INTRODUCTION:

Sparfloxacin, a synthetic fluoroquinolone antibacterial agent acts on DNA gyrase and toposiomerase IV, enzymes which, like human topoisomerase, prevents the excessive super coiling of DNA during replication or transcription. By inhibiting their function, the drug thereby inhibits normal cell division.

Increasing the aqueous solubility of insoluble and slightly soluble drugs (Sparfloxacin is practically insoluble in water) has been achieved by various methods to avoid the usage of organic solvents. Because of corrosive, toxicity, volatility, hazards to environment and also high cost of organic solvents, alternative method has been developed.

Hydrotropic solubilization is one of the methods to enhance the aqueous solubility of less water-soluble drugs with eco-friendly.

Sparfloxacin is a synthetic broad-spectrum antimicrobial agent for oral administration. Sparfloxacin has activity against a wide range of gram-negative and gram-positive microorganisms. Mostly Sparfloxacin is used in acute bacterial exacerbations of chronic bronchitis and community-acquired Pneumonia, uncomplicated skin infections, acute pelvic inflammatory disease, Uncomplicated cystitis, Complicated urinary tract infections, Prostatitis. Chemically, Sparfloxacin, an amino difluoro quinolone, is 5-Amino-1-cyclopropyl-7-(cis-3,5-dimethyl-1- piperazinyl)-6,8-difluoro-1,4-dihydro-4-oxo-3-quinolinecarboxylic acid^1,2. The chemical structure (Fig. 1) is:

Fig 1: Chemical Structure of Sparfloxacin

Few RP-HPLC and UV-Visible spectroscopic methods were employed for the analysis of Sparfloxacin in solid dosage form and also in combined dosage form with other drugs^3-15. Many organic solvents are used as the mobile phase and to dissolve the drug in above said methods. Hydrotropic solubilizing agent may be a proper choice to preclude theuse of organic solvents and to enhance the aqueous solubility of drug. Maheswari et.al^16-18 has developed various analytical techniques using hydrotropic solubilization for analyzing poorly water-soluble drugs in bulk and pharmaceutical formulations.

In this present communication, UV spectroscopic method was developed for the analysis of Sparfloxacin in solid dosage forms using 20% sodium benzoate solution as solubilizing agent and proposed method was validated as per ICHQ2(R1) - 2005 guidelines^19-21.

MATERIAL AND METHODS:

Preliminary solubility studies of pure drug:

Solubility of Sparfloxacin was determined at 28±1ºC. Accurately about 100mg of pure drug was taken in screw capped 30mL glass vials containing different solvent systems likes distilled water, buffer of pH 4.0, buffer of pH 9.2, 10% sodium salicylate solution, 20% sodium salicylate solution, 10% sodium benzoate solution, 20% sodium benzoate solution, 20% sodium citrate solution, 20% sodium acetate solution. The vials were shaken for 12 hours in rotatory shaker. These solutions were allowed to equilibrate for the next 24hours and then centrifuged for 5 minutes at 2000rpm. The supernatant liquid from each glass vial was filtered through what man filter paper no.41. Then the filtrates were diluted with distilled water to get 10μg/mL concentration and absorbance was measured at 292nm. Drug content present in each solvent system was calculated from the standard linearity graph.

Standard calibration graph:

The standard stock solution of Sparfloxacin (1mg/mL) was prepared in 20% sodium benzoate solution. This standard stock solution was suitably diluted with distilled water, to get the concentration ranges from 2-20 μg/mL. The solution containing 10μg/mL of Sparfloxacin was scanned between 200 and 400nm. The λ max was found at 292nm and recorded graph was shown at Fig.2. The absorbance of the other diluted solutions was measured at same wavelength and standard calibration graph (Fig.3) was plotted against concentration^22-24. The other optical parameters were displayed on Table 1.

