Method Development For Simultaneous Estimation Of Acebrophylline, Montelukast And Desloratadin

Method Development for Simultaneous Estimation of Acebrophylline, Montelukast and Desloratadin KAPIL KUMAR PATIDAR Supervisor: Dr. Ravindra K. Rawal

DEPARTMENT OF PHARMACEUTICAL ANALYSIS ISF COLLEGE OF PHARMACY, MOGA 1

Introduction UV Spectroscopy: • UV spectroscopy is type of absorption spectroscopy in which light of ultra-violet region (200-400 nm) is absorbed by the molecule. Absorption of the ultra-violet radiations results in the excitation of the electrons from the ground state to higher energy state. • The energy of the ultra-violet radiation that are absorbed is equal to the energy difference between the ground state and higher energy states.

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Conti…… High

Performance

Liquid

Chromatography (HPLC) • High-performance

liquid

chromatography (HPLC; formerly referred to as high-pressure liquid chromatography) is a technique in analytical

chemistry

used

to

separate, identify and quantify each component in a mixture. •

HPLC is a form of liquid chromatography used to separate compounds that are dissolved in solution.

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Conti…… • COLUMN • Column forms the heart of any HPLC system. • The quality of its packing materials are essential for good separation and minimum dispersion. • The stainless steel column of internal diameter 4.0 to 4.6 mm and length 10-30cm are used. • Examples: -Si-CH2-CH3 – C-2 Ethyl Silyl – C-8 Octyl Silyl – C-18

Octadecyl Silyl

– CN Cyanopropyl Silyl

-Si-(CH2)7-CH3 -Si-(CH2)17-CH3 -Si-(CH2)3-CN

 

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Conti…… Advantage of UV

Advantage of HPLC

1. Qualitative & Quantitative Analysis

1. Determination of all the compounds present in mixtures, ultimately by both quantifying and characterizing them.

2. Measuring concentration of solution, absorption band can also used to determine the concentration of compounds in a solution

2. High quality information from less resolved data.

3. Detection of functional groups, it 3. These methods are used to is possible to detect the presence of resolve the drug mixtures contain certain functional groups with the up to six drugs. help of UV Spectrum.

4. Detection of impurities

4. Cost effectiveness 5

Conti…… METHODS OF MULTICOMPONENT ANALYSIS USING UV- VISIBLE SPECTROPHOTOMETER AND HPLC   • • • •

Method A: Simultaneous equation method Method B: First order derivative method Method C: Q-analysis/Absorption ratio method Method D: Reversed phase-high performance liquid chromatography SIMULTANEOUS ESTIMATION

 Simultaneous estimation is required when the active pharmaceutical ingredients are more than two in single dosage form.  Simultaneous estimation is an accurate and less time consuming approach for determination of many API’s in single dosage form. 6

Conti…… Combinations available for Multi Drugs Therapy 1.Acebrophylline[A] + Montelukast Sodium[M]+ Desloratadin[D] 2. Acebrophylline[A] + Montelukast Sodium[M] 3. Montelukast Sodium[M] + Desloratadin[D] Combination for research work: Acebrophylline[200mg] + Montelukast Sodium(10mg) + Desloratadin[5mg] Manufacturer by:-1.vivid biotek (India) Pvt. Ltd. New Delhi india

2. Mescot Health Series Pvt. Ltd. Sidcul, Haridwar- 2494043 7

Conti……

Acebrophylline:

OH

O N

N \

O

COOH

N

N

Br

N H NH2 Br

Acebrophylline mol.wt. 616.31 Solubility Category

Methanol Mucoregulator with anti inflammatory

Acebrophylline:- chemically 4-[(2-amino-3,5-dibromo-phenyl methylamino] cyclohexan-1-ol; 2-(1,3-dimethyl- 2,6-dioxo-purin- 7-yl)acetic acid

is a

hydroxy naphthaquinone derivative used for treatment of asthma Its mechanism of action is reducing inflammation in the airways and helping to boost ciliary clearance.

PATIL, S. S., et al. (2009).

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Conti…… UV conditions: Conditions Chemical used

Descriptions Methanol, Ethanol and water

Detection wavelength (λmax)

273nm

HPLC condition:

WANG, L., et al. (2011).

Conditions

Descriptions

Chemical used

Phosphate buffer solution(pH 7) : Acetonitrile (50:50%v/v)

Flow rate

1mL/min

Column used

SB- C18

UV detection (λmax)

273nm

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Conti…… Montelukast sodium: S Cl

COONa

N HO

Montelukast sodium

mol.wt. 608.2

UV conditions:

Condition Solubility

Descriptions Methanol, Ethanol(95%)and Water

UV detection (λmax)

 254 nm

• Montelukast is used to prevent wheezing, difficulty breathing, chest

tightness, and coughing caused by asthma. ALSARRA, I. A.,et al(2004).

