Skip to main content

DETERMINATION OF SYNTHETIC INTERMEDIATE OF DICLOFENAC SODIUM IN REACTION MIXTURES BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Pharma courses

pharma courses

About Authors:
1) Mr Jaesh N.Jadhav
M.Pharm,Sinhagad Institute of pharmacy,Pune.
2) Mr. J.G. Chandorkar*.
Head Analytical development laboratory,
Indofil Industries limited,Thane
*Jayant.chandorkar@rediffmail.com

Abstract
A present work describes a simple & accurate reversed phase HPLC Method for the simultaneous estimation of Ortho Chlorophenol [OCP], 2,6 – Dichlorophenol [DCP], 1-(2,6-Dichlorophenyl) indolin-2-one [VTCL], 1(2,6-Dichlorophenyl) N-Phenyl, N-Chloro Acetyl, 2,6-Dichloro Aniline [VTDL], 1(2,6-Dichlorophenyl) ether [DCPE], 1(2,6-Dichlorophenyl) amine [DCPA] in Diclofenac Sodium bulk manufacturing. This paper describes a new rapid, easy Isocratic reversed phase HPLC method for the separation and estimation of six intermediates and Diclofenac Sodium. The primary purpose of this study is to compile HPLC data on the determination of these seven products, the compilation of such HPLC data being useful as reference guide.

Reference Id: PHARMATUTOR-ART-1378

Introduction
A new simultaneously Isocratic high performance liquid chromatographic method of determination of six intermediate, viz., Ortho Chlorophenol [OCP], 2,6 – Dichlorophenol [DCP], 1-(2,6-Dichlorophenyl) indolin-2-one [VTCL], 1(2,6-Dichlorophenyl) N-Phenyl, N-Chloro Acetyl, 2,6-Dichloro Aniline [VTDL], 1(2,6-Dichlorophenyl) ether [DCPE], 1(2,6-Dichlorophenyl) amine [DCPA] and one drug Diclofenac Sodium, using ion-pairing chromatography is presented1. Quantitative analysis was carried out by the in house method.

The mobile phase used was Water [400 ml], Methanol [300 ml], Acetonitrile [300 ml] and Acetic acid [10 ml] with pH adjusted to 7.0. the separation were carried out using fine pack silica. RP C18 column [25 cm x 4.6 mm) 05 m. The minimum quantifiable concentrations of all the drugs have been determined as well as the linearity range. The calculation revealed that any one of the seven drugs can be used as an internal standard for the determination of the other six drugs. Calculations pertaining to Diclofenac Sodium as internal standard have been cited in the paper, together with a detailed statistical analysis of the data.

These above intermediates getting manufactured during synthesis of Diclofenac Sodium 2, 3. It is necessary to develop a method which can analyze all the precursor of the synthetic route will get analysed by a single analytical method using Isocratic High Performance Liquid Chromatography.

Several methods are available for the estimation of the individual intermediates in Bulk Drug and formulations using spectrometric, HPLC method etc4, 5. The analytical chemistry of Diclofenac Sodium also has been extensively studied references on individual / combinations to drug among the three are numerous6. Only some of them are cited here7.

Experimental: -
Apparatus –
The apparatus used was a Perkin Elmer and UV- Detector 785 A UV/VIS Detector set at 254 nm. The integration was done on a PC based Delta 5.0 software. The column comprised fine pack silica C18 [25 cm X 4.6 mm] 05 m.

Reagents –
Water, Methanol, Acetonitrile and Acetic acid were HPLC grade, from S.D. Fine Chem. & E. Merck respectively.

Drug standard –
All the drug standards were obtained from M/s. Aarti Drugs Ltd., Tarapur with certified copies of analysis. The stock solutions were prepared by dissolving 100 mg of each drug in 100 ml of mobile phase. Another stock solution was prepared mixing the stock solution of the all the seven drugs. The stock solutions were serially diluted in ten fold dilutions to the required concentration.

Mobile phase –
To prepare 1 litre of mobile phase, place 400 ml Water, 300 ml Methanol, 300 ml Acetonitrile and 10 ml of Acetic Acid into a suitable container. Mix the mobile phase thoroughly, and filter and degas under vacuum.

Assay procedure: –
The chromatographic conditions are listed below –
Chromatograph - Perkin Elmer System
Mobile phase – Water: Methanol: Acetonitrile: Acetic acid (40: 30: 30: 01 v/v)
Column - Lechrosphere 25 cm x 4.6 mm i .e. - Stainless steel column containing C-18, 5mODS packing.
Detector - 785 A UV / VIS Detector.
Integrator – Delta 5.0
Flow Rate – 1.0 ml/min.
Injection Volume – 20 ml.
Temperature – Ambient.

Retention time
Ortho Chlorophenol [OCP] = 4.0 Min.
2,6 – Dichlorophenol [DCP] = 5.2 Min.
1-(2,6-Dichlorophenyl) indolin-2-one [VTCL] = 7.5 Min.
1(2,6-Dichlorophenyl) N-Phenyl, N-Chloro Acetyl, 2,6-Dichloro Aniline [VTDL] = 9.0 Min.
Diclofenac Sodium = 11.5 Min.
1(2,6-Dichlorophenyl) ether [DCPE] = 12.5 Min.
1(2,6-Dichlorophenyl) amine [DCPA] = 14.2 Min.
Run time = 30 min.

