:
SRINIVAS COLLEGE OF PHARMACY,
VALACHIL, MANGALORE.
MASTER OF PHARMACY
(QUALITY ASSURANCE)
31ST MAY 2010
‘‘NEW ANALYTICAL METHODS AND THEIR VALIDATION FOR
THE ESTIMATION OF VORICONAZOLE AND LOPERAMIDE IN
BULK AND MARKETED FORMULATIONS’’
2
6.0
BRIEF RESUME OF THE INTENDED WORK:
NEED FOR STUDY:
Analytical Method Development for Pharmaceutical Formulations:
Analytical methods are essential to characterize drug substances and drug
products composition during all stages of pharmaceutical development. For routine
analytical purpose it is always necessary to establish methods capable of analyzing huge
number of samples in a short time period with high accuracy and precision
The number of drugs, which may be either new entities or partial structural
modification of the existing ones, introduced into the market is increasing every year.
Very often there is a time lag from the date of introduction of a drug into the market to
the date of its inclusion in pharmacopoeias. Hence, standards and analytical procedures
for these drugs may not be available in the pharmacopoeias. It becomes necessary,
therefore to develop new analytical methods for such drugs. These products can present
challenges to the analytical chemist responsible for the development and validation of
analytical methods.
Basic criteria for new method development of drug analysis:
1.
The drug or drug combination may not be official in any pharmacopoeias.
2.
A proper analytical procedure for the drug may not be available in the literature
due to patent regulations.
3.
Analytical methods may not be available for the drug in the form of a
formulation due to the interference caused by the formulation excipients.
4.
Analytical methods for a drug in combination with other drugs may not be
available.
5.
The existing analytical procedures may require expensive reagents and solvents.
It may also involve cumbersome extraction and separation procedures and these
may not be reliable.
Analytical method development provides the support to track the quality of the
3
product from batch to batch. Estimation can be performed by the following two
methods:

Titrimetric methods and

Instrumental methods.

Spectrophotometric Methods

Chromatographic Methods
Methods for analyzing drugs in dosage forms can be developed, provided one has
knowledge about the nature of the sample, its molecular weight, polarity, ionic character
and the solubility parameter. Method development involves considerable trial and error
procedures. The most difficult problem usually is where to start, what type of column is
worth trying with what kind of mobile phase.
The following is a suggested method development scheme for a typical HPLC-UV
related substance method.
1. To define the goals for method development (e.g., what is the intended use of the
method?) and to understand the chemistry of the analytes and the drug product.
2. To develop preliminary HPLC conditions to achieve minimally acceptable
separations. These HPLC conditions will be used for all subsequent method
development experiments.
3. To develop a suitable sample preparation scheme for the drug product
4. To determine the appropriate standardization method and the use of relative
response factors in calculations.
5. To identify the “weaknesses” of the method and optimize the method through
experimental design. Understand the method performance with different
conditions, different instrument set ups and different samples.
6. To complete method validation according to ICH guidelines as mentioned in
Q2 (R1).
6.1 BRIEF INTRODUCTION ABOUT VORICONAZOLE.
VORICONAZOLE is prepared by Pfizer pharma as a formulation and is used a
Triazole anti fungal medication that is generally used to treat serious, invasive fungal
infections as under brand name Vfend.
4
STRUCTURE :
IUPAC NAME : (2R,3S)-2-(2,4-difluorophenyl)-3-(5-fluoropyrimidin-4yl)-1-(1H1,2,4-triazol-1-yl)butan-2-ol.
MOLECULAR FORMULA : C16H14F3N5O
MOLECULAR WEIGHT : 349.311
CATEGORY: Anti-fungal drug
PROTEIN BINDING : 58%
METABOLISM : Hepatic cytochrome P450 enzymes CYP2C19, CYP2C9, CYP3A4
HALF-LIFE : Dose –dependent
MELTING POINT : 127-130 ˚C
5
DISSOCIATION CONSTANT : 2.71 , 11.54
STORAGE : store at controlled room temperature 20-25˚c
SOLUBILITY:
It is insoluble in aqueous solution of pH 4 and below. It very slightly
soluble in distilled water and acetonitrile, slightly soluble in ethanol, and freely soluble
in methanol.
PHARMACOLOGY:
MECHANISM OF ACTION:
Voriconazole is well absorbed orally with a bioavailability of 96%, allowing
patients to be switched between intravenous and oral administration. Being metabolized
by hepatic cytochrome P450, voriconazole interacts with some drugs. Administration is
contraindicated with some drugs (such as sirolimus, rifampin, rifabutin, and ergot
alkaloids) and dose adjustments and/or monitoring when coadministered with others
(including cyclosporine, tacrolimus, omeprazole, and phenytoin). Voriconazole may be
safely administered with cimetidine, ranitidine, indinavir, macrolide antibiotics,
mycophenolate, and prednisolone. Because voriconazole is metabolized by the liver, the
dose should be halved in patients with mild to moderate hepatic impairment (Child-Pugh
score A or B). There is no data available for patients with severe hepatic impairment
(Child-Pugh C). No dose adjustment is necessary for renal impairment or advanced age,
but children seem to clear voriconazole faster than adults and drug levels may need
monitoring.
ADVERSE EFFECT:
The most common side effects associated with voriconazole include transient
visual disturbances, fever, rash, vomiting, nausea, diarrhea, headache, sepsis, peripheral
edema, abdominal pain, and respiratory disorder.
6
Unlike most adverse effects, which are similar to other azole antifungal agents,
visual disturbances (such as blurred vision or increased sensitivity to light) are unique to
voriconazole. These have been reported by more than 30% of patients in clinical trials.
6.1.1 OBJECTIVES OF THE STUDY:
In the proposed work, attempt shall be made :
 To develop a new instrumental method for estimation of Voriconazole.
 To developed a validated method according to ICH guidelines.
 To apply validated method for the estimation of voriconazole in pharmaceutical
formulation.
6.1.2 REVIEW OF LITERATURE:
A literature survey was carried out for the estimation of Voriconazole in bulk and
marketed dosage forms. It was found that very few methods have been reported for this
drug. The collection of references are reproduced below:

