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Simple Spectrophotometric Methods For Determination Of Ambroxol

Hydrochloride From Pharmaceutical Formulation

Article  in  International Journal of PharmTech Research · July 2012

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 International Journal of PharmTech Research
CODEN (USA): IJPRIF ISSN : 0974-4304
Vol.4, No.3, pp 994-998, July-Sept 2012

Simple Spectrophotometric Methods For

Determination Of Ambroxol Hydrochloride From
Pharmaceutical Formulation
Rele Rajan V.*, Gurav Pankaj J.

D.G. Ruparel College, Mahim, Mumbai 400 016,MH,India.

*Corres.author : [email protected], Phone no. 9870125925, 9757226296

Abstract: Simple sensitive and accurate spectrophotometric methods have been developed for the estimation
of ambroxol hydrochloride in pharmaceutical dosage form. The drug was diazotised with sodium nitrite in
presence of acetic acid and then it was coupled with catechol or resorcinol or β-naphthol in alkaline medium.
The resulting coloured chromogenic species in solution were directly measured at their maximum absorption at
425 nm respectively. The proposed methods were validated statistically. The linearity was found to be 1.0-16
μg/ml, 10-60 μg/ml, and 1.0-20 μg/ml for method I, II and III respectively. The low values of standard
deviation and percentage RSD indicate high precision of methods. Hence these methods are useful for routine
estimation of ambroxol hydrochloride in tablets.
Keywords: Ambroxol Hydrochloride, Glacial acetic acid, Sodium nitrite, Catechol ,Resorcinol,β-naphthol,
Sodium hydroxide, Pharmaceutical dosage form.

INTRODUCTION Glacial acetic acid, sodium nitrite, catechol,

Ambroxol Hydrochloride is trans-4- resorcinol, β-naphthol, resorcinol, and sodium
[(2Amino-3,5-dibromobenzyl)amino] cyclohexanol. hydroxide of AR grade were used in the study.
It shows molecular formula as C13H18Br2N2O.HCl
with molecular weight 414.57. It is official in BP1 Preparation of standard solution and reagents:-
and IP2 . Ambroxol is a metabolite of bromhexine. A standard solution of 1000 µg/ ml of ambroxol
It is an expectoration improver and mucolytic agent hydrochloride was prepared by dissolving 100 mg of
used in the treatment of acute and chronic disorders pure drug in 20 ml alcohol and then diluting the
characterized by the production of excess or thick solution to 100 ml with distilled water. Working
mucus. A literature survey reveals a spectrophoto - standard solution of ambroxal hydrochloride of 100
metric3-6, HPLC7-12 and miscellaneous13-19 methods. µg /ml concentration was prepared by diluting 10 ml
of 1000 µg /ml to 100 ml with distilled water.
MATERIAL AND METHODS A 10% (v/v) solution of acetic acid, 0.5%
A SHIMADZU -UV1800 double beam uv-visible (w/v) solution sodium nitrite, 0.1% (w/v) of
recording spectrophotometer with pair of 10 mm catechol, resorcinol and β-naphthol and 4% (w/v)
matched quartz cell was used to measure absorbance solution sodium hydroxide were prepared in distilled
of solutions. water.
A SHIMADZU analytical balance with 0.01 mg was
used.
Rele Rajan V. et al /Int.J.PharmTech Res.2012,4(3) 995

EXPERIMENTAL naphthol and 1.0 ml of sodium hydroxide were

Method I (with Catechol ) successively added to each flask and volume was
Aliquots of the working standard solution of made upto the mark with distilled water. The
ambroxol hydrochloride (10-160 μg/ml ) were solutions were allowed to stand for 5 minutes to
transferred in a series of 10 ml volumetric flask. complete the reaction. The absorbance was
Then 0.5 ml of acetic acid, 2.0 ml of sodium nitrite, measured at 425 nm against reagent blank prepared
2.0 ml of catechol and 0.3 ml of sodium hydroxide in similar manner.
were successively added to each flask and volume Estimation from tablets
was made upto the mark with distilled water. The Twenty tablets of labelled claim 30 mg of
solutions were allowed to stand for 5 minutes to ambroxol hydrochloride were weighed accurately.
complete the reaction. The absorbance was Average weight of each tablet was determined.
measured at 425 nm against reagent blank prepared Tablets were crushed into fine powder. An
in similar manner. accurately weighed quantity of powder equivalent to
100 mg of ambroxol hydrochloride was transferred
Method II (with Resorcinol) into a beaker and it was shaken with 20 ml of ethyl
Aliquots of the working standard solution of alcohol and filtered. The filtrate and the washing
ambroxol hydrochloride (100-600 μg/ml) were were collected in a 100 ml volumetric flask. This
transferred in a series of 10 ml volumetric flask. filtrate and the washing were diluted up to the mark
Then 0.5 ml of acetic acid, 2.0 ml of sodium nitrite with distilled water to obtain concentration as 1000
and 1.0 ml of resorcinol were successively added to μg /ml. A 10 ml of 1000 μg /ml solution was further
each flask and volume was made upto the mark with diluted to 100 ml with distilled water to give final
distilled water. The solutions were allowed to stand concentration of 100 μg/ml.
for 5 minutes to complete the reaction. The Appropriate aliquots of drug solution were
absorbance was measured at 425 nm against reagent taken. The individual assay procedures was carried
blank prepared in similar manner. out for the estimation of drug contents in tablets.
The concentration of the drug in the tablets was
Method III (with β-naphthol) Aliquots of the calculated using calibration curve. The recovery
working standard solution of ambroxol experiment was carried out by standard addition
hydrochloride (10-200 μg/ml ) were transferred in a method. The values of optical and regression terms
series of 10 ml volumetric flask. Then 0.5 ml of of analysis are given in table no I.
acetic acid, 1.5 ml of sodium nitrite, 0.6 ml of β-

