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Salicylic Acid
Salicylic Acid
Introduction
Experimental
Aspirin (acetylsalicylic acid) is extensively used in the treatment Apparatus
of mild to moderate pain, fever, and inflammatory diseases. Liquid chromatograph
Aspirin belongs to the groups of medicines known as salicylates The liquid chromatograph used in this study was a
and anti-inflammatory analgesics and is a nonsteroidal anti- PerkinElmer system (Norwalk, CT). The PerkinElmer system
inflammatory agent (1,2). Salicylic acid is the hydrolysis product consisted of a Series 200 LC pump, Series 200 Autosampler
of aspirin and is usually present in small amounts of aspirin– equipped with a 150-µL loop and PerkinElmer Series 200 Peltier
caffeine effervescent tablets. Salicylates are not as therapeutically sample tray, Series 245C diode-array detector with a 600 Series
effective as aspirin (1,2). Caffeine, like theobromine and theo- link interface, Turbochrom software, and HP LaserJet 5000
phylline, is a xanthine derivative. Caffeine belongs to the group of printer (Hewlett-Packard, Palo Alto, CA).
medicines called central nervous system stimulants. It is used to
help restore mental alertness when unusual tiredness, weakness, Column
or drowsiness occurs (1,2). Both of these drugs are classified as The columns used were commercially available stainless steel
“Generally Regarded as Safe” and are widely used throughout the HPLC with a column length of 15 cm and inside diameter of 4.6
over-the-counter medicines (3,4). mm packed with Hypersil C18 (5 µm) or Keystone BDS C18
At present, there is not a United States Pharmacopeia (USP) (Thermo Hypersil-Keystone, Cheshire, U.K.).
compendial monograph specific for the simultaneous assay of
aspirin, salicylic acid, and caffeine (Figure 1) (5). A literature Mobile phase
The mobile phase was a mixture of water–methanol–glacial
* Author to whom correspondence should be addressed: email MaryJean.Sawyer.B@Bayer.com. acetic acid (690:280:30). This mobile phase maintained an
Reproduction (photocopying) of editorial content of this journal is prohibited without publisher’s permission. 393
Journal of Chromatographic Science, Vol. 41, September 2003
apparent pH of 4.7 for several weeks. The mixture was degassed Analytical sample
using helium sparge. The flow rate was 1.5 mL/min, autosampler The sample tablets were provided by Bayer Consumer Care
was chilled to 4°C, and column temperature was ambient. Manufacturing (Elkhart, IN). The effervescent tablets were pre-
pared to contain 500 mg aspirin and 60 mg caffeine. The analyt-
Chemicals ical effervescent placebo was provided by Bayer Consumer Care
HPLC-grade methanol and glacial acetic acid were purchased Formulation Development (Morristown, NJ).
from VWR Scientific Products (South Plainfield, NJ) and used
without further purification. Water was purified using a Milli-Q Standard preparation
filter system (Millipore, Milford, MA). Formic acid was purchased Salicylic acid reference standard solution (free salicyclic acid)
from Fisher Scientific (Springfield, MA). Caffeine and salicylic Approximately 50 mg of salicylic acid (accurately weighed) was
acid standards were purchased from USP Convention (Rockville, transferred to a 50-mL volumetric flask, dissolved in and diluted
MD). Aspirin reference standard was prepared as a pharmaceu- to volume with methanol–formic acid (95:5), and mixed well.
tical active ingredient (Bayer, Spain), and qualified for use as an
Preparation of samples
Assay
Ten tablets were ground to a fine powder using a glass mortar
and pestle. An accurately weighed portion equivalent to 250 mg
aspirin and 30 mg caffeine was quantitatively transferred to a 250-
mL volumetric flask. Approximately 150 mL of methanol–formic
acid (95:5) was added to the flask and sonicated (at least 5 min) to
(A)
(B)
394
Journal of Chromatographic Science, Vol. 41, September 2003
dissolve. The flask was brought to volume with methanol–formic Results and Discussion
acid (95:5) and mixed well. An aliquot of the sample was then fil-
tered through a 0.45-µm nylon filter directly into an HPLC vial Precision
and sealed. The HPLC vial was placed immediately into an The system precision (system suitability) was determined by
autosampler previously chilled to 4°C. chromatographing six replicate injections of the working stan-
dard solution at the 100% level. Calculated were the relative
Recovery retention times (void time = 1), coefficient of variation (%RSD) of
Aliquots of spiking solutions (containing both caffeine and the peak area responses, the tailing factors, and the resolution.
aspirin) were pipetted in triplicate into 250-mL volumetric flasks The range for the coefficient of variation was from 0.2% to 1.3%.
containing placebo at levels corresponding to approximately The tailing factors were less than or equal to 2.0, and the resolu-
50%, 75%, 100%, 125%, and 150% of the label claim for one tion between peaks was greater than 4 (Table I, Figure 2A).
tablet. The samples were then prepared according to the assay
1 0.00420 0.00426 101.4 Amount added (mg) 0.004 0.100 0.125 0.150
2 0.00420 0.00423 100.7 1. Peak area response 5064 50999 63121 74063
3 0.00420 0.00415 98.8 2. Peak area response 5029 54007 59775 72196
Mean recovery Overall = 99.2% 2. Peak area response 4927 54399 65855 77466
%RSD Overall = 3.4% Correlation coefficient = 0.95
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Journal of Chromatographic Science, Vol. 41, September 2003
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Journal of Chromatographic Science, Vol. 41, September 2003
ment Group of Bayer Health Care for their support in providing for the determination of pyridostigmine bromide, acetaminophen,
authentic samples and working placebo. acetylsalicylic acid and caffeine in rat plasma and urine. J. Pharm.
Biomed. Anal. 26: 939–47 (2001).
7. N. Ramos-Marots, F. Agurre-Gomez, A. Molina-Diaz, and L.F.
Captia-Valvery. Application of liquid chromatography to the simulta-
neous determination of acetylsalicylic acid, caffeine, codeine, parac-
References etamol, pyridoxine and thiamin in pharmaceutical preparations.
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