To perform Analytical method validation of Paracetamol and Ibuprofen in pure and tablet form by UV spectrometric method.

Drug Regulation & Regulatory Authorities
Dept. Of Quality Assurance & Regulatory Affairs
L. J. Institute of Pharmacy, Ahmedabad.
EXPERIMENT NO.: DATE:
AIM: To perform Analytical method validation of Paracetamol and Ibuprofen in
pure and tablet form by UV spectrometric method.
REFERENCES:
1. http://en.wikipedia.org/wiki/Paracetamol.
2. http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/
Q2_R1/Step4/Q2_R1__Guideline.pdf.
3. A.H.Beckett & J.B.Stenlake, Practical Pharmaceutical Chemistry, Fourth Edition
(part two),282,284 &285
4. Anurekha Jain, Practical’s In Modern Pharmaceutical Instrumental Analysis, By
Nirali Prakashan, 4.13 to 4.16.
REQUIREMETNS:
Apparatus: 100ml & 10ml volumetric flasks, pipettes, beakers.
Reagents: Paracetamol API, Ibuprofen API, Paracetamol & Ibuprofen Tablet
(Combiflam®
), Methanol, Distilled Water.
Equipments:
1) Shimadzu UV-1800
Sr. No.: A114550/08677
2) Shimadzu UV-1800
Sr. No.: A114549/08780
THEORY:
 General Description of Paracetamol:
Name: Paracetamol INN or Acetaminophen USAN.
Dose: 325 mg.
Chemical name: N-Acetyl-p-aminophenol.
Structure:

Class: mild analgesic.
Use: over-the-counter analgesic (pain reliever) and antipyretic (fever reducer).It is
commonly used for the relief of headaches and other minor aches and pains and is a major
ingredient in numerous cold and flu remedies. In combination with opioid analgesics,
Paracetamol can also be used in the management of more severe pain such as post-surgical
pain and providing palliative care in advanced cancer patients. Though Paracetamol is
used to treat inflammatory pain, it is not generally classified as an NSAID because it
exhibits only weak anti-inflammatory activity.
Half-life: 1–4 hours.
Adverse effects: causes gastrointestinal problems or allergic skin reactions. Blood
dyscrasia (e.g. thrombocytopenia), methaemoglobinemia, and hemolytic anemia are very
rare.
 General Description of Ibuprofen:
Name: Ibuprofen INN
Dose: 400mg
Chemical Name: (RS)-2-(4-(2-methylpropyl) phenyl) propanoic acid
Structure:
Class: Anti inflammatory.
Use: Ibuprofen is used primarily for fever, pain, dysmenorrhea and inflammatory diseases
such as rheumatoid arthritis. It is also used for pericarditis and patent ductus arteriosus.
Half-life: 1.8–2 h
Adverse effects: Common adverse effects include: nausea, dyspepsia, gastrointestinal
ulceration/bleeding, raised liver enzymes, diarrhea, constipation, epistaxis, headache,
dizziness, rash, salt and fluid retention, and hypertension.

 General Description of Tablet (Combiflam®
):
Generic Name Paracetamol + Ibuprofen
Indication Analgesic
Strength 400 ibuprofen & 325 PCM
 Principle of Simultaneous Equation method:
In simultaneous equation method, when no region can be found free from overlapping
spectra of two chromophores, it is still possible to device a method based on
measurements at two wavelengths. Two dissimilar chromophores must necessary have
different powers of light absorption at some point or in linear absorption spectra. If
samples contain two absorbing drugs (X and Y), each of which absorbs at the λmax of the
other, it may be possible to determine both drugs by the technique of simultaneous
equations.
Figure: Wavelengths for the assay of substances X and Y in admixture by the
method of simultaneous equations.
For further calculation following information is required
a) The absorptivity of X at λ1 and λ2, ax1 and ax2 respectively.
b) The absorptivity of Y at λ1 and λ2, ay1 and ay2 respectively.
c) The absorbance of diluted sample at λ1 and λ2, A1 and A2 respectively.
Let Cx and Cy be the concentration of drugs in the diluted sample.
The concentration of given mixture can be determined by simultaneous equation,
if following criteria are satisfied.
A) The λmax of both the compound should be distinct.