Fig 2: UV scan spectrum for Sparfloxacin in 20% sodium benzoate

Fig 3: Linearity graph for Sparfloxacin

Table 1: Optical Characteristics

Parameters	Values
λ max	292 nm
Linearity	2-20μg/mL
Regression equation	y = 0.5186x – 0.0132
Correlation coefficient	r²= 0.9992
Slope	0.05138
Standard Error on Slope	0.0003432
95% CI of Slope	0.0506 – 0.05216
Intercept	0.004818
Standard Error on Intercept	0.004061
95% CI of Intercept	-0.004367 – 0.01400
Standard Deviation of Residuals	0.007199

Method validation as per ICH guidelines:

Accuracy of the method was determined by the recovery studies in the tablet formulations of the Sparfloxacin^25-27. Recovery studies were carried out by addition of known quantities of standard drug to pre-analyzed sample at three different levels. Also, the experiment was repeated six times in a day to determine intra-day precision (repeatability) and on six different days to determine inter-day precision (ruggedness)^28,29. The percentage relative standard deviation was calculated at each concentration level. The values of method validation are given in Table 2.

Table 2: Validation data for proposed method

Level of standard drug added	Repeatability				Ruggedness
Level of standard drug added	Mean % Recovery	% RSD	SEM	95%CI	Mean % Recovery	% RSD	SEM	95%CI
10 %	99.10	0.3100	0.1255	98.77-99.42	99.46	0.4083	0.1658	99.03-99.89
20 %	99.90	0.4793	0.1955	99.40-100.41	99.67	0.5013	0.1889	99.21-100.14
30 %	99.88	0.4963	0.2013	99.36-100.40	99.75	0.4245	0.1729	99.31-100.20

*(n=6)

Table 3: Results for percentage drug content of marketed formulation

Label claim (mg)	Amount of drug estimated (mg)	Percentage drug content (%w/w)	Mean percentage content (%w/w)	%RSD	SEM	95% CI
200	197.71	98.85	99.55	0.5389	0.2190	98.99-100.10
	199.62	99.81
	199.21	99.60
	198.83	99.41
	198.46	99.23
	200.80	100.42

Content analysis of Sparfloxacin tablet formulations by proposed method:

Twenty tablets of Sparfloxacin marketed formulation was weighed and ground to fine powder. An accurately weighed powder sample equivalent to 100mg of Sparfloxacin was transferred in to a 100mL volumetric flask. Then 50mL of 20% sodium benzoate solution was added and the flask was shaken for about 20minutes to dissolve the drug. Then the volume was made up to the mark with distilled water. The solution was filtered through what man filter paper No.41. The above sample stock solution was diluted with distilled water to get the concentration of 10μg/mL. The absorbance was measured at 292 nm and the drug content of the tablet formulation was then calculated (Table 3).

RESULTS AND DISCUSSION:

From the solubility studies, the drug was satisfactorily soluble in 20% sodium benzoate solution as compared to its solubility in other solvents. Beer’s law is obeyed in concentration range of 2-20μg/mL with correlation coefficient of 0.9992 and regression equation of y = 0.5186x – 0.0132. The percentage drug content was estimated with marketed formulation using proposed UV method and results were ranged from 98.85% - 100.42% w/w with %RSD of 0.5389. Then the method was validated as per ICH guidelines in terms of % recovery and precision (repeatability and ruggedness). Percentage recovery study was performed in terms of inter-day and intraday where as repeated for six consecutive days and also repeated for six times in same day. Percentage recovery greater than 99% and %RSD less than 1% at each level clearly indicates that the method is accurate and precise enough for the content analysis of drug in pure and pharmaceutical dosage forms^30,31.

CONCLUSION:

The proposed method for content analysis of Sparfloxacin was found to be simple, accurate, precise enough, environment friendly and economic. It is concluded that the method is highly reproducible and reliable and has good agreement with label claim of drug.

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Received on 21.03.2023 Modified on 29.08.2023

Asian J. Pharm. Ana. 2024; 14(1):17-20.

DOI: 10.52711/2231-5675.2024.00004