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Conti……

Montelukast is also used to prevent bronchospasm (breathing difficulties) during exercise. Montelukast is in a class of medications called leukotriene receptor antagonists (LTRAs). It works by blocking the action of substances in the body that cause the symptoms of asthma and allergic rhinitis. HPLC conditions: Condition Chemical used

Detection wavelength (λmax) Al Omari,et al. (2007)

Descriptions Potassium Dihydrogen Phosphate: phosphoric acid(70:30, % v/v) 254nm 11

Conti…… Desloratadine:

Desloratadine

mol.wt. 310.84

Solubility

Methane, Ethanol and water

Category

Anti-Histaminic

• Desloratadine (DLT) is a non sedative, long acting antihistamine with selective peripheral H1 receptor antagonistic activity.

• Chemically desloratadine is 8-chloro-6, 11-dihydro-11-(4piperidinylidene)-5H-benzo [5,6]cycloheptane [1,2-b pyridine] Bondili, et al.(2011).

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CONT..... UV conditions Conditions

Description

Chemical used

Methanol

Detection wavelength (λmax)

242nm

HPLC condition

Patel, R. B.,et al. (2012).

Condition

Description

Chemical used

Phosphate buffer : acetonitril:methanol(50:4 0:10)

Flow rate

0.8mL/min

Column

C-18

UV detection (λmax)

247nm

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National & International Status Drug Name

Equipment

Year

Journal

A

UV

2013

GENESIS JOURNAL

HPLC

2011

JOURNAL OF NORTHWEST UNIVERSITY

UV

2009

JOURNAL OF PHARMACY RESEARCH

HPLC

2007

JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS

D

UV HPLC

2011 2012

IJPIR JOURNAL ARABIAN JOURNAL OF CHEMISTRY

A+M

UV

2012

INDO AMERICAN JOURNAL OF PHARMACEUTICAL RESEARCH

M+D

HPLC

2015

AMERICAN JOURNAL OF ANALYTICAL CHEMISTRY

M

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Hypothesis Method Development  Method validation is the process used to confirm that the analytical procedure employed for a specific test is suitable for its intended use.  Results from method validation can be used to judge the quality, reliability and consistency of analytical results, it is an integral part of any good analytical practice.

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Conti….. Parameters Performed in Analytical Method of Validation: 1.Specificity/Selectivity 2.Linearity 3.Range 4.Accuracy 5.Precision 6.Limit of Detection(LOD) 7.Limit of Quantification(LOQ) 8.Ruggedness 9.Robustness 10.System Suitability Testing Thus, the present study is designed to investigate the procedures involved for testing of marketed formulation which is not designed yet. 16

Aim & objectives • Aim of proposed research project is to develop and validate a multipurpose method for simultaneous estimation of Acebrophylline[A]+Montelukast Sodium[M]+Desloratadin[D] in marketed formulations. • Till date, there is no UV/VIS and HPLC method reported for this combination so, we will provide a new, cheaper and less time consuming method.

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Rationale & Novelty To develop method for the drug since method has not been developed on UV/VIS Spectrophotometer or HPLC i.e. ABROPHYYL-DM .

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Plan of work 2016-17 Work

Aug, Sep, 16 16

Oct, 16

Nov, 16

Dec, 16

Jan, 17

Feb, 17

Mar, Apr, 17 17

May, 17

Literature survey and Procurement of drugs

UV Method Development and validation HPLC Method Development and validation Comparison of UV and HPLC Method Results Publication and Thesis writing

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Facility required

(a) Chemicals • • • • • •

Methanol Ethanol Phosphate buffer HCl Acetonitrile NaOH

(b) Instruments In- House • IR •

UV

• HPLC

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Expected outcome



Development and validation of multipurpose method for simultaneous estimation of Acebrophylline[A]+Montelukast Sodium[M]+Desloratadin[D]

formulations

on

UV/VIS

spectrometer and HPLC which has not been developed till date.  Review/ Research paper Publication

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References •

• •

•

•

•

PATIL, S. S., et al. (2009). "Development and statistical validation of spectrophatometry method for estimation of Montelukast in bulk and tablet dosage form." Journal of Pharmacy Research Vol 2(4). WANG, L., et al. (2011). "Determination of the content of acebrophylline capsules by RP-HPLC." Journal of Northwest University (Natural Science Edition) 4: 020. ALSARRA, I. A.,et al(2004). "Development of a stability-indicating HPLC method for the determination of Montelukast in tablets and human plasma and its applications to pharmacokinetic and stability studies." Saudi Pharmaceutical Journal 12(4): 136-143. Al Omari,et al. Effect of light and heat on the stability of montelukast in solution and in its solid state. Journal of pharmaceutical and biomedical analysis, 2007. 45(3), pp.465-471. Bondili, et al.(2011). "Spectroscopic method for determination of Desloratadine in bulk and its tablet dosage forms." International Journal of Pharmaceutical & Industrial Research 1.2 (2011): 131-34. Patel, R. B.,et al. (2012). "Validation of stability indicating high performance liquid chromatographic method for estimation of desloratadine in tablet formulation." Arabian Journal of Chemistry .

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Thank you !!!....

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