B) Both compound should absorb at λmax of one-another.
C) There is no reaction between solutes.
Absorbance is an additive property.
Now, simultaneous equation,
C x = A2ay1-A1ay2 C y = A1ax2-A2ax1
ax2ay1-ax1ay1 ax2ay1-ax1ay1
Where,
Cx = Concentration of drug x (PCM) in the mixture.
Cy = Concentration of drug y (Ibuprofen) in the mixture.
λ1 = Maximum absorption wavelength for drug x
λ2 = Maximum absorption wavelength for drug y
ax1 = Absorptivity of drug x at λ1.
ax2 = Absorptivity of drug x at λ2.
ay1 = Absorptivity of drug y at λ1.
ay2 = Absorptivity of drug y at λ2.
A1 = Absorption of sample at λ1.
A2 =Absorption of sample at λ2.
 Validation Of Analytical Methods
Validation of an analytical method is documented evidence which provide a high
degree of assurance that the given method will consistently produce a product meeting its
predetermined specifications and quality attributes.
Validation parameters as per ICH
1) Accuracy
The accuracy of an analytical method may be defined as the closeness of the test
results obtained by the method to the true value. It is the measure of the exactness of the
analytical method developed. Accuracy may often be expressed as percent recovery by the
assay of a known amount of analyte added. Accuracy may be determined by applying the
method to samples or mixture of excipients to which known amount of analyte have been

added both above and below the normal levels expected in the samples. Accuracy is then
calculated from the test results as the percentage of the analyte recovered by the assay.
Dosage form assays commonly provide accuracy within 3-5% of the true value. The ICH
documents recommend that accuracy should be assessed using a minimum of nine
determinations over a minimum of three concentration levels, covering the specified range
(i.e. three concentrations and three replicates of each concentration).
2) Precision
The precision of an analytical method is the degree of agreement among individual
test results when the method is applied repeatedly to multiple sampling of homogenous
samples. This is usually expressed as the standard deviation or the relative standard
deviation (coefficient of variation). Precision is a measure of the degree of reproducibility
or of the repeatability of the analytical method under normal operating circumstances.
Repeatability:
Repeatability can be defined as the precision of the procedure when repeated by
same analyst under the same operating conditions (same reagents, equipments, settings
and laboratory) over a short interval of time. Repeatability is also termed as intra-assay
precision.
The ICH documents recommend that repeatability should be assessed using:
a) A minimum of 9 determinations covering the specified range for the procedure (e.g., 3
concentrations/3 replicates each); or
b) A minimum of 6 determinations at 100% of the test concentration.
It is normally expected that at least six replicates be carried out and a table showing each
individual result provided from which the mean, standard deviation and co-efficient of
variation should be calculated for set of n values.
The RSD values are important for showing degree of variation expected when the
analytical procedure is repeated several times in a standard situation. (RSD below 1% for
bulk drugs and RSD below 2% for assays in finished product).
Intermediate precision:
Intermediate precision expresses within-laboratories variations: different days,
different analysts, different equipment, etc.
The extent to which intermediate precision should be established depends on the
circumstances under which the procedure is intended to be used. The applicant should
establish the effects of random events on the precision of the analytical procedure. Typical
variations to be studied include days, analysts, equipment etc. It is not considered