NOW YOU CAN ALSO PUBLISH YOUR ARTICLE ONLINE.

SUBMIT YOUR ARTICLE/PROJECT AT articles@pharmatutor.org

Subscribe to PharmaTutor Alerts by Email

FIND OUT MORE ARTICLES AT OUR DATABASE

Discussions
The said method is studied for its ruggedness, reproducibility and Linearity studies. Lowest Limit of detection of all the impurities are also studied. Recovery study shows- Almost 98 % recovery of all the impurities along with the product (Diclofenac Sodium)
Ruggedness of the method is also shown in Table –2.

Lowest limit of detection is also indicated in Table –3

Table– I:
Results of Analysis of Synthetic Mixtures of Ortho Chlorophenol [OCP], 2,6 – Dichlorophenol [DCP], 1-(2,6-Dichlorophenyl) indolin-2-one [VTCL], 1(2,6-Dichlorophenyl) N-Phenyl, N-Chloro Acetyl, 2,6-Dichloro Aniline [VTDL], Diclofenac Sodium [DFS], 1(2,6-Dichlorophenyl) ether [DCPE], 1(2,6-Dichlorophenyl) amine [DCPA] Average of Triplicate Determinations.

Name of

Mixture No. – 1

Mixture No. – 2

Mixture No. – 3

Compounds

Taken

Found

Recovery %

Taken

Found

Recovery %

Taken

Found

Recovery %

OCP

15.8

15.62

98.89

25.0

24.64

98.56

28.7

28.22

98.34

DCP

42.6

42.04

98.70

13.4

13.11

98.89

17.0

16.59

97.59

DCPE

9.3

9.11

98.01

9.78

9.66

98.85

12.82

12.59

98.26

DCPA

22.8

22.46

98.52

19.2

18.99

98.94

27.6

27.21

98.62

VTCL

12.0

11.99

99.92

10.6

10.45

98.59

20.6

20.29

98.53

VTDL

10.7

10.48

98.01

8.7

8.59

98.85

15.7

15.42

98.26

DFS

98.96

97.96

98.99

161.0

160.0

99.38

124.7

123.3

98.88

Table – II:

Area Counts of Replicate Runs in Standard Mixture.

Con.

OCP

DCP

DCPE

DCPA

VTCL

VTDL

DFS

1

99.36

99.43

99.63

98.32

100.71

99.00

100.35

2

99.82

99.65

99.50

99.24

99.74

99.12

100.33

3

99.22

99.47

99.88

99.38

98.57

99.44

100.23

4

99.99

100.47

99.88

99.05

99.43

100.73

99.99

5

99.68

99.66

99.61

98.91

99.92

99.24

99.74

6

100.02

100.41

99.79

99.97

99.95

100.53

99.59

Average

99.68

99.84

99.71

99.14

99.72

99.67

100.03

C.V. [%]

0.3312

0.4686

0.1582

0.5605

0.7062

0.7577

0.3193

Conclusion:
A rapid easy & accurate HPLC method has been developed for determination of all the probable impurities viz. OCP, DCP, VTCL, VTDL, DCPE, DCPA by external Std. Method. Calculations have revealed the validity of using any one of the impurities for the estimations of other impurities.

Acknowledgements:
The author thanks Management of Aarti Drugs for encouragement in carrying out this work.

Reference: -
1) Borenstein, M. R., Y. Xue, S. Cooper und T. B. Tzeng. 1996. Sensitive capillary gas chromatographic-mass spectrometric-selected-ion monitoring method for the determination of diclofenac concentrations in human plasma. J. Chromatogr., 685:59-66
2) Brogden, R. N., R. C. Heel, G. E. Pakes, T. M. Speight und G. S. Avery. 1980. Diclofenac sodium: A review of its pharmacological properties and therapeutic use in rheumatic diseases and pain of varying origin. Drugs, 20:24-48.
3) Davies, N. M., und K. E. Anderson. 1997. Clinical pharmacokinetics of diclofenac. Clin.Pharmacokinet., 33:184-213.
4) Goa, K. L., und P. Chrisp. 1992. Ocular diclofenac. A review of its pharmacology and clinical use in cataract surgery, and potential in other inflammatory ocular conditions. Drugs-Aging, 2:473-486.
5) Godbillon, J., S. Gauron und J. P. Metayer. 1985. High-performance liquid chromatographic determination of diclofenac and its monohydroxylated metabolites in biological fluids. J. Chromatogr., 338:151-159.
6) Gonzales, G., R. Ventura, A. K. Smith, R. de la Torre und J. Segura. 1996. Detection of non-steroidal anti-inflammatory drugs in equine plasma and urine by gas chromatography-mass spectrometry. J. Chromatogr., 719:251-264.
7) Kuhlmann, O., G. Stoldt, H.-G. Struck und G.-J. Krauss. 1998. Simultaneous determination of diclofenac and oxybuprocaine in human aqueous humor with HPLC and electrochemical detection. J. Pharm. Biomed. Anal., 17:1351-1356.

NOW YOU CAN ALSO PUBLISH YOUR ARTICLE ONLINE.

SUBMIT YOUR ARTICLE/PROJECT AT articles@pharmatutor.org

Subscribe to PharmaTutor Alerts by Email

FIND OUT MORE ARTICLES AT OUR DATABASE

Search this website