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Journal of Chemical Technology

Vol. 13, July 2006, pp. 353-359

Sensitive bromatometric methods for the assay of metaprolol tartrate


in dosage forms
K Basavaiah1*, B C Somashekar1 & V Ramakrishna2
1
Department of Chemistry, University of Mysore, Manasagangotri, Mysore 570 006, India
2
Department of Pharmaceutical Chemistry, Govt. College of Pharmacy, Bangalore 560 027, India
Email: basavaiahk@yahoo.co.in
Received 25 July 2005; revised received 12 April 2006; accepted 1 May 2006

One titrimetric and two spectrophotometric methods are described for the determination of metaprolol tartrate (MPT)
using bromate-bromide mixture and two dyes, methyl orange and indigo carmine, as reagents. In titrimetry, an acidified
solution of MPT is reacted with a known excess of bromate-bromide mixture and after a pre-determined time, the unreacted
bromine is determined by iodometric titration. The spectrophotometric methods involve the addition of a known excess of
bromate-bromide mixture to MPT in acidic medium followed by determination of the residual bromine by reacting with
fixed amount of either methyl orange and measuring the absorbance at 520 nm (Method A) or indigo carmine and measuring
the absorbance at 610 nm (Method B). In all the methods, amount of bromine reacted corresponds to the amount of MPT.
The working conditions of the methods have been optimised. Titrimetry allows the determination of MPT in 2.5 - 7.5 mg
range and the calculations are based on a 1:1(MPT : KBrO3) reaction stoichiometry. In the spectrophotometric methods,
Beer’s law is valid over the concentration ranges 0.5 -5.0 and 1.5 - 15.0 μg mL-1 MPT for method A and method B,
respectively. Method A with a molar absorptivity of 8.17 × 104 L mol-1 cm-1 is more sensitive than method B (€ = 2.70 × 104
L mol-1 cm-1) . The limits of detection and quantification are reported for both the methods. The methods could usefully be
applied to routine quality control of tablets containing MPT. No interference was observed from common pharmaceutical
adjuvants. Statistical comparison of the results with those obtained by an established UV-spectrophotometric method
showed excellent agreement and indicated no significant difference in accuracy and precision. The reliability of the methods
was further ascertained by recovery studies.

Keywords: Metaprolol tartrate, Assay, Bromate-bromide, Dyes, Dosage forms


IPC Code: A61K9/08

Beta blockers have been in clinical use for over 30


years and have an accepted role in the treatment of
high blood pressure, the secondary prevention of
myocardial infarction and in the treatment of
arrhythmias. Metaprolol tartrate (MPT), (±) -
(isopropylamino-3-[4-(2-methoxyethyl) phenoxy]-2-
propanol tartrate (Fig. 1), is known as a cardio
selective beta adrenergic receptor blocker1. Nearly
200 articles have appeared since the early 1980s
describing the detection and determination of this Fig 1⎯ Structure of MPT
drug using a variety of analytical techniques, but a
large number of them are devoted to the analysis of high performance liquid chromatography13-16, high
biological samples. The drug is official in the United performance thin layer chromatography17, gas
States Pharamcopoeia2 which describes a non- chromatography-mass spectroscopy18, densitometry19,
aqueous titrimetric procedure for bulk drug and a high ion-selective electrode based potentiometry20, and
performance liquid chromatographic method for assay voltammetry21 have been reported. But most of the
in tablets. For the determination of this drug in dosage above techniques are tedious, time-consuming and
forms, several techniques including UV- difficult to perform besides involving expensive
spectrophotometry , near infrared spectroscopy7,
3-6
instrumental setup, and are applicable for assay in
fluorimetry8,9, AAS10,11, liquid chromatography12, combined dosage forms.
354 INDIAN J CHEM. TECHNOL., JULY 2006