necessary to study these effects individually. The use of an experimental design (matrix) is
encouraged.
Reproducibility:
Reproducibility means the precision of the procedure when it is carried out under
different conditions-usually in different laboratories-on separate, identical samples taken
from the same homogenous batch of material. Comparisons of results obtained by
different analysts, by the use of different equipments, or by carrying out the analysis at
different times can also provide valuable information.
3) Linearity
The linearity of an analytical procedure is its ability (within a given range) to
obtain test results which are directly proportional to the concentration (amount) of analyte
in the sample.
A linear relationship should be evaluated across the range of the analytical procedure. It
may be demonstrated directly on the drug substance (by dilution of a standard stock
solution) and/or separate weighing of synthetic mixtures of the drug product components
using the proposed procedure.
Linearity should be evaluated by visual inspection of a plot of signals as a function of
analyte concentration or content. If there is a linear relationship, test results should be
evaluated by appropriate statistical methods, for example, by calculation of a regression
line by the method of least squares. The correlation coefficient, y-intercept, slope of the
regression line and residual sum of squares should be submitted. A plot of the data should
be included. In addition, an analysis of the deviation of the actual data points from the
regression line may also be helpful for evaluating linearity.
The linear range of detectability that obeys Beer’s law is dependent on the compound
analyzed and the detector used. The working sample concentration and samples tested for
accuracy should be in the linear range. The claim that the method is linear is to be justified
with additional mention of zero intercept by processing data by linear least square
regression. Data is processed by linear least square regression declaring the regression co-
efficient and b of the linear equation y= ax + b together with the correlation coefficient of
determination r. For the method to be linear the r value should be close to1.
For the establishment of linearity, a minimum of 5 concentrations is recommended. Other
approaches should be justified.
4) Range
The range of an analytical procedure is the interval between the upper and lower
concentration (amounts) of analyte in the sample (including these concentrations) for

which it has been demonstrated that the analytical procedure has a suitable level of
precision, accuracy and linearity.
The specified range is normally derived from linearity studies and depends on the intended
application of the procedure. The following minimum specified ranges should be
considered:
For the assay of a drug substance or a finished (drug) product: normally from 80 to 120%
of the test concentration;
For content uniformity, covering a minimum of 70 to 130% of the test concentration,
unless a wider more appropriate range, based on the nature of the dosage form (e.g.,
metered dose inhalers), is justified;
For dissolution testing: +/-20 % over the specified range;
e.g., if the specifications for a controlled released product cover a region from20%, after 1
hour, up to 90%, after 24 hours, the validated range would be 0-110% of the label claim.
5) Limit of Detection and Limit of Quantitation
Limit of Detection (LOD): - The limit of detection is the parameter of limit tests. It is the
lowest level of analyte that can be detected, but not necessarily determined in a
quantitative fashion, using a specific method under the required experimental conditions.
The limit test thus merely substantiates that the analyte concentration is above or below a
certain level.
Limit of Quantitation (LOQ): - Limit of quantitation is a parameter of quantitative
assays for low levels of compounds in sample matrices such as impurities in bulk drugs
and degradation products in finished pharmaceuticals.
The limit of quantitation is the lowest concentration of analyte in a sample that may be
determined with acceptable accuracy and precision when the required procedure is
applied. In many cases, the limit of quantitation is approximately twice the limit of
detection.
Several approaches for determining the detection limit are possible, depending on whether
the procedure is a non-instrumental or instrumental.
I. Based on Visual Evaluation
Visual evaluation may be used for non-instrumental methods but may also be used
with instrumental methods. The detection limit is determined by the analysis of
samples with known concentrations of analyte and by establishing the minimum
level at which the analyte can be reliably detected.
II. Based on Signal-to-Noise

