1. SwethaBhavani N, HimaBindu S, Venkateswara Rao A, Panikumar D Anumolu
Simultaneous Quantification of Lornoxicam and Paracetamol Combination By Application of First Derivative UV- Spectroscopy
SwethaBhavani N, HimaBindu S, Venkateswara Rao A, Panikumar D Anumolu

2. contents Introduction Objective References Drug profile1
Introduction
Objective
Drug profile
Results & discussions
References
Conclusion
Materials & methods

3. DERIVATIVE SPECTROSCOPY2
Derivatization of actual spectra with respect to wavelength
Zero order
First order
Second order
Zero, first and Second-order UV derivative spectrum

4. 3
Advantages of First derivative spectroscopy: (1) Precise determination of the λmaxcan be obtained from the zero crossing of the first derivative. (2) Improved spectral resolution (3) Discrimination of broad bands
Resolution enhancement in derivative spectroscopy

5. OBJECTIVES 4
Development and validation of UV derivative spectrophotometric method for simultaneous estimation of Lornoxicam and Paracetamol
Application of developed method for simultaneous estimation of Lornoxicam and Paracetamol in marketed tablets

6. Drug profile of Lornoxicam5
Category: NSAID Chemical name: 6-Chloro-4-hydroxy-2-methyl-N-2pyridinyl-2H-thieno[2,3-e]- 1,2thiazine-3-carboxamide1,1-dioxide Molecular formula:C13H10Cl N3O4S2 Molecular weight: g/mol Melting point: c Solubility: Slightly soluble in chloroform & 0.1mol/LNaOH, and very slightly soluble in methanol and acetonitrile, hardly soluble in water. pKa : 4.7 Partition coefficient : Log P (octanol/pH 7.4 buffer):1.8 ,octanol-water- 3.15(calc)

7. Drug profile of Paracetamol:6
Category: Analgesic and antipyretic Chemical name: N-(4-Hydroxyphenyl)acetamide Molecular formula:C8H9NO2 Molecular weight:151.2 Melting point: 169.0° to 170.5° Solubility: Very slightly soluble in cold water, considerably more soluble in hot water; soluble in ethanol, methanol, dimethylformamide, ethylene dichloride, acetone, and ethyl acetate; very slightly soluble in chloroform; slightly soluble in ether; practically insoluble in petroleum ether, pentane, and benzene pKa : 9.5 (25°).Partition coefficient : Log P(octanol/water), 0.5. Appearance: White crystals or crystalline powder

8. Instrument : Shimadzu UV-1800 Spectrophotometer7
Materials and methods
Instrument : Shimadzu UV-1800 Spectrophotometer
Chemicals Used –0.01N NaOH

9. Preparation of standard stock solutions
Method development 8
Preparation of standard stock solutions
LOR(10mg) and PAR (10mg) were weighed and transferred into two separate 10ml volumetric flasks
Dissolved in 1ml 0.01N NaOH and volume was made up to 10ml These solutions have concentration
1000mcg/ml-
stock A
From stock A 5ml each of LOR and PAR are taken into two separate 50ml volumetric flasks and volume makeup was done with 0.01N NaOH
100mcg/ml stock B
From stock B 10mcg/ml solutions of LOR and PAR were prepared with 0.01N NaOH and scanned in the range of nm using 0.01N NaOH as blank
S.No.

10. 9 A B S O R N C E
WAVE LENGTH(nm)
Zero-order UV overlaid spectrum of Lor(10mcg/ml) and Par(10mcg/ml) in 0.01N NaOH

11. 10
Since the zero order spectra of two drugs are overlapping, zero order spectra were transformed into corresponding first derivative spectra
גmax for LOR
zcp for LOR
dA/dג
zcp for PAR
גmax for PAR
WAVELENGTH
First Order UV Overlaid Spectrum of Lor (10mcg/ml) and Par (10mcg/ml) in 0.01N NaOH

12. Calibration curve for of LOR & PAR11
Preparation of calibration curve for Lornoxicam
From stock Solution - B (LOR) 0.2,0.6,1,1.4,1.8,2.2ml transferred into 10ml volumetric flasks and volume was adjusted to 10ml with 0.01N NaOH
(2-22mcg/ml)
First derivative spectra were recorded and derivative absorbance was measured at 347 nm
Preparation of calibration curve for Paracetamol :
From stock Solution-B (PAR) 0.1, 0.6, 1.2, 1.8, 2.4 , 3, 3.6ml transferred into 10ml volumetric flasks.
(1-36mcg/ml)
First derivative spectra were recorded and derivative absorbance measured at 272.5nm

13. Calibration plot of LOR & PAR12
zcp for LOR
גmax for LOR
dA/dג
zcp for PAR
ג max for PAR
WAVELENGTH (nm)
UV first-derivative linearity range of LOR (2-22mcg/ml) and PAR
(1-36mcg/ml) in 0.01N NaOH

14. Concentration (mcg/ml) Derivative absorbance (AM ± SD) (n=3)13
Data for standard plot of LOR in 0.01N NaOH at 347nm
Concentration(mcg/ml)
S.No.
Concentration (mcg/ml)
Derivative absorbance (AM ± SD) (n=3) 1 2 ± 3 6 ± 4 10 ± 5 14 ± 18 ± 7 22 ±
Derivative absorbance
Standard plot of LOR in 0.01N NaOH at 347 nm

