Simple UV Spectrophotometric Assay of Bromazepam

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ISSN: 0976 - 5700

IntRJPharmSci

Research article

Simple UV Spectrophotometric Assay of Bromazepam Safila Naveed*1, Fatima Qamar1 and Syeda Zainab2 of Pharmacy Jinnah University for women Karachi, Pakistan 2Faculty of Pharmacy University of Karachi, Pakistan

1Faculty

ABSTRACT Objective: A sample of the drug was dissolved in water to produce a solution containing Bromazepam six brands. Method: Different dilutions were made ranging from 0.937ppm to 15ppm. The absorbance of dilutions was measured at 233nm against the solvent blank to determine the assay. In our study a rapid and simple U.V spectrophotometric assay was developed. The assay included measuring the absorbance of six different of Bromazepam brands dilutions at the wavelength of 233 nm. Five different dilutions of each brand of Bromazepam ranging from 0.937ppm to 15ppm were prepared. Results: Their percent assay was calculated and regression equation and regression line were obtained to predict further availability of drugs. The correlation coefficient was found to be 0.9993, 0.996, 0.9993, 0.9996, 0.9981, 0.9997 for Sedonil, Bromalex, Lexidium, Anxolite, Lexotanil and Relaxin respectively. Conclusion: All brands were within the range. Keywords- UV spectrophotometry, Bromazepam, Assay, Sedonil, Bromalex, Lexidium, Anxolite, Lexotanil, Relaxin.

Wavelength selection Bromazepam solution was prepared in water. The solution was scanned in the UV regions. The wavelength maximum (λmax) was observed at 233 nm and this wavelength was adopted for absorbance measurement. Standard stock solution

Fig. 1: Structure of Bromazepam

1.5 mg of Bromazepam standard was accurately measured and transferred to a volumetric flask. Volume was made up by water to produce 50 ml.

INTRODUCTION

Sample preparation

Bromazepam (figure 1) is a benzodiazepine derivative. In 1963 this drug was patented by Roche1. It is an anti-anxiety agent having similar side effects of diazepam (Valium). In spite of treating panic states or anxiety, this drug additionally used as premedicant prior to minor surgery. It typically comes in doses of 3 mg and 6 mg tablets2. Bromazepam metabolize in to 3hydroxybromazepam through oxidative biotransformation in liver3.

Six different Bromazepam brands (Sedonil, Bromalex, Lexidium, Anxolite, Lexotanil and Relaxin) were purchased from different pharmacies located in Karachi, Pakistan. The tablets of each brand of Bromazepam (Sedonil, Bromalex, Lexidium, Anxolite, Lexotanil and Relaxin) were of same batch. The all tablets were labeled to claim Bromazepam 1.5mg per tablet. All the brands have 5 year shelf life.

P. Richter reported A method using second derivative absorption spectrophoto-metry for the determination of bromazepam in pharmaceutical formulations4 Salah M. Sultan et al developed spectrophotometric method for the assay of bromazepam by making a complex of the drug with iron (II) and measuring the absorbance at λmax.5 In our research using U.V spectrophotometer assay method of bromazepam have been developed. The superiority of this method over the other is that this method is cost effective and require a shorter period for analysis. We have successfully performed such types of assay for different drugs for the measurement of active substances6-12. EXPERIMENTAL

Average weight of 20 tablets of each brand of Bromazepam was determined. Tablets of each brand of Bromazepam were then crushed uniformly with the help of a mortar and pestle. Sample powder equivalent to 1.5 mg of Bromazepam of each brand of Bromazepam was transferred into a volumetric flasks containing 10mL water. After 5 minute of sonication period volume was made up to 50 ml with water. Procedure After preparation of standard and tablet solutions, strength of solution 15 ppm in 50 ml, absorbance of the sample preparation and standard preparation at the wavelength of maximum absorbance at about 233nm were measured. Quantity in mg, of Bromazepam per tablet was calculated.

