Indian Journal of Chemical Technology

Vol. 16, January 2009, pp. 74-78

Simple and facile methods for the determination of carvedilol in pharmaceuticals

T V Sreevidya & B Narayana*
Department of Post Graduate Studies and Research in Chemistry, Mangalore University, Mangalagangothri 574 199, India
Email: n_badiadka@yahoo.co.uk
Received 23 November 2007; revised 6 October 2008

Simple and sensitive methods for the determination of carvedilol (CAR) are described. The methods are based on the
reaction of CAR with ninhydrin (NIN) in basic medium to form coloured iminium salt having absorption maximum at
402 nm and also with acetaldehyde (AA) in presence of sodium nitroprusside in basic medium to form coloured product
having absorption maximum at 558 nm. The calibration graphs are found to be linear over 0.2 - 1.2 and 0.6 - 2.0 µg mL-1
with molar absorptivity values of 2.571 × 10 4 and 1.617 × 10 4 L mol-1 cm-1 for CAR-NIN and CAR-AA respectively.
The optimum reaction condition and other analytical parameters were evaluated. The statistical evaluations of the
methods were examined by determining intra-day and inter-day precision. The methods were successfully applied to
the assay of CAR in tablet formulations. The accuracy and reliability of the methods were further ascertained by
parallel determination by a reference method and by calculating the Student’s t-test and F-test values at 95%
confidence level.

Keywords: Carvedilol (CAR), Ninhydrin, Sodium nitroprusside, Spectrophotometry

Carvedilol, 1-(9H-carbazol-4-yloxy) - 3-[2-(2- spectrophotometric methods reported recently10,11 are

methoxyphenoxy) ethylamino] propan-2-ol, is a non -
selective β - adrenergic blocking agent with α1 -
blocking activity indicated for the treatment of mild to
moderate congestive heart failure (CHF). It is the first
β-blocker labeled in United States especially for the
treatment of heart failure of ischemic or
cardiomyopathic origin with significant antioxidant
acitivity1-3. Relative to other β-blockers, carvedilol
(CAR) has minimal ‘inverse agonism’ indicating a
reduced negative chronotropic and inotropic effect,
which decreases its potential to worsen symptoms of
heart failure4. At high dosages, it exerts calcium
channel blocking activity5. The benefits of using CAR
in patients with CHF in both single-center and
multicenter trials have been reported in the
literature6-8. It prevents vitamin E, gluthation and SH
protein depletion induced by oxidative stress, the
main defense mechanisms against tissue injury caused
by free radicals9.
Several analytical methods such as
10,11 12-16
spectrophotometry , HPLC , capillary
electrophoresis17,18, fluorometry19,20, synchronous
fluorimetry21, HPLC-MS/MS22, differential pulse
voltammetry23, GC-MS24 have been widely used for
the determination of CAR. Recently, a second
polymorph25 of CAR has also been reported. It is
worthwhile to mention here that the two
in the UV region and thus are not a selective methods,
since excipients present in the dosage form of the
drug can interfere with the estimation method. In view
of the importance of CAR in the treatment of CHF,
simple methods for the determination of CAR have
been developed. The proposed methods when
compared with the reported spectrophotometric
methods take the advantage over the UV-
spectrophotometric method in terms of selectivity
(Table 1). The proposed methods are simple, accurate
and easy to apply in routine analysis of the drug.
Experimental Procedure
A Shimadzu UV-2550 UV-VIS Spectrophotometer
with 1 cm matched quartz cells was used for
absorbance measurements. All reagents used were of
analytical reagent grade and distilled water is used for
the preparation of all solutions. Ninhydrin (0.3%),
NaOH (1%), Na2CO3 (2%), sodium nitroprusside
(1%) and CH3CHO of A R grade were used.
Preparation of carvedilol (CAR) solution
A 1000 µg mL-1 standard drug solution was
prepared by dissolving 0.1 g of CAR in 50 mL
ethanol and diluting to the mark in a 100 mL
calibrated flask. This stock solution was diluted
appropriately to get the working concentration.
 SREEVIDYA & NARAYANA: DETERMINATION OF CARVEDILOL IN PHARMACEUTICALS 75

