doi: 10.2306/scienceasia1513-1874.2005.31.207
ABSTRACT: Three simple, rapid, reliable and cost-effective methods based on titrimetry and spectrophotmetry
in non-aqueous medium are described for the determination of ranitidine in pharmaceuticals. In titrimetry,
the drug dissolved in glacial acetic acid was titrated with acetous perchloric acid with visual and potentiometric
end point detection, crystal violet being used as indicator for visual titration. Spectrophotometry involved
adding different amounts of the drug to a fixed amount of perchloric acid-crystal violet mixture followed by
measurement of absorbance at 570 nm. The absorbance was found to increase linearly with the concentration
of the drug and formed the basis for quantification. The titrimetric methods are applicable over 1-15 mg
range of ranitidine, and in spectrophotometry, calibration graph was linear from 10 to 70 µg ml-1. The
apparent molar absorptivity is calculated to be 2.2 × 103 l mol-1 cm-1 and the calculated Sandell sensitivity is
161.7 ng cm-2. The limits of detection and quantification are found to be 1.07 and 3.58 µg ml-1, respectively.
The procedures were used to determine ranitidine in pharmaceutical products and the results were found to
be in good agreement with those obtained by the reference method. Associated pharmaceutical materials did
not interfere. The accuracy and reliability of the methods were further ascertained by recovery studies via
standard-addition technique with percent recoveries in the range 96.3 to 102.5 %.
(CH3 ) (CH2)
2 NCH2 CH2 S 2 NH
NO2
O C CH
CH3 NH
Fig 1. Structure of RNH.
208 ScienceAsia 31 (2005)
chromatography (HPTLC)18 and differential pulse at 570 nm. The increase in absorbance is related to the
adsorptive stripping voltammetry19 have also been drug concentration. The methods, in addition to being
applied for the determination of ranitidine in biological rapid, sensitive, accurate and precise, gave satisfactory
samples. results when applied to formulations containing
HPLC20-26 continues to occupy a prominent position ranitidine. Additionally, the methods can be used in
among the methods used for the assay of ranitidine in laboratories where modern and expensive instruments
pharmaceutical preparations. Several electroanalytical such as GC-MS, HPLC, capillary electrophoresis,
techniques including polarography27,28, differential voltammetry etc, are not available.
pulse polarography29, oscillopolarography30, and linear
sweep-voltammetry31 have been employed for the MATERIALS AND METHODS
sensitive determination of ranitidine in
pharmaceuticals. Other reported techniques include Apparatus
combined gas chromatography-mass spectroscopy32, All absorbance measurements were made with a
proton magnetic resonance spectroscopy 33, near Systronics Model 106 spectrophotometer provided
infrared reflectance spectrophotometry 34 , UV with matched 1-cm quartz cells. Potentiometric titration
spectrophotometry35-38, automated fluorimetry39, and was performed with an Elico 120 digital pH meter
capillary electrophoresis40. provided with a combined glass-SCE system. The KCl
Titrimetric methods with potentiometric41 and of the salt bridge was replaced with 0.1 M lithium
coulometric42 end-point detection are applicable for perchlorate in glacial acetic acid.
300 mg and 5-90 mg of ranitidine, respectively.
Potentiometric methods based on ion-selective Reagents and Solutions
electrodes43-45 proposed by various workers, although All chemicals used were of analytical reagent grade.
applicable in µM to mM concentration range, require All solutions were made in glacial acetic acid unless
strict pH control for accurate and precise results. specified otherwise.
Visible spectrophotometry because of its simplicity,
sensitivity, speed and reliability is next only to HPLC in Per chloric Acid (0.01 M): To 4.5 ml of 70 %
Perchloric
terms of its application to the analysis of ranitidine in perchloric acid (S.d. Fine Chem., Mumbai, India) was
pharmaceuticals. Methods based on redox 46-48, added 150 ml of glacial acetic acid, mixed well; added
oxidative coupling49, nitrosation50, charge-transfer 10.5 ml of acetic anhydride and allowed the solution
complexation 51,52 and ion-pair complexation 53,54 to cool for 30 min; finally diluted to 500 ml with glacial
reactions have been proposed for the assay of ranitidine acetic acid and allowed to stand overnight. This
in its dosage forms. Hassan and Belal55 have recently perchloric acid (~0.1 M) was diluted to 0.01 M with
developed a kinetic spectrophotometric method for glacial acetic acid and standardized with pure
ranitidine. Procedures based on redox, oxidative potassium biphthalate and crystal violet indicator.
