SIMULTANEOUS QUANTIFICATION OF LORNOXICAM/ PARACETAMOL TABLETS BY APPROACH OF FIRST DERIVATIVE UV-SPECTROSCOPY
Swetha Bhavani N
Department of Pharmaceutical Analysis, Gokaraju Rangaraju College of Pharmacy, Osmania University, Hyderabad, Andhra Pradesh-500090, India
Keywords: Lornoxicam, paracetamol, derivative spectrophotometry.
Abstract

Background: Derivative spectroscopy provides a greater selectivity and spectral discrimination than common spectroscopy. It is the dominant approach for resolution of one analyte whose peak is hidden by a large overlapping peak of another analyte in multi component analysis. Hence, this technique we have been successfully applied for simultaneous quantification of lornoxicam and paracetamol in combined tablets. Materials and Methods: The method is based on the derivative spectrophotometric method at zero-crossing wavelengths. Two wavelengths 347 nm (zero crossing point for paracetamol) and 272.5nm (zero crossing point for lornoxicam) were selected for the quantification of lornoxicam and paracetamol respectively, using 0.01 M sodium hydroxide as solvent and Shimadzu (Japan) UV-Visible spectrophotometer (UV-1800) instrument. Results: The first derivative amplitude-concentration plots were rectilinear over the range of 2-22 µg/mL and 1-75 µg/mL with detection limits of 0.06 and 0.08 µg/mL and quantification limits of 0.2 and 0.26 µg/mL for lornoxicam and paracetamol respectively. The proposed method was statistically validated as per ICH guidelines. The percentage recovery was within the range between 97-101 and % relative standard deviation for precision and accuracy of the method was found to be less than 2. Conclusion: The proposed method was effectively applied to routine quality control analysis of studied drugs in their tablet formulations.

Article Information

Identifiers and Pagination:
Year:2013
Volume:5
First Page:50
Last Page:58
Publisher Id:JAppPharm (2013 ). 5. 50-58
Article History:
Received:January 5, 2013
Accepted:March 3, 2013
Collection year:2013
First Published:April 1, 2013

INTRODUCTION

Lornoxicam chemically known as 6-Chloro-4-hydroxy-2-methyl-N-2pyridinyl-2H-thieno [2,3-e]-1,2thiazine-3-carboxamide1,1-dioxide, [Figure 1] is a novel non-steroidal anti inflammatory agent with marked analgesic properties [1]. It is not official in any pharmacopeia. Paracetamol chemically N-(4-Hydroxyphenyl) acetamide, [Figure 2] is a centrally and peripherally acting non-opoid analgesic and antipyretic. It is official in Indian Pharmacopoeia [IP] [2], British Pharmacopoeia [BP] [3] and United States Pharmacopoeia [USP] [4].  Lornoxicam [8 mg] and paracetamol [500 mg] have been formulated in a fixed dose tablet dosage form for the treatment of severe pains.

A detailed literature survey revealed that the few analytical methods include UV-spectrophotometry [5], high performance liquid chromatography (HPLC) [6] and HPTLC [8,9] methods available for simultaneous quantification of lornoxicam and paracetamol.

To the best of our knowledge, no method reported on the use of derivative spectrophotometry for the simultaneous quantification of lornoxicam and paracetamol in 0.01N sodium hydroxide as solvent. Moreover there was no simple, eco-friendly and economical method available for estimation of lornoxicam and paracetamol either in bulk drug or in formulation by first derivative spectroscopy using 0.01N NaOH as solvent.

                  

Figure 1. Chemical structure of lornoxicam           

Figure 2. Chemical structure of paracetamol

 

Derivative spectrophotometry  more superior than normal spectroscopy by minimizing  number of analytical problems like  resolution of multicomponent systems, removal of sample turbidity, matrix back ground and enhancement of spectral details. It is the dominant approach for resolution of one analyte whose peak is hidden by a large overlapping peak of another analyte in multi component analysis [10-16]. Hence, an attempt has been made to develop a simple, eco-friendly and economical first –derivative spectrophotometric method for simultaneous quantification of lornoxicam and paracetamol either in bulk drug or in tablet dosage form using 0.01N NaOH as solvent and method was statically validated as per ICH guidelines.

