“DEVELOPMENT OF NEW ANALYTICAL METHODS AND THEIR VALIDATION

FOR THE DETERMINATION OF ETODOLAC AND ANALGIN IN BULK AND

MARKETED FORMULATIONS”

MASTER OF PHARMACY

DISSERTATION PROTOCOL

SUBMITTED TO THE

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA,BANGALORE

BY

BHALANI MONABEN CHIMANBHAI

Under The Guidance ofDr.E.V.S. Subrahmanyam. M.PHARM. Ph.D

P.G. DEPARTMENT OF QUALITY ASSURANCE,SRINIVAS COLLEGE OF PHARMACY, MANGALORE – 574143

2013-2015

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

BANGALORE, KARNATAKA

ANNEXURE –II

REGISTRATION OF SUBJECT FOR DISSERTATION

1.0 NAME AND ADDRESSOF

THE CANDIDATE

BHALANI MONABEN CHIMANBHAI

DEPARTMENT OF QUALITY

ASSURANCE,

SRINIVAS COLLEGE OF PHARMACY,

VALACHIL,POST PARENGIPETE,

MANGALORE TQ-574143

2.0 NAME OF THE

INSTITUTION

SRINIVAS COLLEGE OF PHARMACY,

VALACHIL, MANGALORE.

3.0 COURSE OF STUDY &

SUBJECT

MASTER OF PHARMACY

(QUALITY ASSURANCE)

4.0 DATE OF ADMISSION 25th JULY 2013

5.0 TITLE OF THE TOPIC:

“DEVELOPMENT OF NEW ANALYTICAL METHODS AND THEIR

VALIDATION FOR THE DETERMINATION OF ETODOLAC AND ANALGIN

IN BULK AND MARKETED FORMULATIONS”

6.0 BRIEF RESUME OF THE INTENDED WORK:

6.1 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 large 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.

6.2 Basic criteria for new method development of drug analysis:

The drug or drug combination may not be official in any pharmacopoeias.

A proper analytical procedure for the drug may not be available in the literature due

to patent regulations.

Analytical methods may not be available for the drug in the form of a formulation

due to the interference caused by the formulation excipients.

Analytical methods for a drug in combination with other drugs may not be

available.

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 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.

Method development scheme for a typical HPLC-UV related substance describe below,

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 an 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).

6.3 A BRIEF INTRODUCTION ABOUT ETODOLAC1,2:

Chemical structure of Etodolac:

Chemical name: (RS)-2-(1,8-Diethyl-4,9-dihydro-3H-pyrano[3,4-b]indol-1-yl)acetic acid

Empirical formula: C17H21NO3

Molecular weight: 287.35

Solubility: Methanol and Ethanol

Protein binding: 100%

Half-life: 7.3±4.0 hours

Excretion: renal 

Melting point: 145-148ºC

Characteristics: White crystalline compound

PHARMACOLOGY1,2:

DRUG CATEGORY:

Anti-inflammatory Agents

Anti-rheumatic Agents

MECHANISM OF ACTION:

NSAIDs are used for the management of mild to moderate pain, fever, and inflammation.

They work by reducing the levels of prostaglandins, which are chemicals that are

responsible for pain and the fever and tenderness that occur with inflammation. Etodolac

blocks the enzyme that makes prostaglandins (cyclooxygenase), resulting in lower

concentrations of prostaglandins. As a consequence, inflammation, pain and fever are

reduced.

Post-marketing studies demonstrated that etodolac inhibition of cyclooxygenase is

somewhat COX-2 selective [2] similar to celecoxib and other "COX-2 inhibitors." Unlike

rofecoxib, both etodolac and celecoxib can fully inhibit COX-1 and are designated as

having "preferential selectivity" toward COX-2. The (inactive against COX) r-enantiomer

of etodolac inhibits beta-catenin levels in hepatoma cells.