The determination of the limit of detection of instrumental procedures is carried
out by determining the signal-to-noise ratio by comparing test results from the
samples with known concentration of analyte with those of blank samples and
establishing the minimum level at which the analyte can be reliably detected. A
signal-to-noise ratio of 2:1 or 3:1 (For LOD) and 10:1(For LOQ) is generally
accepted. The signal-to-noise ratio is determined by dividing the base peak by the
standard deviation of all data points below a set threshold. Limit of detection is
calculated by taking the concentration of the peak of interest divided by three times
the signal-to-noise ratio.
III. Based on the Standard Deviation of the Response and the Slope
The detection limit may be expressed as:
LOD = 3 * σ / S
LOQ = 10 * σ / S
Where, σ = the standard deviation of the intercept
S = the slope of the calibration curve
6) Selectivity and Specificity
The selectivity of an analytical method is its ability to measure accurately and
specifically the analyte of interest in the presence of components that may be expected to
be present in the sample matrix.
If an analytical procedure is able to separate and resolve the various components of a
mixture and detect the analyte qualitatively the method is called selective. On the other
hand, if the method determines or measures quantitatively the component of interest in the
sample matrix without separation, it is said to be specific.
Hence one basic difference in the selectivity and specificity is that, while the former is
restricted to qualitative detection of the components of a sample, the latter means
quantitative measurement of one or more analyte.
Selectivity may be expressed in terms of the bias of the assay results obtained when the
procedure is applied to the analyte in the presence of expected levels of other components,
compared to the results obtained on the same analyte without added substances. When the
other components are all known and available, selectivity may be determined by
comparing the test results obtained on the analyte with and without the addition of the
potentially interfering materials. When such components are either unidentified or
unavailable, a measure of selectivity can often be obtained by determining the recovery of
standard addition of pure analyte to a material containing a constant level of the other
components.

7) Robustness and Ruggedness
Robustness: - The robustness of an analytical method is a measure of its capacity to
remain unaffected by small but deliberate variation in method parameters and provides an
indication of its reliability during normal usage.
The evaluation of robustness should be considered during the development phase and
depends on the type of procedure under study. It should show there liability of an analysis
with respect to deliberate variations in method parameters.
Ruggedness: - The ruggedness of an analytical method is the degree of reproducibility of
test results obtained by the analysis of the same samples under a variety of normal test
conditions such as different laboratories, different analysts, using operational and
environmental conditions that may differ but are still within the specified parameters of
the assay.
For the determination of ruggedness, the degree of reproducibility of test result is
determined as function of the assay variable. This reproducibility may be compared to the
precision of the assay under normal condition to obtain a measure of the ruggedness of the
analytical method.
PROCEDURE:
 Linearity
1) For PCM:
For Stock solution of Paracetamol:
Stock solution (100g/ml) of PCM was prepared by dissolving 0.01 g PCM in 100 ml
volumetric flasks and completing the volume with methanol:water(30:70).
Selection of analytical wavelength for PCM:
The solution of PCM was prepared in methanol:Water(30:70) at a concentration of 15 µg
/ml from the stock solution. It was scanned in the wavelength range of 200-400 nm.
Maximum absorbance was obtained at 245 nm. This analytical wavelength was selected
for determination of PCM.
For calibration curve:
An aliquots of stock solution of PCM (0.5, 1, 1.5,2, and 2.5 ml) were pipettes out in 10ml
volumetric flasks and further diluted to attain concentration of about 5, 10, 15, 20, 25
µg/ml.

Now plot the graph of Absorbance Vs Concentration. Find out the r2
value from the
Graph.
2) For Ibuprofen:
For Stock solution of Ibuprofen:
Stock solution (100g/ml) of IBU was prepared by dissolving 0.01 g IBU in 100 ml
volumetric flasks and completing the volume with methanol:water(30:70)
Selection of analytical wavelength for IBU:
The solution of IBU was prepared in methanol:Water(30:70) at a concentration of 15 µg
/ml from the stock solution. It was scanned in the wavelength range of 200-400 nm.
Maximum absorbance was obtained at 222 nm. This analytical wavelength was selected
for determination of IBU.
For calibration curve:
An aliquots of stock solution of IBU (0.5, 1, 1.5,2, and 2.5 ml) were pipettes out in 10ml
volumetric flasks and further diluted to attain concentration of about 5, 10, 15, 20, 25
µg/ml.
value from the
Graph.
For Mixture:
An aliquots of mixture solution following mls were pipettes out in 10ml volumetric flasks
and further diluted to attain concentration of about 5, 10, 15, 20, 25 µg/ml.
Concentration(µg/ml) ml of PCM ml of IBU
5 0.5 0.615
10 1.0 1.230
15 1.5 1.845
20 2.0 2.460
25 2.5 3.075
value from the
Graph.
 Accuracy
From the stock solution (100µg/ml) pipette out 1 ml of PCM stock solution and 1.23ml of
IBU stock solution in 3 volumetric flasks and add following ml of standard stock solution
of PCM and IBU to make 80,100 & 120% of solution to each volumetric flasks
respectively. Similarly 2 other sets are prepared.

