Acta Chromatographica, No.

7, 1997

ANALYSIS OF TABLETS AND CAPLETS CONTAINING

KETOPROFEN BY NORMALAND REVERSED-PHASE HPTLC WITH ULTRAVIOLET
ABSORPTION DENSITOMETRY
ON PREADSORBENT PLATES

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J. Sherma and C. D. Incarvito

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Department of Chemistry, Lafayette College, Easton, PA 18042, USA

SUMMARY
An HPTLC method has been developed for the quantitative
determination of ketoprofen in analgesic tablets and caplets using
preadsorbent normal phase silica gel and reversed phase C-18 chemically
bonded silica gel plates with fluorescent indicator and scanning
densitometry of fluorescence-quenched zones of samples and standards.
Pharmaceutical tablets and caplets were analyzed and the results compared
to the label declarations. Precision was evaluated by performing replicate
analyses and accuracy validated by recovery analysis of a sample fortified
with a known concentration of ketoprofen.
INTRODUCTION
Quantitative thin layer chromatography methods were described earlier
for the determination of aspirin, phenacetin, and caffeine [1], aspirin,
acetaminophen, and caffeine [2], and naproxen and ibuoprofen [3] in
analgesic tablets. In this paper, the quantitative TLC methodology is
extended to the determination of the newest analgesic product, ketoprofen.
Methods available in the literature for quantitatively determining
ketoprofen and metabolites in pharmaceutical dosage forms involve high
performance liquid chromatography (HPLC) with UV detection [4, 5] and
gas chromatography [6], and qualitative TLC screening procedures have
been reported [7, 8]. This paper describes a simple, sensitive, and fast
quantitative HPTLC method for determination of ketoprofen employing
direct densitometry of fluorescence-quenched sample and standard zones
on high performance preadsorbent silica gel and C-18 chemically bonded
silica gel plates containing fluorescent indicator. The method was applied
to the two dosage forms available in the United States market, 12.5 mg
tablets and caplets.
EXPERIMENTAL
Preparation of standard solutions
Ketoprofen (2-[3-benzoylphenyl]propionic acid; Sigma, St. Louis, MO,
USA) stock standard solution was prepared at a concentration of 1.00
mg/mL in methanol. The TLC standard solution was prepared by 1:10

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dilution with methanol to a concentration of 0.100 g/L A 6.25 mg/mL

spiking solution in methanol was also prepared.
Preparation of sample solutions
A tablet or caplet with a label values of 12.5 mg of ketoprofen was
placed in a 100 mL volumetric flask and crushed into numerous pieces
with a flattened glass rod. Methanol was added to the line, and the solution
was magnetically stirred for 15 minutes to completely dissolve the active
ingredient. Some excipients remained undissolved and were removed by
filtering through glass wool packed in a small disposable pipet in order to
prepare a clear solution prior to spotting onto the TLC plate.
Thin layer chromatographic analysis
Analyses were performed on Whatman (Clifton, NJ, USA) LHPKDF
10 x 20 cm high performance silica gel layers with 19 lanes, preadsorbent
spotting area, and fluorescent phosphor, and on Whatman LKC18F 20 x
20 cm C-18 chemically bonded silica gel preadsorbent plates with
fluorescent phosphor. Plates were prewashed by development with
dichloromethane-methanol (1 + 1). For analysis of the samples, 2.50, 5.00,
and 7.50 l aliquots of the TLC standard solution (0.250-0.750 g) and
duplicate 4.00 l aliquots of the sample solution (containing 0.500 g of
ketoprofen based on the label value) were spotted in adjacent lanes using a
10 L Drummond (Broomall, PA, USA) digital microdispenser. Silica gel
plates were developed for a distance of 6.5 cm beyond the preadsorbentsilica gel interface with ethyl acetate-glacial acetic acid (95 + 5) mobile
phase in a vapor-equilibrated, paper-lined Camag (Wilmington, NC, USA)
HPTLC twin-trough chamber. The development time was ca. 11-14
minutes. C-18 plates were developed in a twin-trough TLC chamber for a
distance of 13 cm (ca. 120 min) with methanol-0.5 M NaCl (55 + 45). The
plates were air dried and inspected under 254 nm UV light in a viewing
box, and standard and sample zones scanned at 264 nm with a Shimadzu
CS-930 densitometer in the single-beam, UV reflectance mode. This
wavelength was found to provide maximum absorption by measurement
of the in situ spectrum of a ketoprofen standard zone on a silica gel plate
using the spectral mode of the densitometer. A calibration equation
relating standard weights and scan areas was determined by use of a linear
regression program on a personal computer, and the weight of ketoprofen