Adams AIH, Bergold AM.1 developed and validated high performance liquid
chromatographic method for determine voriconazole content in tablets by using
stationary phase was merck Li chrospher 100 RP-8 (125mm× 4.6mm I.D., 5µm
partical size) column. Mobile phase was methanol: triethylamine solution o.6 %,
PH 6.0 (50:50 v/v). Flow rate was 1 ml/min. range was 20.0-100.0 µg/ml. and
detected at UV range 255 nm.

Srinubabu G, Raju AI, Sarath N, Kirankumar P, Seshagiri RJVLN.2 developed
and validated high performance liquid chromatographic method for determine
voriconazole in pharmaceutical formulation using an experimental design by
using stationary phase was a reversed phase C 18 column (25cm × 4.6mm I.D.,
particle size 5µm). Mobile phase was acetonitrile: water (50:50, v/v). flowrate
was 1 ml/min. range was 5.0-50.0 µg/ml. and detected at UV range 260 nm.
7

Stopher DA, Gage RG.3 developed a rapid high performance liquid
chromatographic assay for voriconazole in human plasma, by using stationary
phase was a Kromasil C 18, 5mm, 250_4.6mm with a 10× 3.2 mm guard
cartridge
packed
with
same
material.
Mobile
phase
was
acetonitrile: ammonium phosphate buffer. (pH 6.0.,0.04M) (50:50v/v). Flow rate
was 0.8ml/min. range was 0.2-10µg/ml.and detected at UV range 255nm.

Lei Z, Glickman RD, Nancy C, Sponsel WE, Graybill JR, Kwokwai LD.4
voriconazole in aqueous humor by liquid chromatography –
determine
electrospray ionization – mass spectrometry , by using stationary phase was delta
PAK C 18 analytical column , (15- mm particle size ,30033.9mm). mobile phase
was used acetonitrile : water: TFA mixture(70:29.99:0.01v/v/v).flow rate was
0.5 ml/minute.range was 0.2 to 10 µg/ml.then it was detected in UV range
255nm.

