| Research Article |
Open Access |
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| Method Development and Validation for estimation of
Moxifloxacin HCl in tablet dosage form by RP-HPLC method |
| P Rama Subbaiah, M V Kumudhavalli*, C Saravanan, M Kumar and R Margret Chandira |
| Department of Pharmaceutical Analysis, Vinayaka Mission’s College of Pharmacy, Vinayaka Missions University, Salem-636008, Tamilnadu, India |
| *Corresponding author: |
Dr. M.V.Kumudhavalli
3D/58, Babu Nagar
Salem–636001,
Tamilnadu, India
Tel: 919942574759
E-mail: Kumudhu27@ymail.com |
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| Received August 30, 2010; Accepted September 27, 2010 Published September
30, 2010 |
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| Citation: Subbaiah PR, Kumudhavalli MV, Saravanan C, Kumar M, Chandira RM (2010) Method Development and Validation for estimation of Moxifloxacin HCl in
tablet dosage form by RP-HPLC method. Pharm Anal Acta 1:109. doi:10.4172/2153-
2435.1000109 |
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| Copyright: © 2010 Subbaiah PR, et al. This is an open-access article distributed
under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the
original author and source are credited. |
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| Abstract |
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| A rapid, sensitive and specific RP-HPLC method involving UV detection was developed and validated for
determination and quantification of Moxifloxacin HCl in tablet dosage form. Chromatography was carried out on a
pre-packed Luna C-18, 5µ (250 x 4.6) mm column using filtered and degassed mixture of Buffer: Methanol (55:45)
as mobile phase at a flow rate of 1.0 ml/min and effluent was monitored at 293 nm. The pH of the mobile phase was
adjusted with acetic acid to 6.3±0.4. The method was validated in terms of linearity, precision, accuracy, and specificity,
limit of quantification and limit of detection. The assay was linear over the concentration range of 20 mcg-60 mcg/
ml. Accuracy of the method was determined through recovery studies by adding known quantities of standard drug to
the pre analysed test solution and was found to be 99.3 %-100.2 % within precision RSD of 0.58 for Moxifloxacin HCl.
The system suitability parameters such as theoretical plates, retention time factor and tailing factor were found to be
7968, 5.855 and 1.207 respectively. The method does require only 10 min as run time for analysis which proves the
adoptability of the method for the routine quality control of the drug. |
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| Keywords |
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| Moxifloxacin HCl; RP-HPLC; Estimation; Validation |
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| Moxifloxacin HCl is chemically 1-cyclopropyl-7-((s,s)-2,8-
diazabicyclo(4.3.0)non-8-yl)-6- fluoro-8-methoxy-1,4-dihydro-4-oxo-3
quinoline carboxylic acid. Moxifloxacin is an antibiotic used to treat
respiratory infections, including acute sinusitis, acute exacerbations
of chronic bronchitis, and community-acquired pneumonia, as well
as dermatological infections, as a second-line agent in tuberculosis.
The literature survey [1-7] reveals that there is some HPLC methods
have been reported. In this paper we describe a simple, inexpensive,
sensitive and validated HPLC [8-12] method for the determination of
Moxifloxacin HCl in bulk and pharmaceutical formulation. |
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| Working standard of Moxifloxacin HCl was obtained from well
reputed research laboratories. HPLC grade Methanol, Merck grade
Orthophosphoric acid and Triethylamine and Milli-Q water were
procured from the market. The separation was carried out on
isocratic HPLC system with pre-packed Luna C-18, 5µ (250 x 4.6)
mm column using filtered and degassed mixture of Buffer: Methanol
(55:45) as mobile phase. |
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| Buffer Preparation |
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| Transferred 2 ml of orthophosphoric acid into 1000 ml of water
and the PH were adjusted to 2.5 with triethylamine, filtered through
0.45µm nylon membrane filter and degassed. |
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| Mobile phase |
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| Buffer and methanol were mixed in the ratio of 55:45 and
sonicated to degas. |
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| Standard preparation |
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| Accurately Weighed and transferred Moxifloxacin HCl equivalent
to 20 mg of Moxifloxacin Working Standard into a 50 ml clean dry
volumetric flask, and 30 ml of mobile phase was added, sonicated for
5 minutes, and diluted to volume with mobile phase. Further diluted
5 ml to 50 ml with mobile phase. |
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| Flow rate 1.0 ml/min; detection wavelength 293 nm; injection
volume 10µl; column used Luna C18 (5µm, 250x4.6 mm); column
temperature: 25oC; mobile phase: Buffer: Methanol (55:45). |
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| Working standard of various concentrations was prepared by
taking aliquots of standard solution and diluted to get required
concentration for calibration plot and which was injected. |
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| Sample preparation |
