Analytical Method Development and Stability Studies of Carvedilol E-Book

Kishanta Kumar Pradhan

• Herausgeber: GRIN Verlag
• Sprache: Englisch

Veröffentlichungsjahr: 2015

Beschreibung

Master's Thesis from the year 2011 in the subject Medicine - Pharmacology, Pharmacy, grade: 8.0, , course: B.Pharm.,M.Pharm, language: English, abstract: A reverse phase high performance liquid chromatographic method (HPLC) has been developed for the method development validation of Carvedilol in bulk and pharmaceutical formulation by using YMC PACK PRO 4.6 X 150 mm (5µm Particle size). The mobile phase was Buffer: Acetonitrile: (70:30) and pH was adjusted to 2 pumped at a flow rate of 1 ml/min and the eluents were monitored at 320nm. Linearity was obtained in the concentration range of 10-90 μg/ml. The retention time of Carvedilol was found to be 3.2 minute. The method was validated for specificity, accuracy, precision, linearity, and limit of detection, limit of quantification, robustness and solubility stability. LOD and LOQ were found to be 0.001 μg/ml and 0.011μg/ml respectively. The method was statistically validated and RSD was found to be less than 2% indicating high degree of accuracy and precision of the proposed HPLC method. Stability study report revealed that the drug is susceptible for acidic, alkaline, oxidative, photolytic and UV degradation. The drug is stable to thermal degradation. More over the degradants were well separated from its API. Due to its simplicity, rapidness, high precision and accuracy, the proposed HPLC method may be used for determining Carvedilol in bulk drug samples or in pharmaceutical dosage forms.

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CONTENTS

1. Introduction

1.1 Spectrophotometric methods

1.2 ULTRA VIOLET SPECTROSCOPY [3-10]

1.2.1METHODS CARRIED OUT:

1.3. HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

1.3.1Modes of CHROMATOGRAPHY [12-29]

1.3.2 Elution TECHNIQUES [30]

1.4 METHOD DEVELOPMENT [31-36]

1.4.1. Completing the HPLC Method Development

1.5VALIDATION OF ANALYTICAL METHODS [37-47]

1.5.1. Specificity

1.5.2. Accuracy

1.5.3. Precision

1.5.4. Linearity

1.5.5. RANGE

1.5.6. LIMIT of Detection

1.5.7. limit of QUANTIficaTION

1.5.8. robustness

1.5.9. RUGGEDNESS

1.6 GUIDELINES OF FORCED DEGRADATION [48-56]

1.6.1. HYDROLYTIC Degradation:

1.6.2 Thermal and Thermal/Humidity Degradation

1.6.3 Oxidative Degradation

1.6.4 Photolytic Degradation

1.7 DRUG PROFILE OF CARVEDILOL [57-62]

1.7.1 Structure:

1.7.2 Description:

1.7.3 Mechanism of Action:

1.7.4 Pharmacology:

1.7.5 Dosing:

2. LITERATURE SURVEY

3. PROLOGUES TO THE EXPERIMENTAL WORK

3.1 AIM AND Objective OF PURPOSED Work

3.1.1scope of the propesed work

3.2 Plan of Purposed Work

3.3REQUIREMENT

3.3.1. Instruments used

3.3.2 Chemicals and reagents used

3.3.3Glass wares

4. EXPERIMENTAL

4.1 UV-Vis Spectrophotometric Method for The Estimation of Carvedilol

4.1.1Optimization:

4.2.METHOD

4.2.1Preparation of Stock Solution

4.2.2Preparation of working standard solution and construction of standard graph

4.2.3VALIDATION

4.3 RP-HPLC Method for the Estimation of Carvedilol

4.3.1 Optimization Chromatographic Conditions

4.3.2 Method

4.3.3 Validation

4.4 FORCED DEGRADATION STUDIES

4.4.1 Degradation in Acidic Condition

4.4.2 Degradation in Basic Condition

4.4.3 Oxidative Degradation

4.4.4 Photolytic Degradation

4.4.5 UV- Degradation

4.4.6 Thermal Degradation

5. RESULTS AND DISCUSSION

5.1 UV-VISIBLE SPECTROPHOTOMETRIC METHOD

5.1.1 Method

5.1.2 VALIDATION:

5.2 RP-HPLC METHOD DEVELOPMENT

5.2.1 Optimization

5.2.2 VALIDATION:

5.3FORCED DEGRADATION STUDIES

5.3.1. Acid Hydrolysis:

5.3.2 Alkaline Hydrolysis:

5.3.3. Photolytic Degradation:

5.3.4. UV-Degradation:

5.3.5 Thermal Degradation:

5.3.6 Oxidative Degradation

5.3.7 SUMMARY OF STRESS DEGRADATION

CONCLUSIONS

REFERNCES

1. Introduction

1.1 Spectrophotometric methods

Spectrophotometric method are a large group of analytical methods that are based on atomic and molecular spectroscopy i.e. the integration between electromagnetic radiation and matter when electromagnetic radiation passes through a layer of analyte certain frequencies may be selectively removed by absorption, a process in which electromagnetic energy is transferred to the atom, ion or molecules composing the sample. Absorption promotes these particles from their normal ground state, to one or higher exited states.