% of
solution
80% 100% 120%
PCM(ml) IBU(ml) PCM(ml) IBU(ml) PCM(ml) IBU(ml)
1.0 1.23 1.0 1.23 1.0 1.23
0.8 0.98 1.0 1.23 1.2 1.47
Total 1.8 2.21 2.0 2.46 2.2 2.70
Measure the absorbance of each set i.e. total 3 concentrations of 3 sets. Take the Mean of
each concentration of each set. Find out the % recovery, Standard Deviation & % Relative
Standard Deviation.
 Precision
Repeatability: Select the middle concentration i.e. 15µg/ml and carry out the repeatability
by taking the absorbance of the solution 6 times.
Calculate the mean of the absorbance and find the Standard Deviation and % Relative
Standard Deviation.
Intraday precision: Prepare 3 concentrations from the stock solution (i.e. 5, 15 &
25µg/ml), make another 2 sets similarly. Measure the absorbance of each concentration of
each set at the time interval of 2 hours.
Calculate the Mean, Standard Deviation & % Relative Standard Deviation.
Interday precision: Prepare 3 concentrations from the stock solution (i.e. 5, 15 &
25µg/ml), make another 2 sets similarly. Measure the absorbance of each concentration of
each set for 3 days.
Calculate the Mean, Standard Deviation & % Relative Standard Deviation.
 LOD & LOQ
LOD & LOQ are found by using the following equation.
LOD = 3.3 * σ / S
LOQ = 10 * σ / S
Where, σ = the standard deviation of the intercept
S = the slope of the calibration curve
 Robustness and Ruggedness
Robustness: It is carryout by doing deliberate variation in method parameters is done (i.e.
change in wavelength). Absorbance of any one concentration (i.e. 15µg/ml) is measured at

2 different wavelengths i.e. 244.8&245.2 nm. And calculate the % Assay or % Drug
Recovery. Repeat the procedure three times.
Ruggedness: It is carried out by the analysis of the sample (i.e. 15µg/ml) under a variety of
normal test conditions such as different laboratories, different analysts, using operational
and environmental conditions that may differ but are still within the specified parameters
of the assay.
Instruments used:
1. Shimadzu UV-1800
Sr. No.: A114550/08677
2. Shimadzu UV-1800
Sr. No.: A114549/08780
Measure the Absorbance and calculate Mean, Standard Deviation, % Relative standard
deviation & % Assay.
CALCULATION:
 Linearity
PCM 245
Conc.
(µg/ml)
Abs Mean SD %RSD Absorptivity
1 2 3
5 0.403 0.405 0.401 0.403 0.002 0.496278 806
10 0.755 0.749 0.761 0.755 0.006 0.794702 755
15 1.103 1.112 1.094 1.103 0.009 0.815956 735
20 1.477 1.468 1.486 1.477 0.009 0.609343 738
25 1.861 1.859 1.863 1.861 0.002 0.107469 744
743
y = 0.0728x + 0.0284
R² = 0.9995
0
0.5
1
1.5
2
0 10 20 30
absorbance
concentration(mcg/ml)
linearity of PCM at 245nm(Mean)

