Original

Chromatography, Vol.22 No.2(2001)

Original

Analysis of the Active Ingredient Cimetidine inAcid Reduction Tablets By High Performance Thin Layer Chromatography withUltraviolet Absorption Densitometry
Stacey D. Wagner and Joseph Sherma*
Department of Chemistry, Lafayette College, Easton PA 18042, USA
Received for review April 9, 2001.

Accepted May 21, 2001.

Abstract
A quantitative method using silica gel HPTLC plates with fluorescent indicator, automated sample application, and UV absorption densitometry was developed for the determination of cimetidine in acid reduction tablets used to treat ulcers and other hyperacidity stomach disorders.
Four pharmaceutical tablet products containing cimetidine as the active ingredient were analyzed to test the applicability of the new method.
Precision was evaluated by replicate analyses of samples and accuracy by simultaneous analysis of unfortified and fortified samples on a single plate (standard addition). The percent cimetidine in the tablets analyzed ranged from 102% to 109% compared to label values, precision
ranged from 1.2% to 2.2% relative standard deviation, and the error in the standard addition analysis was 0.985% compared to the fortification level. These validation levels are within the guidelines of the International Conference on Harmonization for pharmaceutical analysis.
Keywords: thin layer chromatography, cimetidine, densitometry.

method described below, for which excellent accuracy and preci-

1. Introduction
Cimetidine is a histamine H2−receptor antagonist that mark-sion are demonstrated, is faster and more convenient and uses lesssolvent compared to HPLC.

edly inhibits gastric secretion. It is widely prescribed throughout
the world in tablet form as a therapeutic agent to reduce stomach

2. Experimental

acidity and treat ulcers. The standard method for determining cimetidine pharmaceutical drug substance and tablets involves highPreparation of Standard Solutions

performance liquid chromatography (HPLC) using a C-18 chemi-

The TLC standard solution of cimetidine (Sigma, St. Louis,

cally bonded silica gel column; a mobile phase composed ofMO, USA; catalog no. C 4522; CAS registry no.51481-61-9) was

methanol, phosphoric acid,1-hexanesulfonate, and water; and UV

prepared at a concentration of 2.50 mg/ml in absolute ethanol. Aabsorption detection at 220 nm [1]. A computer-based literaturestock solution for fortification in the standard addition analysis was

search of Chemical Abstracts located 19 papers describing thinprepared at a concentration of 20.0 mg/ml in absolute ethanol.

layer chromatography (TLC) analysis of cimetidine, including
separation and study of UV spectra [2]; detection on layers by

Preparation of Sample Solutions

chemical derivatization [3], qualitative screening [4,5], qualitative

Four brands of cimetidine tablets with label values of 200 mg

identification [6], determination of drug release from tablets [7],were obtained from pharmacies. Test solutions were prepared byand detection by TLC/fast atom bombardment-mass spectrometry

grinding a tablet into a fine powder with a mortar and pestle ; the[8]. No previous publications on the quantitative analysis of ci-powder was quantitatively transferred through a funnel into a 100-

metidine in pharmaceutical dosage forms were found. The newmL volumetric flask by washing with absolute ethanol. The solu-

quantitative high performance thin layer chromatography (HPTLC)

tion was stirred magnetically at high setting for 30 min, after which*Corresponding author: Tel, +1-610-330-5220; fax, +1-610-330-5714;
e-mail, shermaj@latayetle.edu―９７―Chromatography, Vol.22 No.2(2001)

curacy of the method.

the stir bar was removed by use of a magnetic rod. The solution
was diluted to volume with absolute ethanol and shaken to mix

3. Results and Discussion

thoroughly. The undissolved solid excipients were allowed to settle
for 60 minutes prior to analysis. The theoretical concentration ofDevelopment with the mobile phase described above on theeach tablet test solution was 2.00 mg/ml based on the label declara-HPTLC silica gel layers containing fluorescent indicator producedtions.

compact, flat, dark fluorescence-quenched bands of cimetidine (Rf0.43) against a bright green background when viewed under a 254

Thin Layer Chromatographic Analysis

nm UV light. The excipients in the tablets analyzed included corn-Analyses were performed on Merck 20 cm × 10 cm high per-starch, cellulose, magnesium stearate, polyethylene glycol, polysor-formance silica gel 60F254 GLP plates (EM Separations Technol-bate 80, povidone, sodium lauryl sulfate, sodium starch glycolate,ogy, Gibbstown, NJ, no. 5613/6). Sample and standard solutions

and titanium dioxide. No additional zones representing these ex-were applied by means of a Camag (Wilmington, NC, USA) Lino-

cipients were detected in chromatograms. Calibration curves had

mat IV automated spray-on band applicator equipped with a 100-µl

linearity correlation coefficient (r) values of 0.997-0.999 for 10.0 tosyringe and operated with the following settings: band length 640.0 µg of cimetidine.

mm, application rate 4 s/µl, table speed 10 mm/s, distance between

Four brands of tablets (a name brand and three generic brands)

bands 4 mm, distance from the plate edge 0.7 cm, and distancewere analyzed by the described procedure. The recoveries com-from the bottom of the plate 1.5 cm. The volumes applied for eachpared to the label value of 200 mg are shown in Table 1. To assess

analysis were 4.00 µl, duplicate 8.00 µl, and 16.00 µl of the TLC

precision, two tablets of different brands were analyzed four timesstandard (10.0-40.0 µg) of cimetidine and duplicate 10.00 µl ali-each, and recoveries were 106%±2.2% and 106%±1.8%.One tablet

quots of the sample solutions (20.0 µg theoretical content).

