Simultaneous High-Performance Liquid Chromatographic Determination of Paracetamol, ppt
SIMULTANEOUS ESTIMATION OF PARACETAMOL, IBUPROFEN … · SIMULTANEOUS ESTIMATION OF PARACETAMOL,...
Transcript of SIMULTANEOUS ESTIMATION OF PARACETAMOL, IBUPROFEN … · SIMULTANEOUS ESTIMATION OF PARACETAMOL,...
www.iajpr.com
Pag
e29
64
Indo American Journal of Pharmaceutical Research, 2015 ISSN NO: 2231-6876
SIMULTANEOUS ESTIMATION OF PARACETAMOL, IBUPROFEN AND FAMOTIDINE
BY USING RP-HPLC METHOD
Allabasha Mumtaz, Mrs. Iffath Rizwana Department of Pharmaceutical Analysis & QA, Deccan School of Pharmacy (Affiliated to Osmania University),Nampally,Hyderabad-
500001.
Corresponding author
Allabasha Mumtaz
Department of Pharmaceutical Analysis & QA,
Deccan School of Pharmacy(affiliated to osmania university),
Nampally,Hyderabad-500001.
Copy right © 2015 This is an Open Access article distributed under the terms of the Indo American journal of Pharmaceutical
Research, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
ARTICLE INFO ABSTRACT
Article history
Received 03/09/2015
Available online
30/09/2015
Keywords
Simultaneous Estimation,
RP-HPLC,
Ibuprofen,
Famotidine & Paracetamol.
A new simple, accurate, precise and reproducible RP-HPLC method has been developed for
the simultaneous estimation of ibuprofen and famotidine and paracetamol in tablet dosage
forms using Kromosil ODS column,C18(250x4.6 ID) 5µm in isocratic mode. The mobile
phase consisted of Solution used – Phosphate buffer: CAN (40+60) pH 4.0. The flow rate was
1.0 ml/min and detection wavelength was carried out at 230nm. The retention times of
ibuprofen and famotidine & paracetamol were 6.477 min and 8.217 min & 2.703 min,
respectively. The method was linear over the concentration range for ibuprofen 2-10 μg/ml
and for and famotidine 2-10 μg/ml & for paracetamol 2-10 μg/ml. The recoveries of
ibuprofen and famotidine were found to be in the range of 99.2% and 99.85% & 100.49%
respectively. The validation of method was carried out utilizing ICHguidelines. The described
HPLC method was successfully employed for the analysis of pharmaceutical formulations
containing combined dosage form.
Please cite this article in press as Allabasha Mumtaz et al. Simultaneous Estimation of Paracetamol,Ibuprofen And Famotidine By
Using RP-HPLC Method. Indo American Journal of Pharmaceutical Research.2015:5(09).
www.iajpr.com
Pag
e29
65
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
INTRODUCTION
DRUG PROFILE
Name: famotidine
Description: A competitive histamine H2-receptor antagonist. Its main pharmacodynamic effect is the inhibition of gastric
secretion. [PubChem]
Structure:
Fig 1: chemical structure of famotidine.
Brand Names: Amfamox,Antodine,Digervin,Dispromil,Famosid,Famosan
Weight: Average: 337.445
Monoisotopic: 337.044934829
IUPAC name: 3-[({2-[(diaminomethylidene)amino]-1,3-thiazol-4-yl}methyl)sulfanyl]-N'-sulfamoylpropanimidamide
Name: Ibuprofen
Description: Ibuprofen, a propionic acid derivative, is a prototypical nonsteroidal anti-inflammatory agent (NSAIA) with analgesic
and antipyretic properties
Structure:
Fig 2:Chemical structure of ibuprofen.
Brand Names: Actiprofen,Betagesic,Dalsy,Dolormin,Tefin,Trendar
Weight: Average: 206.2808
Monoisotopic: 206.13067982
IUPAC name: 2-[4-(2-methylpropyl)phenyl]propanoic acid
Name: Paracetamol
Description: Acetaminophen, also known as paracetamol, is commonly used for its analgesic and antipyretic effects. Its
therapeutic effects are similar to salicylates, but it lacks anti-inflammatory, antiplatelet, and gastric ulcerative effects
Structure:
Fig 3:Chemical structure of paracetamol.
www.iajpr.com
Pag
e29
66
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Brand Names: Abenol,Acephen,Tapanol,Tempra,Salzone,Rounox.