Pehourcq F, Jarry C, Bannwarth B.5 developed direct injection HPLC micro
method for the determination of voriconazole in plasma using an internal surface
reversed
phase
column.by using chromolith
RP
18e(100mm× 4.6mm)
monolithic silica rod HPLC column as a stationary phase. Mobile phase used was
acetonitrile:
potassium
di
hydrogen
phosphate
buffer,
100mm,
PH
6.0(17:83V/V), Flow rate was 1µl/minute at a range of 0.5 to 10µg/ml at 255nm
in UV.

Shihabi ZK.6 performed HPLC of voriconazole assay in serum. for that he used
chromrlith
RP
18
e(250mm× 4.6mm)
monolithic
silica
rod
HPLC
column.mobile phase was ammonium di hydrogen carbonate buffer(PH
6.5):acetonitrile(70:30v/v) at a flow rate 1 ml/minute at a range of 0.1 to
5µg/ml.at a 255nm in UV.

Brian K, Sheila GN, Stephen I, Susan HJ, Caroline MB, David DW.7 validate
an assay for voriconazole in serum sample using liquid chromatography-tandem
mass spectrometry in that C 18 cartridge ( 2 mm × 4 mm ) as stationary phase .
Mobile phase was step gradient of 50% to 100% methanol containg 2mM
(mili_molar) ammonium acetate and 0.1% v/v formic acid. Flow rate was
8
0.6ml/min at range 0.38 to 15.3 mg/lit, detection of UV range was at 254 nm.

Cheng S, Qui F, Huang J, He J.8 developed and validate simple and rapid HPLC
method for quantitative determination of voriconazole in rat and dog plasma by
using diamonsil C18 column (250×4.6-mm) as stationary phase. Mobile phase
was Acetonitrile : water: acetic acd (55:45:0.25, v/v/v) with PH of 4.0 with flow
rate was 1.0 ml/min.; range was 0.1 to 50.0µg/ml and detected at256 nm UV
range.

Michael C, Teichert J, Preiss R.9 developed determination of voriconazole in
human plasma and saliva using HPLC with fluorescence detection by using luna
C 18 column as a stationary phase , mobile phase was used aceto nitrile :
methanol : water (50:30:20, v/v/v) at 1.0 ml/min flow rate with range of 0.1 to
10 µg/ml . Detection, fluorescence with range 254 nm and 372 nm.

Pennick GJ, Clark M, Sutton DA, Rinaldi MG.10 develop validate HPLC assay
for voriconazole by using reverse phase Luna 5µm C 18 column as a stationary
phase. Mobile phase was methanol: water (50:50 v/v) with flow rate of 1.0
ml/min, range was 0.2 to10 µg/ml, detect at UV range 254 nm.
6.2 BRIEF INTRODUCTION ABOUT LOPERAMIDE.
Loperamide is a synthetic piperidine derivative, is an opioid drug effective against
diarrhea resulting from gastroenteritis or inflammatory bowel disease. In most
countries it is available generically and under brand names such as Lopex, Imodium,
Dimor, Fortasec and Pepto Diarrhea Control. It was developed at Janssen
Pharmaceutical.
9
STRUCTURE :
IUPAC NAME:
4-[4-(4-chlorophenyl)-4-hydroxypiperidin-1-yl]- N,N-dimethyl-2,2Di-phenylbutanamide.
MOLECULAR FORMULA : C29H33Cl N2O2
MOLECULAR WEIGHT :
477.037
CATEGORY: Anti-diarrheal drug.
BIOAVAILABILITY : Not significantly absorbed from the gut
PROTEIN BINDING : 97%
METABPLISM : hepatic
SOLUBILITY: It is soluble in methanol.
ROUTES : oral, insufflations
10
PHARMACOLOGY :
MECHANISM OF ACTION :
Loperamide is an opioid-receptor agonist and acts on the μ-opioid receptors in the
myenteric plexus of the large intestine; by itself it does not affect the central nervous
system like other opioids.
It works by decreasing the activity of the myenteric plexus, Loperamide also decreases
colonic mass movements and suppresses the gastrocolic reflex. Loperamide molecules
do not cross the blood-brain barrier in significant amounts, and, thus, it has no analgesic
or euphoric properties. Any that do cross the blood-brain barrier are quickly exported
from the brain by P-glycoprotein also known as multidrug resistance protein. Tolerance
in response to long-term use has not been reported. However, loperamide has been
shown to cause a mild physical dependence during preclinical studies, specifically in
mice, rats, and rhesus monkeys. Symptoms of mild opiate withdrawal have been
observed following abrupt discontinuation of long-term therapy with loperamide.
ADVERSE EFFECT :
Adverse drug reactions (ADRs) associated with Loperamide includes abdominal pain
and bloating, nausea, vomiting and constipation. Rare side-effects associated with
Loperamide are paralytic ileus, dizziness and rashes.
6.2.1 OBJECTIVES OF THE STUDY:
In the proposed work, attempt shall be made :
 To develop a new instrumental method for estimation of Loperamide.
 To developed a validated method according to ICH guidelines.
 To apply validated method for the estimation of Loperamide in pharmaceutical
formulation.
11
6.2.2 REVIEW OF LITERATURE:
A literature survey was carried out for the estimation of Loperamide in marketed
dosage forms. It was found that very few methods have been reported for this drug. The
collection of references are reproduced below:

El-sherif ZA, Mohamed AO, Walash MI, Tarras FM.11 was developed
Spectrophotometric determination of loperamide hydrochloride by acid-dye and
charge-transfer complexation methods in the presence of its degradation products
by Two simple, sensitive and accurate spectrophotometric methods for the
determination of loperamide hydrochloride are described. The first method is
based on the formation of ion-pair association complex (1:1) with bromothymol
blue (BTB), bromophenol blue (BPB) and naphthol blue black B (NBB). The
coloured
products
are
extracted
into
chloroform,
and
measured
spectrophotometrically at 414 (BTB), 415 (BPB) and 627 nm (NBB). The second
method is based on the reaction of the basic loperamide with iodine in
chloroform to give molecular charge-transfer complex with intense bands at 295
and 363 nm. The proposed methods have been applied successfully for the
analysis of the drug in pure form and in its dosage forms.

Hewalla II.12 was developed simple, sensitive and rapid spectrofluorimetric and
derivative absorption spectrophotometric procedures are described for the
accurate determination of loperamide hydrochloride. The spectrofluorimetric
method involves the measurement of the fluorescence of the compound in an
ethanol-sulphuric acid mixture (90:10, v/v). The derivative spectrophotometric
method involves the measurement of either the second derivative peak amplitude
(crest to trough, i.e. maximum to minimum) between 258 and 263 nm or the
second derivative peak height (i.e. maximum to zero line) at 224 nm of an
ethanolic solution of the drug. The proposed methods have been used for the
determination of loperamide in pharmaceutical formulations.

Streel B, Ceccato A, Klinkenberg R, Hubert PH, Galephar MF.13 was developed
and Validated
A sensitive and selective method based on liquid
chromatography-tandem mass spectrometry (LC-MS/MS) has been developed
12
for the quantitative determination of loperamide in human plasma.After
conditioning, the plasma sample is loaded on the DEC filled with endcapped
ethyl silica (C2(EC)) and washed twice with water. The analytes are therefore
eluted by dispensing methanol. The eluate is then collected and added with
ammonium acetate solution in order to inject an aliquot of this final extract in the
LC-MS/MS system. On-line LC-MS/MS system using atmospheric pressure
chemical ionization (APCI) has been developed for the determination of
loperamide. The separation is obtained on a octadecylsilica based stationary
phase using a mobile phase consisting in a mixture of methanol and 5mM
ammonium acetate solution (25:75, v/v).The limit of quantitation (LOQ) was
around 50 pg/ml for loperamide.

Jin HY, Hye JK, Sibeum L, Sung JH, Won K, Cheol JM.14 developed a simple
rapid and selective LC-MS method
was developed and validated for the
determination of loperamide hydrochloride in human plasma, Methyl tertbutylether (MTBE) was used to extract loperamide hydrochloride
from an
alkaline plasma sample. LC separation was performed on a Zorbax RX C18
column (5 microm, 2.1 mm x 150 mm) using acetonitrile-water-formic acid
(50:50:0.1 (v/v)) as a mobile phase. The retention time of loperamide
hydrochloride was 1.2. The developed method was successfully used to study
the bioavailability of a low dose (8 mg) of loperamide hydrochloride.