was analyzed six times, and the recovery was 108%±1.2%. As anPlates were developed to a distance of 6 cm beyond the origin

additional measure of reproducibility, in addition to the RSD of du-with ethyl acetate-methanol-conc. ammonium hydroxide (75 : 20 :plicate analyses, the percent difference between the scan areas for5) in a vapor-equilibrated Camag HPTLC twin trough chamber

duplicate sample aliquots was calculated for each analysis, and thelined with a saturation pad (Analtech, Newark, DE, no. 81-12). Therange was 0.90-2.9%. All tablets analyzed throughout the method

development time was 17 min. After development, the plates weredevelopment and validation studies assayed within the 90-110%

air-dried for 5 min in a fume hood, and sample and standard zones

specification range in the USP 24/NF 19 for cimetidine tablets [1].

were quantified by linear scanning at 254 nm with a Camag TLC
Scanner I with a deuterium source, slit dimension settings of length
4 and width 4, and a scanning rate of 4.0 mm/s. The wavelength

Table 1. Recoveries of Cimetidine from Tablets

used was 254 nm. The CATS-3 software produced a linear regres-SampleRecoverysion calibration curve relating standard zone weights to their optiBrand 1

mized scan areas. The analyte weights in the sample zones were
determined from their areas by automatic interpolation from theTablet 1
Tablet 2
Tablet 3

calibration curve. The percent recovery was calculated for eachtablet analysis by comparing the theoretical weight predicted by thelabel value to the mean experimental weight of the duplicate sam-

105%
107%
109%

Brand 2ple zones.

Tablet 1
Tablet 2
Tablet 3

The accuracy of the method was validated by a standard addition analysis. A tablet test solution was prepared according to theprocedure described above. An 800 µl aliquot of this solution was

106%
102%
106%

Brand 3

mixed with 80.0 µl of the stock solution to double the concentra-

Tablet 1
Tablet 2
Tablet 3

tion of cimetidine based on the label value. Volumes were measured with 1000 µl and 100 µl Drummond (Broomall, PA, USA)
microdispensers, respectively. The original and fortified sample so-104%
102%
105%

Brand 4lutions were analyzed on the same plate by application of duplicate
10.0 µl and 5.00 µl volumes, respectively, and the four standards

Tablet 1
Tablet 2
Tablet 3

described above. The difference between the mean experimental
weights and the added weight was calculated to determine the ac-

―９８―

109%
107%
108%

Chromatography, Vol.22 No.2(2001)

Stacey D. Wagner and Joseph ShermaInc., Rockville, MD, USA, 2000, pp.412-414.
[2]The accuracy of the new method was validated by a standard

Gyeresi, A.; Gergely, M.; Vamos, J. J. Planar Chromatogr.
-Mod. TLC 2000, 13, 296-300.

addition method in which unfortified and fortified sample solutions
were analyzed on the same plate. The analysis of the unfortified[3]

Ogoda Onah, J. Acta Pharm (Zagreb) 1999, 49, 217-220.

sample yielded a 98.0% recovery relative to the label value. The[4]

Brzezinka, H.; Dallakian, P.; Budzikiewicz, H. J. Planar

theoretical fortification weight was 17.9 µg of cimetidine, and theChromatogr. -Mod . TLC 1999, 12, 96-108.

analysis of the fortified sample yielded 18.0 µg, representing a per-[5]Steinberg, D.M.; Sokoll, L.J.; Bowles, K.C.; Nichols, J.H.;
Roberts, R.; Schultheis, S.K.; O’Donnell, C.M. Clin. Chem.

cent error of 0.985% or a percent recovery of 101%.

(Washington, D.C.) 1997. 43, 2099-2105.

It has been shown that the new HPTLC method achieved re[6]coveries as a percentage of tablet label value, precision for replicate

Romano, G.; Caruso, G. ; Masumarra, G.; Pavone, D.; Cru-analyses, and accuracy of analyte analysis from a fortified standard

ciani, G. J. Planar Chromatogr. -Mod . TLC 1994, 7, 233-

addition sample that compare favorably with those reported regu-241.

larly in the literature for HPTLC and HPLC pharmaceutical dosage[7]

Chyla, A.; Kaniewska, T. Biul. Inst. Lekow 1990, 37, 37-43.

forms. The results also meet the guidelines of the International[8]

Bolton, G.C.; Allen, G.D.; Nash, M.; Proud, H.E. Methodol.
Surv. Biochem. Anal . 1990, 20, 353-354.

Conference on Harmonization (ICH) for validation of pharmaceuti[9]cal assays of drug products, which are a precision of 2-3% RSD

Ferenczi-Fodor, K.; Vegh, Z.; Nagy-Turak, A.; Renger, B.;
Zeller, M. J. AOAC Int. 2001, 84, (in press).

and recovery (accuracy) of 95-105% [9]. Previous papers describe[10] Muller, E.; Sherma, J. J. Liq. Chromatogr. & Relat. Tech-

the overall advantages of quantitative HPTLC relative to HPLC fornol . 1999, 22, 153-159.

assay of pharmaceutical dosage forms [10-12].

[11] Greshock, T.; Sherma, J. J. Planar Chromatogr. -Mod. TLC
1997, 10, 460-463.

References[1][12] Lippstone, M.B.; Grath, E.K.; Sherma, J. J. Planar Chroma-The United States Pharmacopeia / The National Formularytogr. -Mod. TLC 1996, 9, 456-458.

(USP 24/NF 19), United States Pharmacopeial Convention,―９９―