Weight: Average: 151.1626
Monoisotopic: 151.063328537
IUPAC name: N-(4-hydroxyphenyl)acetamide
LABEL CLAIM
Trade Name: IBU C
Manufacturer: Winston Pharmaceuticals Ltd
Unit: 20mg/325mg/400mg
Type: Tablet
Significance for the choice of drug
The drugs (paracetamol and ibuprofen and famotidine) is selected for study because on this combination of drug not
much work has been carried out. No analytical work has been reported so far on paracetamol and ibuprofen and famotidine by
using rp-hplc. Hence by observing the existing literatures, the present work is carried out to develop a economical, and less
time consuming, precise method.
LITERATURE REVIEW
Review of literature
Literature survey was carried out on the proposed topic with the facility internet and refering scientific journals. The survey
reveals that, no work has been undertaken on the proposed topic and some related research work are coated below.
1.S S Chitlange, *et al., has developed a High performance thin layer chromatographic method is developed for simultaneous
estimation of ibuprofen and pseudoephedrine hydrochloride in tablets.
Narasimha Swamy Lakka, *et al., has developed a reverse phase high performance liquid chromatographic (RP-HPLC)
method suitable for simultaneous determi-nation of Ibuprofen and Paracetamol in dissolution of solid dosage forms in pharmaceuticals
has been developed.
Dimal A. Shah,*et al., has developed this method. isocratic, reversed phase-liquid-chromatographic assay method was
developed for the quantitative determination of ibuprofen and famotidine in combined-dosage form.
Prasanna Reddy Battu and MS Reddy has developed A simple, selective, accurate high Performance Liquid Chromatographic
(HPLC) method was developed and validated for the analysis of Paracetamol and Ibuprofen.
Gnana Raja M1
Geetha G2 and Sangaranarayanan A1 has developed a simple, precise, accurate, simultaneous and stability-indicating RPLC
method developed with an effective resolution for active pharmaceutical ingredients and marketed drug products. This method
effectively separate all the related substances of Ibuprofen and Paracetamol along with impurities. This method is using in the
estimation assay of Ibuprofen and paracetamol in drug substance also.
CRITICAL APPRAISAL OF LITERATURE REVIEW
The existing literature review shows that mostly pharmacological work has been done & only one analytical work has been
reported on paracetamol and ibuprofen and famotidine. Hence by observing literature, this work is carried out to obtain a low retention
time.
AIM & OBJECTIVE
AIM AND OBJECTIVE OF THE WORK
Pharmaceutical industries rely upon quantitative chemical analysis to ensure that the raw materials used and the final products
obtained meet the required specification.
Existing literature reveals that analytical methods like HPLC of Famotidine, Ibuprofen and Paracetamol reported with different
drug combinations.
Therefore, the present study has been undertaken in order to develop a simple, rapid, efficient and reproducible RP-HPLC method
for the analysis of Famotidine, Ibuprofen and Paracetamol in pharmaceutical dosage forms.
OBJECTIVES
Following are the objectives of present work
To perform method development for the drugs
To study effect of various mobile phases used in method development
To determine the drug content in given pharmaceutical dosage form
To validate analytical methods as per ICH guidelines
To demonstrate that it is suitable for its intended purpose
To establish identity, detect and quantitate impurities and to assess characteristics
www.iajpr.com
Pag
e29
67
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Plan of work
Solubility determination of paracetamol and famotidine & ibuprofen in various solvents and buffers.
Determine the absorption maxima of both the drugs in UV–Visible region in different solvents/buffers and selecting the solvents
for HPLC method development.
Optimize the mobile phase and flow rates for proper resolution and retention times.
Validate the developed method as per ICH guidelines.
EXPERIMENTAL INVESTIGATION
INSTRUMENTS USED
The following are the details of instrument used on which the work is carried out.
Table 1 Details of instrument.