Huizhong z, Fei N, Jiuru L.15 was established for the determination of
loperamide
hydrochloride.
The
possible
mechanism
for
the
Post-
chemiluminescence reaction was discussed via the investigation of the CL
kinetic characteristics, the CL spectra, the fluorescence spectra. The PCL
intensity responded linearly to the concentration of loperamide hydrochloride in
the range 8.0 10-10 to 6.0 10-7 g · ml-1 with a linear correlation of 0.9995. The
detection limit was 4 10-10 g · ml-1. The relative standard deviation was 2.4% for
4.0 10-8 g · ml-1 loperamide hydrochloride (n=11). This method has been
applied to the determination of loperamide hydrochloride in human plasma and
pharmaceutical samples with satisfactory results.
13

Huaibing HE, Abu S, John CLE, Alastair JJW, David LH.16 Development and
validation of an LC–MS method for quantitation of loperamide (LOP) and its Ndemethyl metabolite (DMLOP) in human plasma. O-Acetyl-loperamide (A-LOP)
was use as an internal standard in the assay. After addition of the internal
standard, the compounds of interest were extracted with methyl tert.-butylether
and separated by HPLC on a C18 reversed-phase column using an acetonitrile–
water gradient containing 20 mM ammonium acetate. The three compounds were
well separated by HPLC and no interfering peaks were detected at the usual
concentrations found in plasma. Analytes were quantitated using positive
electrospray ionization in a triple quadrupole mass spectrometer operating in the
MS–MS mode. The intra- and inter-assay variability of LOP and DMLOP ranged
from 2.1 to 14.5% for the low, medium and high quality control samples. The
method has been used successfully to study loperamide pharmacokinetics in
adult humans.
7.0
MATERIALS AND METHODS:
 All experiments will be carried out in the Department of Quality Assurance.
Srinivas college of Pharmacy, Valachil, Mangalore.
 Pure sample of voriconazole and loperamide will be procured from Industries
involved in bulk manufacture of this drug.
 Dosage formulation will be procured from local market.
 The methods will be developed and validated in Q.A. lab of Srinivas college
of Pharmacy.
 The methods will be first developed, then Validated as per ICH guidelines,
then the method will be applied to the formulations.
 UV spectrophotometer Shimadzu-UV1700 with spectral band width of 2nm
and 10nm and matched quartz will be used for measuring absorbance for
voriconazole solutions and loperamide solution.
14