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| Weighed and powered 20 tablets. Transferred the powder
equivalent to 400 mg of Moxifloxacin into 100 ml of clean, dry,
volumetric flask and, to this added 70 ml of mobile phase and
sonicated for about 15 minutes, further made up the volume with
mobile phase and then filtered through 0.45 micron filter. Further
diluted 1 ml of the filtrate to 100 ml with mobile phase. 10µl of the
standard preparation and assay preparation were separately injected
and chromatographed. |
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| Linearity [13-18] was demonstrated by analysing six different
concentrations of active compound. Peak areas were recorded for all
the peaks and calibration plot was constructed by plotting peak area
vs. concentrations of Moxifloxacin HCl which were found to be linear
in the range of 20 mcg/ml-60 mcg/ml. Coefficient of correlation was
0.9990 (Figure 1). Accuracy [13-18] was done by recovery study using
standard addition method, known amount of standard Moxifloxacin
HCl in to pre-analysed samples and subjected to proposed HPLC
method. The results of recovery studies are shown in (Table 1). |
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Figure 1: Linearity curve of Moxifloxacin HCl. |
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Table 1:Analysis of tablet containing moxifloxacin HCL. |
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| To demonstrate agreement among results, a series of
measurements are done with Moxifloxacin HCl six replicate injections
of the specific standard at various time intervals on the same day
were injected into the chromatograph and the value of % RSD was found to be 0.58. In inter-day precision [13-18] same standard was
injected on different days and the found % RSD was found to be 0.95
%. The results were showed in the (Table 2). |
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| The regression value was found to be 0.9990 for Moxifloxacin
HCl, which shows the response is linear from 20µg/ml to 60µg/
ml. Selectivity experiment showed that there is no interference or
overlapping of the peaks either due to excipients or diluents with the
main peak of Moxifloxacin HCl. The percentage RSD for precision
is < 2 which confirms that method is sufficiently precise and the
total run time required for the method is only 10 min for eluting
Moxifloxacin HCl. The proposed method is simple, fast, accurate, and precise and can be used for routine analysis in quality control of
Moxifloxacin HCl. |
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| References |
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- Motwani SK, Khar RK, Ahmad FJ, Chopra S, Kohli K, et al. (2007) Application of
a validated stability-indicating densitometric thin-layer chromatographic method
to stress degradation studies on moxifloxacin. Anal Chim Acta 582: 75-82.
- SA Shah, IS Rathod, BN Suhagia, MV Baladaniya (2005) An HPTLC method
was developed and validated for the estimation of Moxifloxacin in its tablet
formulation. Indian J Pharm Sci 67: 112-115.
- Motwani SK, Chopra S, Ahmad FJ, Khar RK (2007) A Simple UVSpectrophotometric
method was developed for the estimation of Moxifloxacin
both as bulk drug and pharmaceutical formulation. Spectrochim Acta A Mol
Biomol Spectrosc 68: 250-256.
- Guerra FL, Paim CS, Steppe M, Schapoval EE (2005) Microbiological assay
method was developed and validated for the estimation of Moxifloxacin in
Tablets. J AOAC Int 88: 1086-1092.
- Wen D, Lin Z, Milosavlsev S, Shum L An LC-MS/MS method was developed
for the estimation of Moxifloxacin in human plasmaAvantix Laboratories, Inc,
2Novartis Pharmaceuticals Corporation.
- Boubakar B Ba, Etienne R, Ducint D, Quentinc, Sauxmc (2001) An HPLC
method was developed and validated for the estimation of Moxifloxacin in
growth media. J Chromatogr B Biomed Sci Appl 754: 107-112.
- Taylor, Francis (2007) Developed Differential pulse polarographic method for
the determination of Moxifloxacin in pharmaceutics, serum and urine. Analytical
letters 40: 529-546.
- Mendum J, Denny RC,Thomas MN (2004) Vogel’s Text book of Quantitative
Analysis, 6th Edn, Pearson education ltd.
- Beckett AH, Stanlake JB (2002) Practical Pharmaceutical Chemistry, 4th Edn,
Part 2, CBS Publishers and Distributors.
- Kasture AV, Wadodkar SG, Mahadik KR, More HN (1996) Textbook of
Pharmaceutical Analysis – II, 11th Edn, Nirali Prakashan publication.
- Chatwal GR, Anand SK (2004) Instrumental Methods of Chemical Analysis,
Himalaya Publishing House.
- Swartz ME, Krull IS (1997) Analytical Method Development and Validation,
Marcel Decker, Inc., New York, 25-91.
- Sethi PD (2006) High Performance Liquid Chromatography: Quantitative
Analysis of Pharmaceutical Formulations, CBS Publishers and Distributors,
New Delhi, 1st Edn, 7, 57-63, 116-120.
- Snyder LR, Kirkland JJ, Glajch (1996) Practical HPLC Method Development,
John Wiley & Sons, Inc., New York; 2nd Edn 406-415.
- Green JM (1996) A Practical Guide to Analytical Method Validation, Analytical
Chemistry 68: 305A-309A.
- Riley CM, Rosanke TW (1996) Development and Validation of Analytical
Methods, Biddle Ltd., Guildford; 8-11, 46-60.
- Hong D, Shah M (2000) Development and Validation of HPLC Stability
Indicating Assays in Drug Stability Principles & Practice, Marcel. Decker, New
York, 338-340.
- ICH (1996) Validation of Analytical Procedure, International Conference on
Harmonization, IFPMA, Geneva.
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