UV-Visible Spectrophotometer Nicolet evolution 100
HPLC Shimadzu(LC 20 AT VP)
HPLC Agilent 1200 series
Ultra sonicator Citizen, Digital Ultrasonic Cleaner
pH meter Global digital
Electronic balance Shimadzu
Syringe Hamilton
HPLC Column Kromosil ODS
column,C18(250x4.6 ID) 5µm
METHODOLGY
Preparation of standard stock solution of Paracetamol:
32.5mg of Paracetamol was weighed and transferred in to 100ml volumetric flask and dissolved in methanol and then make
up to the mark with methanol and prepare 10 µg /ml of solution by diluting 0.3ml to 10ml with methanol.
Preparation of standard stock solution of Iboprofen:
40mg of Iboprofen was weighed in to 100ml volumetric flask and dissolved in Methanol and then dilute up to the mark with
methanol and prepare 10 µg /ml of solution by diluting 0.25ml to 10ml with methanol.
Preparation of standard stock solution of Famotidine:
2mg of Famotidine was weighed in to 100ml volumetric flask and dissolved in Methanol and then dilute up to the mark with
methanol and prepare 10 µg /ml of solution by diluting 5ml to 10ml with methanol.
RESULTS AND DISCUSSIONS
Results
The wavelength of maximum absorption (λmax) of the drug, 10 μg/ml solution of the drugs in methanol were scanned using
UV-Visible spectrophotometer within the wavelength region of 200–400 nm against methanol as blank. The resulting spectra are
shown in the fig. no. 8.1, 8.2 and 8.3 and the absorption curve shows characteristic absorption maxima at 243nm for Paracetamol, 221
nm for Ibuprofen and 265nm for Famotidine and 230nm for the combination.
UV-VIS spectrum of Paracetamol
Obeservation: λmax was found to be 243nm for Paracetamol
UV-VIS spectrum of Ibuprofen
Observation: λmax was found to be 221nm for Ibuprofen
UV-VIS spectrum of Famotidine
Observation: λmax was found to be 265nm for Famotidine
UV-VIS spectrum of Paracetamol, Ibuprofen and Famotidine and the isosbestic point was 230nm.
Optimised Chromatographic conditions
Mobile phase : Phosphate Buffer: Acetonitrile
Ratio : 40:65
Column : Zodiac, C18 (250×4.6× 5µ)
Wavelength : 230nm
Flow rate : 1ml/min
pH :4
www.iajpr.com
Pag
e29
68
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Preparation of mixed standard solution
Weigh accurately 325mg of Paracetamol and 400mg of Ibuprofen and 20mg of Famotidine in 100 ml of volumetric flask and
dissolve in 10ml of mobile phase and make up the volume with mobile phase From above stock solution 325µg/ml of Paracetamol and
400µg/ml of Ibuprofen and 20µg/ml of Famotidine is prepared by diluting 1ml to 10ml with mobile phase. This solution is used for
recording chromatogram.
Fig. 4: Chromatogram of Paracetamol, Ibuprofen and Famotidine.
Observation:Efficiency and resolution and shape of the peaks were good. Hence the method was optimized.
Table 2 : Optimized chromatographic conditions.
Mobile phase Phosphate buffer : Acetonitrile 40:60
pH 4.0
Column INERTSIL column,C18(150x4.6 ID) 5µm
Flow rate 1.0 ml/min
Column temperature Room temperature(20-25oC)
Sample temperature Room temperature(20-25oC)
Wavelength 230nm
Injection volume 20 µl
Run time 10min
Retention time About 2.703min for Paracetamol and 6.477min for
Ibuprofen and 8.217min for Famotidine.
VALIDATION
Specificity by Direct comparison method There is no interference of mobile phase, solvent and placebo with the analyte peak and also the peak purity of analyte peak
which indicate that the method is specific for the analysis of analytes in their dosage form.
Preparation of mixed standard solution
Weigh accurately 325mg of Paracetamol and 400mg of Ibuprofen and 20mg of Famotidine in 100 ml of volumetric flask and
dissolve in 10ml of mobile phase and make up the volume with mobile phase From above stock solution 325µg/ml of Paracetamol and
400µg/ml of Ibuprofen and 20µg/ml of Famotidine is prepared by diluting 1ml to 10ml with mobile phase. This solution is used for
recording chromatogram.