PCM 222
Conc.
(µg/ml)
1 2 3
5 0.192 0.194 0.19 0.192 0.002 1.041667 384
10 0.37 0.365 0.375 0.37 0.005 1.351351 370
15 0.542 0.536 0.548 0.542 0.006 1.107011 361
20 0.71 0.703 0.717 0.71 0.007 0.985915 355
25 0.903 0.895 0.911 0.903 0.008 0.885936 365
362.75
IBU 222
Conc.
(µg/ml)
1 2 3
5 0.289 0.293 0.297 0.293 0.004 1.365188 586
10 0.542 0.544 0.546 0.544 0.002 0.367647 544
15 0.816 0.821 0.819 0.818667 0.002517 0.307404 545.73
20 1.039 1.041 1.046 1.042 0.003606 0.346022 521
25 1.297 1.292 1.295 1.294667 0.002517 0.194383 516
531.6825
y = 0.0352x + 0.0148
R² = 0.9995
0
0.2
0.4
0.6
0.8
1
0 10 20 30
absorbance
linearity of PCM at 222nm(Mean)

IBU 245
Conc.
(µg/ml)
Abs Mean SD % RSD Absorptivity
1 2 3
5 0.225 0.229 0.232 0.228667 0.003512 1.53581 456
10 0.382 0.389 0.39 0.387 0.004359 1.12633 387
15 0.514 0.526 0.529 0.523 0.007937 1.517639 348.6
20 0.68 0.693 0.698 0.690333 0.009292 1.345955 349
25 0.821 0.84 0.845 0.835333 0.012662 1.515836 338
355.65
y = 0.05x + 0.0481
R² = 0.9992
0
0.2
0.4
0.6
0.8
1
1.2
1.4
0 5 10 15 20 25 30
absorbance
linearity of IBU at 222nm (Mean)
y = 0.0303x + 0.0779
R² = 0.9993
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
0 5 10 15 20 25 30
absorbance
linearity of IBU at 245nm(Mean)

Conc.
(µg/ml)
Abs(222nm)
5 0.302
10 0.535
15 0.777
20 0.995
25 1.245
Conc.
(µg/ml)
Abs(245nm)
5 0.279
10 0.558
15 0.844
20 1.094
25 1.376
y = 0.0469x + 0.067
R² = 0.9997
0
0.2
0.4
0.6
0.8
1
1.2
1.4
0 5 10 15 20 25 30
absorbance
Mixture at 222nm
y = 0.0546x + 0.0112
R² = 0.9996
0
0.5
1
1.5
0 5 10 15 20 25 30
absorbance
Mixture at 245 nm

Intra day - PRECISION
Wavelength = 222 nm
Concentration
(µg/ml)
1 2 3 Mean Sd %RSD
5 0.302 0.302 0.297 0.300333333 0.002886751 0.961182468
15 0.777 0.775 0.769 0.773666667 0.004163332 0.538129944
25 1.245 1.243 1.239 1.242333333 0.00305505 0.245912299
Wavelength = 245
Concentration
(µg/ml)
1 2 3 Mean SD %RSD
5 0.279 0.277 0.274 0.276666667 0.002516611 0.909618607
15 0.842 0.837 0.835 0.838 0.003605551 0.430256715
25 1.376 1.374 1.368 1.372666667 0.004163332 0.303302477
Inter day - PRECISION
Wavelength = 222 nm
Concentration
(µg/ml)
1 2 3 Mean SD %RSD
5 0.302 0.298 0.291 0.297 0.005567764 1.874668136
15 0.777 0.762 0.756 0.765 0.010816654 1.413941677
25 1.245 1.231 1.227 1.234333333 0.009451631 0.76572762
Wavelength = 245
Concentration
(µg/ml)
1 2 3 Mean SD %RSD
5 0.276 0.271 0.268 0.271666667 0.003535534 1.301423524
15 0.842 0.837 0.831 0.836666667 0.003535534 0.422573774
25 1.376 1.369 1.362 1.369 0.004949747 0.361559348
Accuracy
For API Binary Mixture
Sr. No. Concentration Abs @ 222 nm Abs @ 245 nm % Recovery
1 80% 0.765 0.650 111.93
2 100% 0.847 0.754 105.93
3 120% 0.923 0.827 101.88
For Tablet Formulation
Sr. No. Concentration Abs @ 222 nm Abs @ 245 nm %Recovery
1 80% 0.624 0.505 97.5
2 100% 0.645 0.521 90.92
3 120% 0.714 0.572 92.46