coupling and ion-pair complexation reactions, although
sensitive, involved unstable reagents46, a contact time Crystal Violet Indicator: Prepared by dissolving
of 30 min49 or extraction step53,54, besides lacking in 0.1 g of dye (S.d. Fine Chem., Mumbai, India) in 100 ml
selectivity46-48. Methods based on nitrosation50 reaction glacial acetic acid.
is least sensitive (0.3-12 µg ml-1) and even the charge-
transfer complexation methods are not sufficiently Perchloric Acid-Crystal Violet Mixture (1.5 mM
sensitive (50-250 µg ml-1). The only titrimetric method HClO4 - 0.25 mM Crystal Violet): Prepared by mixing
based on neutralization reaction41 in aqueous medium 15 ml of 0.01 M perchloric acid and 10 ml of 1000 µg
is applicable for 300 mg of drug. ml-1 crystal violet solutions and diluting to 100 ml with
The methods based on modern instrumental glacial acetic acid in a drug calibrated flask.
techniques19-39 although sensitive require expensive
instrument and maintenance. The present paper Standard Drug Solution
describes three methods based on the basic property Pharmaceutical grade ranitidine hydrochloride
of the drug molecule. In the titrimetric procedures, the (RNH) was procured from Glaxo Smithkline, Nashik,
drug solution in glacial acetic acid is titrated directly India, as a gift, and was used as received. A stock
with acetous perchloric acid, the end point being standard solution containing 2 mg ml-1 RNH was
determined either visually using crystal violet indicator prepared by dissolving 500 mg of pure drug in glacial
or potentiometrically using modified glass-saturated acetic acid and diluting to the mark in a 250 ml calibrated
calomel electrode system. Spectrophotometry involves flask. This solution (2 mg ml-1) was used for titrimetric
treating a fixed amount of perchloric acid-crystal violet work, and for spectrophotometric work, the same was
mixture with the drug and measuring the absorbance diluted appropriately with glacial acetic acid to get 100
ScienceAsia 31 (2005) 209
0.15 0.5
In a preliminary study, 20 µg ml-1 crystal violet in the
base form was found to exhibit a convenient absorbance
0.12
0.4 at 570 nm. In the presence of 2 ml of 1.5 mM perchloric
acid in a total volume of 10 ml, this absorbance
Potential (Volts)
DE/DV, (V/ml)
0.09
0.3 decreased to a constant minimum. Hence, different
0.06 amounts of drug were treated with a fixed amount of
0.03
0.2 acid-dye mixture i.e., 2 ml of 1.5 mM HClO4 - 0.25 mM
crystal violet (100 µg ml-1 of crystal violet) to determine
0 0.1 the concentration range of the drug that could be
0 1 2 3 4 5 6 7
ml of perchloric acid determined by the method of absorbance transitions
of the dye accompanying the pH changes. The dye
Fig 2. Potentiometric end point. colour even in the presence of drug was found to be
stable for several hours, and the order of addition of
8
reactants was not critical.
The increasing absorbance values at 570 nm were
6 plotted against the increasing concentration of drug to
obtain a calibration graph. Beer’s law is obeyed over the
10 -pH
0.3
the pH of the solution and that the colour change is not
sudden but occurs continuously as the pH changes
over a definite range. To a fixed amount of acid-dye 0.2
mixture where the dye is in the acid form (emerald
green), different amounts RNH were added. This caused 0.1
a progressive increase in pH of the solution because of
neutralization of acid by the added drug (base), and as
0
a result, the concentration of the base form of the dye
0 20 40 60 80
increases. This is shown by the proportional increase
Concentration of RNH, µg ml
-1
in the absorbance of the solution at 570 nm (Fig 4)