From a pharmaceutical analysis stand point, it is MPT with 1-fluoro-2,4-dinitro benzene (FDNB) in
highly desirable if the assay methods are simple, rapid tetraborate - HCl medium was used by Shingbal and
and affordable by small-scale industrial laboratories, Bhangle27. In a method presented by one of the same
without a compromise on the requirements of authors28, MPT was reacted with acetaldehyde and
sensitivity, selectivity, and accuracy and precision. In chloranil in the presence of Ag2O to produce a blue
this respect, titrimetry and spectrophotometry chromogen measurable at 680 nm. There are two
continue to be used in industrial quality control reports on the use of charge-transfer complex
laboratories because of their low cost, versatility of reactions employing 4-chloro-7-nitro-2,1,3-
applications and ease of operation. However, there is benzoxadiazole (NBD-Cl)29 and tetracyano ethylene
only one report22 on the titrimetric determination of (TCNE) or chloranilic acid(CAA)10 as π - acceptors
MPT and the method uses metavanadate as the for the determination of MPT. A method10 has been
oxidimetric reagent which is performed in high H2SO4 proposed whereby the chelate formed with copper(II)
concentration. in the presence of CS2 was extracted into isobutyl
MPT has rendered itself to analysis by visible methyl ketone before absorbance measurement. Other
spectrophotometry through several chemical method determining MPT involved the formation of
reactions. Nitration of MPT to produce a yellow copper(II)dithiocarbamate complex11 by derivatisation
derivative in H2SO4 medium was introduced as a the secondary amino group of the drug with CS2 and
quantitative method by Sanghavi and Vyas23. CuCl2 before extracting into chloroform, and
Formation of a yellow colour on reacting the drug measurement. But, most of the currently available
with iron(III) chloride24 in HCl medium has been the spectrophotometric methods suffer from one or the
basis of assay in pharmaceutical preparations. Ersoy deficiency such as poor sensitivity, narrow linear
and Kocaman25 have used bromothymol blue as ion- range of response, stringent experimental conditions
pair reagent for the sensitive determination of MPT. like heating or extraction step, and/or use of
Based on a similar reaction but using benzyl orange as expensive chemicals (Table 1).
a chromogenic reagent, a quantitative assay method In the present study, two techniques, titrimetry and
has been proposed by Vujic et al.26. A stable greenish spectrophotometry have been employed for the assay
- yellow chromophore resulting from the reaction of of MPT using bromate-bromide mixture as the
Table 1⎯Comparison of the existing spectrophotometric methods with the proposed methods for MPT
Sl No. Reagent/s used* λmax, nm Linear range, μg ml-1 Remarks Ref.
1 KNO3 440 <120 Less sensitive 23
2 Iron (III) choride 380 40 - 200 Less sensitive 24
3 Bromothymol 410 2 - 14 Requires rigid pH control, involves liquid-liquid 25
blue extraction
4 Benzyl orange 401 3.4 - 34.2 Requires rigid pH control, involves liquid-liquid 26
extraction
5 FDNB 380 Involves heating a 75° C for 40 min and uses an 27
------- expensive chemical
6 Actaldehyde- 680 25 - 75 Requires heating at 75° C for 10 min, narrow 28
chloranil- Ag2O linear range of response and less sensitive
7 NBD - Cl 470 0.4 - 60.0 Requires partially non-aqueous medium and 29
expensive chemical
8 a) TCNE 415 10 -170 Requires non-aqueous medium; less sensitive 10
b) CAA 510 (3.49 × 103)
20 -230
(1.73 × 103)
9 CS2-CuCl2 435.4 Upto 60 (1.08 × 104) Involves liquid-liquid extraction; less sensitive 10
10 CS2-CuCl2- 440 11 - 55 Involves liquid-liquid extraction and less 11
dithiocarbamate sensitive
11 a) KBrO3-KBr/ 520 0.5 - 5.0 No heating or extraction step, long linear range of Present
methyl orange 610 (8.17 × 104) response, highly sensitive and use inexpensive methods
b) KBrO3-KBr/ 1.5 - 15.0 chemicals
indigo carmine (2.7 × 104)
Figures in the parentheses are molar absorptivities in l mol-1 cm-1
BASAVAIAH et al.: SENSITIVE BROMATOMETRIC METHODS FOR THE ASSAY OF METAPROLOL TARTRATE 355