SIDE EFFECTS:

Etodolac may cause side effects, or adverse drug reactions. It is advised to contact a

physician if any of these symptoms are severe or persistent: Constipation, diarrhoea, gas or

bloating, vomiting, headache, dizziness, ringing in the ears, running nose, sore throat and

blurred vision.

Some side effects can be serious. It is advised if any of the following symptoms occur to

immediately contact a physician, and to discontinue use until such a time.

Unexplained weight gain, swelling of the eyes, face, lips, tongue, throat, hands, feet,

ankles, or lower legs, fever or chills, blisters, rash, itching, hives, hoarseness, difficulty

breathing or swallowing, yellowing of the skin or eyes, excessive tiredness, unusual

bleeding or bruising, lack of energy, loss of appetite, pain in the upper right part of the

stomach, flu-like symptoms, pale skin, fast heartbeat, cloudy, discolored, or bloody urine,

difficult or painful urination, back pain.

6.4 REVIEW OF LITERATURE:

A great deal of work has been done by the scientist about the current application and future

possibilities for altering the drug activities and evaluation with new method development

by instrumental methods.

1. Ayman A Gouda and Wafaa S Hassan3 have reported Spectrophotometric

determination of Etodolac in pure form and pharmaceutical formulations. The

determination was based on the oxidation of etodolac by Fe3+ in the presence of o-

phenanthroline(o-phen) or bipyridyl(bipy) and/or Fe+3-bipy mixtures in acetate

buffer solution at optimum pH at 510 & 520 nm with o-phen&bipy. Standard

deviations were ≤ 0.76% with recoveries of 99.87% - 100.21%.

2. Mit J. Patel, R. Badmanaban and C. N. Patel4 have reported A reversed-phase

liquid chromatographic (RP-HPLC) method was developed for the simultaneous

determination of tolperisone hydrochloride (TOLP) and etodolac (ETD) in a

combined fixed dose oral formulation. The analysis was carried out using a

phenomenax C-18, pre-packed column. A mobile phase containing a phosphate

buffer (pH 5.5) : Methanol : Acetonitrile : Tri-ethylamine (40 : 40 : 20 : 1.5), with

the pH adjusted to orthophosphoric acid, was pumped at a flow rate of 1.0 ml min1

with a UV-detector and PDA detection at 257 nm. Retention time was 3.91 minutes

and 6.89 minutes for TOLP and ETD, respectively. The method was validated for

linearity, accuracy, precision, sensitivity, and specificity. The method showed good

linearity in the range of 3 – 21 μg ml for TOLP μg / ml and 8 – 56 μg / ml for ETD.

The detection limit of the proposed method was 0.16 μg / ml and 0.58 μg / ml for

TOLP and ETD, respectively. The quantification limit of the proposed method was

0.51 μg / ml and 1.7 μg / ml for TOLP and ETD, respectively. The % recovery was

within the range of 99.42 – 101.15 for TOLP and 98.63 – 100.94 for ETD. The

percentage RSD for precision of the method was found to be less than 2%. The

method was validated as per the International Conference on Harmonization (ICH)

guidelines. The developed method could be applied for routine analysis of TOLP

and ETD in tablet dosage form.

3. Vaijanath G. Dongre, Sweta B. Shah, Gunaji S. Bayer, Manisha Phadke,

Vivek K. Jadhav5 have reported Simultaneous Determination of Etodolac and

Acetaminophen in Tablet Dosage Form by RP-LC. A simple, specific, precise and

accurate reverse phase liquid chromatographic (RP-LC) method has been developed

for the simultaneous determination of etodolac and acetaminophen in tablet dosage

form. The chromatographic separation was achieved on a BDS Hypersil C18, 100

mm × 4.6 mm, 5 μm column at a detector wavelength of 274 nm using an isocratic

mobile phase consisting of a mixture of 0.05% aqueous orthophosphoric acid and

acetonitrile in the ratio of 50:50 (v/v) at a flow rate of 1.0 mL min−1. The retention

times for etodolac and acetaminophen were found to be 1.32 and 4.24 min,

respectively. The method was validated for the parameters like specificity, linearity,

precision, accuracy and robustness. The method was found to be specific and

stability indicating as no interfering peaks of impurities, degradant and excipients

were observed. The square of correlation coefficients (R 2) for etodolac and

acetaminophen were 0.9996 and 0.9998 while percentage recoveries were 101.32

and 100.94%, respectively. Intra- and inter-day relative standard deviations for both

the components were <2.0%. The proposed RP-LC method can be applied for the

routine analysis of commercially available formulations of these drugs either as

such or in combination.