Table 1 Comparison table for reported method and proposed method

Reagents Range (µg mL-1) λ max(nm) Molar absorptivity Reaction Ref
(L mol-1 cm-1) conditions
- 4.0 – 36.0 285 1.539 × 104 Methanol (as solvent) 14
- 2.0 – 20.0 242 2.193 × 104 Methanol (as solvent) 15
- 1. 0 – 10.0 245 4.453 × 104 PEG-400+ Water (as solvent) 15
Proposed method
Ninhydrin 0.2 – 1.2 402 2.571 × 104 NaOH medium -
Acetaldehyde + SNP 0.6 – 2.0 558 1.617 × 104 Na2CO3 medium -
PEG – Polyethylene
glycol
SNP – Sodium
nitroprusside
Determination of CAR CAR with ninhydrin in presence of a base to form
Using ninhydrin as reagent yellowish-orange coloured iminium salt, which has an
Aliquots containing (0.2 – 1.2 mL) 10 µg mL-1 of absorption maximum at 402 nm (Scheme 1). Method
CAR were transferred into a series of 10 mL standard II is based on the reaction of CAR with acetaldehyde
flasks using a micro burette. To this 2 mL of 0.3% in presence of sodium nitroprusside in basic medium
ninhydrin was added followed by 0.5 mL of 1% to form an adduct, which has an absorption maximum
NaOH solution. The contents were shaken well and at 558 nm (Scheme 2).
were set aside for 5 min and diluted up to the mark
with distilled water and mixed well. The absorbance Optimization of experimental conditions
of each solution was measured at 402 nm against the Preliminary experiments were performed to fix the
corresponding reagent blank. concentration of the reagents that could be measured
spectrophotometrically and were found to be 0.3 %
Using acetaldehyde as reagent
(2 mL), 10 % (1 mL), and 1% (1 mL) for ninhydrin,
Aliquots containing (0.6 – 2.0 mL) 10 µg mL-1 of
acetaldehyde and sodium nitroprusside respectively.
CAR were transferred into a series of 10 mL standard
For method I, NaOH medium was found to be ideal
flasks using a micro burette. To this 1 mL of freshly
and 0.5 mL of 10 % NaOH in a total volume of
prepared acetaldehyde solution (10%) and 1 mL of
10 mL was adequate for the reaction to take place and
sodium nitroprusside (1%) were added followed by
1 mL of Na2CO3 (2%) solution. The contents were
shaken well and were set aside for 15 min and was
heated on a water bath for about 5 min and diluted up
to the mark with distilled water and mixed well. The
absorbance of each solution was measured at 558 nm
against the corresponding reagent blank.
Assay of formulations
Two different CAR dietary supplement products
were purchased and a sample stock solution of each
was prepared by grinding an amount of powdered
tablets equivalent to 0.1 g of CAR to a fine powder
using a mortar and pestle and transferring into a
100 mL volumetric flask by washing with methanol
and made upto the mark with methanol. A convenient
aliquot was then subjected to analysis by each
method.
Results and Discussion
The determination of CAR is achieved by two
simple methods. Method I involves the reaction of Scheme 1  Reaction of CAR with ninhydrin in presence of base
76 INDIAN J. CHEM. TECHNOL., JANUARY 2009

at five levels was determined each day for a week,

preparing all solutions fresh. The relative standard
deviation values were less than 2 % which represent
the best appraisal of the procedures in daily use.

Table 2 Analytical parameters for the determination of CAR

Parameters CAR-NIN CAR-AA

λmax (nm) 402 558

Beer’s law limits 0.2 – 1.2 0.6 – 2.0

-1
(µg mL )
Molar Absorptivity 2.571 × 104 1.617 × 104
(L mol-1 cm-1)
Sandell Sensitivity 1.58 × 10-2 2.51 × 10-2
(µg cm-2)
Limit of detection** 0.139 0.157
-1
(µg mL )
Limit of quantification** 0.418 0.222
-1
(µg mL )
Scheme 2  Reaction of CAR with acetaldehyde and sodium
nitroprusside in presence of base Regression Equation* Y= bX + a Y= bX + a