Spectra were measured by UV visible 1601 Shimadzu double beam spectrophotometer using water as a solvent for the assay of Bromazepam drug. IntRJPharmSci.2014; 05(01); 0013

Naveed et al_______________________________________________________________________________________________________ISSN: 0976 – 5700

RESULT AND DISCUSSION On different brands (Sedonil, Bromalex, Lexidium, Anxolite, Lexotanil and Relaxin) of Bromazepam tablets Pharmaceutical assay was carried out by using a spectrophotometer. Table-1 shows the average weight of the tablet, different concentration, absorbance at 233nm and percent assay of Bromazepam. Ranges of dilutions from 0.937 ppm to 15 ppm solution of Bromazepam brands were prepared (Sedonil, Bromalex, Lexidium, Anxolite, Lexotanil and Relaxin). Their percent assay is calculated. Percent assay of different brands (Sedonil, Bromalex, Lexidium, Anxolite, Lexotanil and Relaxin) of Bromazepam tablets is shown in Fig-2. Lexidium shows a highest percent assay of 142.48%. Relaxin, Sedonil, Anoxilite, Lexotanil shows a percent assay of 128.1%, 119.61%, 107.19%, 100% respectively. While among all the brands Bromalex show at least percent assay of 91.5%. Regression equation and regression line are obtained to predict further availability of drugs. To detect linearity, solutions of 15ppm, 7.5ppm, 3.75ppm, 1.875ppm and 0.937ppm of each brand of Bromazepam was prepared. Absorbance was taken thrice in triplicate at each level in spectrophotometric analysis. Table-2 shows the absorbance of different dilutions. For

linearity plot concentration vs. absorbance at level 15ppm, 7.5ppm, 3.75ppm, 1.875ppm and 0.937ppm is shown in Fig.38. The correlation coefficient was found to be 0.9993, 0.996, 0.9993, 0.9996, 0.9981, 0.9997 for Sedonil, Bromalex, Lexidium, Anxolite, Lexotanil and Relaxin respectively. The correlation coefficient value should not be less than 0.99. Squared correlation coefficient value for all the brands (Sedonil, Bromalex, Lexidium, Anxolite, Lexotanil and Relaxin) was well within the limit. CONCLUSION A good linear relationship was observed for different concentrations ranges of 15ppm, 7.5ppm, 3.75ppm, 1.875ppm and 0.937ppm. The correlation coefficient was found to be 0.9993, 0.996, 0.9993, 0.9996, 0.9981, 0.9997 for sedonil, Bromalex, Lexidium, Anxolite, Lexotanil and Relaxin respectively. Correlation coefficient value should not be less than 0.99.Squared correlation coefficient value for all the brands (Sedonil, Bromalex, Lexidium, Anxolite, Lexotanil and Relaxin) was well within the limit.

Table 1: Evaluation of bromazepam tablets

Adamjee

Brand Name Sedonil

Average wt of tablet g 0.246

wt for 15 ppm in 50 ml 0.246

Indus

Bromalex

0.276

Sami

Lexidium

Epla

Pharma

Abs at 233 nm

% assay

0.183

119.61

0.276

0.14

91.5

0.2

0.2

0.218

142.48

Anxolite

0.198

0.198

0.164

107.19

Martin Dow

Lexotanil

0.201

0.201

0.153

100

Macter

Relaxin

0.205

0.205

0.196

128.1

Figure 2: Assay of different brands

IntRJPharmSci.2014; 05(01); 0014

Naveed et al_______________________________________________________________________________________________________ISSN: 0976 – 5700

Table-2 Absorbance of different dilution of different brands Conc 15 7.5 3.75 1.875 0.937