4. Jadhav Alpa V., Gohel A., Bhavika., Sondagar Mital M., Patel Bhavna A.,

Parmar Shraddha J.6 have reported Method Development And Validation For

The Simultaneous Estimation Of Paracetamol And Etodolac By Derivative UV

Spectroscopic Method. A simple, novel, sensitive, precise and specific validated

Spectrophotometric method was developed for simultaneous determination of

Paracetamol and Etodolac in synthetic mixture and its dosage form. Methanol:

Water (60:40) was selected as a common solvent for estimation of Paracetamol and

Etodolac with λmax at 247 nm and 280 nm respectively in methanol: water (60:40

v/v). Derivative method was selected for the estimation of both the drug

simultaneously. The linearity was obtained in the concentration ranges of 5-25

µg/ml for Paracetamol and 2-18 µg/ml for Etodolac. The Zero Crossing Point

(ZCP) of Paracetamol was 219.27 nm and Etodolac was 224.28 nm. The correlation

coefficient was found to be 0.9994 and 0.9983 for Paracetamol and Etodolac

respectively. The detection limit and quantification limit were found to be 0.34 and

1.02 µg/ml for Paracetamol and 0.37 and 1.11 µg/ml for Etodolac respectively. The

method was validated as per the International Conference on Harmonization (ICH)

guidelines.

5. Thankappan Surya; Parmar Ashok; Sailor Bhavika; Vekariya Kinjal;

Khasia Vasant7 have reported Development and Validation of Spectroscopic

method for Simultaneous Estimation of Etodolac and Thiocolchicoside in tablet

formulation. Simple, precise and economical spectrophotometric method has been

developed for simultaneous estimation of Etodolac and Thiocolchicoside in

combined tablet dosage form. The first method is based on the use of simultaneous

equation method(Method A) and the second method is based on Absorbance

correction method(Method B). In method A, absorbance is measured at two

wavelengths, one being λmax of etodolac 223nm and the other being λmax of

thiocholchicoside at 260nm. In method B, one wavelength is the λmax of etodolac

223nm and other wavelength is 360nm(where ETD does not interfere in absorbance

of TCD). Both the drugs obey the Beer's law in the concentration ranges employed

for these methods. The methods were validated by following the analytical

performance parameters suggested by International Conference on Harmonization.

All validation parameters were within the acceptable range. These developed

methods can be applied for routine analysis.

6. Rohit Shah, Chandrakant Magdum, Shitalkumar Patil, Dhanya Kumar

Chougule and Nilofar Naikwade8 have reported Validated Spectroscopic Method

for Estimation of Aceclofenac from Tablet Formulation. Aceclofenac is a non

steroidal anti-inflammatory drug with good analgesic and anti-rheumatic properties.

Various methods for analysis of the same are available but are time consuming and

expensive. Here they have developed a new, precise and simple UV

spectrophotometric method for estimation of aceclofenac from tablet formulation.

The drug obeyed the Beer’s law and showed good correlation. It showed absorption

maxima at 273 nm; in phosphate buffer pH 7.4. The linearity was observed between

0 – 20 mcg/mL. The results of analysis were validated by recovery studies. The

recovery was more than 99%. The method was found to be simple, accurate,

precise, economical and robust.