Preparation of sample solution:
5tablets (each tablet contains 325mg of Paracetamol and 400mg of Ibuprofen and 20mg of Famotidine) were weighed and
taken into a mortar and crushed to fine powder and uniformly mixed. Tablet stock solutions of Paracetamol (325μg/ml) and Ibuprofen
(400μg/ml) and Famotidine (20µg/ml) were prepared by dissolving weight equivalent to 325mg of Paracetamol and 400mg of
Ibuprofen and 20mg of Famotidine and dissolved in sufficient mobile phase. After that filtered the solution using 0.45-micron syringe
filter and Sonicated for 5 min and dilute to 100ml with mobile phase. Further dilutions are prepared in 5 replicates of 325μg/ml of
Paracetamol and 400μg/ml of Ibuprofen and 20µg/ml of Famotidine was made by adding 1ml of stock solution to 10 ml of mobile
phase.
www.iajpr.com
Pag
e29
69
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Fig. 5: Chromatogram for specificity of Paracetamol, Ibuprofen and Famotidine sample and standard.
Linearity and range
Preparation of mixed standard solution
Weigh accurately 325mg of Paracetamol and 400mg of Ibuprofen and 20mg of Famotidine in 100 ml of volumetric flask and
dissolve in 10ml of mobile phase and make up the volume with mobile phase From above stock solution 325µg/ml of Paracetamol and
400µg/ml of Ibuprofen and 20µg/ml of Famotidine is prepared by diluting 1ml to 10ml with mobile phase. This solution is used for
recording chromatogram.
Table 3: Linearity Preparations.
Preparations
Volume from
standard stock
transferred in
ml
Volume made up in
ml (with mobile
phase)
Concentration of solution(µg /ml)
Paracetamol Ibuprofen Famotidine
Preparation 1 1 10 32.5 40 2
Preparation 2 1.5 10 48.75 60 3
Preparation 3 2 10 65 80 4
Preparation 4 2.5 10 81.25 100 5
Preparation 5 3 10 97.5 120 6
www.iajpr.com
Pag
e29
70
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Fig 6:Chromatogram of Paracetamol, Ibuprofen and Famotidine preparation-1(50%),2(75%),3(100%),4(125%),5(150%).
Table 4: linearity of Paracetamol &Table 5: linearity of Ibuprofen.
S.No. Conc.(µg/ml ) Area S.No. Conc.(µg/ml ) Area
1 32.5 1238.342 1 40 1624.112
2 48.75 1819.191 2 60 2437.545
3 65 2297.198 3 80 3084.379
4 81.25 3001.861 4 100 4053.050
5 97.50 3474.678 5 120 4731.806
Table 6: linearity of Famotidine.
S.No. Conc.(µg/ml ) Area
1 2 639.476
2 3 961.082
3 4 1224.443
4 5 1643.461
5 6 1874.468
Fig. 7: Linearity graph of Paracetamol. Fig.8: Linearity graph of Ibuprofen.
www.iajpr.com
Pag
e29
71
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Fig. 9: Linearity graph of Famotidine.
The relationship between the concentration of Paracetamol, Ibuprofen and Famotidine and area of Paracetamol, Ibuprofen
and Famotidine should be linear in the specified range and the correlation should not be less than 0.99.
Accuracy Accuracy of the method was determined by Recovery studies. To the formulation (pre analyzed sample), the reference
standards of the drugs were added at the level of 50%, 100%, 150%. The recovery studies were carried out three times and the
percentage recovery and percentage mean recovery were calculated for drug is shown in table. To check the accuracy of the method,
recovery studies were carried out by addition of standard drug solution to pre-analyzed sample solution at three different levels 50%,
100% & 150%.
Fig. 10: Chromatogram of 50% recovery (injection 1,2,3).
www.iajpr.com
Pag
e29
72
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Fig. 11: Chromatogram of 100% recovery (injection 1,2,3).
www.iajpr.com
Pag
e29
73
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Fig. 12: Chromatogram of 150% recovery (injection 1,2,3).
Table 7: Recovery results for Paracetamol.
Table 8: Recovery results for Ibuprofen.
