International Journal of Trend in Scientific

Research and Development (IJTSRD)

International Open Access Journal
ISSN No: 2456 - 6470 | www.ijtsrd.com | Volume - 2 | Issue – 5

UV-Visible
Visible Spectrophotometric Method of Paracetamol Tablet
Formulation
Selvi S R
Lecturer Senior
enior Grade, Department of Physics, Muthiah Polytechnic College,
Annamalai Nagar
Nagar, Chidambaram, Tamil Nadu, India

ABSTRACT
A novel, safe and sensitive method of however less ss frequently as an in-line
in analytical tool
spectrophotometric estimation in UV-region
region has been [16]. For some applications, these spectroscopic
developed for the assay of Paracetamol in its tablet techniques are not feasible, such as those that require
formulation. The method have been developed and the quantification of low-dosed
low analytics.
validated for the assay of Paracetamol using Methanol Fluorescence spectroscopy can be an alternative to the
and water as diluents. Which does not shows any conventional
nal spectroscopic techniques for these
interference in spectrophotometric estimations. All the applications because of its high sensitivity and
parameters of the analysis were chosen according to detection sensitivity. A drawback of fluorescence
ICH [Q2 (R1)] guideline and validated statistically spectroscopy is that the analytic needs to be a native
using RSD and %RSD along long with neat chromate fluorophore in order to detect it, which limits the
grams. number of possible
ossible applications for this technique.
UV spectroscopy is a widely used quantitative
Keyword: UV Spectrography, Paracetamol, analytical technique that finds its application in many
Concentration, Tablets research domains and is capable of quantifying very
low concentrations (<0.01%) [20e24]. Nevertheless,
I. INTRODUCTION
studies describing on-line
line and in-line
in applications of
Spectroscopic techniques are increasingly proposed as
UV/VIS spectroscopy with fibre-optic
fibre probes are
alternative methods for the quantification of APIs in
limited. O'Keeffeet al. monitored the ozone
pharmaceuticals. This is due to their advantages over
concentration of a gas in an aluminium glass cell with
the traditional techniques, such as fast, in
in-line, non-
a fibre-based
based UV/VIS spectroscopy system . Quinn et
invasive and non-destructive
destructive measuremen
measurements without
al. followed the reaction of a nucleoside with trityl
the need of sample preparation. Near infrared (NIR)
chloride in pyridine in a liquid environment, using a
and Raman spectroscopy have been identified as
fibre-optic
optic transmission probe. The concentration of
effective PAT-tools for real-time
time measurements of
starting material and product was predicted via a PLS
critical process and product attributes during
regression model. Furthermore, a mixing study using
pharmaceutical processing. Raman spectroscopy is
a fibre-optic
optic UV/VIS monitoring technique was
until now mostly applied for solid dosage forms [1e6].
reported by Ng and Assirelli. In this paper,
Some in-line
line quantitative applications for hot
hot-melt
bromophenol blue sodium salt was used as a non- non
extrusion processes have also been reported [7e9].
reactive tracer in distilled water. A good agreement
Raman spectroscopy has an added value for
between the UV/VIS technique and the traditional
quantification purposes of pharmaceutical
conductivity technique
nique was found. Other examples of
formulations where
here water is present, such as in semi
semi-
on-line and in-line
line UV spectroscopic applications in
solid and liquid formulations, since water produces
literature are drug dissolution tests, where the drug
almost no Raman signal. Research has already been
release was monitored in real-time. However, the use
conducted to investigate the opportunity offered by
of UV spectroscopy for in-line line monitoring of critical
Raman spectroscopy for these formulations [10e15],
quality attributes during pharmaceutical