6.5 A BRIEF INTRODUCTION OF ANALGIN9:

Chemical structure ofAnalgin:

Iupac name: sodium [(2,3-dihydro-1,5-dimethyl-3-oxo-2-phenyl-1H-pyrazol-4-yl)methylamino] methanesulfonate

Synonyms: Metimazole, Novalgin, Dipyrone

Formula:C13H16N3O4S.Na

Molecular weight: 333.337 Solubility : Water and Ethanol

Bioavailability: 85%

Protein binding: 58-60%

Half-life:1-4 hours

Excretion: renal

Melting point : 222-226 °C

Characteristics: A white or yellowish crystalline powder.

PHARMACOLOGY10,11:

MEDICAL USES:Analgin is used for the treatment of pains of different origin and variable intensity:

toothache, headache, arthralgia, neuralgia, myositis, mild to moderate visceral pain, high

fever, not responding to other drugs.

MECHANISM OF ACTION:

The exact mechanism and location of action of metamizole are not explained in detail. It is

assumed that it operates in a combined central and peripheral effects. The central action is

explained by inhibition of prostaglandin synthesis, secretion of histamine and serotonin

from mast cells of thalamus. Peripheral effect is explained by the inhibition of

prostaglandin synthesis which sensitize nociceptors to the effect of algogen mediators. In

addition, metamizole has a direct blocking effect on the inflammatory hyperalgesia.

Analgin is a medicine of the pyrazolone group, possessing hard analgesic and antipyretic

effects and moderate anti inflammatory activity. Blocking of the synthesis of endogenous

pyrogens - prostaglandins D and E - is the cause for the antipyretic activity and also for the

analgesic action of this drug. The decrease of the prostaglandins production in the

periphery (and respective decrease of nerve endings sensitivity) plays a relatively smaller

role. In contrast to the other nonnarcotic analgesic drugs, Analgin stimulates the release of

β-endorphins, explaining its activity in cases of visceral pain. Analgin has a slight

spasmolytic activity on the smooth muscle cells of the biliary and urinary tracts and also on

the muscle of the uterus.

SIDE EFFECTS:

The serious adverse reactions after administration might include a shock. Signs of the

starting shock are cold sweat, dizziness, nausea, skin discoloration, shallow breathing. It is

accompanied by precordial tightness, rapid pulse, cold extremities and a sharp drop in

blood pressure. Immediately after the onset of first symptoms the administration should be

discontinued and the intense shock therapy must be applied.

Another serious side effect is blood dyscrasias (agranulocytosis, leukopenia,

thrombocytopenia).

Uncommon side effects: allergic rash, low blood pressure.

Rare side effects: spotty pustel-rashes, white-blood cell-deficiency.

Very rare side effects: Schmerzmittel-asthma, severe skin reactions (Stevens-Johnson

Syndrome, Lyell Syndrome), circulatory shock, lack of granulocytes, lack of platelets,

deterioration of kidney function, protein in the urine, urinary excretion deficiencies,

Harnsperre, inflammation of the blood vessels (for injection), emotional disorders (anxiety,

agitation, delirium, depression, delusions, apathy).

Specialties: Vereinzelt particularly serious cases of skin reactions can occur such as the

Toxic Epidermal Necrolysis (TEN). Called this also staphylogenes Lyell Syndrome

cutaneous reaction leads to the large blasigen skin separation and is a severe, life-

threatening disease.

During the application of Analgin there may be agranulocytosis and lack of platelets to a

life-threatening blood-formation disturbance, therefore the blood count should be

controlled regularly by doctors. The patient gets fever after application of the active

substance or he noticed subcutaneous bleeding, immediately discontinue treatment and the

doctor is to ask.

When injections, pain in the injection site and local reactions at the injection site may occur

in some cases. Also, there can be a strong drop in blood pressure in an injection into the

veins in rare cases.

6.6 REVIEW OF LITERATURE:

1. Dr. K. Raghubabu and B. Kalyana Ramu12 have reported Development of New

Visible Spectrophotometric Determination of Analgin in Bulk & Formulations using

p-Anisidine –ferric chloride as Oxidative Coupling Reagent. A simple and sensitive

visible spectrophotometric method has been developed for the determination of

analgin from its bulk drug and formulations. This method is based on the oxidative

coupling reaction with p-anisidine in the presence of iron (III) to yield the violet-red

colored product having maximum absorbance at 550nm. Regression analysis of

Beer's law plot showed good correlation in the concentration range of 4.0-40.0μg/ml.