Recovery
level
Accuracy PARACETAMOL Average %
Recovery Amount
taken(mcg/ml)
Area Average
area
Amount recovered
(mcg/ml)
%Recovery
50% 32.5 1230.121 1229.964 32.53 100.10
100.49%
32.5 1233.330
32.5 1226.442
100% 65 2307.790 2309.946 64.74 99.60
65 2304.415
65 2317.632 150% 97.5 3482.522 3484.028 99.23 101.77
97.5 3489.291 97.5 3480.271
Recovery
level
Accuracy IBUPROFEN Average %
Recovery Amount
taken(mcg/ml)
Area Average
area
Amount
recovered(mcg/ml)
%Recovery
50% 40 1630.644 1633.280 40.01 100.03
99.92%
40 1634.598
40 1634.598
100% 80 3108.923 3105.724 79.82 99.78
80 3091.955
80 3116.294
150% 120 4744.754 4744.464 119.97 99.97
120 4734.981
120 4753.656
www.iajpr.com
Pag
e29
74
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Table 9: Recovery results for famotidine.
Precision
Method precision
Prepared sample preparations of Paracetamol, Ibuprofen and Famotidine as per test method and injected 5 times in to the column.
The % Relative standard deviation of Assay preparations of Paracetamol, Ibuprofen and Famotidine should be not more than 2.0%.
Fig. 13: Chromatogram of precision injection 1,2,3,4,5.
Recovery level Accuracy Famotidine Average
%
Recovery Amount
taken(mcg/ml)
Area Average
area
Amount
recovered(mcg/ml)
%Recovery
50% 2 620.403 637.150 1.99 99.47
99.85%
2 646.847
2 644.200
100% 4 1226.821 1233.068 4.00 99.96
4 1221.586
4 1220.796
150% 6 1874.678 1876.592 6.01 100.13
6 1879.899 6 1875.198
www.iajpr.com
Pag
e29
75
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Table 10: Results for Method precision of Paracetamol & Table 11: Results for Method precision of ibuprofen.
PARACETAMOL IBUPROFEN
S.No. Rt Area S.No. Rt Area
1 2.703 2312.214 1 6.477 3124.917
2 2.677 2265.782 2 6.470 3094.066
3 2.703 2304.778 3 6.477 3096.574
4 2.677 2291.044 4 6.470 3106.287
5 2.677 2274.485 5 6.470 3085.496
avg 2.699 2289.661 avg 6.471 3101.468
stdev 0.0150 19.619 stdev 0.003 15.058
%RSD 0.56 0.86 %RSD 0.05 0.49
Table 12: Results for Method precision of famotidine.
FAMOTIDINE
S.No. Rt Area
1 8.217 1226.539
2 8.217 1215.054
3 8.217 1203.559
4 8.217 1208.404
5 8.217 1227.891
avg 8.217 1216.289
stdev 0.00 10.787
%RSD 0.00 0.89
ASSAY
Preparation of samples for Assay
Preparation of mixed standard solution
Weigh accurately 325mg of Paracetamol and 400mg of Ibuprofen and 20mg of Famotidine in 100 ml of volumetric flask and
dissolve in 10ml of mobile phase and make up the volume with mobile phase From above stock solution 325µg/ml of Paracetamol and
400µg/ml of Ibuprofen and 20µg/ml of Famotidine is prepared by diluting 1ml to 10ml with mobile phase. This solution is used for
recording chromatogram.
Preparation of sample solution:
5tablets (each tablet contains 325mg of Paracetamol and 400mg of Ibuprofen and 20mg of Famotidine) were weighed and
taken into a mortar and crushed to fine powder and uniformly mixed. Tablet stock solutions of Paracetamol (325μg/ml) and Ibuprofen
(400μg/ml) and Famotidine (20µg/ml) were prepared by dissolving weight equivalent to 325mg of Paracetamol and 400mg of
Ibuprofen and 20mg of Famotidine and dissolved in sufficient mobile phase. After that filtered the solution using 0.45-micron syringe
filter and Sonicated for 5 min and dilute to 100ml with mobile phase. Further dilutions are prepared in 5 replicates of 325μg/ml of
Paracetamol and 400μg/ml of Ibuprofen and 20µg/ml of Famotidine was made by adding 1ml of stock solution to 10 ml of mobile
phase.
www.iajpr.com
Pag
e29
76
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Fig. 14: Chromatogram of Assay standard preparation-1,2,3,4,5.
www.iajpr.com
Pag
e29
77
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Fig. 15: Chromatogram of Assay sample preparation-1,2,3,4,5.
www.iajpr.com
Pag
e29
78
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Table No. 13: Assay Results.