We are supposed to use following type of reagents
1) 1,10- phenanthroline
2) (MBTH), 3- methyl, 2- benzothiazoline hydrazine
3) 4- amino phenazene
4) Para di methyl amino benzaldehyde- orthophosphoric acid
5) (FCR) Folin ciocaltaeau reagent
In combinations,
 3-methyl 2-benzothiazoline hydrazine and cerric ammonium sulphate
 3-methyl 2-benzothiazoline hydrazine and ferric ammonium sulphate etc.
Can be used as reagents for Spectrophotometric method development.
7.1 Sources of data:
 References from library – Srinivas College of Pharmacy, Valachil,
Mangalore.
 www.pharmainfo.net.
 www.google.com
 www.sciencedirect.com
 www.rxlist.com
 www.pubmed.com
 www.medline.com
 www.Wikipedia.com
 www.springerling.com
 www.ncbi.nlm .nih.gov/pmc/articles.com
15
7.2 Does the study require any investigation to be conducted on
patients or animals?
No
7.3 Has the ethical clearance been obtained from your institution in
case of 7.2?
Not applicable
8.0
REFERENCES:
1.
Adams AIH, Bergold AM. Development and validation of a high performance
liquid chromatographic method for the determination of voriconazole in tablet
content. Chromatographia 2005; 62(7-8):429-34.
2.
Srinubabu G, Rraju AI, Sarath N, Kirankumar P, Seshagiri RJVLN.
Development and validation of a high performance liquid chromatographic
method for the determination of voriconazole in pharmaceutical formulation
using an experimental design. Talanta 2007; 71:1424-29.
3.
Stopher DA, Gage RA. A rapid high performance liquid chromatography assay
for voriconazole in human plazma. J Pharm Biomed Anal 1998; 17:1449-53.
4.
Lei Z, Glickman RD, Nancy C, Sponsel WE, Graybill JR, kwok-wai LDJ.
Determination of voriconazole in aqueous humor by liquid chromatographyelectrospray ionization-mass spectrometry. J Chromatogr B 2002; 776:213-20.
5.
Pehourcq F, Jarry C, Bannwarth B. Direct injection high performance liquid
chromatographic micro method for the determination of voriconazole in plasma
using an internal surface reversed-phase column. Biomed chromatogr 2004; 18
(9):719-22.
6.
Shihabi ZK. Simple assay for voriconazole in serum by high performance liquid
chromatography. J Liq Chromatogr Tech 2008; 31(2):263-68.
7.
Brian K, Sheila GN, Stephan I, Susan HJ, Caroline MB, David DW. Validation
of an assay for voriconazole in serum sample using liquid chromatographytandem mass spectrometry. Therap Drug Monitor 2004; 26(6):650-57.
16
8.
Cheng S, Qiu F, Huang J, He J. Development and validation of a simple and
rapid high performance liquid chromatographic method for the quantitative
determination of voriconazole in rat and dog plasma. J Chromatogr Sci 2007;
45(7):409-14.
9.
Michael C, Teichert J, Preiss R. Determination of voriconazole in human
plasma and saliva using high performance liquid chromatography with
fluorescence detection. J Chromatogr B 2008; 865(1-2):74-80.
10.
Pennick, GJ, Clark M, Sutton DA, Rinaldi MG. Development and validation of
a high performance liquid chromatography assay for voriconazole. Antimicrob
agents Chemother 2003; 47: 2348-50.
11.
El-Sherif ZA, Mohamed AO, Walash MI, Tarras FM. Spectrophotometric
determination of loperamide hydrochloride by acid-dye and charge-transfer
complexation methods in the presence of its degradation products. J Pharm
Biomed Anal 2000; 22(1):13-23.
12.
Hewalla II. Spectrofluorimetric and derivative absorption spectrophotometric
techniques for the determination of loperamide hydrochloride in pharmaceutical
formulations. J Pharm Biomed Anal 1995;13(6):761-67.
13.
Streel B, Ceccato A, Klinkenberg R, Hubert PH, Galephar MF. Developed and
Validation of a liquid chromatographic-tandem mass spectrometric method for
the determination of loperamide in human plasma. J Chromatogr B Analyt
Technol Biomed Life Sci 2005; 814(2):263-73.
14.
Jin HY, Hye JK, Sibeum L, Sung JH, Won K, Cheol JM.LC-MS Determination
and bioavailability study of loperamide hydrochloride after oral administration
of loperamide capsule in human volunteers. J Pharm Biomed Anal 2004; 36
(2):421-27.
15.
Huizhong Z, Fei N, Jiuru L. Developmend and validation of loperamide
hydrochloride by Post-chemiluminescence spectra and fluorescence spectra.
Analytical letter 2007;40(18):3405–16.
16.
Huaibing H, Abu S, John CLE, Alastair JJW, David LH. Development and
validation of an LC–MS method for quantitation of loperamide in human
plasma. J Chromatogr B Biomed Sci App 2000; 744(2): 323-31.
17
9.0
SIGNATURE OF THE CANDIDATE
10
REMARKS OF THE GUIDE
11
12
NAME AND DESIGNATION OF
GUIDE
Makvana Kevin kumar R.
Forwarded for Approval
Dr. E.V.S. Subrahmanyam,
Professor and Head,
Dept of Quality Assurance
SIGNATURE
Dr. E.V.S. Subrahmanyam,
13
HEAD OF THE DEPARTMENT
Professor and Head,
Dept of Quality Assurance
14
15
16
SIGNATURE
REMARKS OF THE PRINCIPAL
Forwarded for approval
SIGNATURE
Dr.Ramakrishna Sharbaraya A.
Principal,Srinivas college of
Pharmacy, Valachil, Mangalore.
18