Table I : Optical and regression of drug in different methods

Parameter Methods

I II III
λ max (nm) 425 425 425
Beer Law Limits (µg/ml) 1-16 10-60 1-20
Molar absorptivity(L/mol.cm)
Sandell’s sensitivity
Correlation coefficient(r2) 0.999 0.9999 0.999
Regression equation (y=b+ac)
Slope (a) 0.060 0.009 0.045
Intercept -0.002 -0.002 -0.002
Rele Rajan V. et al /Int.J.PharmTech Res.2012,4(3) 996

Table II : Results of recovery of drug for Catechol.

Method I
Amount Amount Total Percent Standard Percentage Mean Mean
of drug of amount recovery deviation of relative standard of
added standard recovered (%) standard deviation C.O.V.
μg/ml added deviation
μg/ml. C.O.V.
1 0 1.0214 102.14 0.04243 4.1539

2 1 2.0000 100.00 0.06801 3.4007 0.05771 2.7570

3 2 3.0119 100.39 0.05748 1.9085

4 3 4.0238 100.59 0.06297 1.5649

Mean of percent (%) recovery = 100.78

Table III : Results of recovery of drug for Resorcinol

Method II

Amount Amount Total Percent Standard Percentage Mean Mean

of drug of amount recovery deviation of relative standard of
added standard recovered (%) standard deviation C.O.V.
μg/ml added deviation
μg/ml. C.O.V.
10 0 10.1429 101.429 0.28305 2.79064

20 10 20.1587 100.794 0.41999 2.08343 0.41630 1.9035

30 20 30.1587 100.529 0.41999 1.39260

40 30 40.2381 100.595 0.54218 1.34744

Mean of percent (%) recovery = 100.586

Table-IV : Results of recovery of drug for β-naphthol

Method III
Amount Amount Total Percent Standard Percentage Mean Mean
of drug of amount recovery deviation of relative standard of
added standard recovered (%) standard deviation C.O.V.
μg/ml added deviation
μg/ml. C.O.V.
1 0 1.0159 101.59 0.07667 7.54663

2 1 2.0000 100.00 0.09071 4.53563 0.08018 4.1333

3 2 3.0159 100.53 0.07667 2.54207

4 3 4.0159 100.398 0.07667 1.90906

Mean of percent (%) recovery = 100.629
Rele Rajan V. et al /Int.J.PharmTech Res.2012,4(3) 997

RESULT AND DISCUSSION interference from the common excipient used in the
The spectrophotometric methods are popular formulation. The reproducibility, repeatability and
due to their sensitivity in assay of the drug and accuracy of these methods were found to be good,
hence spectrophotometric methods have gain which is evidenced by low values of standard
considerable attention for quantitative determination deviations.
of many pharmaceutical preparations. These The proposed methods suggested in literature
proposed methods are spectrophotometric methods were applied need costly reagent for development of
for the determination of ambroxol hydrochloride chromogen and useful in higher concentration. The
from its formulations i.e. tablets. proposed methods are simple, sensitive, accurate,
The colour chromogens are formed and are precise, and reproducible applicable to even very
stable. The working conditions of these methods low concentration as compare to previous methods
were established by varying one parameter at time suggested in literature. They are directly applied to
and keeping the other parameters fixed by observing drug to form chromogen. Hence they can be
the effect produced on the absorbance of the colour successfully applied for the routine estimation of
species. The various parameters involved for ambroxol hydrochloride in bulk and pharmaceutical
maximum colour development for these methods dosage form even at very low concentration in
were optimized. The proposed methods were formulation such as tablets.
validated statistically and by recovery studies. The The strong recommendation is made here for the
molar absorptivity and Sandell’s sensitivity values proposed methods for determination of ambroxol
(Table no. I) show the sensitivity of methods while hydrochloride from its formulation viz. tablets.
the precision was confirmed by % RSD (relative
standard deviation). The optical characteristics such
as absorption maxima (nm), molar absorptivity ( L ACKNOELEDGEMENT
mole-1 cm-1), correlation coefficient (r) and sandell Authors express sincere thanks to the Principal Dr.
sensitivity ( μg/cm2/0.001) were calculated and are T.M. Desai of D. G. Ruparel college for providing
also summarized in table I. Assay results of necessary facilities work , Dr.Atul Pusalkar for
recovery studies are given in table no. II, III and IV. providing gift sample of ambroxol hydrochloride.
Results are in good in agreement with labelled
value. The percent recovery obtained indicates non

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