oxidimetric reagent and methyl orange and indigo by diluting 112 mL of concentrated acid (S. d. Fine
carmine as the spectrophotometric reagents. The Chem., Mumbai, India, Sp gr 1.18) to 250 mL with
methods, when applied to the assay of MPT in dosage water. This was further diluted to 2M with water.
forms, have been found to be convenient and are free Pharmaceutical grade MPT certified to be 99.7 %
from difficulties encountered in many of the existing pure was gifted by Astra - Zeneca, Bangalore, India,
methods. and used as received. A stock standard solution
containing 1 mg mL-1 drug solution was prepared by
Experimental Procedure dissolving 250 mg of pure drug in water diluting to
Reagents and materials the mark with water in a 250 mL calibrated flask. This
A Systronics Model 106 digital spectrophotometer solution was used in titrimetric work and the same
provided with matched 1-cm quartz cells was used for was diluted stepwise to yield working concentrations
all absorbance measurements. All chemicals used of 10 and 30 μg mL-1 for spectrophotometric
were of analytical reagent grade and distilled water investigations.
was used to prepare all solutions. Bromate-bromide
mixture (5×10-3 M KBrO3 - 50×10-3 M KBr) was Methods
Titrimetry
prepared by dissolving accurately weighed 0.8 g of A 10 mL aliquot of pure drug solution containing
KBrO3 (Sarabhai M Chemicals, Baroda, India) and 2.5-7.5 mg of MPT was accurately measured and
6 g of KBr (Indian Drugs and Pharmaceuticals Ltd, transferred into a 100 mL Erlenmeyer flask. The
Hyderabad, India) in water and diluting to 1 litre in a solution was acidified by adding 5 mL of 2 M
calibrated flask, and the reagent was used in hydrochloric acid. Ten mL of bromate-bromide
titrimetric work. A 0.03 M sodium thiosulphate
reagent (5×10-3 M w. r. t. KBrO3) was pipetted into
solution was prepared by dissolving about 7.45 g of
the flask, the flask was stoppered, the contents mixed
compound (Sisco Chem Industries, Mumbai, India) in
and let stand for 10 min with occasional swirling.
1 litre of water and standardized30. A 10% potassium
Finally, 5 mL of 10 % potassium iodide solution was
iodide solution was prepared by dissolving 10 g of
added, and the liberated iodine was titrated against
salt (Merck Chemicals, Mumbai,India) in 100 mL of
0.03 M thiosulphate solution using starch as indicator
water. To prepare 1% starch indicator 1 g of soluble
towards the end point. A blank titration was
starch (S. d. Fine Chem., Mumbai, India) was made
performed, and the amount of drug in the measured
into paste in water and poured into 100 mL boiling
aliquot was calculated from the amount of KBrO3
water, boiled for 1 min and cooled. A stock standard
reacted with drug.
solution of bromate-bromide equivalent to 1000 μg
mL-1 KBrO3 containing 10-fold excess of KBr was Spectrophotometric method A
prepared by dissolving 100 mg of KBrO3 and 1 g of Different aliquots (0.5, 1.0…….5.0 mL) of
KBr in water and diluting to 100 mL with water in a standard 10 μg mL-1 MPT solution were accurately
calibrated flask. This was diluted appropriately to get measured into a series of 10 mL calibrated flasks by
10 and 30 μg mL-1 KBrO3 solutions for method A and means of a micro burette and the total volume was
method B, respectively. A stock solution equivalent to adjusted to 5 mL by adding water. To each flask were
500 μg mL-1 methyl orange was prepared by added, 2 mL of 5 M hydrochloric acid and 1 mL of
dissolving 58.8 mg of the dye (S. d. Fine Chem., bromate-bromide reagent (10 μg mL-1 w. r. t KBrO3)
Mumbai, India, dye content 85%) in water and in succession. The flasks were stoppered immediately,
diluting to 100 mL in a calibrated flask, and filtered contents mixed, and allowed to stand for 5 min with
using glass wool. It was diluted 10-fold to get 50 μg occasional shaking. Lastly, 1 mL of 50 μg mL-1
mL-1 dye solution for use in method A. stock solution methyl orange solution was added to each flask, the
containing 1000 µg mL-1 indigo carmine was first volume was diluted to the mark with water, mixed
prepared by dissolving 111 mg of dye (S. d. Fine well and absorbance measured at 520 nm against a
Chem., Mumbai, India, dye content 90 %) in water reagent blank after 10 min.
and diluting to 100 mL in a calibrated flask, and
filtered. A working concentration of 200 μg mL-1 dye Spectrophotometric method B
solution was obtained by 5-fold dilution with water Varying aliquots (0.5-5.0 mL) of standard 30 μg
for method B. A 5 M hydrochloric acid was prepared mL-1 MPT solution were accurately transferred into a
356 INDIAN J CHEM. TECHNOL., JULY 2006

