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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
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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
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US patent 3100770, Rodney Ian Friar, "5-PYRIDYL-1, 4Benzodiazepine Compounds", published 1961-11-7, issued 196313-7.
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