Simultaneous Estimation of Atorvastatin, Clopidogrel and Aspirin in Capsule Dosage forms using UV-Spectroscopy

 

Sunil Singh*, Nitin Dubey and D.K. Jain

College of Pharmacy, IPS Academy, Rajendar Nagar, Indore [Madhya Pradesh], India

*Corresponding Author E-mail: rssunil29@rediffmail.com

 

ABSTRACT:

Atorvastatin [R-(R*,R*)]-2-(4-fluorophenyl)-ß, δ-dihydroxy-5-(1-methylethyl)-3-phenyl-4-[(phenyl amino)carbonyl]-1H-pyrrole 1-heptanoic acid an Anti-hyperlipidemia and Clopidogrel (2-Chloro phenyl, 6-7 dihydro4H thieno [3,2-c]pyridine-5-yl)acetic acid is an Anti-platelet drug  and Aspirin (2-Acetoxy benzoic acid) an NSAIDs. This paper presents two methods i.e. first derivative spectrophotometery and multicomponent spectrophotometery for simultaneous estimation of these three drugs combination in pharmaceutical formulations. The first derivative amplitudes at 276, 226 and 222 nm were utilized for simultaneous estimations. The multicomponent amplitudes at 247, 220 and 235 nm were utilizatied. Proper selection of wavelength for estimations of one drug by other in derivative spectra and multicomponent lead to successful development of methods for simultaneous estimation. The results of analysis were validated statistically that included parameters such as precision, LOD, LOQ, recovery and robustness. Both methods were simple, economical, accurate, reproducible and precise.

 

KEYWORDS: Simultaneous determination; Derivative spectroscopy; Multicomponent method

 

 

INTRODUCTION:

A combination of atorvastatin1,2,9 (ATOR) clopidogrel2,3 (CLO) and aspirin1,2,3 (ASP) in the form of a tablet or capsule formulations is widely used for the moderate to serve antihyperlipidimic not controlled by a single agent. The official monographs describe the procedure for individual assay of ATOR, CLOP and ASP. 

 

RP-HPLC4,5 methods have been reported for quantative simultaneous determination of ATOR and ASP, ASP and CLOP combinations. Clopidogrel and Aspirin in combined dosage form has been reported to be analyzed and quantified by simultaneous eqution method11,15 of UV-Spectrophotometery. But till no specrophotometeric method has been reported for the quantitative determination of ATR, CLO and ASP in combined dosage form10,12.

 

In recent years, derivative spectrophotometry6,7,8 with zero crossing technique8 has been reported to be a useful method in resolving overlapping spectra of multiple components. The method is also reported for successful determination of active ingredients in presence of matrix16,17. In the present work derivative spectroscopy and multi-component methods has been exploited for quantitative determination of ATR, CLO and ASP in combined dosage form13,14.

 

MATERIALS AND METHODS:

Instrumental:

Analysis carried out on Shimadzu UV 1601 UV-VIS spectrophotometer, a double beam high speed scanning spectrophotometer with a photomultiplier tube detector and having spectral bandwidth of 1nm (190-900nm).

 

Chemicals and reagents:

ATOR, CLOP and ASP were received as gratis sample by Trester Formulation Pvt. Ltd., India, Commercial tablets containing ATOR (10mg), CLOP (75mg), and ASP (75mg), Ecospein Gold (Trester Formulation Pvt. Ltd., India) were used for study. All the chemicals used analytical grade (E. Merck, India).

 

Method-I:

UV-Derivative spectroscopic method:

Standard stock solution:

To prepare stock solution of Atorvastatin, (100 µg/ml) 100mg of Atorvastatin was placed in 100 ml volumetric flask and dissolved in 75 ml of methanol and the volume was made up to the mark with methanol, to obtain the solution of 1000 µg/ml. 10 ml of the solution was diluted up to 100ml with methanol to produce final stock solution of 100 µg/ml of Atorvastatin. Standard stock solution of Clopidogrel and aspirin was prepared similarly as that of Atorvastatin, the equmolar solution containing 100µg/ml of solution and prepared appropriate dilutions; all chemicals used were of analytical grade.

