Validated Derivative Spectrophotometric method for simultaneous estimation of Levocetirizine Dihydrochloride and Phenylephrine Hydrochloride from tablet formulations

 

G. K. Dyade

 

SVPM’s College of Pharmacy, Malegaon (BKII), Baramati, Dist. Pune-413115

 

ABSTRACT:

A simple, precise and accurate spectrophotometric method was developed for simultaneous estimation of LevocetirizineDiHCl and Phenylephrine HCl in bulk and tablet dosage form. This method is based on use of derivative method for simultaneous estimation of both the drug from tablet using distilled water as solvent. Phenylephrine HCl shows maximum absorption at 273 nm and LevocetirizineDiHCl shows absorption at 231 nm. Phenylephrine HCL and LevocetirizineHCl obeys Beer’, s law in the concentration range 0-40 µg/ml and 0-30µg/ml respectively. During estimation of Tablet no interference was found from excipients. The dosage form contains 10 mg Phenylephrine HCl and 5 mg LevocetirizineDiHCl. The suitability of this method for estimation of both drugs was proved by validation. Statistical analysis of data shows that the developed method is sound under analytical condition and can be used for routine analysis of both drugs.

 

 

 

INTRODUCTION:

LevocetirizineDiHCl^[1-5] is a second generation antihistaminic agent, chemically it is R-[2-[4-[(4-chlorophenyl) phenylmethyl]-1-piperazinyl] ethoxy] acetic acid dihydrochloride. This R-enantiomer has a 30-fold higher affinity than the S-enantiomer and dissociates more slowly from H1 receptors. Levocetrizine is indicated for the relief of symptoms associated with allergic rhinitis in adults and children 6 years of age and older.

 

Phenylephrine^[1-5] is selective direct acting α agonist differs from epinephrine only in lacking a para OH group; chemically it is (R)-3-Hydroxy-alpha [(methyl amino) methyl] benzene methanol hydrochloride.

It is direct acting sympathomimetic agent. It is mainly used as a nasal decongestant in both oral and topical preparations. When applied to mucous membrane it reduces congestion and swelling by constricting the blood vessels of the membrane. 

 

Literature survey revealed that there are reported methods for simultaneous estimation of levocetirizine diHCl alone or with other drug by RPHPLC^[6-7], UV spectrophotometric^[8], HPTLC methods. Similarly phenylephrines HCl alone or with other drugs were also determined by RPHPLC, UV spectrophotometric methods. Both drugs are official in BP^[9] and IP^[10]. The chemical structure of drug is shown in Fig. 1.

 

MATERIALS AND METHOD^[11-16]:

Instrument:

A Shimadzu model UV-VIS 1700 a double beam UV-VIS spectrophotometer with spectral bandwidth of 2 nm and wavelength accuracy of ± 1 nm with 10 mm matched Quartz cells was used. Electronic balance Afcoset FX 300 was used.

 

Pure Drugs and Lab Reagents:

Gift sample of Phenylephrine HCl and Levocetirizine DiHCl were procured from Cure Pharma Ltd. Pune.Double distilled water available in laboratory and reagents of AR grade were used. Formulation containing Phenylephrine HCl 10 mg and LevocetirizineDiHCl 5 mg Brand Name Relent (Dr. Reddy’s Lab) was procured from local market.

 

Preparation of Standard stock solution:

Accurately weighed pure drug powder equivalent to 20 mg of Phenylephrine HCl and 20 mg of Levocetirizine DiHCL separately and transferred to separate 100 ml volumetric flask. Dissolved in water and volume was made up to 100 ml with water which produced 200µg/ml conc. of each drug. Aliquots of stock solution were further diluted to obtain 10 µg/ml conc. of each drug.

 

First order Derivative method:

Standard solutions each of 10ug/ml of Phenylephrine HCL and LevocetirizineDiHCL were prepared in 10 ml volumetric flask and scanned in 200 to 400 nm wavelength range against water as blank. Absorption spectra of both drugs were recorded and the λ_max of Phenylephrine HCl and LevocetirizineDiHCl was found at 273 and 231nm respectively. Series of standard solutions were prepared in conc. range of 5-40 µg/ml. and scanned in 200 to 400 nm range in spectrum mode on the spectrophotometer. Absorbance was recorded at 231 and 273nm wavelength for CET and PHE respectively and calibration graph was plotted.

