Determination of Prochlorperazine Maleate and Pyridoxine Hydrochloride in Combined Tablet Dosage form by using first order derivative Spectrophotomeric method

 

G. B. Bhagwat1,2*, P. T. Sonawane2 and V. V. Bopinwar2

1Department of Pharmaceutical Chemistry, Sharad Pawar College of Pharmacy, Wanadongri, Hingna Road, Nagpur 441 110 (M.S.), India.

2Department of Pharmaceutical Chemistry, Kasturi Shikshan Sanstha’s College of Pharmacy, Shikrapur, Pune 412 208 (M. S.), India

 

 

ABSTRACT:

A simple, accurate, economical and reproducible procedure for simultaneous estimation of Prochlorperazine Maleate and Pyridoxine Hydrochloride in their tablet dosage form is described. In this method 0.3M HCl was utilized as solvent. In first order derivative spectroscopic method, two wavelengths 252nm for prochlorperazine maleate (zero cross for pyridoxine hydrochloride) and 228.5nm for pyridoxine hydrochloride (zero cross for prochlorperazine maleate) were selected. Both drugs obey Beer-Lambert’s law in concentration range of 1-5 μg/ml (prochlorperazine maleate) and 5-25 μg/ml (pyridoxine hydrochloride) respectively in 0.3M HCl. The suggested method is validated by using ICH validation parameters like accuracy, precision, specificity, ruggdness and linearity. The results of recovery study were found to be within the prescribed limit of 98-102%. During the linearity study it was observed that absorbance values of Prochlorperazine Maleate and Pyridoxine Hydrochloride in marketed formulation were linear in the range of 80% to 120% of test concentration with R2 close to 1 for this method of analysis.

 

KEYWORDS: Prochlorperazine Maleate, Pyridoxine Hydrochloride, first order derivative spectroscopic method

 

INTRODUCTION:

Prochlorperazine maleate (PCM), 2-Chloro-10-[3-(4-methylpiperazin-1-yl)propyl] 10 H phenothiazine bis[hydrogen(Z)-butenedioate] (fig. 1), is an antiemetic and antipsychotic drug [1] while pyridoxine hydrochloride (PDH), 5-Hydroxy-6-methyl-3,4-pyridine dimethanol hydrochloride (fig. 2), is a nutritional component  [2]. Both the drugs are official in IP, BP and USP. Literature survey revealed that HPLC [3, 4] methods have been reported for pyridoxine hydrochloride with other drugs. However, no method is reported for simultaneous estimation of these two drugs in combined dosage form. This prompted us to develop simple, rapid, accurate, economical and sensitive simultaneous estimation method for their determination in combination.

 

 

Figure 1: Structure of prochlorperazine maleate

 

Figure 2: Structure of pyridoxine hydrochloride

 

MATERIAL AND METHODS:

Instrumentation:

UV-Visible double beam spectrophotometer, model 1700 Pharmaspec with 1 cm quartz cells was used. Electronic balance (Modal AUW-220D) was used for weighing.

 

Reagents and chemicals:

The chemicals used were of analytical grade. Commercially available tablets of prochlorperazine maleate and pyridoxine hydrochloride in combination were procured from local market. Standard prochlorperazine maleate and pyridoxine hydrochloride were received as gift sample from FDA Mumbai.

 

Selection of solvent:

A 0.3M HCl solution was selected as the suitable solvent for the estimation of PCM and PDH.

 

Preparation of standard stock solution:

Standard stock solutions of PCM and PDH were prepared separately by dissolving 25 mg each of standard PCM and PDH in 0.3M HCl and making up the volume up to 100 ml (250 μg/ml).

 

Preparation of mixed standard solution:

An accurately weighed quantity of 5 mg PCM and 25 mg of PDH was transferred to a 100 ml volumetric flask. The drugs were dissolved in 50 ml of 0.3M HCl and finally Volume was made upto the mark with 0.3M HCl. This solution was diluted further with 0.3M HCl to get the concentration of five mixed standards as shown in table-1.

 

Table 1: Concentration of PCM and PDH in mixed standard solution

 Name of drugs

Concentration in µg/ml

Std.       Std.2            Std.3         Std.4              Std.5

PCM

01         02                03             04                         05

PDH

05        10                 15            20                                   25

 

Selection of scanning range and sampling wavelength:

The standard solutions of PCM and PDH were diluted with 0.03 M HCl individually so as to get the concentration of 10µg/ml for each drug and were scanned individually in the UV range 400-200 nm. The spectral data thus obtained was then processed to obtain first derivative spectrum at wavelength interval 0.1 nm. From the Spectrum of PCM showed zero crossing point at 228.5 nm (fig.3) while PDH showed zero crossing at 252.0 nm (fig.4). The two spectra were overlain is shown in fig. 5.