 

Sample preparation:

The content of twenty capsules were taken and weighed. Powder weight equivalent to 75mg of CLOP (corresponding amount of ATOR 10mg and 75mg ASP) was accurately taken and transferred to a 50 ml of volumetric flask and 20ml of methanol added to the same and flask was sonicated for the 30 min. The flask was shaken, and the volume was diluted to the mark with the same mixture. The above solution was filtered using whatman filter paper no.1. Appropriate volume of the aliquot was transferred to a 50 ml volumetric flask and the volume was made up to the mark with methanol. The first derivative spectra were recorded and then measured at 276, 226 and 220 nm for ATOR, CLOP, and ASP respectively. The derivative spectra is shown in Fig 1 and analytical data is Tab: 1.

 

Figure 1. Overlay Derivative Spectra of ATOR, CLOP and ASP

 

Table 1: Statistical Validation of dosage form by derivative method

S. No.

Drug

%Mean ± S.D.

1

2

3

ATOR

CLOP

ASP

99.4± 1.140

100± 0.273

99.7± 0526

 

Method- II: UV – Multicomponent method:

Standard stock solution:

Three stock solutions were prepared by dissolving of 1mg, 7.5mg and 7.5mg of ATOR, CLOP and ASP in 100ml of methanol, respectively. Six mixed standerd solution were prepared from the stock solution with different concentration ranging from 2-6, 15-52.5, and 15-52.5 µg/ml of ATOR, CLOP and ASP, respectively. All the mixed standard solution were scanned over the range of 190-390nm in the multicomponent mode using three sampling point, 247, 220 and 235 nm that is the λmax of ATOR, CLOP and ASP respectively. These solutions were used to calculate the linear dynamic range and for the relative quantification of capsules.

 

Sample preparation:

About twenty capsules were taken and weighed. Powder weight equivalent to 75mg of CLOP (corresponding amount of ATOR 10mg and 75mg ASP) was accurately weighed and transferred in 100ml volumetric flask and 40ml methanol added then sonicated for 30 min, after that volume is made-up at the 100ml. The solution was further diluted and run in the range from 190-390 nm. UV measurement was recorded. The multicomponent spectrum is shown in Fig.2 and analytical data present in Table 2.

 

Figure 2. Overlay Multicomponent Spectra of ATOR, CLOP and ASP

 

Table 2: Statistical Validation of dosage form by multicomponent method

S. No.

Drug

%Mean ± S.D.

1

2

3

ATOR

CLOP

ASP

100.08±0.722

100.01±0.121

99.99±0.100

 

RESULT AND DISCUSSION:

In UV-Derivative Spectroscopic method the zero crossing spectra were utilized for analysis. In contrast the first derivative spectra of each pure drug was found to be shone zero crossing point (Fig.1) and assist in there simultaneous estimation. The first derivative wavelengths were utilized 276nm for ATOR, 226 nm for CLOP and 222nm for ASP. The percentage recovery value obtained lia with in standard limit of 98% to 101 % for the method. Which conforms the method is accurate and free from any interference of excipients. The low value of standard deviation obtained conforms precision of the method. In the multicomponent method were considered 247nm, 220nm and 235nm for estimation of ATOR, CLOP and ASP in combined dosage form. The reproducibility, repeatability and accuracy of the proposed method were found to be satisfactory (Table.3) which is evidenced by low values of standard deviation, percent relative standard deviation and standard error.

 

 

Table 3: Validation parameters for UV-Spectroscopic methods

S.N.

Validation

Parameter

Method (Mean %+ S.D.)