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Aug 2018 Page: 1749
 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470
manufacturing processes of semi-solids and liquids is does not exhibit significant antiinflammatory activity
not yet described in literature. This work deals with (it is a weak COX inhibitor). This is despite the
the validation of the developed method for the assay evidence that Paracetamol and NSAIDs have some
of Paracetamol from its dosage form (tablets). Hence, similar pharmacological activity.
the method can be used for routine quality control
analysis and also stability. To date, the mechanism of action of Paracetamol is
not completely understood. The main mechanism
Paracetamol or acetaminophen is a widely used over- proposed is the inhibition of cyclooxygenase (COX),
the-counter analgesic (pain reliever) and antipyretic and recent findings suggest that it is highly selective
(fever reducer). It is commonly used for the relief of for COX-2. While it has analgesic and antipyretic
headaches and other minor aches and pains and is a properties comparable to those of aspirin or other
major ingredient in numerous cold and flu remedies. NSAIDs, its peripheral anti-inflammatory activity is
In combination with opioid analgesics, Paracetamol usually limited by several factors, one of which is the
can also be used in the management of more severe high level of peroxides present in inflammatory
pain such as post-surgical pain and providing lesions. However, in some circumstances, even
palliative care in advanced cancer patients [5]. The peripheral anti-inflammatory activity comparable to
onset of analgesia is approximately 11 minutes after other NSAIDs can be observed. An article [18] in
oral administration of Paracetamol [6], and its half- Nature Communications from researchers in London,
life is 1-4 hours. Though acetaminophen is used to UK and Lund, Sweden in November 2011 has found a
treat inflammatory pain, it is not generally classified hint to the analgesic mechanism of acetaminophen
as an NSAID because it exhibits only weak anti- (Paracetamol), being that the metabolites of
inflammatory activity. While generally safe for use at acetaminophen e.g. NAPQI, act on TRPA1-receptors
recommended doses (1,000 mg per single dose and up in the spinal cord to suppress the signal transduction
to 4,000 mg per day for adults), acute overdoses of from the superficial layers of the dorsal horn, to
Paracetamol can cause potentially fatal liver damage alleviate pain.
and, in rare individuals, a normal dose can do the
same; the risk is heightened by alcohol consumption. Paracetamol is metabolised primarily in the liver, into
Paracetamol toxicity is the foremost cause of acute non-toxic products. Three metabolic pathways are no
liver failure in the Western world, and accounts for table:
most drug overdoses in the United States, the United
Glucuronidation is believed to account for 40% to
Kingdom, Australia and New Zealand [7-10]. It is the two-thirds of the metabolism of paracetamol.
active metabolite of Phenacetin, once popular as an
Sulfation (sulfate conjugation) may account for
analgesic and antipyretic in its own right, but unlike 20-40% .
Phenacetin and its combinations, Paracetamol is not
N-hydroxylation and rearrangement, then GSH
considered carcinogenic at therapeutic doses [11]. The conjugation, accounts for less than 15%. The
words acetaminophen (used in the United States, hepatic cytochrome P450 enzyme system
Canada, Japan, South Korea, Hong Kong, and Iran metabolizes paracetamol, forming a minor yet
[12] and Paracetamol (used elsewhere) both come significant alkylating metabolite known as NAPQI
from a chemical name for the compound: para- (N-acetyl-p-benzo-quinone imine). NAPQI is then
acetylaminophenol and para-acetylaminophenol. In irreversibly conjugated with the sulfhydryl groups
some contexts, it is simply abbreviated as APAP, for of glutathione [20]. All three pathways yield final
acetyl-paraaminophenol. products that are inactive, nontoxic, and
eventually excreted by the kidneys. In the third
pathway, however, the intermediate product
NAPQI is toxic. NAPQI is primarily responsible
for the toxic effects of Paracetamol; this
constitutes an example of toxication.