which is corroborated by the correlation coefficient of
0.9892. Fig 4. Calibration curve.
ScienceAsia 31 (2005) 211
3.00 3.02 0.67 1.31 3.00 3.03 0.98 0.85 20.00 19.87 0.65 1.24
8.00 7.92 1.00 0.91 8.00 8.10 1.25 1.10 40.00 39.82 0.45 1.11
13.00 12.94 0.46 0.08 13.00 12.84 1.23 0.79 60.00 58.84 1.93 0.92
different levels. The relative error (%) which is a measure test for precision with those of the reference method
of accuracy, is < 2% revealing high accuracy of the at 95 % confidence level as summarized in Table 2. The
methods. The relative standard deviation (RSD), which results showed that the calculated t- and F- values did
is an indicator of precision, is less than 1.5 % and not exceed the tabulated values inferring that proposed
speaks of excellent precision of the methods. The methods are as accurate and precise as the reference
results of the study are compiled in Table 1. method.
Accuracy and reliability of the methods were further
Application ascertained by performing recovery experiments. To a
The proposed methods were successfully applied fixed amount of the drug in formulation (pre-analysed),
to determine RNH in tablets and injections. The same pure drug at three different levels was added, and the
batch tablets and injections were analysed by an total was found by the proposed methods. Each test
established procedure35 for comparison. The results was repeated three times. The results compiled in Table
obtained by the proposed methods agree well with 3 show that the recoveries were in the range 96.30 -
those of reference the method35 and with the label 102.50% indicating that commonly added excipients
claim. to tablets such as talc, starch, gelatin, sodium alginate,
The results were also compared statistically by a magnesium stearate, calcium gluconate and calcium
Student’s t-test for accuracy and by a variance ratio F- dihydrogen orthophosphate, did not interfere in the
Table 2. Results of analysis of tablets containing RNH by the proposed methods and comparison with the established
methods*
Ranitina 300 98.86 ± 0.91 99.50 ± 1.25 98.24± 1.68 100.36 ± 1.74
t=0.94; F=1.88 t=0.65;F=3.40 t=1.41; F=3.66
Histac b 300 99.32 ± 1.60 100.37 ± 1.26 100.42 ± 1.80 98.63 ± 0.85
t=1.16; F= 1.61 t=0.88;F=1.26 t=0.89;F=3.54
Zinetac c 150 102.28 ± 0.93 101.62 ± 1.36 101.36 ± 0.68 101.28 ± 1.16
t=0.91; F=2.13 t=1.56;F=1.87 t=1.51;F=1.55
Aciloc d 150 101.26 ± 0.84 100.63 ± 0.71 100.69 ± 1.28 102.28 ± 1.64
t=1.28; F= 1.40 t=0.73;F=2.32 t=1.30;F=2.47
Injections
Ranitina 25 101.26 ± 0.44 101.76 ± 0.62 100.88 ± 0.74 101.86 ± 0.85
t=1.49; F=1.98 t=0.88;F=2.83 t=1.47;F=3.73
Histac b 25 100.48 ± 0.96 99.74 ± 1.42 100.44 ± 0.78 99.76 ± 1.28
t=0.98; F= 2.19 t=0.06;F=1.51 t=1.01;F=1.78
RNHc 25 101.38± 1.04 102.04± 1.58 100.86± 0.72 101.71 ± 0.46
t=0.70; F=2.31 t=0.81;F=2.09 t=0.69;F=5.11
Aciloc d 25 97.68 ± 0.52 98.16 ± 1.22 98.74 ± 1.08 98.13 ± 0.66
t=0.85; F=3.21 t=1.81;F=4.31 t=1.20;F=1.61
determination.
