DEVELOPMENT AND VALIDATION OF HIGH PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR ANALYSIS OF GATIFLOXACIN & ITS IMPURITY

Pharma courses

pharma courses



{ DOWNLOAD AS PDF }

 

ABOUT AUHTOR
Narendra M.Petha, J.G.Patil,Mr J.G.Chandorkar

Indofil Industries Ltd, Gujarat
jchandorkar-icc@modi.com

ABSTRCT 
Gatifloxacin is a antibacterial agent, Rapid, sensitive and selective analytical method is essential for monitoring the different reactions steps involved in process development of Gatifloxacin. A simple isocratic reverse phase High Performance Liquid Chromatographic [HPLC] method was developed for simultaneous separation of different intermediates and other impurities. The method was utilized successfully in analyzing the reaction streams, related substances in final product and for the assay in drug.

REFERENCE ID: PHARMATUTOR-ART-2468

PharmaTutor (ISSN: 2347 - 7881)

Volume 5, Issue 3

Received On: 08/11/2016; Accepted On: 22/11/2016; Published On: 01/03/2017

How to cite this article: Petha NM, Patil JG, Chandorkar JG;Development and Validation of High Performance Liquid Chromatography method for analysis of Gatifloxacin & its Impurity; PharmaTutor; 2017; 5(3); 37-41

 

INTRODUCTION: -
Impurity profiling is the common name of a group of analytical activities, the aim of which is the detection, identification/structure elucidation and quantitative determination of organic and inorganic impurities, as well as residual solvents in bulk drugs and pharmaceutical formulations. Since this is the best way to characterize the quality and stability of bulk drugs and pharmaceutical formulations, this is the core activity in modern drug analysis. Due to the very rapid development of the analytical methodologies available for this purpose and the similarly rapid increase of the demands as regards the purity of drugs it is an important task to give a summary of the problems and the various possibilities offered by modern analytical chemistry for their solution.

Gatifloxacin level in biological fluids, in different pharmaceutical formulations and as a raw material for related substances have previously been determined by spectrophotometric, gas chromatographic [HPLC] techniques. Literature surveys reveals hardly any method for the analysis of reaction mixture obtained in the preparation of Gatifloxacin. There is an increasing need for rapid and sensitive method for the determination of raw materials, intermediates and finished products in reaction stream during process development of Gatifloxacin. The HPLC system consisted of Jasco make UV/VIS detector model 1575, along with Borwin software (Integrator) were used. Analysis were performed on Stainless steel column containing C-18 packing,

EXPERIMENTAL: -

CHROMATOGRAPHIC CONDITION: -
          
1) COLUMN: 250 mm x 4.6 mm i.e. - Stainless steel column containing C18, 5 u
2) MOBILE PHASE: BUFFER: ACETONITRILE (80: 20)
3) FLOW RATE: 1.0 ml / minute.
4) DETECTOR WAVELENGTH: 210 nm.
5) SAMPLE SIZE: 20 ul

The approximate Retention Times that should be obtained using these chromatographic conditions are:

- GATIFLOXACIN approximate retention time  = 1.85 Minutes
- 2-Methyl Piperazine approximate retention time = 04.46 Minutes.

PREPARATION OF MOBILE PHASE:

Buffer [0.025 gm Ortho Phosphoric acid] 80 v & 20 v of Acetonitrile. Adjust pH 3.0 by TEA.  

PREPARATION OF THE TEST SOLUTION FOR ANALYSIS OF GATIFLOXACIN:

Amber glassware must be used when preparing these solutions because GATIFLOXACIN is Photosensitive.

ANALYTICAL STANDARD SOLUTION 
[A] Accurately weigh 100 mg [u 5 mg] of Working standard of GATIFLOXACIN  & transfer to a 50 ml volumetric flask. Add 10 ml of mobile phase & sonicate until dissolved. Allow to cool to room temperature & dilute to volume with mobile phase.  
[B] Accurately weigh 5 mg [u1 mg] of the working standard of 2-Methyl Piperazine &  transfer to 50 ml volumetric flask. Add 10 ml of mobile phase & dilute to volume with mobile phase.
[C] Transfer 4.0 ml of solution [B], in solution [A] in volumetric flask. & Dilute to volume with Mobile
phase & mix thoroughly.