series of 10 mL calibrated flasks by means of a micro hydrochloric acid concentration was maintained.
burette, and the total volume was brought to 5 mL by Hence, 0.4 M acid concentration for the
adding water. Two mL 5 M hydrochloric acid and bromination/oxidation step and the iodometric back
1.5 mL of bromate-bromide reagent (30 μg mL-1 w. r. titration was used in the assay. Reaction was complete
t KBrO3) were added to each flask, the flasks were in 10 min and yielded stoichiometry of 1:1 (MPT :
immediately stoppered, the contents mixed well and KBrO3), and contact times up to 20 min had no effect
let stand for 10 min with occasional shaking. Then, on the stoichiometry of the reaction. A constant
1 mL of 200 μg mL-1 indigo carmine solution was molar-ratio was obtained when excess of reagent was
added to each flask, the volume was completed to the not more than 2 times the theoretical amount. Under
mark with water, mixed well and the absorbance of the optimum conditions, 2.5 - 7.5 mg of MPT could
each solution was measured at 610 nm against a be determined with good accuracy and precision with
reagent blank after 10 min. reaction stoichiometry of 1:1.
In either method, a standard graph was prepared by
Spectrophotometric methods
plotting the absorbance as a function of concentration
The ability of bromine to effect
of MPT. The concentration of the unknown was read
oxidation/bromination of MPT and bleach the
from the calibration graph or deduced from the
colour of methyl orange and indigo carmine dyes
respective regression equation derived from the
has been used for the indirect spectrophotometric
Beer’s law data.
assay of MPT. In both methods, the drug was
Method for tablets reacted with a measured excess of in situ generated
Twenty tablets were weighed and powdered. An bromine in acid medium and the unreacted bromine
amount of powder equivalent to 100 mg of MPT was was determined by reacting with either methyl
accurately weighed into a 100 mL calibrated flask, orange or indigo carmine followed by absorbance
60 mL of water added and shaken for 20 min. Then, measurement at 520 or 610 nm. In either method,
the volume was diluted to the mark with water, mixed the absorbance increased linearly with increasing
well and filtered using a Whatmann No. 42 filter concentration of MPT.
paper. A suitable aliquot was then analysed by MPT, when added in increasing amounts to a fixed
titrimetry. The tablet extract (1000 μg mL-1 MPT) was amount of bromine, consumed the latter and there
diluted with water to obtain working concentrations of occurred a concomitant fall in its concentration. When
10 and 30 μg mL-1 for analysis by spectrophotometric a fixed amount of either dye was added to decreasing
methods. amounts of bromine, a concomitant increase in the
concentration of dye resulted. This was observed as a
Results and Discussion proportional increase in absorbance at the respective
The acidified solution of bromate and bromide λmax with increasing concentration of MPT and Beer’s
behaves as an equivalent solution of bromine and has law is obeyed in the concentration range given in
been widely used for the determination of many Table 2.
organic and inorganic substances31,32. The present Preliminary experiments were performed to
methods make use of oxidising/brominating ability, determine the maximum concentration of each dye
and bleaching action of in situ generated bromine. spectrophotometrically, and these were found to be 5
and 20 μg mL-1 for methyl orange and indigo
Method development
Titrimetry carmine, respectively. A bromate concentration of 1.0
Direct titration of MPT with in situ generated μg mL-1 (in the presence of excess KBr) was found to
bromine was not successful. However, the reaction destroy the red colour due to 5 μg mL-1 methyl orange
between the two was found to occur when the two whereas in the case of 20 μg mL-1 indigo carmine,
were allowed to stand for some time, thus enabling 4.5 μg mL-1 KBrO3 was sufficient to bleach the blue
the indirect titrimetric determination of MPT. Hence, colour in acid conditions. Hence, different amounts of
several factors like nature of acid and its MPT were reacted with 1.0 mL of 10 μg mL-1 KBrO3
concentration, reaction time, and the excess of reagent in method A and 1.5 mL of 30 μg mL-1 KBrO3 in
were optimized. Reproducible and stoichiometric method B before determining the residual bromine as
results were obtained when 0.24 to 0.56 M described under the respective procedures.
BASAVAIAH et al.: SENSITIVE BROMATOMETRIC METHODS FOR THE ASSAY OF METAPROLOL TARTRATE 357