The proposed method is applied to commercial available tablets and the results are

statistically compared with those obtained by official method and validated by

recovery studies. The results are found satisfactory and reproducible. The method is

applied successfully for the estimation of the analgin in formulations without the

interference of excipients. The method offers the advantages of rapidity, simplicity

and sensitivity, normal cost and can be easily applied to resource-poor settings

without the need for expensive instrumentation and reagents.

2. V. Vlasova, A. V. Shilova and Yu. S. Fokina13 have reported Spectrophotometric

Assay of Active Components in Medicinal Formulations Using the Vierordt Method

for Analysis of Analgin-quinine and Panadol Extra. A method for the

spectrophotometric assay of analgin, caffeine, quinine hydrochloride and

paracetamol in two-component medicinal formulations was developed using a new

criterion for selecting analytical wavelengths (AWL). Content calculations were

performed using several sets of AWL. Model solutions were used to show that use

of three sets of AWL decreases estimation errors for components in mixtures

withboth low and high content. This method was used to analyze the medicinal

formulations Panadol Extra and Analgine-quinine.

3. Saidul Zafar Qureshi, Ahsan Saeed, Tausiful Hasan14 have reported

Spectrophotometric Determination of Novalgin in tablets by use of Potassium

Iodate.An indirect colour reaction has been studied for determination of novalgin in

tablets. The method is simple, rapid and reproducible with a relative standard

deviation of 0.2%. Novalgin is determined spectrophotometrically by means of its

colour reaction with potassium iodate. Beer's law is obeyed over the range 1–10 mg

of drug. A tentative reaction mechanism has been proposed.

4. Hamide Z. Senyuva, Sureyya Ozcan, Burak Veli Kabasakals15 have reported

Validated Simple, Rapid and Accurate HPLC Methods for determination of

Metamizole Sodium in Solid and Liquid Dosage Forms. The aim of the study was to

develop and validate high performance liquid chromatography (HPLC) assay for the

rapid determination of Metamizole Sodium in solid and liquid dosage forms. The

experimental procedure involved reversed-phase- HPLC with a Zorbax SB C18

column (5 µm particle size, 4.6 ID x 250 mm), metanol- water volumetric solution

(80: 20, v/v) mobile phase, UV detection at 254 nm for Metamizole Sodium. The

flow rate of the, mobile phase was 1 mL/min. The retention time of Metamizole

Sodium is ca. 3.1-3.3 min. Formulations components did not give rise to any

interfering peaks. Calibration curve was linear over the range 0.5-100 µg/mL for

analyte. The metamizole recovery range is at three level (10µg/mL, 15µg/mL and

25µg/mL) 93-100 %. The method was validated with respect to linearity, precision,

accuracy, specificity and robustness. Due to its simplicity and accuracy, the assay

method is suitable for routine analysis of both solid and liquid formulations.

5. Piotr Jedziniaka, Konrad Pietruka, Edyta Sledzinskaa, Malgorzata Olejnika,

Teresa Szprengier, Juszkiewicza & Jan Zmudzkia16 have reported Rapid method

for the determination of metamizole residues in bovine muscle by LC-MS/MS.

Metamizole is a pyrazolone non-steroidal anti-inflammatory drug allowed for use in

food-producing animals. According to Council Directive 96/23, residues of this drug

have to be monitored because of the potential risk to consumer’s health. Metamizole

is hydrolysed to its marker residue 4-methylaminoantypyrine.This compound is

further metabolised to three main metabolites: 4-formylaminoantipyrine, 4-

aminoantipyrine and 4-acetylaminoantipyrine. The MRL of 4-

methylaminoantipyrine in animal tissues is 100 µg kg−1. Considering the above

points, a method for the detection of four metamizole metabolites in bovine muscles

was developed. Analytes were extracted from muscle by a mixture of acetonitrile

and sodium acetate buffer. After centrifugation, the supernatant was passed through

alumina cartridges, diluted with mobile phase and analysed by using LC-MS/MS.

