“DEVELOPMENT OF NEW ANALYTICAL METHODS AND THEIR VALIDATION FOR THE DETERMINATION OF METOCLOPRAMIDE HYDROCHLORIDE AND LORATADINE IN BULK AND MARKETED FORMULATIONS”

MASTER OF PHARMACY DISSERTATION PROTOCOL

SUBMITTED TO THE

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES KARNATAKA, BANGALORE

BY

SOLANKI MEGHABEN JAGDISHBHAI

Under The Guidance of

DR. E.V.S. SUBRAHMANYAM. M.PHARM. Ph.D

P.G. DEPARTMENT OF QUALITY ASSURANCE,

SRINIVAS COLLEGE OF PHARMACY, MANGALORE – 574143

2011 – 2013

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

BANGALORE, KARNATAKA

ANNEXURE – II

REGISTRATION OF SUBJECT FOR DISSERTATION

1.0 / NAME OF THE CANDIDATE
ADDRESS / SOLANKI MEGHABEN JAGDISHBHAI
DEPARTMENT OF Q.A.,
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 / 31st OCTOBER,2011
5.0 / TITLE OF THE TOPIC:
“DEVELOPMENT OF NEW ANALYTICAL METHODS AND THEIR VALIDATION FOR THE DETERMINATION OF METOCLOPRAMIDE HYDROCHLORIDE AND LORATADINE IN BULK AND MARKETED FORMULATIONS”
6.0
7.0
8.0 / 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 andvalidation of analytical methods.
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).
6.2 A BRIEF INTRODUCTION ABOUT METOCLOPRAMIDE
HYDROCHLORIDE[1]
MOLECULAR STRUCTURE:

IUPAC NAME: 4-amino- 5-chloro-N-(2-(diethylamino)ethyl)-2- methoxybenzamide
Hydrochloride
MOLECULAR FORMULA: C14H22ClN3O2·HCl
MOLECULAR WEIGHT: 336.26 g/mol
CATEGORY: Antiemetic and gastroprokinetic agent
PHYSICAL APPEARANCE: white, crystalline, odourless powder
SOLUBILITY: very soluble in water, freely soluble in alcohol, sparingly soluble in
methylene chloride
MELTING POINT: 147˚C (297˚F)
BIOAVAILABILITY: 80±15%(oral)
PROTEIN BINDING : 30%
METABOLISM: Hepatic
HALF-LIFE: 5 to 6 Hours
EXCRETION: 70-85% renal, 2% faecal
ROUTE: Oral, Intravenous, Intramuscular
PHARMACOLOGY:[1]

MEDICAL USES:

Metoclopramide hydrochloride is commonly used to treat nausea including that which is due to chemotherapy and that occurring post operatively. Evidence also supports its use for gastroparesis (poor stomach emptying) and gastroesophageal reflux disease.

Antiemetic: Metoclopramide hydrochloride commonly treats nausea and vomiting associated with conditions such as uremia, radiation sickness, malignancy, labor, infection, migraine headaches, and emetogenic drugs. In the setting of painful conditions such as migraine headaches, Metoclopramide hydrochloride may be used in combination with paracetamol (acetaminophen) (available in the UK as Paramax, and in Australia as (Metomax) or in combination with aspirin (Migramax).

Gastroprokinetic: Metoclopramide hydrochloride increases peristalsis of the jejunum and duodenum, increases tone and amplitude of gastric contractions, and relaxes the pyloric sphincter and duodenal bulb. These gastroprokinetic effects make Metoclopramide useful in the treatment of gastric stasis (e.g. after gastric surgery or diabetic gastroparesis ), as an aid in gastrointestinal radiographic studies by accelerating transit through the gastrointestinal system in barium studies, and as an aid in difficult intubation of the small intestine. It is also used in gastroesophageal reflux disease (GERD).

Other Indications: By inhibiting the action of dopamine, Metoclopramide hydrochloride

has sometimes been used to stimulate lactation. It can also be used in the treatment of migraines in the setting of allodynia, where it is more effective than triptans.
Veternary Use: Metoclopramide hydrochloride is also use in animals. It is commonly used to prevent vomiting in cats and dogs. It is also used as a gut stimulant in rabbits.