At the zero crossing point of PCM (228.5 nm), PDH showed a measurable dA/dλ where as zero crossing point of PDH (252.0 nm), PCM showed measurable dA/dλ. Hence the wavelengths 228.5 nm and 252.0 nm were selected as analytical wavelengths for determination of PDH and PCM respectively

.

Figure 3: UV Spectrum of PCM

 

Figure 4: UV Spectrum of PDH

 

Figure 5: Overlain first order derivative spectrum of PCM and PDH

 

Study of Beer-Lambert’s law at selected wavelength:

PCM obeyed Beer Lambert’s law in concentration range of 1-5 µg/ml whereas PDH obeyed the law in concentration range of 5-25 µg/ml. The dA/dλ of each solution was measured at 252.0 nm for PCM and 228.5 nm for PDH, respectively. The dA/dλ of each solution was measured at 252.0 nm for PCM and 228.5 nm for PDH respectively.

 

Application of proposed method for the estimation of PCM and PDH in tablet:

Marketed tablets Emidoxyn Forte (Shreya Life Science Pvt Ltd., Mumbai) were used for the simultaneous estimation of PCM and PDH. Twenty tablets were accurately weighed.  Average weight of tablet was calculated. A quantity of tablet powder equivalent to 5 mg of PCM and 25 mg of PDH was transferred to 100 ml volumetric flask and dissolved in 0.3M HCl and volume was made to 100 ml. The solution was filtered through Whatman filter paper no.41.                          

The aliquot portion of filtrate was further diluted with 0.3M HCl to get final concentration 5 µg and 25 µg of PCM and PDH, respectively. The absorbance of resulting solutions was measured at 252.0 nm and 228.5 nm in 1 cm cell against blank. The content of PCM and PDH in tablet was calculated using the formula-

 

Where, As = Absorbance of sample,

Astd = Absorbance of standard,

Wstd = Weight of standard,

Ws = Weight of sample,

A = Average weight,

LC = Label claim

 

Method Validation:

Accuracy of an analytical method is the closeness of test results obtained by that method to the true value. It was ascertained on the basis of recovery studies performed by standard addition method. The preanalysed tablet powder equivalent to about 5 mg PCM was taken in 100 ml volumetric flask, to it standard solutions of PCM and PDH were added in the different proportions. The mixture was shaken for 15 minutes and volume was adjusted to the mark with 0.3M HCl and the solutions were filtered through Whatman filter No.41. An aliquot portion of the resultant solutions were appropriately diluted with 0.3M HCl to get final concentration within the range of mixed standard.

 

 

Table 2: Results for estimation of PCM and PDH in marketed formulation

Wt. of Sample (g)

% of drug Estimation*

SD

RSD

SE

PCM

PDH

PCM

PDH

PCM

PDH

PCM

PDH

       0.1380                                          

100.03              

100.10      

0.0438         

0.1862       

0.0019              

0.0346             

0.0015     

0.0277

*Results are mean of five replicates. SD is standard deviation of the mean, RSD is relative standard deviation of the mean,

SE is the standard error of the mean.

 

 

Table 3: Results and statistical data for recovery study

Sr. No.

Amount of pure drug added (μg/ml)

% Recovery*

PCM

PDH

PCM

PDH

1

5

25

99.84

100.20

2

5

25

99.99

99.85

3

5

25

100.06

100.33

*Results are mean of five replicates.

 

 Table 4: Results of specificity studies

    Sr. No

Sample

% Label claim*

PCM

PDH

      1

      2

      3

      4

Normal

Alkali

Acid

oxide

100.11

98.05

99.89

1 01.01

100.23

99.03

100.00

101.21

*Results are mean of five replicates.

 

Table - 5: Results of Ruggedness Study

Condition

% label claim*

SD

RSD

PCM

PDH

PCM

PDH

PCM

PDH

Intraday

100.01

100.43

0.0305

0.6658

0.0093

0.4433

Interday

99.93

99.93

0.0989

0.0472

0.0009

0.0004

Different analyst

99.86

99.96

0.0989

0.0742

0.0009

0.0004

*Results are mean of five replicates.

 

 

Ruggedness was carried out by repeating the procedure under different conditions

A. Days (Intraday and Interday)  

B. Different analysts

 

Precision of analytical method is the degree of agreement among individual results when the method is applied repeatedly to multiple readings of homogenous sample. It is expressed as the SD or RSD of series of measurements. It was ascertained by replicate estimation of the drugs by proposed method.