Multi-component

Derivative spectroscopy

ATOR

CLOP

ASP

ATOR

CLOP

ASP

1

Linearity (R square)

0.998

0.999

0.998

0.999

0.999

0.998

2

Accuracy

100.08± 0.0072

100.01± 0.0121

99.99 ± 0.019

99.4±  1.140

100± 0.27

99.7± 0526

3

Precision

Intraday

Interday

 

99.98± 0.089

 

99.99± 0.012

99.78±0.410

99.89±0.024

4

Recovery

80%

100%

120%

 

99.56 ± 0.523

 

99.62 ± 0.088

 

99.66 ± 0.210

 

100.3±0.585

 

99.76±0.215

 

100.1± 0.509

99.80 ± 0.213

99.76 ± 0.080

99.69 ± 0.023

99.89± 0.441

99.92± 0.070

99.80± 0.224

100.01 ± 0.594

99.73 ± 0.317

99.35 ±0.161

100.01±0.594

100.04±0.489

100.05±0.721

5

LOD (mg/ml)

0.12

0.67

0.67

0.55

0.74

0.69

6

LOQ (mg/ml)

0.37

2.09

2.01

1.69

2.27

2.1

7

Robustness (%)

99.7

99.53

99.30

98.9

99.38

99.58

 

 

ACKNOWLEDGEMENTS:

The authors express their gratitude to Treaster Formulation Pvt. Ltd., Puducherry, for the sample of pure atorrvastatin, clopidogrel and aspirin.

 

REFERENCES:

1.       Indian Pharmacopoeia, Vol. II, New Delhi: the controller of publications, Govt. of India; 1996, pp. 347

2.       Goodman and Gilman: The Pharmacological basis of therapeutics, 10th edition, New York, McGraw Hill Book, 1701; 2001

3.       US Pharmacopoeia, 23 edition us Pharmacopial convention, Rockville MD.1995, pp77

4.       Shah DA, Bhatt KK .Mehta RS: developed and validated a RP-HPLC method for Determination of Atorvastatin and Aspirin in capsule dosage form, Indian Journal of Pharmaceutical Science 2007; 69: 546- 549

5.       Anandkumar K, Ayyappan T, Raghu Raman V: RP-HPLC analysis of Aspirin and Clopidogrel In combination. Indian Journal of Pharmaceutical Science 2007; 69:597-599

6.       Rao GD. (2002). A Text book of Pharmaceutical Analysis. (1stEdn.), Vol-I, pp 1-2

7.       ICH, Q3B Validation of Analytical Procedures: Methodology, International Conference on Harmonization, November 1996

8.       Backett AH, Stenlake JB Eds: In practical Pharmaceutical Chemistry (4th edn), part II CBS publisher, New Delhi, 1997, 275-279

9.       British Pharmacopoeia. Vol.2 HMSO Publication centre, London, 1993 pp.55

10.     M.Parrissi-poulou, V Reizopoulou, M. Kouppans P.Macheras, Int.J. Sharma 51 (1989) 169-174

11.     Morel B, Analyst113 (1983) 1077 – 1082.

12.     Savitzky A, MJE Golay, Journal of Analytical Chemistry 36; 1964:1627 – 1639.

13.     Ahuja S, Scypinski SIn: Hand book of Modern Pharmaceutical Analysis, Vol-3, Academic Press; 2001: 1-22.

14.     Kealey K, and Haines PJ:  Analytical Chemistry, 1st Edn. Bios scientific Publisher Limited; 2002

15.     Sharma BK: Instrumental methods of chemical analysis, 24th Edn. Goel Publishing house, Meerut, C-286 to C-311; 2005

16.     Sahu LK, JKGupta: determine derivative spectrophotometric analysis of Psudoephedrine, chlorpheneramine and bromhexine. Indian Journal of Pharmaceutical Science: 2008;66(6):790

17.     Prasad CVN, Parihar C, Sunil P, Parimoo P: Simultaneous determination of amlodipine HCL, Hydrochlorothiazide and atenolol in combined formulation derivative spectroscopy; 2007: 258

 

 

 

Received on 12.04.2010        Modified on 10.05.2010

Accepted on 22.05.2010        © AJRC All right reserved

Asian J. Research Chem. 3(4): Oct. - Dec. 2010; Page 885-887