MECHANISM OF ACTION:

Metoclopramide hydrochloride appears to bind to dopamine D2 receptors where it is a receptor antagonist, and is also a mixed 5-HT3 receptor antagonist / 5HT4 receptor agonist. The antiemetic action of Metoclopramide hydrochloride is due to its antagonist activity at D2 receptors in the chemoreceptor trigger zone (CTZ) in the central nervous system (CNS)- this action prevents nausea and vomiting triggered by most stimuli. At higher doses, 5-HT3 antagonist activity may also contribute to the antiemetic effect. The gastroprokinetic activity of Metoclopramide hydrochloride is mediated by muscarinic activity, D2 receptor antagonist activity and 5-HT4 receptor agonist activity. The gastroprokinetic effect itself may also contribute to the antiemetic effect. Metoclopramide hydrochloride also increases the tone of the lower esophageal sphincter.

ADVERSE EFFECTS:

Common adverse drug reaction (ADRs) associated with Metoclopramide hydrochloride therapy include restlessness, drowsiness, dizziness, fatigue, and focal dystonia. Infrequent ADRs include hypertension, hypotension, hyperprolactinaemia, leading to galactorrhea, constipation, depression, headache, and extrapyramidal effects such as oculogyric crisis. Rare but serious ADRs associated with therapy include agranulocytosis, supraventricular tachycardia, hyperaldosteronism, neuroleptic malignant syndrome, akathisia and tardive dyskinesia. Recent research suggests that Metoclopramide hydrochloride may be the most common cause of drug-induced movement disorders. The risk of extrapyramidal effects is increased in people under 20 year of age and with high dose of prolonged therapy. Tardive dyskinesia may be persistent and irreversible in some patients. The majority of reports of tardive dyskinesia occur in people who have used Metoclopramide hydrochloride for more than three months. Consequently, the USFDA recommends that Metoclopramide hydrochloride be used for short term treatment, preferably less than 12 weeks. In 2009, the USFDA required all manufacturers of Metoclopramide hydrochloride to issue a black box warning regarding the risk of tardive dyskinesia with chronic or high-dose use of the drug. Dystonic reactions may be treated with benztropine, diphenhydramine, trihexyphenidyl or procyclidine.

CONTRAINDICATIONS AND PRECAUTION:

Metoclopramide hydrochloride is contraindicated in phaeochromocytoma. It should be used with caution in Parkinson’s disease since, as a dopamine antagonist, it may worsen
symptoms. Long-term use should be avoided in patients with clinical depression as it may worsen mental state. Also contraindicated with a suspected bowel obstruction.
Use in pregnancy: Metoclopramide hydrochloride has long been used in all stages of pregnancy with no evidence of harm to the mother or unborn baby. A large cohort study of babies born to Israeli women exposed to Metoclorpamide hydrochloride during pregnancy found no evidence that the drug increases the risk of congenital malformations, low birth weight, preterm birth, or perinatal mortality. Metoclopramide hydrochloride is excrete into milk.
6.3 REVIEW OF LITERATURE:
A literature survey was carried out for the estimation of Metoclopramide hydrocloride in bulk and marketed dosage forms. It was found that a very few methods have been reported for this drug. The collection of references are reproduced below:
1.  Hosakere D. Revanasiddappa and Malligere A. Veena[2] have developed sensitive spectrophotometric determination of Metoclopramide hydrochloride and Dapsone in bulk sample and dosage forms. The method is based on the diazo – coupling reaction of the studied drugs with a new coupling agent, imipramine hydrochloride, in an acid medium. The resulting violet – colored azo dyes exhibit maximum absorption at 570 nm for both Metoclopramide hydrochloride and Dapson.
2.  N. P. Dudhane, M. J. Umekar, R. T. Lohiya,[3] have developed a validated RP-HPLC method for estimation of Metoclopramide hydrochloride and Paracetamol in solid dosage form. RP-HPLC estimation of drugs in selected combination was done using Phenomenex ODS 5μ, C18 column (250×4.6mm) and Acetonitrile: Methanol: (0.5%) TEA Buffer (18.5:6.5:75) as mobile phase which shows sharp and resolved peak when detected at 273nm.
3.  Dudhane N.P., Vidhate S.S., Borkar B.H., Lohiya R.T., Umekar M.J[4]. have developed simultaneous UV spectrophotometric estimtion of Metoclopramide hydrochloride and Paracetamol in solid dosage form.
Method I: Simultaneous Equation method, wavelengths selected were
243.0 nm and 273.5 nm for estimation of Metoclopramide
hydrochloride (MET) and Paracetamol (PAR) respectively.
Method II: Absorbance Ratio method, wavelengths selected were 243.0
nm, 262.0nm Isoabsorptive point of Paracetamol and
Metoclopramid hydrochloride.
Method III: Correction method,309.0 nm
4.  Vinay Wamorkar, Manjunath S.Y., M.Mohan Varma[5], have done development and validation of UV spectroscopic method for determination of Metoclopramide hydrochloride in bulk and tablet formulation. Drug concentrations in various tests were determined spectrophotometrically (SL‐164 Double beam UV spectrophotometer, Elico, India.) at 272 nm using 1 cm quartz match cells.
5.  Nawal. A. Al-Arfaj[6] has developed flow-injection chemiluminescent determination of Metoclopramide hydrochloride in pharmaceutical formulations and biological fluids using the [ru(dipy)32+]–permanganate system. The method is based on the CL reaction of Metoclopramide with Ru(dipy)32+ and KMnO4 in a sulfuric acid medium. Under the optimum conditions, a calibration graph was obtained over the concentration range 0.005–3.5 _gml−1 with a limit of detection (S/N = 2) of 1 ngml−1. The correlation coefficient was 0.99993 (n = 8) with a relative standard deviation of 0.48% for 10 determinations of 1 _gml−1 of drug. The method was successfully applied to the determination of Metoclopramide in pharmaceutical preparations and biological fluids after IP administration of 25 mg kg−1 dose to rats. The elimination half-life was 2.5 ± 0.4 h.
6.  Ahmad Khan, Syed Baqir Shyum Naqvi, Muhammad Harris Shoaib, Rabia Ismail Yousaf, Jallat Khan, Muhammad Hanif and Asadullah Madni[7] have done validation and application of RP-HPLC method for the quantification of Metoclopramide hydrochloride in oral formulations prepared for IVIVC studies. A reverse chromatographic method was used with the mobile phase of Acetonitrile, 20m M.Potassium dihydrogen phosphate buffer solution (pH 3 adjusted with orthophosphoric acid) in the ratio of 40:60.The column used was Waters C18 3.9×300mm μBondapak (RP). The flow rate of the mobile phase was 2ml/minute. The detector was set at the wavelength of 275nm.
7.  S R Patel, Dr. L J Patel[8] have done development and validation of first derivative spectroscopy method for simultaneous determination of Ondansetron and Metoclopramide in combined dosage form. The first derivative spectroscopy was performed on a double beam UV Visible spectrophotometer using methanol as a solvent. Absorbances were recorded at 266 nm (ZCP of ONDA) and 253 nm (ZCP of METO) for Metoclopramide hydrochloride and Ondancetron, respectively.
8.  Gaikwad Shubhangee, Kondawar Manish, Nazarkar Swapnil, Phase Sheetal and Narkhede Harshal[9] have developed RPHPLC method for
the simultaneous determination of Metoclopramide hydrochloride and Paracetamol in tablet dosage form. The method was carried out on a HiQsil
C8, (4.6×250mm) column with a mobile phase consisting of acetonitrile:acetate buffer (pH 6.78) (50:50v/v) at a flow rate of 1ml/min. Detection was carried out at 308 nm. The retention time of Paracetamol and Metoclopramide hydrochloride was 3.2 and 5.5 min respectively.

6.4 A BRIEF INTRODUCTION ABOUT LORATADINE[10][11]

MOLECULAR STRUCTURE:

IUPAC NAME: ethyl 4-(8-chloro-5,6-dihydro-11 H-benzo[5,6]cyclohepta[1,2-b]pyridin-
11-ylidene)-1-piperidinecarboxylate
MOLECULAR FORMULA: C22H23ClN2O2
MOLECULAR WEIGHT: 382.88 g/mol
CATEGORY: H1 histamine antagonist ,drug used to treat allergies
PHYSICAL APPEARANCE: white to off white powder
SOLUBILITY: insoluble in water, soluble in acetone, alcohol and chloroform
BIOAVAILABILITY: Almost 100%
PROTEIN BINDING: 97-99%
METABOLISM: Hepatic(CYP2D6 and 3A4 mediated)
HALF LIFE: 8 Hours
EXCRETION: 40% as conjugated metabolites into urine, similar amount into the feces
ROUTES: ORAL
PHARMACOLOGY:[11][12]
MECHANISM OF ACTION: Loratadine competes with free histamine and exhibits specific, selective peripheral H1 antagonistic activity. This blocks the action of endogenous histamine, which subsequently leads to temporary relief of the negative symptoms (eg. nasal congestion, watery eyes) brought on by histamine. Loratadine has low affinity for chlolinergic receptors and does not exhibit any appreciable alpha-adrenergic blocking activity in-vitro. Loratadine also appears to suppress the release of histamine and leukotrienes from animal mast cell, and the release of leukotrienes from human lung fragments, although the clinical importance of this is unknown.

ADVERSE EFFECTS: