1. “SPECTROSCOPIC METHOD DEVELOPMENT
FOR LOSARTON POTASSIUM IN TABLET”
A project report in Pharmaceutical Analysis
By
Ninad .M.Phadke
Under the guidance of :
Mrs. Dabhade M.P.
(M.PHARM)
S.N.D. COLLEGE OF PHARMACY,YEOLA

2. INTRODUCTION TO ANALYTICAL
CHEMISTRY
 “Analytical chemistry deals with methods for determining the
chemical composition of samples of matter.”
 The drug or dosage form for human use must have excellent quality
and purity, free from any impurity, because these dosage forms directly
affects the life, hence their analysis is important which is carried out
using analytical methods.
 The analytical method development for new API and dosage form
involves development and validation of method for testing those API
and dosage form in QC laboratories routinely as part of regular Quality
control sample testing using the methods of Analytical Chemistry.
 Analytical Chemistry involves two branches Qualitative and
Quantitative.

3. BRANCHES OF ANALYTICAL
CHEMISTRY
 QUALITATIVE ANALYSIS : Qualitative analysis deals with
the identification of elements, ions or compounds present in a sample.
 QUANTITATIVE ANALYSIS : Quantitative analysis deals
with the determinations of how much of one or more constituents are
present.

4. Classification of Analytical Methods
A) CLASSICAL METHODS:
For classical qualitative analysis the separated compounds were treated with reagents
that could be recognized by either color, by their boiling or melting points, their
solubility in a series of solvents or their optical activities.
 Advantages of Classical Methods:
• Procedure is simple and accurate.
• The equipment needed is cheap.
• Methods are based on absolute measurements.
• Specialized training is not required.
 Limitations of Classical Methods:
• Chemical environment is critical.
• There is a lack of versatility.
• There is a lack of specificity.
• Accuracy decreases with decreasing amount.

5. B) INSTRUMENTAL METHODS :
Measurements of physical properties of analyte such as conductivity, electrode
potential, light absorption or emission, mass to charge ratio and fluorescence
forms the basis of Instrumental Qualitative Analysis .
 Advantages of Instrumental Methods:
 Small Samples can be used.
 High sensitivity is obtained.
 Measurements obtained are reliable.
 The determination is very fast.
 Complex samples can be handled.
 Limitations of Instrumental Methods:
 Skilled person is required.
 The sensitivity and accuracy depends on the instruments or wet chemical
methods.
 Cost of equipment is high.
 Sizable space is required.

6. APPLICATIONS OF ANALYTICAL
CHEMISTRY
 In medicine, analytical chemistry is the basis for clinical laboratory
tests, which helps physicians, in diagnosis of various diseases and
better recovery of patients.
 In industry, it provides the means of testing raw materials and for
assuring quality of finished products, like fuel, paints, and other
pharmaceuticals products .
 Suspected environmental contaminant testing is great achievement by
analytical chemistry.

7. PRINIPLE OF UV VISIBLE
SPECTROSOPY
 The ultraviolet-visible spectroscopy is a common analytical technique
for qualitative and quantitative analysis of solid, liquid or gas samples
 The intensity of the absorbance is detected by Beer-Lambert’s law.
 Lambert’s Law: At a given concentration (C) of a homogenous
absorbing system the intensity of transmitted light decreases
exponentially with increasing path length.
-dI / dT = KI
 Beer’s Law: For a layer of defined path length the intensity of
transmitted light decreases exponentially with increase in
concentration (C) of a homogenous absorbing system.
It = Io e –ac

8.  The combination of these laws gives the Beer-
Lambert Law,
Log Io/ It = A = abc
Where,
A = Absorbance of the solution at a particular wavelength.
Io = Intensity of the light beam.
It = Intensity of beam after passing through solution.
a = Absorptivity of molecule at the wavelength of beam.
b = Path length of cell in cm.
c = Concentration of solution in gm/lit.

9. CHOICE OF SOLVENT
Minimum wavelength
Solvent
( nm )
Acetonitrile 190
Water 190
Cyclohexane 195
Hexane 200
Methanol 200
Ethanol 200
Ether 215
Methylene chloride 220
chloroform 240

10. Methods Of Quantitative Spectrophotometric
Assay of Medicinal Substances
 The concentration of a single component in a sample may be determined
by a simple Spectrophotometric methods of absorbances, provided that
the other components have a sufficiently small absorbance at the
wavelength of measurements.
 The single component & multicomponent mixture containing the API can
be analyzed by either of following method :
 Simultaneous equation method.
 Multicomponent analysis.
 Absorption ratio method.
 The assay of an absorbing substance may be quickly carried out by
preparing solution in a transparent solvent and measuring its absorbance
at a suitable wavelength {i.e.maximum absorption (λmax)}.

11. INSTRUMENTATION OF U.V.
Diagram of double beam UV Spectrophotometer

12. An instrument for measuring the absorption of
UV or visible radiation contains following
important components :
 Source of light –Hydrogen or deuterium lamp, tungsten
lamp.
 Prism or Gratings monochromator.
 Sample container-Quartz cuvettes.
 Detectors
 Signal processor and readout.