for method II, Na2CO3 was found ideal and 1 mL of Slope (b) 0.024 0.021
2% Na2CO3 was found to be optimum. A 5 min Intercept (a) 0.047 0.045
standing time was found necessary for the complete Correlation Coefficient 0.9996 0.9990
reaction to take place. Under these conditions, the
system was stable for a period of over 8 h. * Y is the absorbance and X is the concentration in µg mL-1
** Calculated using ICH - Guidelines
Analytical data
Table 3  Evaluation of accuracy and precision
A linear correlation was found between
absorbance and concentration of the drug. The Using ninhydrin as reagent
optical parameters such as molar absorptivity, Beer’s
Amount Amount SD RE (%) RSD
law limits and Sandell sensitivity values were taken found* (%)
calculated and are given in Table 2. The table also
has values for correlation coefficients, intercepts and (µg mL-1) (µg mL-1) (µg mL-1)
slopes. The limit of detection and quantification 0.4 0.401 0.002 0.150 0.454
calculated according to ICH guidelines26 are also 0.6 0.601 0.003 0.067 0.479
incorporated. 0.8 0.801 0.002 0.125 0.234
1.0 1.006 0.011 0.560 1.131
Method validation 1.2 1.205 0.009 0.383 0.718
Accuracy and precision
Using acetaldehyde as reagent
To evaluate the accuracy and precision of the
methods, pure drug within the working limits was 0.6 0.603 0.001 0.433 0.189
0.8 0.801 0.002 0.100 0.185
analyzed, each determination being repeated five
1.0 1.008 0.008 0.800 0.830
times. The RE (%) and RSD (%) values were less 1.2 1.206 0.009 0.500 0.742
than 1 which indicate the high accuracy and precision 1.4 1.405 0.011 0.343 0.796
for the methods (Table 3). For a better analysis of * Mean value of five determinations
reproducibility on a day-to-day basis, a series of RE- Relative Error; SD- Standard Deviation; RSD-Relative
experiments was run in which standard drug solution Standard Deviation
 SREEVIDYA & NARAYANA: DETERMINATION OF CARVEDILOL IN PHARMACEUTICALS
Interference study
In the pharmaceutical analysis, it is important to
test the selectivity towards the excipients and fillers
added to the pharmaceutical preparations. Several
species that can occur in the real samples together
with drug were investigated. To investigate the effect
of tablet fillers on the measurements involved in the
methods, standard addition method was carried out. A
mixture containing lactose, starch and glucose was
extracted with water and filtered. The filtrate was then
added to a known amount of pure drug solution and
subjected to analysis by the proposed methods. An
error of less than 1.5 % in the absorbance readings
was considered tolerable. From this study it is
apparent that the usual co-formulated substances
seldom interfere in the proposed methods.
Application
The proposed methods were applied to the
determination of CAR in two commercial dietary
supplements (tablets). An aliquot containing 0.8 mL
(10 µg mL-1) drug solution was taken and assayed
according to the proposed methods. The content of
the tablet formulation was calculated by applying
suitable dilution factor. The accuracy of the
proposed method was checked by a thorough
analysis of each spiked sample (Table 4). The results
were also compared statistically with those of the
reference method at 95% confidence level. The
calculated student’s t- test and F-test values
(Table 4) did not exceed the tabulated value,
indicating that there was no significant difference
between the proposed methods and the reference
method in respect to accuracy and precision.
Table 4  Result of assay of formulations by the proposed
methods
Brand name Labeled Found* ± Found* ± SD Reference
amount SD Using Using AA method10
(mg) NIN
Carloc 25 25.0 24.88 ± 0.08 24.98 ± 0.17 24.72 ± 0.14
a a
t = 2.22 t = 2.63
b
F = 3.14 b
F = 1.47
Carvidas 25 25.0 24.89 ± 0.08 24.92 ± 0.09 24.78 ± 0.07
a
t = 2.25 a
t = 2.69
b b
F = 1.44 F = 1.77
* Mean value of five determinations
a
Tabulated t-value at 95% confidence level is 2.78
b
Tabulated F-value at 95% confidence level is 6.39
77
Conclusion
Simple spectrophotometric methods for the
determination of CAR have been developed and
validated according to ICH guidelines. The methods
compared to other existing methods do not entail any
rigorous experimental variables which affect the
reliability of the results. The ingredients usually
present in the pharmaceutical formulations of these
drugs seldom interfere in the proposed methods. The
proposed methods are simple, accurate and easy to
perform and can be used for the routine determination
of CAR in bulk and in dosage forms.
Acknowledgement
Authors thank Department of Science &
Technology, Government of India, for financial
support through FIST programme.
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