Km. Babita¹*, Kapil Kalra¹, Neetu Rajpoot¹, Poonam Joshi² and Vandana Pokhriyal¹

 ¹Alpine college of management and technology, Dehradun Uttarakhand India.

²Uttaranchal Institute of Pharmaceutical Sciences, Uttaranchal University, Dehradun Uttarakhand India.

Corresponding Author E-mail: babitanegi495@gmail.com

DOI : https://dx.doi.org/10.13005/bpj/2621

Abstract

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 (λmax 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.

Keywords

absorbance ratio method; hydrotropic agent; Simultaneous estimation method

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Copy the following to cite this article: Babita K, Kalra K, Rajpoot N, Joshi P, Pokhriyal V. Simultaneous Estimation of Ciprofloxacin and Tinidazole by U.V Spectrophotometer using a Hydrotropic Solubilization Technique. Biomed Pharmacol J 2023;16(1)

Copy the following to cite this URL: Babita K, Kalra K, Rajpoot N, Joshi P, Pokhriyal V. Simultaneous Estimation of Ciprofloxacin and Tinidazole by U.V Spectrophotometer using a Hydrotropic Solubilization Technique. Biomed Pharmacol J 2023;16(1). Available from: https://bit.ly/3DNaY5T

Introduction

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.

Material and Methods 

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 

Double beam UV-Visible Spectrophotometer of Agilent Technologies (Model No- G6860A) take placed. All weighing done on electronic balance.

Selection of solvent 

solubility of 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 (1000μg/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. of CPH (1000µg/ml) and TNZ (1000µg/ml).

Preparation of 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 λmax. 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 independently prepared in D.W and the solutions were scanned under spectrum mode for 200-400nm wavelength range against blank to analyses the λmax .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.

Figure 1: Calibration curve of ciprofloxacin at 270nm Click here to view Figure

 

Figure 2: Calibration curve of Tinidazole at 320nm  Click here to view Figure

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 (λ1) isoabsorptive point for both drugs and other at 270nm(λ2) 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.

Figure 3: Overlay spectra of ciprofloxacin and tinidazole. Click here to view Figure

Derivation of equation ^[13]: In Q analysis method, from overlay spectra of CPH and TNZ absorbances were detected at chosen wavelength i.e. at 270nm (λmax of CPH ) and 288nm (isoabsorptive point). The concentration of each drug in the tablet is determined by substituting the absorbances and absorptivity in the equation

CCPH = (1.97-1.33)/ (4.21-1.337) x A1/ax1

CTNZ = (1.97-4.21)/ (1.337-4.21) x A2/ ay1

A1 and A2 absorbance of samples at 270nm and 288nm. ax1 and ay1 are the absorptivity of CPH at 270nm and TNZ at 288nm.

Analysis of The Tablet Formulation 

On concurrent evaluation of CPH and TNZ in retail tablet dosage form (CIPLOX-TZ) containing retail product 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 λ1 and λ2 and centralization of these two medications in the tablet were determined. The consequences of investigation were in table 3.

Validation of Developed Method 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.

Linearity¹⁵

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.

Precision¹⁶ 

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.

Accuracy¹⁷

Known amount of standard of CPH and TNZ were added to the pre-analyzed sample solutions of tablet dosage forms. Results are shown in table 2.

LOD and LOQ ^18,19: LOD and LOQ can be calculated as:

LOD = 3.3σ/S

LOQ = 10σ/S

Result and Discussion 

Table 1: Absorptivity values (A 1 %, 1 cm) of CPH and TNZ by method A and B. 

METHOD I						METHOD II
Conc. μg/ml	CPH		Conc. μg/ml	TNZ		Conc. μg/ml	CPH		Conc. μg/ml	TNZ
	270nm	320nm		270nm	320nm		270nm	288nm		270nm	288nm
0.2	865	471	10	354	347	0.2	771	180	10	431	161
0.4	680	397	20	226	315	0.4	646	197	20	256	132
0.6	646	433	30	188	262	0.6	590	170	30	223	108
0.8	614	467	40	163	207	0.8	578	133	40	195	146
1.0	598	454	50	159	188	1.0	564	100	50	168	194
1.2	578	469	60	149	204	1.2	562	167	60	150	183
mean	662	445		206	253	mean	618	157	mean	237	154

 

Table 2: Determination of accuracy of CPH and TNZ 

Drug	Level of recovery (%)	Quantity taken (µg/ml)	Quantity added (µg/ml)	Total (µg/m l)	Quantity recovered		% Recovery(n=3)
					Method I	Metho d II	Method I	Method II
CPH	80	5	4	9	9.1	9.2	101.1	102.2
	100	5	5	10	9.8	9.6	98	96
	120	5	6	11	10.8	10.5	98.1	95.45
TNZ	80	6	4.8	10.8	10.7	10.6	99.07	98.1
	100	6	6	12	12.2	12.1	100	100.8
	120	6	7.2	13.2	13.0	13.1	98.4	99.2

 

Table 3: Analysis of tablet formulation for method A & Method 

METHOD	LABEL CLAIM		AMOUNT FOUND		% RECOVERY
	CPH	TNZ	CPH	TNZ	CPH	TNZ
A	500	600	496	598	99.2 %	99.6 %
B	500	600	490	605	98 %	100.8 %

 

Table 4: Optical characteristics Data 

Parameters	Method A		Method B
	CPH	TNZ	CPH	TNZ
Analytical wavelength	270nm and 320nm		270nm and 288nm
Beer’s range	0.2 –1.2µg/ml	10–60µg/ml	0.2 –1.2µg/ml	10- 60µg/ml
Slope	0.52	0.01	0.51	0.009
Intercept	0.06	0.023	0.05	0.06
Correlation coefficient	0.9995	0.997	0.999	0.998
Intra-day precision	0.366	0.951	0.350	0.421
Inter-day precision	0.433	0.853	0.465	0.243
LOD	0.310	0.180	0.101	0.212
LOQ	0.718	0.821	0.103	0.991
Tablet assay	99.2%	99.6%	98%	100.8%

Discussion 

Solvency of ciprofloxine and Tinidazole was viewed as in excess of 15 folds in 2.0M urea solution when compared with dissolvability in D.W. It is apparent from table 3 that the percent drug assessed in tablet definition was 99.2% for CPH and 99.6% for TNZ by technique An and 98% for CPH and 100.8% for TNZ by method II The optical characteristic such as, t , regression equation, beer’s range and Correlation coefficient also determined in table 4.

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 involvement generated by urea in the estimation, therefore we analyzed that two UV spectroscopy techniques 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

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