CONCLUSIONS
drug added,
Although HPLC methods with uv-dtection are
Total Recovery
101.60
102.60
101.10
100.00
98.90
96.20
99.21
97.50
99.20
routinely used for the determination of ranitidine19-25
found, of pure
%
the procedures require maintenance of elevated
Method C
296.4
401.9
498.6
294.2
399.4
491.8
µg
400.0
200.0
300.0
400.0
200.0
300.0
400.0
Amount
µg
101.4
101.8
101.8
101.8
98.6
98.6
98.6
µg
100.75
102.00
100.71
102.50
100.85
100.38
99.86
99.25
99.25
found, of pure
12.80
11.24
12.17
13.08
11.17
12.08
13.05
ACKNOWLEDGEMENTS
Amount
7.0
6.0
8.0
6.0
7.0
8.0
6.0
7.0
8.0
of pure
mg
4.86
4.86
4.86
5.12
5.12
5.12
5.02
5.02
5.02
of drug
mg
REFERENCES
drug added,
Total Recovery
101.00
100.50
102.00
100.25
101.17
97.86
98.83
98.37
99.00
found, of pure
13.05
11.00
12.94
12.07
12.91
12.21
11.01
mg
7.0
7.0
6.0
8.0
6.0
8.0
7.0
8.0
6.0
mg
5.01
5.07
5.07
5.01
5.01
5.07
4.99
4.99
4.99
300mg
form
tablets
tablets
Histac
25 mg
7. Arafat T, Al-Saket M, Awad R, Saleh M, Gharaigeh M and 24. Beaulieu N, Laeroix P M, Sears RW and Lovering E G (1988)
Sallam S (1990) Analysis of ranitidine in plasma using high High performance liquid chromatographic method for the
performance liquid chromatography, Alexandria J Pharm 4, determination of ranitidine and related substances in raw
11-3. materials and tablets, J Pharm Biomed Anal 77, 889-92.
8. Rahman A, Hoffmann N E and Rustum A M (1989) 25. Evans M B, Haywood PA, Johnson D, Martin-Smita M and
Determination of ranitidine in plasma by high performance Munro G (1989) Chromatographic methods for determining
liquid chromatography, J Chromatogr Biomed Appl 7, 747- the identity, strength and purity of ranitidine hydrochloride
53. both in the drug substances and its dosage forms-an exercise
9. Kaka J S (1988) Rapid method for cemetidine and ranitidine in method selection, development, definition and validation,
determination in human and rat plasma by HPLC, J Liq J Pharm Biomed Anal 7, 1-22.
Chromatogr 11, 3447-56. 26. Aboul-Enein H Y and Rafiqul-Islam M (1990) Liquid
10. Rustum A M (1988) Rapid and sensitive HPLC determination chromatographic analysis of ranitidine hydrochloride in
of ranitidine in human plasma: Application to pharma kinetic pharmaceutical preparations, Toxicol Enviorn Chem 29, 47-
study, J Liq Chromatogr 1, 2315-35. 51.
11. Lopez-Calull C, Garcia-Capdevila L and Arroyo C and Bonal 27. Richter P, Ines-Toral M and Munoz-Vargas F (1994)
J (1997) Simple and robust high performance liquid Polarographic behaviour and determination of ranitidine in
chromatographic determination of ranitidine in micro pharmaceutical formulations Analyst (London) 119, 1371-4.
volumes of human serum, J Chromatogr Biomed Appl 693, 28. Abou-Zuhri, Hannoun M, Al-Khalil S I and Hassna H (1988)
228-32. Polarographic assay of ranitidine drugs in pharmaceutical
12. Lloyd T L, Perschy T B, Gooding A E and Tanlinnson J J formulations, Anal Lett 21, 1845-53.
(1992) Robotic solid phase extraction and high performance 29. Sankar P S and Reddy S J (1988) Voltammetric analysis of
liquid chromatographic analysis of ranitidine in serum or ranitidine, Indian J Pharm Sci 51, 263-4.
plasma, Biomed Chromatogr 6, 311-6. 30. Liu K Z, Li R T and Zhang Z Y (1992) Determination of
13. Karnes H T, Openg-Meusah K, Farthing D ans Beightol L A ranitidine by simple-sweep oscillopolarography, Zhongguo
(1987) Robotic solid phase extraction and high performance Yiyao Gongye Zazhie 23, 177-9.
liquid chromatographic determination of ranitidine from 31. Liu K Z, Li Y T and Liu H L (1992) Determination of
urine, plasma and peritoneal dialysis, J Chromatogr. Biomed ranitidine by sweep-derivative linear-sweep voltammetry,
Appl 422, 165-73. Zhongguo Yiyao Gonghye Zazhie 23, 187-90.
14. Prueksritanont T, Sittichai N, Prueksritanont S and Vongsaroj 32. Grinstead G F (1989) Ranitidine and high concentrations of
R (1989) Simultaneous determination of ranitidine and its phenhypropanolamine-methoamhetamine assay, Clin Chem
metabolites in human plasma and urine by HPLC, J (Winsten Salem NC) 35, 1998-9.