 

MATERIALS AND METHODS

Instrumentation

Shimadzu (Japan) UV-Visible spectrophotometer (UV-1800) with 1cm matched quartz cells was used for spectrophotometric analysis and a calibrated electronic single pan balance (Shimadzu, Aux-220) were used during the analysis.

Reagents and chemicals

Lornoxicam and paracetamol bulk drugs were obtained as gift samples from Dr.Reddy’s laboratories Ltd, Hyderabad, India. Tablets (Lornasafe -plus and Lornoxi-P) were procured from local pharmacies. Anhydrous sodium hydroxide was purchased from Hi-media, Mumbai.

Preparation of standard stock solutions

Each of standard lornoxicam and paracetamol (10mg) were weighed and transferred into two separate 10 mL volumetric flasks and dissolved in 0.01N NaOH. The flasks were shaken and volume was made up to the mark with 0.01N NaOH.  From this 5 ml solution was diluted to 50 ml with 0.01N NaOH to obtain a standard solution of lornoxicam and paracetamol having final concentration of 100 µg/mL of each.

 

Figure 3. Normal overlaid UV spectrums of lornoxicam (…) and paracetamol (--)

Figure 4. First order derivative UV overlaid spectrum of lornoxicam (…..) and paracetamol (--)

 

Selection of wavelengths

Standard solution of lornoxicam (LOR) and paracetamol (PAR) were diluted appropriately with 0.01N NaOH to obtain solution containing lornoxicam(10µg/mL) and paracetamol (10µg/mL). Spectra of these diluted solutions were scanned in the spectrum mode between 200 nm to 400 nm using 0.01N NaOH as a blank. The zero-order spectra of lornoxicam and paracetamol were transformed to corresponding first-derivative spectra in the range of 200 - 400 nm. The overlaid spectra (zero and first order) lornoxicam and Paracetamol are shown in Figure 3 and 4.

Derivative conditions

The overlaid zero-order spectra of standard solution of lornoxicam and paracetamol at 10 µg/mL and spectra were found to be similar in nature and overlapping. It was observed that lornoxicam and paracetamol contribute significantly below 300 nm wavelength for absorbance. Hence, the derivative graphical method was used to estimate lornoxicam and paracetamol in presence of each other. First-order derivative spectra of lornoxicam and paracetamol were overlapped. The wavelength 347 nm was selected for the quantification of lornoxicam (where the derivative response for paracetamol was zero). Similarly, 272.5nm was selected for the quantification of paracetamol (where the derivative response for was zero). Characteristic wavelengths (zero-crossing points) for lornoxicam and paracetamol were confirmed by varying the concentrations of both drugs.

Calibration curves for lornoxicam and paracetamol

 The standard solution of lornoxicam and paracetamol were used to prepare two different sets of working standard solutions of lornoxicam (0.2-4.2 µg/mL) and  paracetamol (1-75 µg/mL).The first-derivative spectra were recorded using the prepared solutions against 0.01N NaOH as blank. The values of first-derivative absorbance were plotted against corresponding concentrations to construct the calibration curves.

Determination of Lornoxicam and paracetamol in their combined dosage form (assay)

 Twenty tablets of each marketed formulation (Lornoxi-P and Lornasafe-plus), each containing 8 mg of lornoxicam and 500mg of paracetamol were taken and accurately weighed. Average weight was determined and crushed into fine powder. An accurately weighed quantity of powder equivalent to 8 mg lornoxicam and 500mg paracetamol was transferred to volumetric flask of 10 mL capacity, Volume was made up to the mark with 0.01N NaOH. The above solution was filtered through whatmann filter paper (No.41).The filtrate was further diluted to obtain sample solutions of concentrations within linearity range. The derivative absorbance of sample solutions were measured at selected wavelengths used for the quantification of lornoxicam and paracetamol.