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in the sample was calculated from the calibration equation using the
average area of the duplicate sample aliquots. The experimental weight
was compared to the theoretical label value.
The method was validated by the following recovery experiment. A
ketoprofen tablet test solution was prepared as described above, and the
theoretical 0.125 mg/mL concentration of a 50.0 mL portion was doubled
by addition of 6.25 mg of ketoprofen as 1.00 ml of a 6.25 mg/mL
methanol stock solution, followed by dilution with 49.0 mL of methanol.
Duplicate 4.00 L aliquots of the unspiked and diluted spiked samples
were analyzed against a calibration curve prepared from three TLC
standard aliquots spotted on the same plate. Percentage recovery of the
spike was calculated using the following equation: [(g found in spiked
sample x 2 - g found in unspiked sample)/(theoretical amount of added
spike)] x 100. The factor of two corrects for the dilution of the spiked
sample prior to spotting, and the denominator is 0.500 g.
RESULTS AND DISCUSSION
Development on HPTLC silica gel and C-18 layers containing
fluorescent indicator using the mobile phases described above produced
dark, flat compact zones on a bright green background with respective R f
values of 0.72 and 0.25 when viewed under 254 nm UV light. Detection of
ketoprofen by fluorescence quenching was about equally sensitive on the
two layers, allowing 0.25 g amounts to be visually detected and
reproducibly scanned.
Two individual ketoprofen tablets were analyzed by the proposed TLC
method on silica gel, and the results were 98.4% and 99.4% relative to the
12 mg label value. Triplicate analysis of a third tablet on silica gel gave
97.3+/-2.2% (mean+/-standard deviation) relative to the label value. Two
ketoprofen caplets were analyzed on C-18 layers, and the results were
100.6% and 101.1% relative to the 12 mg label value. Triplicate analysis
of a third caplet gave 99.1%+/-0.88% relative to the label value.
Calibration curves for ketoprofen in the 0.25-0.75 g range calculated
for each plate on which samples were analyzed typically had linear
correlation coefficient (r) values of 0.990-0.999 during these analyses. As
another measure of reproducibility, the percent difference between the
scan areas of the duplicate sample aliquots was in the range of 0.9-6%
with an average of ca. 3.5%. No additional, interfering zones appeared in
any of the sample lanes for tablets or caplets on either layer.

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Analysis of the diluted, spiked sample aliquots on a silica gel plate gave
an average value of 0.496 g ketoprofen while the unspiked sample gave
an average value of 0.504 g. These weights represent a recovery of
97.6% of the added 0.500 g spike, which demonstrates the accuracy of
the proposed method.
The analyses of all tablets and caplets were within the 95-105% range
generally specified in the U.S. Pharmacopeia for active ingredients of
pharmaceutical dosage forms. The recoveries as a percentage of label
value, recovery of the spiked sample, and standard deviations are typical
of those obtained for established pharmaceutical TLC analyses [9] and
satisfactory for routine use of the method in a pharmaceutical company
analytical laboratory. The TLC method involves very low solvent usage on
a per sample basis compared to HPLC, leading to savings in purchase and
disposal costs, and the ability to analyze multiple samples on a single plate
containing only three standard zones permits very high sample throughput.
Use of both NP- and RP-TLC provides excellent confirmation of
qualitative and quantitative results.

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[9]

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