Hydrochloric acid was the ideal medium for compiled in Table 3. To evaluate the inter-day
oxidation/bromination reaction as well as the precision, anaysis was performed over a period of five
determination of residual bromine by using either dye. days preparing all solutions afresh each day. The
The reaction between MPT and bromine (in situ) was accuracy of the methods was determined by
unaffected when 1.0 -5.0 mL of 5 M hydrochloric calculating the percentage deviation observed in the
acid was used in about 8 mL. Hence, 1 mL was used analysis of pure drug solution and expressed as the
for both steps in the assay procedures. For percent relative error (RE). Table 3 summarises the
quantitative reaction between MPT and bromine (in intra-day precision and accuracy data for the assay of
situ), a contact time of 10 min was found sufficient in MPT in pure drug solution by the proposed methods,
both methods and constant absorbance readings were which were within 3%. The inter-day precision was
obtained when contact times were extended up to 30 less than 4%.
min. The standing time of 5 min was necessary for the
bleaching of dye colour by the residual bromine. The Application to analysis in dosage forms
measured colour was stable for several hours even in The Indian pharmaceutical industry has at present 4
the presence of reaction product. different brands of tablets in 25, 50 and 100 mg doses.
Three brands of tablets were assayed by the proposed
Quantation parameters methods, and the results are summarized in Table 4.
A linear correlation was found between absorbance at The results obtained were compared with those
λmax and concentration of MPT in the ranges given in obtained by the established UV-spectrophotometric
Table 2 in both the methods. The graphs obeyed Beer’s methods5 which consisted of the measurement of the
law and can be described by the regression equation: absorbance of tablet extract in 0.1 M HCl at 224 nm.
A close agreement between the results obtained by the
Y = a + bX
proposed methods and the reference method interms
of accuracy and precision was obtained as found from
(where Y = absorbance; a = intercept; b = slope and X
the Student’s t-value and F-value.
= concentration in μg mL-1 ) obtained by the method
of least squares. Correlation coefficients, intercepts Table 2⎯ Quantitative parameters of spectrophotometric
and slopes for the calibration data are summarized in methods
Table 2. Sensitivity parameters such as apparent Parameters Method A Method B
molar absorptivity and Sandell sensitivity values, and
the limits of detection and quantification are also λmax (nm) 520 610
Beer’s law limits (μg mL-1) 0.5 - 5.0 1.5 - 15.0
presented in Table 2 and speak of the excellent Molar absorptivity (l mol-1 cm-1) 8.17 × 104 2.70 × 104
sensitivity of the proposed methods. Sandell sensitivity (μg cm-2) 0.0084 0.025
Limit of detection (μg mL-1) 0.054 0.15
Method validation
Limit of quantification (μg mL-1) 0.165 0.47
Assay precision and accuracy
Regression equation (Y)
Intra-day and inter-day precision were assessed Slope (b) 0.11 0.04
from the results of seven replicate analyses on pure Intercept (a) 0.012 -6.7 × 104
drug solution. The mean values and the relative Correlation coefficient (r) 0.9989 0.9988
standard deviation (RSD) values for replicate analyses Y=a + bX where Y is the absorbance and X concentration in
at three different levels (amounts/concentrations) are μg mL-1.