15. Data for standard plot of PAR in 0.01N NaOH at 272.5 nm14
S.No.
Concentration (mcg/ml)
Derivative absorbance (AM ± SD) (n=3) 1 2 ± 3 6
±0.0146 4 12 ± 5 18 ± 24 ± 7 30 ± 8 36 ±
Derivative absorbance
Concentration(mcg/ml)
Standard plot of PAR in 0.01N NaOH at nm

16. Precision Lornoxicam Intra-day precision Inter-day precision15
Lornoxicam
Concentration (mcg/ml)
Intra-day precision
Inter-day precision
Concentration estimated (mcg/ml)
(AM ± SD) (n=3)
% RSD
%RSD 2
2.06 ± 0.004
0.22
1.98± 0.010
0.54 14
 14.54 ± 0.065
0.45
14.02 ± 0.096
0.69 32
22.56 ± 0.22
0.98
22.34± 0.205
0.92
Paracetamol
Concentration estimated (mcg/ml) (AM ± SD) (n=3) 1
 1.22± 0.003
0.25
 1.12± 0.004
0.357 18
18.14 ± 0.116
0.64
18.02 ± 0.129
0.72 36
36.32± 0.272
0.75
36.24± 0.358
0.99

17. Theoretical content (mg) Amount found (mg) (AM ± SD) (n=3)
Recovery studies 16
Formulation
Recovery level (%)
Recovery of analyte
Theoretical content (mg)
Amount found (mg) (AM ± SD) (n=3)
Recovery (%)
% RSD
Lornoxi-P
LOR 2
2.1± 0.015
105
0.714
PAR
125
125.1 ± 0.145
100.08
0.115 80
3.6
3.5± 0.045
97.2
1.285
225
224.5 ± 0.34
99.77
0.151
100 4
4.2 ± 0.009
0.214
250
± 0.985
100.09
0.393
120
4.4
4.4 ± 0.012
0.272
275
275.12± 1.256
100.04
0.456
Lornasafe -plus
2.2 ± 0.012
110
0.56
± 0.252
100.03
0.201
3.4 ± 0.008
94.4
0.24
± 0.556
100.02
0.247
3.7 ± 0.024
92.5
0.67
± 0.357
0.142
4.3 ± 0.014
97.7
0.34
274.9± 0.852
99.9
0.309

18. System Suitability Parameters17
System Suitability Parameters
S.No
Parameter
Lornoxicam
Paracetamol 1
Absorption maxima (nm)
347
272.5 2
Beer's Law Limit (mcg/ml)
2-22
1- 36 3
slope
0.001
-0.003 4
Intercept 5
Correlation coefficient
0.999
0.995 6
Regression equation
y = 0.001x +0.0
y =-0.003x 7
LOD (mcg/ml)
0.1643
0.0745 8
LOQ (mcg/ml)
0.5477
0.248

19. Amount found (mg) (AM ± SD) (n=3)
Assay 18
Formulation
Lornoxicam
Paracetamol
Label claim (mg)
Amount found (mg) (AM ± SD) (n=3)
% Recovery
% RSD
Lornoxi-p 8
8.4 ± 0.011
105
0.13
500
500.16±0.252
0.05
Lornasafe-plus
8.2 ± 0.102
102
1.243
±0.562
100
0.112

20. ADVANTAGES OF PROPOSED METHOD19
Simple spectra and less interference.
High sensitivity and wide linearity range.
Rapid, economical. Eco-friendly, accurate and precise method.
High spectral discrimination.
Single analytical method for quantification of LOR and PAR in combined tablet dosage forms.

21. conclusion 20
It is a simple, novel, more economic, reliable, precise and accurate method for simultaneous estimation of Lornoxicam and Paracetamol in bulk and formulations. So this method can be successfully applied to routine analysis of studied drugs in their tablet dosage forms.

22. References: 21
ICH Harmonized Tripartite guideline validation of analytical procedures: text and methodology, Q2(R1) Current step 4 version, 27 october 1994.
Mark, H. and Workman, J. Derivatives in spectroscopy. Spectroscopy ,18 (4): 32-37
Beckett AH and Stenlake JB. Practical pharmaceutical chemistry; 4th edition, the athlone press. 2007,

23. 22
Lakshmi Sivasubramanian*, Lakshmi K.S. and TinTu.T. PB Khedikar and G Pawnikar, Absorption Correction Method for Simultaneous Estimation of lornoxicam and paracetamol tablet dosage form by absorbance ratio method , International Journal of pharmacy and pharmaceutical sciences, 2010,vol 2,Issue 4 .
DharaJ.Patel*Vivek Patel., Simultaneous determination of paracetamol and lornoxicam in tablets by thin layer chromatography combined with densitometry, An International Journal of chem tech research, pp ,Vol-2 no.4.
T.Raja*,A.LakshmanaRao,Validated High performance Thin layer liquid chromatography method for simultaneous quantitation of paracetamol and lornoxicam in bulk drug and pharmaceutical formulation , International Journal Pharm Biomed Res 2012,3(3),

24. 23
Thanq…