Four metamizole metabolites were separated on a C8 column in 23 min with a

gradient of methanol:acetonitrile:ammoniumformate solution and analysed by using

positive ionisation. Validation of the method indicated a within- laboratory

reproducibility in the range of 7–30% and recovery in the range of 45–95%. The

7.0

method fulfils the criteria for confirmatory methods and, thanks to its labour

efficiency, may also be used for screening purposes.

6. Dhaneshwar Shep, Rakesh Ojha, Rajeshwari Rathod, Sweta Patel, Manish

Nivsarkar, Sanjay Marooand Harish Padh17 have reported Bioequivalence Study

of Two Oral Formulations of Metamizole500 MG IN Healthy Volunteers.

Metamizole (Dipyrone) is widely used and has effective analgesic, antipyretic, and

antispasmodic properties. After oral or intravenous administration, dipyrone is

rapidly hydrolyzed to the active moiety 4-methylaminoantipyrine.The aim of this

study was to assess the bioequivalence of 2 oral formulations of Metamizole 500

mg. This double blind, randomized, single-dose, 2-period crossover study in healthy

Indian adult volunteers was conducted at PERD Centre, Ahmedabad. Subjects

received Metamizole 500 mg of either test or reference formulation with a washout

period of 7 days. After study drug administration, serial blood samples were

collected over a period of 24 hours. Plasma concentration of 4-

methylaminoantipyrine was measured by pre-validated LC-MS method.

Pharmacokinetic (PK) parameters Cmax, Tmax, t1/2, AUC0-t, AUC0-∞, and kel,

were determined for test and reference formulations. The formulations were to be

considered bioequivalent if the log-transformed ratios of Cmax, AUC0-t, and

AUC0-∞ were within the predetermined bioequivalence range of 80% to 125%. A

total of 14 subjects were enrolled. No significant differences were found based on

analysis of variance, with mean values and 90% confidence intervals of

test/reference ratios for these parameters as follows: Cmax, 18.24 versus 18.44

μg/mL (92.68 - 106.61); AUC0-t, 92.97 Versus 91.37 μg.hr/mL (89.49 - 113.09);

and AUC0-∞, 96.64 Versus 94.65 μg.hr/mL (92.31 - 111.63). Since the 90%

confidence intervals for Cmax, AUC0-t, and AUC0−∞ were within the interval of

80-125%, it was concluded that both formulations were bioequivalent, according to

both the rate and extent of absorption.

6.7 OBJECTIVES OF THE STUDY:

In the proposed work, attempt will be made :

To develop a new instrumental method for estimation of Etodolac and Analgin To develop a validated method according to ICH guidelines.

To apply validated method for the estimation of Etodolac and Analgin in

pharmaceutical formulation.

7.1 MATERIALS AND METHODS:

All experiments will be carried out in the Department of Quality Assurance.

Srinivas college of Pharmacy, Valachil, Mangalore.

Pure sample of Etodolac and Analgin will be procured from Industries involved

in bulk manufacture of this drug.

Dosage formulations will be procured from local market.

The methods will be developed and validated in Quality Assurance lab of

Srinivas college of Pharmacy.

The methodswill 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 shall be used for measuring absorbance for Etodolac

and Analgin solutions.

Reagents supposedto use for the estimation:

3-methyl,2-benzothiazoline hydrazone(MBTH), 1,10-phenanthroline, 4-amino phenazene, Para dimethyl amino benzaldehyde (PDAB) 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.