Paracetamol Ibuprofen Famotidine
Standard
Area
Sample
Area
Standard
Area
Sample
Area
Standard
Area
Sample
Area
Injection-1 2259.197 2300.517 3078.755 3104.836 1182.175 1225.573
Injection-2 2289.669 2304.778 3100.404 3096.574 1215.303 1203.559
Injection-3 2283.417 2289.791 3088.048 3089.354 1202.590 1211.852
Injection-4 2300.800 2307.790 3092.034 3180.923 1203.369 1266.821
Injection-5 2309.033 2309.533 3100.765 3109.450 1226.173 1227.260
Average Area 2280.428 2302.482 3089.067 3116.227 1200.023 1227.015
Tablet average
weight 745.006 745.006
745.006
Standard weight 323.90 397.24 19.50
Sample weight
745.006 745.006
745.006
Label amount 325 400 20
std. purity 99.8% 99.8% 99.8%
Amount found in mg
326.28 399.93
19.90
Assay(%purity) 100.42% 99.98% 99.49%
ROBUSTNESS
To demonstrate the robustness of the method, prepared solution as per test method and injected at different variable
conditions like using different conditions like Temperature and wavelength. System suitability parameters were compared with that of
method precision.
The system suitability should pass as per the test method at variable conditions.
Fig. 16: Chromatogram of Paracetamol, Ibuprofen and Famotidine for Robustness (0.8 ml/min,1.2ml/min).
www.iajpr.com
Pag
e29
79
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Fig. 17: Chromatogram of Paracetamol, Ibuprofen and Famotidine for Robustness (228nm,232nm).
Table 14: Result of Robustness study.
Parameter
PARACETAMOL IBUPROFEN FAMOTIDINE
Retention
time(min)
Tailing
factor
Retention
time(min)
Tailing
factor
Retention
time(min)
Tailing
factor
Flow
0.8ml/min
1.0 ml/min
1.2ml/min
3.357
2.703
2.247
1.860
1.879
1.786
8.050
6.477
5.413
1.531
1.500
1.400
10.183
8.217
6.890
1.370
1.284
1.263 Wavelength
228nm
230nm
232nm
2.703
2.677
2.247
1.794
1.879
1.794
6.477
6.470
6.477
1.434
1.500
1.434
8.217
8.217
8.217
1.318
1.284
1.309
Ruggedness
The ruggedness of the method was studied by the determining the analyst to analyst variation by performing the Assay by
two different analysts.
The % Relative standard deviation of Assay values between two analysts should be not more than 2.0%.
Fig18: Chromatogram of Analyst 01 standard & sample preparation.
www.iajpr.com
Pag
e29
80
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
Fig. 19: Chromatogram of Analyst 02 standard & sample preparation.
Table 15: Results for Ruggedness.
PARACETAMOL %Assay IBUPROFEN %Assay FAMOTIDINE %Assay
Analyst 01 98.45 Analyst 01 99.56 Analyst 01 98.34
Analyst 02 99.87 Analyst 02 98.46 Analyst 02 99.54
%RSD 1.02 0.78 %RSD 0.85
Observation
From the observation the %RSD between two analysts Assay values not greater than 2.0%, hence the method was rugged.
DISCUSSIONS
The wavelength of maximum absorption (λmax) of the drug, 10 μg/ml solution of the drugs in methanol were scanned using
UV-Visible spectrophotometer within the wavelength region of 200–400 nm against methanol as blank. The resulting spectra are
shown in the fig. no. 8.1, 8.2 and 8.3 and the absorption curve shows characteristic absorption maxima at 243nm for Paracetamol, 221
nm for Ibuprofen and 265nm for Famotidine and 230nm for the combination. The Isosbestic point was found to be 230nm for
Paracetamol, Ibuprofen and Famotidine in combination and was shown in figure 8.3
Efficiency and resolution and shape of the peaks were good. Hence the method was optimized
CONCLUSION
It can therefore be concluded that use of the method can save much time and money and it can be used in small laboratories
with high accuracy and wide linear range. The method can be successfully used for routine analysis of paracetamol and ibuprofen &
famotidine in bulk drug formulation without interference. From the above experimental results and parameters it is concluded that, this
newly developed method for the simultaneous estimation of paracetamol and famotidine and ibuprofen was found to be simple,
precise, accurate and high resolution and gives shorter retention time.
