Simultaneous Estimation of Ciprofloxacin and Tinidazole by U.V Spectrophotometer using a Hydrotropic Solubilization Technique

Advanced, easy and accurate UV spectroscopy technique (I & II) was developed for estimation of Ciprofloxacin and Tinidazole in bulk and tablet dosage form and validated using hydrotropic solubilization techniques. In Method I we focused on solving the simultaneous equation on the basis of absorbance measurement at 270nm and 320nm wavelength of Ciprofloxacin and Tinidazole individually in 2M urea solution which results in enhancement of solubility of CPH and TNZ to about 15 folds as compared to distilled water.Strategy II is lay out idea of Q-analysis in which absorbance was estimated at 288nm (iso-absorptive point) 270nm (?max of CPH) in 2M urea. Ciprofloxacin and Tinidazole display linearity at chose frequencies and comply with Beer's regulation in the focus scope of 0.2-1.2µg/mL and 10 to 60µg/ mL separately. Recovery reads up for CPS and TNZ were completed and rate recuperation of the medications gotten in the scope of 99.2 - 99.6% (Method A) and 98.0-100.8% (Method B) affirming the exactness of the proposed strategy. Both the techniques showed great reproducibility and recuperation with %RSD less than 2. Factual approval of the information shows that the proposed strategies can be effectively applied for the standard examination of medications in business tablets.

Hydrotropic is solubilization process, where the adding of huge quantity of a subsequent solute outcome in the rise of aqueous solubility of insoluble and slightly solvable drugs.A poorly water-soluble drug refers to a "practically insoluble" 1 . Poor solubility of drugs is a crucial factor in dosage form designing and due to which drugs show low bioavailability. Hydrotropic agent like, nicotinamide, lysine, ß-cyclodextrinand urea,can be utilized for the solvency upgrade 2 . synthetically ciprofloxacin be in to the gathering of manufactured fluoroquinolone anti-infection agents with strong action against wide range microscopic organisms. Tinidazole is dynamic against both gram+ and gram -microbes.

Chemical and Reagent
Analytical grade of reagents and solvents take place used for the quantitative estimation of Ciprofloxacin and Tinidazole from Himgiri traders, whereas gift sample of Ciprofloxacin and Tinidazole was obtained from east African overseas. 2.0M urea was used as hydrotropic agent.

Instrumentation
D o u b l e b e a m U V -V i s i b l e Spectrophotometer of Agilent Technologies (Model No-G6860A) take placed. All weighing done on electronic balance.

Selection of solvent
Solubilityof the two not set in stone at 28 ± 2C. An immense measure of medications was added to the recepticle containing different hydrotropic arrangement. The measuring glasses were shaken precisely for 12 hrs.' at 280C ± 10 C in a mechanical stirrer. Sample were permitted to get stable within 24 hrs. and afterward agitate for 5min at 2000 rpm. The supernatant fluid taken for proper dilution after separated through filter paper (whatmann) and investigated spectrophotometric partner against reference. After examination it was observed that the expansion in the dissolvability of CPH and TNZ was viewed as in excess of 15 folds in 2.0M urea arrangement when contrasted with solvency in refined water 5,6 .

Preparation of Stock Solution (1000ìg/ml)
Accurately weigh 50mg CPH and TNZ independently and moved to two separate 50ml volumetric, dissolve with the 2M urea arrangement and last volume left up to mark with distilled water get stock soln. ofCPH (1000µg/ml) and TNZ (1000µg/ml).  Preparationof Working Standard (100µg/ml) From the above stock solution, 5ml every one of CPH and TNZ was taken more time, to isolate 50ml volumetric carafe and volume was up to 50ml D.W water to acquire working norm of CPH(100µg/ml) and TNZ(100µg/ml).

Method I: Simultaneous Equation Method
Assuming an example contain two absorbing drug (X and Y) all absorbed at different ëmax. it very well may be practicable to choose the two drugs by the simultaneous equation 7,8 .