 

Specificity study was carried out by accurately weighed quantities of tablet powder equivalent to 5 mg of PCM were taken in different 50 ml volumetric flask and were stored for   24 hr under following conditions.

1.       At room temperature (normal)

2.     At 500C after addition of 1.0 ml of 0.1N NaOH (alkali)

3.     At 500C after addition of 1.0 ml of 0.1N HCl (acid)

4.     At 500C after addition of 1.0 ml of 3% H2O2 (oxide) 

 

The samples were diluted upto the mark with 0.3M HCl and filtered through grade-1 filter paper. Aliquot of the filtrate was diluted with 0.3M HCl to get 10 µg/ml concentration of PCM.

 

Linearity study was carried out by accurately weighed quantities of tablet powder equivalent to 80, 90, 100, 110 and 120 % of  label claim were taken and dilutions were done appropriately to obtain a concentration in the range of 80-120% of the test concentration and absorbance were recorded at 252.0 nm and 228.5 nm. Both PCM and PDH were found to be linear in 80-120% of the test concentration.

 

Figure 6: Plot of Linearity and range study for PCM and PDH

 

RESULTS AND DISCUSSION:

An attempt has been made to develop a fast, sensitive, precise, reproducible and economical analytical method for simultaneous estimation of PCM and PDH in their combined dosage form. In this method drugs obey Beer’s law in the concentration range 1-5 μg/ml for PCM and 5-25 μg/ml for PDH, respectively.

 

The result of % estimation of drugs is shown in Table-2. The method was validated as per the ICH and USP guidelines [5, 6, 7]. The results of recovery study were found to be within the prescribed limit of 98-102%, proving the accuracy and showing that the method is free from interference from excipients. For precision, replicate estimations of both PCM and PDH in the same batch of tablet were done by the proposed method, which yielded quite concurrent results, indicating reliability of the method. The values of SD or RSD are within the prescribed limit of 2%, showing high precision of method.

 

In the specificity study, sample was exposed to different stress condition like acid, alkali, peroxide and heat. For ruggedness, proposed method was repeated under different conditions like at different time, on different days and by different analysts, prove that the method is reproducible. During the linearity study it was observed that absorbance values of PCM and PDH in marketed formulation were linear in the range of 80% to 120% of test concentration with R 2 close to 1 for this method of analysis.

 

From the study of validation parameters namely accuracy, precision (SD and RSD), ruggedness (interday, intraday and different analyst), specificity, linearity and range, it was observed that the method is specific, accurate, precise, reproducible and rugged.

 

CONCLUSION:

The proposed method for simultaneous estimation of PCM and PDH in their combined dosage form is quite accurate, precise, reproducible and rugged. Moreover, the method is economical, simple and rapid. Hence, the method can be employed for routine quantitative analysis of tablet dosage form containing PCM and PDH.

 

ACKNOWLEDGEMENTS:

The authors wish to thank Principal Dr. K. P. Bhusari, Sharad Pawar College of Pharmacy, Rashtrasant Tukadoji Maharaj Nagpur University for providing necessary facilities. They also thank Mr. Kamlesh Shende, FDA Department, Mumbai for providing the authentic sample of drugs.

 

REFERENCES:

[1]   Budavari S., The Merck index: An Encyclopedia of Chemicals, Drugs and Biologicals. 13th ed. Whitehouse Station (NJ): Merck Research Lab, Division of Merck; 2001.7850.

[2]   Budavari S., The Merck index: An Encyclopedia of Chemicals, Drugs and Biologicals. 13th ed. Whitehouse Station (NJ): Merck Research Lab, Division of Merck; 2001.8072.

[3]   Poongothai S, Ilavarasan R and Karrunakaran C. M. Simultaneous and accurate determination of vitamins B1, B6, B12, and alpha-lipolic acid in multivitamin capsule by Reverse-phase high performance liquid chromatographic method. Int J Pharm Pharm 2(4);2010:133-139.

[4]   Khor Swan-Choo and Tee E-Siong (1996). Development of a HPLC method for simultaneous  determination of several B-vitamine and ascorbic acid. Mal J Nutr 2:49-65.

[5]   ICH Q2B; Guidelines on validation of analytical procedure; Definitions and terminology, Federal Register, 1995, 60, 11260.

[6]   ICH Q2B; Guidelines on validation of analytical procedure; Methodololgy, Federal   Register, 1996, 60, 27464.

[7]   The United States Pharmacopoeia 24/ National Formulary 19, The United States Pharmacopoeial Convection, Rockville, 2000, 2151.

 

 

Received on 01.11.2012

Modified on 24.11.2012

Accepted on 04.12.2012              

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Research J. Science and Tech.  4(6): November –December, 2012: 262-266