Chromatogr Biomed Appl 82, 175-85. 33. Ozden T, Ungormus A, Tosun A and Ersan S (1997)
15. Salem M S, Gharaiebh A M, Alkayasi H N and Badwan A Quantitative proton magnetic resonance analysis of ranitidine
(1988) High performance liquid chromatographic analysis in solid dosage forms, Spectrosc Lett 30, 835-41.
of ranitidine in plasma and urine, J Clin Pharm Ther 13, 351- 34. Dreassi E, Ceramelli G, Corti P, Perruccio P L and Lonardi S
7. (1996) Application of near infrared reflectance spectrometry
16. Rustum A M, Rahman A and Hoffmann N E (1987) High to the analytical control of pharmaceuticals; ranitidine
performance liquid chromatographic analysis of ranitidine hydrochloride tablet production, Analyst (Cambridge) 121,
in whole blood and plasma by using a short polymeric 219-22.
column, J Chromatogr Biomed Anal 65, 418-24. 35. De Almeida E and Saferin Martins J L (1993) Determination
17. Vinas P, Campllo N, Lopez-Errroz C and Hernandez- of ranitidine hydrochloride in pharmaceutical preparations
Corboda M (1997) Use of post-column fluorescence by ultra-violet and visible spectrophotometry, Anal Lett 26,
derivatisation to develop a liquid chromatographic assay of 1933-41.
ranitidine and its metabolites in biological fluids, J Chromatogr 36. Raghuver S Srivatsava C M R and Vatsa B K (1989)
Biomed Anal 693, 443-9. Spectrophotometric estimation of ranitidine hydrochloride
18. Mody W D, Satia M C, Gandhi T P, Modi I A, Modi R I and in its pharmaceutical dosage forms, Indian Drugs 23, 177-9.
Chakravarthy B K (1996) Sensitive high performance liquid 37. Chattaraji S C, Das S K and Gupta B K (1989)
chromatographic detection and determination of ranitidine Spectrophotometric determination of ranitidine
in plasma samples, J Chromatogr Biomed Anal 676, 175-9. hydrochloride, Indian Drugs 26, 365-7.
19. Altinoz S, Ozer D, Temizer A ans Bayraktar Y (1992) 38. Borne R and Burgess C (1995) Effect of temperature on a
Determination of ranitidine in biological materials by using multi component ultraviolet spectrometric determination
differential pulse adsorptive-stripping voltammetry, Anal Lett and the development of a temperature-independent assay
25, 111-8. procedure, Analyst (Cambridge) 120
120, 2075-80.
20. Bettermann G, Cabrera K, Heizenroeder S and Lubda D 39. Lopez-Erroz C, Vinas P, Campillo N and Hernandez-
(1998) HPLC analysis of active ingredients of Cordoba M (1996) Analyst (Cambridge) 121, 1043-6.
pharmaceuticals, Labour praxis 22, 32-4. 40. Kelly M A, Altria K D, Grace C and Clark B J (1998)
21. Hoyer G L, Le Doux J and Nolan Jr. P E (1997) A sensitive Optimization, validation and application of capillary
stability indicating assay for the H2 blocker ranitidine, J Liq electrophoresis method for the determination of ranitidine
Chromatogr 18, 1239-49. hydrochloride and related substances, J Chromatogr 798,
22. Lau Cam C A, Rahman M and Roos R W (1994) Rapid 297-306.
reversed-phase high performance liquid chromatographic 41. Atkosar Z and Tuncel M (1989) Potentiometric determination
assay method for ranitidine hydrochloride in dosage forms, of ranitidine hydrochloride, Acta Pharm Turk 31,31,139-42.
J Liq Chromatogr 17, 1039-104. 42. Nikolic K, Stankovic B and Bogavac M (1995) Coulometric
23. Dasgupta V (1988) Quantitation of ranitidine hydrochloride determination of ranitidine hydrochloride, Pharmazie 50,
in tablets and injections using HPLC, Drug Dev Ind Pharm 301-2.
14, 1647-55. 43. Wang N X, Chen J M, Yang X L and Luo C X (1992)
214 ScienceAsia 31 (2005)