Determination of wavelength of maximum absorbance of Ciprofloxacin and Tinidazole
From the working standard, 10µg/ml sol n. of CPH and TNZ independentlyprepared in D.W and the solutions were scanned under spectrum mode for 200-400nm wavelength range against blank to analyses the ëmax .Sharp peaks were detected at 270nm for CPH and 320nm for TNZ.

Preparation of Calibration curve of CPH and TNZ
From the working norms of CPH(50µg/ ml) 0.2, 0.4, 0.6, 0.8, 1.0, 1.2 and TNZ(50µg/ml) 10, 20, 30, 40, 50, 60 ml were moved to 10ml volume made up in volumetric flask to mark with D.W. Calibration curve of absorbance versus concentration was plotted and the regression coefficient was determined as shown in fig 1 and 2.
Both the drugs obey's Beer-Lambert's law in the conc. of0.2-1.2µg/ml for Ciprofloxacin and 10-60µg/ml for Tinidazole. The correlation coefficient were found to be R²>0.999. The absorptivity values of CPH and TNZ are calculated at their respective wavelengths and presented in table 1.
Derivation of equation [9] : from the absorptivity values obtained for CPH and TNZ,  simultaneous equation is derived for determination of these two drugs in combination in their pharmaceutical formulations. A1 = 662CCPH + 206CTNZ A2 = 445CCPH + 253CTNZ A1 and A2 are the absorbance of mixture at chosen wavelength 270nm and 320nm 10 .

Method II: Absorbtion Ratio Method Selection of Iso-absorptive point
From the overlay spectrum of CPH and TNZ two wavelength were selected, one at 288nm (ë1) isoabsorptive point for both drugs and other at 270nm(ë2) wavelength of ciprofloxacin for the study. Overlay spectrum is shown in figure 2 and the absorptivity values for CPH and TNZ at selected wavelength are presented in table 1.

Anaysis of the Tablet Formulation
Onconcurrent evaluation of CPH and TNZ in retail tablet dosage form (CIPLOX-TZ) containing retailproduct of CPH-500 mg and TNZ-600mg was completed. In this assay methodology, 20 tablets were squashed and ground to a fine powder. Powder comparable to 25 mg of CPH and 30 mg of TNZ was moved to a 50 ml volumetric flask containing around 50 ml of urea solution broke up and sonicated for 10 min. The solution was sifted through Whatmann filter paper. The solution was additionally dilution with D.W to get conc. of 10µg/ml for ciprofloxacin and 12µg/ ml for tinidazole. Absorbance of these solution were estimated at 270nm(ciprofloxacin) and 320nm(tinidazole) as ë1 and ë2 and centralization of these two medications in the tablet were determined. The consequences of investigation were in table 3. [14] The developed U.V spectrophotometric was validated as per (International council for harmonization) guidelines in terms of linearity, range, specificity, precision, Accuracy, LOD & LOQ for estimation of ciprofloxacin and tinidazole in bulk & tablet dosage form.

Validation Of Developed Method
Linearity 15 : From working standard of CPH(50µg/ml) 0.2, 0.4, 0.6, 0.8, 1.0, 1.2 and TNZ(50µg/ml) 10, 20, 30, 40, 50, 60 ml transferred to 10ml volumetric flask and volume was made up to mark with distilled water. These solution were then scanned in the range of 400-200nm against solvent as blank at their respective wavelength and then calibration curve was plotted.
Precision 16 Repeatability (inter day) and (intraday). In precision for interday we are required to analyses three samples per day for consecutive three days and for intraday one sample was analysed for three days.

CONCLUSION
The analytical study of Ciprofloxacin and Tinidazole by U.V Visible Spectrophotometry was successfully performed. Developed method enhances solubility of both water insoluble drugs and here was no involvementgenerated by urea in the estimation, therefore we analyzed that two UV spectroscopytechniques were established to be easy, accurate, inexpensive and speedy for concurrent estimation of CPH and TNZ in bulk and tablet dosage forms. So different dosage form containing CPH and TNZ can be easily analyzed by utilizing these techniques of spectroscopy. Ciprofloxine belongs to Class IV and Tinidazole belongs to Class II of BCS Classification.

Conflict of Interest
There is no conflict of interest