Table 3⎯ Intra-day precision and accuracy


Titrimetry Method A Method B
MPT MPT Range, Relative RSD, MPT MPT Range, μg Relative RSD, % MPT MPT Range, μg Relative RSD,
taken, found, mg error, % % taken, μg found, μg mL-1 error, % taken, μg found, μg mL-1 error, % %
mg mg mL-1 mL-1 mL-1 mL-1
3.0 3.03 0.12 1.0 0 2.25 1.50 1.48 0.017 1.33 2.41 3.0 2.95 0.028 1.67 2.54
5.0 5.01 0.35 0.20 2.59 3.00 2.92 0.029 2.67 1.35 6.0 5.91 0.093 1.50 1.94
7.0 6.97 0.04 0.43 1.84 4.50 4.40 0.070 2.22 2.36 12.0 11.78 0.049 1.83 2.24
• Mean value of seven determinations
RSD⎯.Relative standard deviation
358 INDIAN J CHEM. TECHNOL., JULY 2006

Table 4⎯ Assay results of dosage forms


Formulation brand Nominal amount Percent found* ± SD
name ** (mg/tablet )
Tirimetry Method A Method B Reference method
100.10 ± 1.41 101.20 ± 1.39 99.98 ± 0.87
Betaloc 50 g t = 0.79 t = 0.80 t = 1.28 100.65 ± 0.78
F = 3.27 F = 3.17 F = 1.24
99.89 ± 1.62 101.30 ± 1.42 102.30 ± 0.93
100 g t = 0.48 t =1.35 t = 3.38 100.28 ± 0.96
F =2.84 F =2.18 F = 1.06
97.98 ± 1.44 98.50 ± 1.33 98.14 ± 1.77
Metolar 100 g t = 1.84 t =1.24 t = 1.45 99.44 ± 1.07
F = 1.81 F = 1.54 F = 2.74
101.77 ± 1.85 99.20 ± 1.73 102.50 ± 1.39
Metapro 50 g t = 1.47 t = 1.57 t = 2.78 100.50 ± 0.88
F = 4.42 F = 3.86 F = 2.49
* Mean value of five determinations
**Marketed by: a. AstraZeneca
b. cipla
c. Microvascular
Tabulated t-value at 95 % confidence level is 2.77
Tabulated F-value at 95 % confidence level is 6.39
Table 5⎯ Results of recovery studies
Titrimetry Method A Method B
Preparation Amount Amount of Total Recovery∗ Amount Amount of Total Recovery∗ Amount Amount of Total Recovery∗
studies of drug in pure drug found, of pure of drug in pure drug found, of pure of drug in pure drug found, of pure
tablet, mg added, mg mg drug, % tablet, μg added, μg μg drug, % tablet, μg added, μg μg drug, %
Metolar 3.912 1.0 4.92 101.00 9.85 10.00 20.10 102.50 24.53 30.0 53.81 97.60
3.912 2.0 5.93 100.75 9.85 20.00 29.90 100.25 24.53 60.0 85.37 101.40
3.912 3.0 6.98 102.00 9.85 30.00 39.55 99.00 24.53 120.0 143.65 99.27
* Mean value of three determinations

Accuracy and validity of the methods were further precision. Unlike most of the existing
established by performing recovery experiments via spectrophotometric methods, the proposed procedures
standard-addition technique. To a fixed amount of are free from stringent experimental conditions and
drug in tablet powder (pre-analyzed), pure MPT was are characterized by long dynamic linear range of
added at three different levels and the total was found response and high sensitivity, and infact, the methods
by the proposed methods. Each test was repeated are one of the most sensitive ever reported for MPT.
three times. The recovery of pure MPT added to Another significant advantage of the
formulations ranged from 97.60 - 102.50% (Table 5) spectrophotometric methods is that the measurement
indicating that tablet excipients such as talc, starch, is made at longer wavelengths where the interference
acacia, stearate, alginate, lactose, calcium gluconate from co formulated substances is far less compared to
and calcium dihydrogen orthophosphate did not shorter wavelengths used in most currently available
interfere in the assay procedures. procedures. In addition, all the three methods have
demonstrated to be fairly accurate and precise, and
Conclusions may be used as advantageous alternatives in industrial
Three simple, rapid and cost-effective methods for quality control laboratories.
the assay of metaprolol tartrate in bulk drug and tablet
dosage form have been developed and appropriately Acknowledgements
validated. The titrimetric method is applicable over a The authors wish to express their gratitude to the
micro scale and is independent of the experimental Quality Control Manger, Astra - Zeneca,
variables that would often affect the accuracy and Bangalore, India for providing pure metaprolol
BASAVAIAH et al.: SENSITIVE BROMATOMETRIC METHODS FOR THE ASSAY OF METAPROLOL TARTRATE 359

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