Sedonil 0.183 0.094 0.046 0.023 0.016

Bromalex 0.14 0.077 0.047 0.023 0.019

Lexidium 0.218 0.104 0.055 0.026 0.013

Anxolite 0.164 0.088 0.047 0.027 0.016

Lexotanil 0.153 0.077 0.046 0.026 0.013

Relaxin 0.196 0.096 0.047 0.023 0.013

Table-3 Regression equations of different brands Brands Sedonil Bromalex Lexidium Anxolite Lexotanil Relaxin

Regression equations y = 0.0121x + 0.0023 y = 0.0087x + 0.0107 y = 0.0145x - 0.0012 y = 0.0105x + 0.0073 y = 0.0097x + 0.0063 y = 0.0131x - 0.0011

R² 0.9993 0.9996 0.9993 0.9996 0.9981 0.9997

Figure 5: linearity of Lexidium

Figure 3: linearity of Sedonil Figure 6: linearity of Anxolite

Figure 4: linearity of Bromalex Figure 7: linearity of Lexotanil


Naveed et al_______________________________________________________________________________________________________ISSN: 0976 – 5700 _____________________________ For Correspondence: Safila Naveed Faculty of Pharmacy Jinnah University for women Karachi, Pakistan E-mail: [email protected]

Figure 8: linearity of Relaxin REFERENCE 1.

US patent 3100770, Rodney Ian Friar, "5-PYRIDYL-1, 4Benzodiazepine Compounds", published 1961-11-7, issued 196313-7. 2. Bromazepam. Pharmaceutical Benefits Scheme (PBS). Australian Government - Department of Health. Retrieved 23 March 2014. 3. Morton A. Schwartz, Edward Postma,Stanley J. Kolis and Arthur S. Leon: Metabolites of bromazepam, a benzodiazepine, in the human, dog, rat, and mouse. Journal of Pharmaceutical Sciences Volume 62, Issue 11, pages 1776–1779, November 1973. 4. P. Richter. Second-derivative spectrophotometric assay of bromazepam in pharmaceutical formulations. International Journal of Pharmaceutics Volume 72, Issue 3, 17 June 1991, Pages 207– 210. 5. Salah M. Sultan, , Yousif A.M. Hassan, Kamal E.E. Ibrahim Sequential injection spectrophoto-metric assay of bromazepam complexed with iron (II) in hydrochloric acid with chemometric optimization. Talanta Volume 50, Issue 4, 15 November 1999, Pages 841–849. 6. Dilshad, H., Naveed, S. and Naqvi, B. (2013) Assay of New Formulations of Isosorbide Mononitrate by Using UV Spectrophotometer: BPJ0000115. World Research Journal of Medicine, 1, 1-2. 7. Naveed, S., Sultana, N., Arayne, M.S. and Dilshad, H. (2014) A New HPLC Method for the Assay of Levofloxacin and Its Application in Drug-Metal Interaction Studies. Journal of Scientific and Innovative Research, 3, 91-96.www.jsirjournal. com 8. Dilshad, H. and Naveed, S. (2014) Manufacturing of New Formulations of Isosorbide Mononitrate by Dry Granulation Method and Their Comparitive Evaluation with Brands Available in the Market. IRJP, 5, 168-171. 9. Naveed, S., Fatima, Q. and Ghulam, S. (2014) Percentage Assay of Metformin in Different Medium Using UV-Spectrophotometer BPJ0000130. World Research Journal of Organic Chemistry, 2, 2320-5679. 10. Naveed, S. (2014) Simple UV Spectrophotometric Assay of Atorvastatin API Formulation and Their Comparative Study. Global Journal of Medical Research, 14, 35-38. 11. Dilshad, H., Naveed, S. and Waheed, N. (2014) Comparitive Study of Four Different Brands of Captopril Available in Karachi. IRJP, 5, 374-377. http://www.irjponline.com/admin/php/uploads/2170_pdf.pdf. 12. Naveed, S. and Qamar, F. (2014) Comparative Study of Different Brands of Alprazolam. Global Journal of Medical Research, 14, 2528.