PREPARATION OF SAMPLE SOLUTION:

Accurately weigh 100 mg of sample & transfer to 50 ml volumetric flask. Add 10 ml of Mobile phase &
sonicate until dissolved. Allow to cool to room temperature & dilute to volume with mobile phase.

INJECTION PRECISION:
Make duplicate injections of the analytical standard solution. Using a computing Integrator measure the GATIFLOXACIN peak area in the injections made.

The relative Standard deviation must not be greater than + 2.0%.

CALCULATION OF RESULTS:

 

1. GATIFLOXACIN CONTENT:

                                                   ASamp.  x  WStd.

GATIFLOXACIN Content   =  ------------------------------- X   P

                                            AStd.      x  WSamp.

ASamp.= Area of GATIFLOXACIN peak in an injection of sample. 
AStd.    =  Mean Area of GATIFLOXACIN Peak in injection of analytical standard solution. 
WSamp.  =  Weight of the sample taken to prepare relevant sample solution. (in mg) 
WStd.  = Weight of GATIFLOXACIN reference standard taken to prepare analytical standard  Solution. (in mg) 
P   = Known Purity of GATIFLOXACIN Reference standard.

      
2.  2-Methyl Piperazine CONTENT:

                                                      ASamp.  x  WStd.
2-Methyl Piperazine Content  =  ----------------------------  X  P
                                                      AStd.      x  WSamp.

ASamp.    = Area of 2-Methyl Piperazine peak in an injection of sample.
AStd.       = Mean Area of 2-Methyl Piperazine peak in injections of analytical standard solution.
WSamp. =  Weight of the sample taken to prepare relevant sample solution. (in mg)
WStd.    = Weight of 2-Methyl Piperazine reference standard taken to prepare analytical standard solution.(in mg)
P  = Known Purity of 2-Methyl Piperazine reference standard

RESULTS AND DISCUSSIONS: -

SYSTEM SUITABILITY:   
System suitability data as shown in Table No. 1 shows method is accurate.  
Table No. 1: -

Compound

Standard Deviation

RSD

Theoretical Plates

Resolution Factor

Tailing Factor

2-METHYL PIPERAZIN

 

 

5.04619

0.6621

5370

1.6

1.1

GATIFLOXACIN

 

 

302.6325

1.624284

7271

-

-

2.SUPPORTING DATA FOR TEST PROCEDURE REPRODUCIBILITY IN ASSAY TEST

Data in Table No. 2 shows method is rugged & reproducible

GATIFLOXACIN

 

 

 

 

 

2-Methyl Piperazine

 

DAY

WEIGHT

 

X

DAY

WEIGHT

 

X

 

[mg/ml]

 

[%]

 

[mg/ml]

 

[%]

28/09/2004

1

 

99.32

28/09/2004

0.01

 

98.73

 

 

 

98.84

 

 

 

99.12

 

 

 

99.5

 

 

 

99.36

29/09/2004

1

 

 

29/09/2004

0.01

 

 

 

 

 

99.57

 

 

 

98.83

 

 

 

99.24

 

 

 

99.21

 

 

 

99.14

 

 

 

99.08

30/09/2004

1

 

 

30/09/2004

0.01

 

 

 

 

 

99.38

 

 

 

99.33

 

 

 

99.72

 

 

 

98.54

 

 

 

99.71

 

 

 

99.34

 

 

 

 

 

 

 

 

 

AVG. [%]

 

99.38

 

 

 

99.06

 

S.D.

 

0.2844732

 

 

 

0.29580399

 

RSD

 

0.2862479

 

 

 

0.29861093

 

2V

 

0.5724959

 

 

 

0.59722186

                 

NOW YOU CAN ALSO PUBLISH YOUR ARTICLE ONLINE.

SUBMIT YOUR ARTICLE/PROJECT AT editor-in-chief@pharmatutor.org

Subscribe to Pharmatutor Alerts by Email

FIND OUT MORE ARTICLES AT OUR DATABASE


 

Pages

FIND MORE ARTICLES