 

From overlain spectra (Fig.No.2) it was observed that the both drugs have absorption at 231nm wavelengths therefore to overcome the interference Derivative method was applied. Zero order spectra of both drug was converted to first order derivative spectra (Fig No.3a and 3b). From overlain spectra (Fig.No.4) 283nm was selected λ_max of Phenylephrine HCl where no interference of LevocetirizineDiHCl and 241nm was selected as λ_max of LevocetirizineDiHCl as it was zero crossing of Phenylephrine HCl Both drugs obeys Beer’s law in first order derivative mode at the respective wavelength

 

Analysis of Tablet Formulation:

Twenty tablets were weighed and crushed to powder, powder equivalent to 10 mg of PHE and 5 mg of CET were weighed and transferred to 100 ml volumetric flask, powder was dissolved in water and volume was made up to 100 ml with water. Mixed well, resulting solution was filtered through Whatman filter paper. Aliquot of solution was diluted to 10 ml with water into 10 ml volumetric flask and solution was scanned and absorption of solution was measured at 241 and 283 nm (Fig.5). Amount of each drug in solution was calculated.

 

RESULTS:

Validation of the Method:

The method was validated as per ICH guidelines

 

Linearity:

The linearity of an analytical method is its ability to obtain test results which are directly proportional to the conc. of analyte. Series of standard solutions were prepared in conc. range of 4 to 40 ug/ml. and scanned in 200 to 400nm range on the spectrophotometer, absorbance of the standard solutions were recorded at 241 and 283nm in derivative order. Regression equations was summarised in table.

 

Precision:

The precision study was carried out by performing assay of tablet six times. Also the reproducibility in result was studied by interday and intraday precision. SD and RSD show methods precision and summarised in table.

 

Assay:

Formulations assay was performed and results are shown in Table No.1

 

Accuracy:

The accuracy of an analytical method expresses the closeness of an agreement between test result and true result. Accuracy study was performed by recovery study i.e. standard addition method. Diluted standard solutions of PHE and CET were prepared and standard solutions added in 80,100 and 120% proportionate. SD and RSD of triplicate study tabulated in Tab. No II

 

 

Table 1 Results of tablet formulation Assay

Tablet sample	Label claim(mg/Tab)		% of Label claim estimated*		Standard deviation		Coefficient of variance
Name of Drug	PHE	CET	PHE	CET	PHE	CET	PHE	CET
Formulation I	10	5	99.96	101.23	1.1384	1.3959	1.1386	1.3789
Formulation II	10	5	99.12	99.71	1.4571	1.3023	1.4700	1.3061

*Mean of six determinations

 

 

Limit of detection (LOD) and Limit of Quantitation (LOQ):

The LOD and LOQ of PHE and CET by the proposed method were determined using calibration graph method and calculated as 3.3σ/S and 10σ/S. Results are summarised in Table No 2.

 

Robustness and ruggedness:

It is measure of capacity of analytical procedure to remain unaffected by small but deliberate variations in method parameter.

 

Results are shown in Table No II

 

Table II: Optical characteristics and statistical analysis of the Derivative method

Parameters		Phenylephrine Hydrochloride	Levocetirizine Di Hydrochloride
Wavelength (nm)		273 nm	231 nm
Beers law range µg/ml		0-40 µg/ml	0-30 µg/ml
Regression equation		Y= 0.0103X	Y= 0.0321X
Correlation coefficient		0.9996	0.9998
LOD µg/ml		1 ug/ml	1 ug/ml
LOQ µg/ml		2ug/ml	3ug/ml
Precision	Repeatability*	± 1.1384	± 1.3959
	Intraday*	± 1.1386	± 1.3789
	Interday*	±1.6958	± 1.6724
Accuracy	RSD*	1.5487	1.3891
Robustness	± 2 nm	± 0.002	± 0.007
Ruggedness	Analyst 1*	± 1.1386	± 1.3959
	Analyst 2*	± 1.4047	± 1.4571

*Mean of Six determinations

 

DISCUSSION:

The method is precise, accurate and reproducible and routinely used for simultaneous estimation of Levocetirizine dihydrochloride and Phenylephrine hydrochloride from combined formulation.

 

ACKNOWLEDGEMENT:

Author is thankful to Cure Pharma Pune for providing pure drug as a gift sample and SVPM College of Pharmacy Malegaon Dist. Pune for providing facilities for research.

 

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Received on 27.12.2018       Accepted on 20.01.2019     

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Asian J. Pharm. Ana. 2019; 9(1):01-04.