8.0

7.2 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

7.3 Does the study require any investigation to beconducted on patients or animals?

No

7.4 Has the ethical clearance been obtained from your institution in case of 7.3?

Not applicable

REFERENCES:1. en.wikipedia.org/wiki/Etodolac

2. www.drugbank.ca/drug/Etoodolac(DB00749)

3. http://journal.chemistrycentral.com/content/2/1/7

4. Patel MJ, Badmanaban R, Patel CN. A reversed-phase liquid chromatographic (RP-

HPLC) method was developed for the simultaneous determination of tolperisone

hydrochloride (TOLP) and etodolac (ETD) in a combined fixed dose oral

formulations. Pharmaceutical Methods 2011;2(2):124–9.

5. Dongre VG, Shah SB, Bayer GS, Phadke M, Jadhav KV. Simultaneous

determination of etodolac and acetaminophen in tablet dosage form by RP-LC.

Chromatographia 2009;69(9-10):1019-23.

6. Jadhav AV, Gohel A, Bhavika, Sondagar MM, Patel BA, Parmar SJ. Method

development and validation for the simultaneous estimation of paracetamol and

etodolac by derivative UV Spectroscopic Method. Int J of Pharm Tech and Res

2013;5(3):1155-60.

7. Surya T, Parmar A, Sailor B, Vekariya K, Khasiya V. Development and validation

of spectroscopic method for simultaneous estimation of etodolac and

thiocolchicoside in tablet formulation. J of Pharm Res 2012;5(6):3004-7.

8. Shah R, Magdum C, Patil SK, Chougule DK, Naikwade N. Validated spectroscopic

method for estimation of aceclofenac from tablet formulation. Res J of Pharm and

Tech 2008;1(4):430-2.

9. en.wikipedia.org/wiki/Metimazole

10. www.sopharma.com/categories/421/42102analgin.html.

11. www.bbpharma.sk/spasmolytics/analgin-injection

12. Raghubabu K, Kalyana RB. Development of new visible spectrophotometric

determination of analgin in bulk & formulations using p-anisidine –ferric chloride

as oxidative coupling reagent. Int J of Anal and Bioanal Chem 2011;1(2):53-6.

13. Vlasova V, Shilova AV, Fokina YS. Spectrophotometric assay of active

components in medicinal formulations using the Vierordt method for analysis of

analgin-quinine and panadol extra. Pharm Chem J 2008;42(10):49-53.

14. Qureshi SZ, Saeed A, Hasan T. Spectrophotometric determination of novalgin in

tablets by use of potassium iodate. Talania 1989;36(8):869-71.

15. Senyuva HZ, Ozcan S, Kabasakals BV. Validated simple, rapid and accurate HPLC

methods for determination of metamizole sodium in solid and liquid dosage Forms.

Anal Lett 1997;30(6):1201-10.

16. Piotr J, Pietruka K, Sledzinskaa E, Malgorzata O, Teresa S, Juszkiewicza,

Zmudzkia J. Rapid method for the determination of metamizole residues in bovine

muscle by LC-MS/MS. Food Additives & Contaminants:Part A 2013;30(6):977-82.

17. Shep D, Ojha R, Rathod R, Patel S, Nivsarkar M, Maroo S, Padh H. Bioequivalence

study of two oral formulations of metamizole 500 mg in healthy volunteers. Int J of

Pharm Sci and Res 2012;3(6):1749-52.

9.0 SIGNATURE OF THE CANDIDATEBhalani Monaben Chimanbhai

10.0 REMARKS OF THE GUIDE

Forwarded for Approval

10.1 NAME AND DESIGNATION OF GUIDE

Dr. E.V.S. Subrahmanyam,Professor and Head,

Dept of Quality Assurance, Srinivas College of Pharmacy.

10.2 SIGNATURE

11.0 HEAD OF THE DEPARTMENT

Dr. E.V.S. Subrahmanyam,Professor and Head,

Dept of Quality Assurance, Srinivas College of Pharmacy.

11.1 SIGNATURE

12.0 REMARKS OF THE PRINCIPAL

Forwarded for approval

12.1 SIGNATURE Dr. Ramakrishna Shabaraya A.

Principal and Director,Srinivas college of pharmacy,

Valachil, Mangalore.