The discussion of the validation of analytical

procedures is directed to the four most common types
Paracetamol is part of the class of drugs known as
of analytical procedures:
“aniline analgesics”; it is the only such drug still in

Identification tests;
use today. It is not considered an NSAID because it

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 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470

Quantitative tests for impurities’ content; analysis is used to ascertain the quantity of molecular

Limit tests for the control of impurities; - species absorbing the radiation. Spectrophotometric
Quantitative tests of the active moiety in technique is simple, rapid, moderately specific and
samples of drug substance or drug product or applicable to small quantities of compounds. The
other selected component(s) in the drug fundamental law that governs the quantitative
product. spectrophotometric analysis is the Beer -Lambert law.
Beer’s law: It states that the intensity of a beam of
The aim and scope of the proposed work are as under: parallel monochromatic radiation decreases

To develop suitable spectrophotometric exponentially with the number of absorbing
method for assay of Paracetamol tablet. molecules. In other words, absorbance is proportional

Perform the validation for the method. to the concentration. Lambert’s law: It states that the
intensity of a beam of parallel monochromatic
II. Experimental methods radiation decreases exponentially as it passes through
Paracetamol standard of was provided by Torque a medium of homogeneous thickness. A combination
Pharmaceuticals (P) Ltd. (India). Paracetamol tablets of these two laws yields the Beer-Lambert law.
containing 500 mg Paracetamol and the inactive
ingredient used in drug matrix were obtained from Beer-Lambert law: When beam of light is passed
market. Analytical grade methanol and water were through a transparent cell containing a solution of
obtained from Spectrochem Pvt. Ltd., Mumbai anabsorbing substance, reduction of the intensity of
(India). light may occur.

Diluent preparation is the using Methanol and water Validation is concerned with assuring that a
(15:85, v/v) used as a diluent. Standard preparation 10 measurement process produces valid measurements. •
mg drug was dissolved in 15 ml methanol and was Results from method validation can be used to judge
shaken well. Then 85 ml water was added to it to the quality, reliability and consistency of analytical
adjust the volume up to 100 ml (100 ppm). From that results. It is an integral part of any good analytical
5 ml was taken and volume was adjusted up to 50 ml practice. • A measurement process producing valid
with diluents. measurements for an intended application is fit for
purpose
20 tablets were weighed and powdered. Powdered
tablet equivalentto 100 mg of paracetamol was Method validation is the process used to confirm that
weighed and taken into 100 ml volumetric flask then the analytical procedure employed for a specific test is
15 ml of methanol was added and shaken well to suitable for its intended use. Results from method
dissolve it after that 85 ml of water was added to validation can be used to judge the quality, reliability
adjust the volume up to 100 ml. From that 1 ml of and consistency of analytical results; it is an integral
solution was withdrawn and taken in 100 ml part of any good analytical practice.
volumetric flask. The volume was adjusted with
diluent up to 100 ml. UV-Visible double beam III. Results and Discussion
spectrophotometer with matched quartzcells (1 cm) Proper wave length selection of the methods depends
Evolution 201 model is used upon the nature of the sample and its solubility. To
develop a rugged and suitable spectrophotometric
UV-Visible spectrophotometry is one of the most method for the quantitative determination of
frequently employed technique in pharmaceutical paracetamol, the analytical condition were selected
analysis. It involves measuring the amount of after testing the different parameters such as diluents,
ultraviolet or visible radiation absorbed by a buffer, buffer concentration, and other
substance in solution. Instrument which measure the chromatographic conditions. Our preliminary trials
ratio, or function of ratio, of the intensity of two were by using different compositions of diluents
beams of light in the U.V-Visible region are called consisting of water with buffer and methanol. By
Ultraviolet-Visible spectrophotometers. In qualitative using diluent consisted of methanol - water (50:50,
analysis, organic compounds can be identified by use v/v) best result was obtained and degassed in an
of spectrophotometer, if any recorded data is ultrasonic bath (Enertech Electronics Private
available, and quantitative spectrophotometric

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 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470
Limited). Below figures represent the spectrums of
blank, standard and test preparation respectively.

Figure 5: UV Spectrum of Blank.

Figure 1: UV spectrum of Paracetamol in Standard
solution.

Figure 6: Linearity curve for Paracetamol.

Specificity: Resolution of the analyte peak from the

Figure 2: UV spectrum of Paracetamol in Sample nearest peak: Solution of each of the analyte was
solution. injected separately and their retention time is noted.
The standard working solution containing a mixture
of the component being analyze is also injected and
each of analyte peaks is check for its resolution from
the nearest (Figures 2,3,4,5).