Robustness
Wavelength
In nm
Absorbance
1 2 3 Mean SD % RSD
244.8 0.844 0.845 0.842 0.843 0.001527525 0.18105791
245 0.846 0.847 0.845 0.846 0.001 0.11820331
245.2 0.845 0.845 0.844 0.844 0.00057735 0.068352439
221.8 0.777 0.777 0.775 0.776 0.001154701 0.148737725
222 0.78 0.78 0.776 0.778 0.002309401 0.296584042
222.2 0.785 0.785 0.783 0.784 0.001154701 0.147220638
Ruggedness
concentration 15 µg/ml
Instrument:- A114550/08677
sr no. Wavelength (nm)
245 222
1 0.834 0.773
2 0.829 0.773
3 0.831 0.771
4 0.827 0.767
5 0.831 0.773
6 0.834 0.769
MEAN 0.831 0.771
SD 0.00275681 0.002529822
%RSD 0.331746059 0.328122196
Instrument:- A114549/08780
sr no. Wavelength (nm)
245 222
1 0.846 0.78
2 0.846 0.778
3 0.839 0.78
4 0.847 0.78
5 o.845 0.776
6 0.846 0.783
MEAN 0.8448 0.7795
SD 0.002925748 0.002140872
%RSD 0.34632431 0.274646837

 LOD = 3.3 * σ / S
At 222 =3.3*0.00057/0.046
=0.040
At 245=3.3*0.001/0.054
=0.061 µg/ml
 LOQ = 10 * σ / S
At 222=10*0.00057/0.046
=0.12
At 245=10*0.001/0.054
=0.18 µg/ml
 % Assay:
X= PCM
Y= IBU
Cx = Conc. Of PCM
Cy= Conc. Of IBU
ax1 = Absorptivity of drug PCM at 245=743
ax2 = Absorptivity of drug PCM at 222=362.75
ay1 = Absorptivity of drug IBU at 222= 355.65
ay2 = Absorptivity of drug IBU at 245=531.68
A1 = Absorption of sample at λ1.
A2 =Absorption of sample at λ2.
C x = A2ay1-A1ay2 C y = A1ax2-A2ax1
ax2ay1-ax1ay1 ax2ay1-ax1ay1
=0.777*355.65-0.845*531.68 =0.845*362.75-0.777*355.65
362.75*355.65-743*531.66 362.75*355.65-743*531.66
=6.5 µg/ml =11.3 µg/ml
 %Assay
15 µg/ml ----- 100% 15 µg/ml ------- 100%
6.5 µg/ml ------ (?) 11.3 µg/ml ----- (?)
=6.5*100/15=43.3% =11.3*100/15=76.6%

RESULTS:
Sr. No. Parameters Normal range Result
1 Linearity 0.999 0.999
2 Accuracy % Recovery 90-110%
3
% RSD < 2%
Wavelength
222nm 245nm
Precision
Repeatability 0.305 0.486
Intraday 0.5817 0.5473
Interday 1.346 0.6943
4 LOD - 0.061 µg/ml
5 LOQ - 0.18 µg/ml
6 Robustness % RSD < 2% 0.197 0.122
7 Ruggedness % RSD < 2% 0.301 0.338
8 % Assay PCM 44.3%
IBU 76.6%
CONCLUSION:
The method was validated as per ICH Q2 R1 and from the result table, we
conclude that method is linear, precise, accurate, robust and rugged.

To perform Analytical method validation of Paracetamol and Ibuprofen in pure and tablet form by UV spectrometric method.

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To perform Analytical method validation of Paracetamol and Ibuprofen in pure and tablet form by UV spectrometric method.