13. Applications of UV-Visible
Spectroscopy
 Identification of various organic, inorganic molecules and
ions by matching their spectrum with reference spectra.
 For qualitative and quantitative analysis of drugs in
pharmaceutical industry.
 Monitoring of reaction rates (chemical kinetics)
 Enzyme assays
 Environmental remote sensing
 Used as detectors in various systems like HPLC,
electrophoresis etc.

14. STEP BY STEP PROCEDURE FOR UV –VISIBLE
SPECTROSCOPIC ANALYSIS OF DRUG
 Literature Review
 Objective Of Work
 Plan Of Work
 Experimental Work
 Results and discussion
 Method Validation
 Conclusion

15. PLAN OF WORK
Losarton Potassium
Calibration Of
Apparatus
Calibration Of UV
Instrument
Selection of suitable
solvent
Determination of max
Calibration curve
Estimation of drug from
tablet formulation
Recovery studies
Result and conclusion

16. OBJECTIVE OF WORK
The objective of this investigation was to devise
simple, precise, rapid and economical method for
the estimation of Losarton Potassium in bulk drug
and the tablet formulation.
 Tablets procured from the market were analyzed by the
proposed method. In this method, the tablets were crushed
and dissolved in solvent and diluted further. Sufficient
amounts of the samples were withdrawn and their
absorbances were noted at 238.4 nm against reagent blank.

17. Optical characteristics and precision
Absorption maxima(nm)
238.4nm
Absorption maxima(nm) 238.4nm
Beer’s law limit (g/ml) 03-30µg/ml
Correlation coefficient(r2) 0.9871
Regression equation 0.0154x + 0.0198

18. TABLET ANALYSIS
SR. NO. Amt taken in Amt found in Amt found in %
(g/ml) (g/ml)
1. 18 17.99 99.94
2. 18 18.01 100.05
3. 24 23.99 99.95
4. 24 24.01 100.04
5. 30 29.99 99.96
6. 30 30.01 100.03

19. Statistical validation of tablet
formulation
Amt found in MEAN Standard Coeffecient of Standard
(%) deviation variation error
100.03 23.83 ± 0.15379 0.932 0.002124

20. Recovery study of Losarton Potassium
Name of drug Level of Conc. Conc. Total absorban Conc. % MEAN
Percenta used added conc. ce Recovered Recovery
ge µg/ml µg/ml in µg/ml
Recovery
50 12 6 18 0.298 17.99 99.94 99.97
Losarton
Potassium 50 12 6 18 0.297 18.01 100.05
50 12 6 18 0.298 17.99 99.94
100 12 12 24 0.440 23.99 99.95 99.99

21. Statistical validation of recovery studies
of tablet formulation
Level of recovery MEAN % Standard Coeffecient Of Standard
Recovery Deviation Variation Error
50 % 99.97 0.076895 0.466 0.001062
100 % 99.99 0.15379 0.932 0.002124
150 % 99.98 0.30758 1.864 0.004248

22. Statistical Results Of Analysis
Sr. No. Tablet Label claim(mg) S.D.* %Recovery*
1. LOSAR 50 ±0.15379 99.98 %
 Thus the statistical study was performed.
The value obtained for S.D seemed to be within range.
This showed the suitability of procedure.

23. METHOD VALIDATION
 Linearity-Linearity was observed in the range of 03-30g/ml.
The calibration curve yielded coefficient of correlation (r2)
0.9871.
 Assay result- Losarton Potassium tablets of a marketed brand
was analyzed by proposed methods, the percentage in tablet
were determined and presented in the above table.
 Accuracy and precision-The % Recovery was found to be in
the range of 99.80-100.5%,indicates the non-interference from
the formulation excipient and confirm the accuracy and
precision of the method.

24. CONCLUSION
 All the above result indicates that, the method employed
here is a very simple, economical and can be used for
routine analyses of the drug, Losarton Potassium.
 The proposed method was found to be accurate, simple
and rapid for routine analysis of the drug. The recovery
was 100.43% (LOSAR) which is close to 100%
indicating reproducibility and accuracy of the method.

25. References
 Govt. of India, Ministry of Health and Family Welfare. Vol. 2. Delhi: Publication by
Controller of Publication; 2007. Indian Pharmacopoeia; pp. 484–554.
 British Pharmacopoeia. (International ed.) 1993;Vol. 1:429, 483. Published on the
Recommendation of the Medicines Commissions Pursuant to Medicines Act 1968, 1993.
 Martindale, The Extra Pharmacopoeia, 33rd ed., Royal Pharmaceutical
Society, London, 2002; pp 921-922.
 United States Pharmacopoeia 29 NF 24, Published on the Recommendation of the
Medicines Commissions Pursuant to Medicines, page no. 587
 Skoog, West, Holler, Crouch, “Fundamentals of analytical chemistry” , eighth edition,
2009 (Indian edition), cengage learning India pvt ltd , New delhi, pageno. 271-280.
 A.V Kasture, K.R Mahadik, S.G Wadodkar, H.N. More, “A textbook of pharmaceutical
analysis, Instrumental methods” , Nirali Prakashan, vol.2, 9th edition, page no. 5-7, 28-30