Method validation

The method was validated for accuracy, precision, specificity, linearity, LOD and LOQ by the following procedures [17].

Accuracy

 The accuracy of the method was determined by calculating recoveries of lornoxicam and paracetamol by the method of standard additions. Known amounts of lornoxicam and paracetamol (80%, 100% and 120%) levels were added to a pre quantified sample solutions. The recovery was verified by estimation of drug in triplicate preparations at each specified concentration level and calculated % RSD.

Precision

The intra-day and inter-day precision of the proposed first-derivative spectrophotometric simultaneous method was determined by estimating the corresponding response three times on the same day (intra- day) and for three consecutive days (inter-day) for three different concentrations of lornoxicam (0.2, 2.2 and 4.2 µg/mL) and paracetamol (1, 45 and 75 µg/mL). The results are reported in terms of relative standard deviation (% RSD).

 Selectivity

Selectivity is the ability of the method to accurately measure a compound in the presence of other components such as impurities, degradation products and matrix components. The selectivity of the proposed method was evaluated through the analysis of a placebo solution, which was prepared with the common excipients of the pharmaceutical formulation. Thus, the mixture of component inert was prepared in their usual concentration employed in tablets (concentrations were determined based in Handbook of Pharmaceutical Excipients [18] and calculated for medium weight of content). The developed method was applied in order to check if any component of the formulation could generate a response with emission band similar to the drugs.

Sensitivity

The sensitivity of the method was determined with respect to LOD and LOQ. The LOD and LOQ were separately determined based on standard calibration curve.

 

RESULTS AND DISCUSSION

A simple, eco-friendly and economic first derivative spectrophotometric method was developed for the simultaneous quantification of lornoxicam and paracetamol  bulk drug and formulations using 0.01N NaOH as solvent and was also validated as per ICH guidelines. The calibration curves shows that, the developed method was linear in the concentration range of 0.2-4.2 µg/mL and 1-75 µg/mL for lornoxicam and paracetamol. [Figure 5 & 6] Limit of detection and limit of quantification values were indicated that the method shows high sensitivity. The optimized conditions for developed method are shown in Table 1. No significant difference between intra-day and inter-day precision, revealed that the method is reproducible (Table 2). The % recovery was within the range between 97-101 (Table 3) and %RSD for commercial formulation was shown less than 2 (Table 4). This indicates that the method is accurate and reliable.

Figure 5 Calibration plot for lorinoxicam

Figure 6 Calibration plot for paracetamol

 

Table 1: Optimum conditions for proposed method

Parameter

       Lornoxicam

         Paracetamol

Absorption maxima (nm)

              347

272.5

Beer's Law Limit(µg/mL)

0.2-4.2

1 – 75

            Slope

0.001

-0.003

          Intercept

0

0

Correlation coefficient

0.994

0.999

Regression equation

y = 0.001x +0.0

y =-0.003x -0.00005

LOD  (µg/mL)

0.06

0.08

LOQ (µg/mL)

0.2

0.26

     





LOD- Limit of detection; LOQ- Limit of quantification 


Table 2: Precision of the method

Concentration (mcg/ml)

Intra-day precision

Inter-day precision

Concentration estimated (µg/mL)

(AM ± SD) *

% RSD

Concentration estimated (µg/mL)

(AM ± SD) *

%RSD

Lornoxicam       

0.2

0.25 ± 0.004

1.6  

0.26 ± 0.002  

0.76

2.2

       2..24 ± 0.015

0.66  

2.22 ± 0.026

1.17

4.2

4.50 ± 0.05

1.11  

4..34± 0.08  

1.84

Paracetamol

1

1.22± 0.003

0.25

        1.12± 0.004

0.35

45

45.14 ± 0.116

0.25

45.02 ± 0.129

0.28

75

75.32± 0.272

0.36

75.24± 0.358

0.47















*Average of three determinations and % RSD is relative standard deviation.