This makes the method more acceptable and cost effective and it can be effectively applied for routine analysis.
Scope for the future work
Comparative estimation of different brands can be done.
Method development by HPLC.
Method validation by UV & HPLC as per ICH guidelines.
REFERENCES
1. Gurdeepchatwal,k. Anand , Instrumental methods of Chemical Analysis, 5th edition, Himalaya publishing house, New Delhi,
2002, 1,1-1.8, 2.566-2.570
2. William Kemp, Organic Spectroscopy, Palgrave, New York, 2005,7-10,328-330
3. D.A. Skoog.F.J. Holler and T.A. Nieman, principle of Instrumental Analysis, 5th edition, Saunders college Publishing,1998 778-
787
4. P.D. Senthi, HPLC: Quantitative Analysis Pharmaceutical Formulations, CBS Publishers distributors New Delhi(India), 2001, 3-
137
5. Chiral phase chromatography- http://scholar.lib.vt.edu/theses/available/etd32298223814/unrestricted/ch_02.pdf
6. Types of elution- http://chemwiki.ucdavis.edu/@api/deki/pages/402/pdf
7. Types of elution- http://hplc.chem.shu.edu/NEW/HPLC_Book/Rev.-Phase/rp_grad.htmL
www.iajpr.com
Pag
e29
81
Vol 5, Issue 09, 2015. Allabasha Mumtaz et al. ISSN NO: 2231-6876
8. A recent book provides a comprehensive treatment of the theory of high-performance gradient chromatography: Lloyd R. Snyder
and John W. Dolan (2006). High-Performance Gradient Elution: The Practical Application of the Linear-Solvent-Strength Model.
Wiley Interscience.
9. H.H.Willard, L.L. Merritt, J.A.Dean, F.A.Settel, Instrumental Method of Analysis, 7th edition, CBS Publishers distributors New
Delhi(India). 1986.
10. ICH Hormonisedtriparitate guidelines.
11. Pacher, P.; Nivorozhkin, A; Szabó, C (2006). "Therapeutic Effects of Xanthine Oxidase Inhibitors: Renaissance Half a Century
after the Discovery of Allopurinol". Pharmacological Reviews 58 (1):114. doi:10.1124/pr.58.1.6. PMC 2233605. PMID
16507884.
12. "WHO Model List of EssentialMedicines". World Health Organization. October 2013. Retrieved 22 April 2014.
13. S S Chitlange, D M Sakarkar, S B Wankhede, S G Wadodkar High performance thin layer chromatographic method for
simultaneous estimation of Ibuprofen and pseudoephedrine hydrochloride.
14. Narasimha Swamy Lakka, Nishant Goswami, P Balakrishna Development and validation of a RP-HPLC for simultaneous
determination of Ibuprofen and Paracetamol in solid dosage forms: Application to dissolution studies
15. Dimal A. Shah, Dixita J. Suthar, Sunil L. Baldania, Usman K. Chhalotiya, andKashyap K. Bhatt Development and Validation of
Liquid Chromatographic Method for Estimation of Ibuprofen and Famotidine in Combined Dosage Form ISRN Analytical
Chemistry.Volume 2012 (2012), Article ID 674392, 5 pages http://dx.doi.org/10.5402/2012/674392
16. Prasanna Reddy Battu1*and MS Reddy2 RP-HPLC Method for Simultaneous Estimation of Paracetamol and Ibuprofen in Tablets
http://www.ajrconline.org/ Asian J. Research Chem. 2(1): Jan.-March, 2009
17. Gnana Raja( M1 ) and Geetha( G2) and Sangaranarayanan (A1 )Simultaneous, Stability Indicating Method Development and
Validation for Related Compounds of Ibuprofen and Paracetamol Tablets by RP-HPLC Method http://omicsonline.org/ Gnana
Raja et al., J Chromat Separation Techniq 2012, 3:8.
54878478451150904