The absorption of light by analytes by raising an electron or electrons to a higher level and other functional group. Every functional group in a molecule of substances is characterized by the absorption of light in a definite region of the spectra and this property is used for the identification of the substances in a drug. In addition to chromospheres, a molecule may contain one or more functional groups that themselves do not absorb in visible region being scanned , but can affect the behavior of the chromospheres that are conjugated with this group called auxochromes (eg. SH, NH2, OH) usually causes absorption by a chromophores at higher wavelength and at a higher value of absorptive then feature in the given chromophores. [1]

Importance of visible spectrophotometry in Pharmaceutical Analysis

Spectrophotometry is generally preferred especially by small scale industries as the cost of the equipment is less and the maintain problems are minimal. The method of analysis is based on measuring the absorption of monochromatic light by colored compound in the visible path of the spectrum .if the analytes are colorless compound they are converted into colored compounds by reaction with suitable compounds .in case majority of the compound are complex and complex legends. The later must be stable and have a constant composition and high color intensity. The photometric methods of analysis are based on the Bouger-Lambert-Beer’s law, which establishes the absorbance of a solution is directly proportional to the concentration of the analyte. The fundamental principle of operation of spectrophotometer covering UV region consists in that light of definite interval of wavelength passes through a cell with solvent and falls on to the photoelectric cell that transforms the radiant energy into electrical energy measured by a galvanometer.[2].

Important applications of spectrophotometer:

Identification of many types of organic, inorganic molecules and ions.

Quantitative determination of many biological, organic and inorganic species.

Quantitative determination of mixtures of analyte.

Monitoring and identification of chromatographic effluents.

Determination of equilibrium constants.

Determination of stoichiometry and chemical reactions.

Monitoring of environmental and industrial process.

Monitoring of reaction rates.

Typical analysis times range from 2 to 30 min for sample.

1.2 ULTRA VIOLET SPECTROSCOPY [3-10]

 This technique of ultra violet spectroscopy is one of most frequently employed method in pharmaceutical analysis. It involves the measurement of the amount of UV radiation (190-380 nm) or visible (380-800 nm) radiation absorbed by a substance in solution. Ultraviolet spectroscopy involves the promotion of electrons (σ, π, n electrons) from the ground state to higher energy state. It is useful to measure the number of conjugated double bonds and also aromatic conjugation with the various molecules.

 The ultraviolet region of the electromagnetic spectrum is frequently subdivided into as follows:

Far vacuum Ultraviolet region (10-200 nm).

Near ultraviolet region (200-400 nm).

Visible region (380-780 nm).

Diagram of an analytical instrument showing the stimulus and measurement of response.

1.2.1METHODS CARRIED OUT:

a) SIMULTANEOUS EQUATION METHOD

b) DERIVATIVE SPECTROSCOPY METHOD

a) Simultaneous equation method

 If a sample contains two absorbing drugs (X and Y) each of which absorbs atλmaxof the others it may be possible to determine both drugs by the technique of simultaneous equation ( Vierodt’s method) provided that criteria apply.

Information required is

1. The absorptivities of X atλ1 and λ2 and ax2, respectively

2.The absorptivities of Y at λ1 and λ2 and ay2, respectively

3.The absorbance’s if the diluted sample at λ1 and λ2, A1 and A2 respectively.

Let cx and cy be the concentrations of X and Y respectively in the diluted sample. Two equations are constructed based upon the fact that at λ1 and λ2

Criteria for obtaining maximum precision, based upon the absorbance ratios, have been suggested (Glenn, 1960) that place limits on the relative concentrations of the components of the mixture. The criteria are the ratios

It should lie outside the range 0.1-2.0 for the precise determination of X and Y respectively. These criteria are satisfied only when theλmaxof the two components is reasonably dissimilar. An additional criterion is that the two components do not interact chemically, thereby negating the initial assumption that the total absorbance is equal to sum of the individual absorbance’s.

b) Derivative Spectrophotometric method

 This method involves the conversion of the normal spectrum into first, second or higher derivative spectrum. The transformation that occurs in the derivative spectrum are understood byreference to a Gaussian band which represents an idealabsorption band.