 

 Table 3: Accuracy of the method (Recovery studies)

 

Formulation

Recovery level (%)

Recovery of analyte

Theoretical content (mg)

Amount found (mg)* (Mean ± SD)

Recovery (%)

% RSD

Lornoxi-P

0

LOR

2

2.1± 0.015

105

0.714

PAR

125

125.1 ± 0.145

100.08

0.115

80

LOR

3.6

3.5± 0.045

97.2

1.285

PAR

225

224.5 ± 0.34

99.77

0.151

100

LOR

4

4.2 ± 0.009

105

0.214

PAR

250

250.24 ± 0.98

100.09

0.393

120

LOR

4.4

4.4 ± 0.01

100

0.272

PAR

275

275.12± 1.25

100.04

0.456

Lornasafe -plus

0

LOR

2

2.2 ± 0.01

110

0.56

PAR

125

125.04 ± 0.25

100.03

0.201

80

LOR

3.6

3.4 ± 0.00

94.4

0.24

PAR

225

225.05 ± 0.55

100.02

0.247

100

LOR

4

3.7 ± 0.02

92.5

0.67

PAR

250

250.12 ± 0.35

100

0.142

120

LOR

4.4

4.3 ± 0.01

97.7

0.34

PAR

275

274.9± 0.85

99.9

0.309

LOR-lornoxicam; PAR- paracetamol; * Average of three determinations and SD- standard deviation. 

 

Table 4: Analysis of commercial tablets (assay)

Formulation

Lornoxicam

Paracetamol

Label claim (mg)

Amount found (mg) (AM ± SD)

% Recovery

% RSD

Label claim (mg)

Amount found (mg) (AM ± SD)

% Recovery

% RSD

Lornoxi-p

8

8.4 ± 0.011

105

0.13

500

500.16±0.25

100.03

0.05

Lornasafe-plus

8

8.2 ± 0.102

102

1.24

500

500.04 ±0.56

100

0.11

AM- Arthemetic mean

 







CONCLUSION

A simple, eco-friendly, sensitive and economic first derivative spectrophotometric method has been proposed for simultaneous quantification lornoxicam and paracetamol in pure form and in tablet dosage forms by using 0.01N NaOH as solvent. The assay values were in good concurrence with their respective labeled claim, which suggested no interference of formulation excipients in the estimation and obtained results from validation proved the proposed method was scientifically sound. Therefore, the developed method can be readily adopted by pharmaceutical quality control laboratory for routine analysis. 

 

ACKNOWLEDGEMENT

The authors are thankful to the management and Prof.C.V.S.Subrahmanyam, Principal, Gokaraju Rangaraju College of Pharmacy

 

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Welcome to the research group of Prof. Dr. Cornelia M. Keck in Marburg. Cornelia M. Keck is a pharmacist and obtained her PhD in 2006 from the Freie Universität (FU) in Berlin. In 2009 she was appointed as Adjunct Professor for Pharmaceutical and Nutritional Nanotechnology at the University Putra Malaysia (UPM) and in 2011 she obtained her Venia legendi (Habilitation) at the Freie Universität Berlin and was appointed as a Professor for Pharmacology and Pharmaceutics at the University of Applied Sciences Kaiserslautern. Since 2016 she is Professor of Pharmaceutics and Biopharmaceutics at the Philipps-Universität Marburg. Her field of research is the development and characterization of innovative nanocarriers for improved delivery of poorly soluble actives for healthcare and cosmetics. Prof. Keck is executive board member of the German Association of Nanotechnology (Deutscher Verband Nanotechnologie), Vize-chairman of the unit „Dermocosmetics“ at the German Society of Dermopharmacy, active member in many pharmaceutical societies and member of the BfR Committee for Cosmetics at the Federal Institute for Risk Assessment (BfR).

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