Linearity: Six points calibration curve were obtained

in a concentration range from 0-150 ppm for
Paracetamol. The response of the drug was found to
be linear in the investigation concentration range and
the linear regression equation was y = 0.004x+0.007
with correlation coefficient 0.998 (Figure 6).
Figure 3: UV Spectrum of Placebo.
Precision: Precision of the analytical method is
ascertained by carrying out the analysis as per the
procedure and as per normal weight taken for
analysis. Repeat the analysis six times. Calculate the
% assay, mean assay, % Deviation and % relative
standard deviation and %RSD. The developed method
was found to be precise as the %RSD values for the
repeatability and intermediate precision studies were
<0.98% and <0.79%, respectively.

Accuracy
Accuracy of the method is ascertained by standard
Figure 4: UV Spectrum of Placebo+analyte. addition method at 3 levels. Standard quantity

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 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470
equivalent to 50%, 100% and 125% is to be added in 2. ChatwalGR, Anand S (2002) Instrumental
sample. The result shown that best recoveries (98.54- Methods of Chemical Analysis. (5thedn),
99.13%) of the spiked drug were obtained at each Himalaya Publishing House, New Delhi.
added concentration, indicating that the method was 3. Davidson AG (2002) Ultraviolet-visible
accurate. Composition of Placebo Added for absorption spectrophotometry. In BeckettAH,
Accuracy Study. Stenlake JB, (4thedn), Practical Pharmaceutical
chemistry. CBS Publishers and distributors, New
Solution stability study Delhi, 275-278.
Results obtain in the solution stability study at
different time intervals for test preparation. It was 4. Patil KM, Bodhankar SL (2005) High-
concluded that the test preparation solution was found performance thin-layer chromatographic
stable up to 8 hr at room temperature, as during this determination of lamotrigine in serum. J
time the result was not decrease below the minimum Chromatogr B AnalytTechnol Biomed Life Sci
percentage. 823: 152-157.
5. Scottish Intercollegiate Guidelines Network
Robustness (SIGN) (2008) Control of pain in adults with
The evaluation of robustness should be considered cancer-A national clinical guideline. Scotland:
during the development phase and depends on the National Health Service (NHS).
type of procedure conditions, assay value of the test
preparation solution was not affected and it was in 6. Moller P, Sindet-Pedersen S, Petersen CT, Juhl
accordance with that of actual. System suitability GI, Dillenschneider A, et al. (2005) Onset of
parameters were also found satisfactory; hence the acetaminophen analgesia: comparison of oral and
analytical method would be concluded as robust. intravenous routes after third molar surgery. Br J
Anaesth 94: 642-648.
System suitability 7. Daly FF, Fountain JS, Murray L, Graudins A,
A system suitability test of the spectrophotometric Buckley NA (2008) Guidelines for the
system was performed before each validation run. Six management of paracetamol poisoning in
replicate reading of standard preparation were taken Australia and New Zeal and explanation and
and %RSD of standard reading were taken for same. elaboration. A consensus statement from clinical
Acceptance criteria for system suitability, %RSD of toxicologists consulting to the Australasian
standard reading not more than 2.0%, were full fill poisons information centres. Med J Aust 188: 296-
during all validation parameter. 301.

IV. Conclusion 8. Khashab M, Tector AJ, Kwo PY (2007)

The present analytical method was validated as per Epidemiology of acute liver failure.
ICH Q2(R1) guideline and it meets to specific CurrGastroenterol Rep 9: 66-73.
acceptance criteria. It is concluded that the analytical 9. Hawkins LC, Edwards JN, Dargan PI (2007)
method was specific, precise, linear, accurate, robust Impact of restricting paracetamol pack sizes on
and having stability indicating characteristics. The paracetamol poisoning in the United Kingdom: a
present analytical method can be used for its intended review of the literature. Drug Saf 30: 465-479.
purpose.
10. Larson AM, Polson J, Fontana RJ, Davern TJ,
Lalani E, et al. (2005)Acetaminophen-induced
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