Simultaneous Estimation of Chlorpheniramine Maleate and Glyceryl Guaiacolate in Pure and Capsule Dosage form by using different Spectrophotometric Methods

Nada Ghaaeb Hussein, Ahmed Mahdi Saeed*

College of Education for Pure Science, Chemistry Department, Diyala University.

*Corresponding Author E-mail: dr.ahmedalanbakey@yahoo.com

ABSTRACT:

Chlorpheniramine Maleate (CPM) and Glyceryl Guaiacolate (GUA) are the β‐lactum antibiotic drug. Sensitive, precise, accurate and simple, UV spectrophotometric methods have been developed for the simultaneous estimation of Chlorpheniramine Maleate (CPM) and Glyceryl Guaiacolate (GUA) in dosage form. Two spectrophotometric methods (simultaneoue equations and Q‐Absorbance ratio) were applied for the determination of the drugs as mixture. The maximum absorbance of drug in solvent mixture composed of water – acetontrile – methanol in a ratio of (80% H₂O – 10% ACN – 10% MOH) was found to be at (261.4 nm and 273 nm) for CPM, GUA respectively, and the Q – isosbestic point was found at 270.4 nm. These wavelengths were selected for the analysis of drugs as mixtures standard and in the manufactured samples using the two developed methods. The methods were linear in the range of (1- 100) μg/mL for (CPM, GUA), with an R² of (0.9996) for CPM and GUA respectively in the mixture. Recovery means were found to be (99.79 % - 100.30 %) for the standard drugs CPM and GUA respectively and in formulating drugs was found to be (99.71 – 100.41 %). LOD and LOQ were established and found to be (0.1 and 0.33) for CPM and GUA respectively. The method was applied for the estimation of the active gradient of the drugs in different samples of manufactured dosage. The accuracy of method was validated by mean percentage recovery, which was found to be in the acceptable range.

KEYWORDS: Determination, Spectrophotometric, Simultaneous equation, Q‐Absorbance ratio.

INTRODUCTION:

 Cough can be caused by air pollution, smoking, allergic and etc. Cough medicines are used for relieve of cough. They are made up of one or more pharmaceutical active drugs. Cough preparations consist of antihistamines, antitussives, decongestants, expectorants and sleep aids¹. In the recent years, combination dosage forms of Chlorpheniramine maleate (CPM) and Glyceryl Guaiacolate (GUA) as an agent to cure of cough is introduced. There is no analgesic agent to treat all forms of pain and there is no ideal analgesic factor, but each worker has advantages and disadvantages to distinguish him from the  rest of the painkillers².

Chlorpheniramine maleate (CPM) IUPAC name [(RS)-3-(4-chlorophenyl)-3-(2- pyridyl) propyl methyl amine hydrogen maleate is a histamine H1 antagonist used in allergic reactions, hay fever, rhinitis, urticaria, and asthma. It is also effective against nausea and motion sickness, with its primary mechanism of action being its ability to reduce acetylcholine levels in the brain³. Glyceryl Guaiacolate (GUA) IUPAC name (RS)-3-(2-methoxyphenoxy) propane-1, 2-diol is an It is is an expectorant drug sold over the counter and usually taken orally to assist the bringing up (expectoration) of phlegm from the airways in acute respiratory tract infections. It is the component of numerous cough cold preparations available worldwide. It is soluble in water, dimethyl formamide and slightly soluble in ethanol. Guaifenesin has not been approved by the FDA for the treatment of fibromyalgia. Based on a small, non-blinded study, Glyceryl Guaiacolate has been promoted to facilitate conception, by thinning and increasing cervical mucus, during the few days before ovulation.⁴ Many analytical methods were reported for the determination of these drugs in pharmaceuticals such as HPLC^5-8, GC⁹, UV/Vis spectrophotometry^10-14, LC/MS¹⁵ and LC-MS/MS^16-19. The aim of this work was to develop the ease and accurate spectrophotometric method for the determination of the drug content in tablet samples from different pharmaceutical companies available in Iraqi pharmaceutical market, to give information about these products, which may or may not comply with the requirements of the standard method or other official methods, as shown in Fig. 1- a, b.

Fig:1: A: Chlorpheniramine maleate (CPM) B: Glyceryl Guaiacolate (GUA)

MATERIALS AND METHODS:

Materials:

CPM and GUA were provided from Samara Drug Industries (SDI), Iraq. Different tablets samples were used as marketed formulation as in Table 3. Methanol HPLC grade (BDH) and freshly prepared deionized water was used throughout the experiment.

Apparatus:

UV - VIS spectrophotometer (UV-1700 Shimadzu – Japan), (Ultrasonic - China), sartorius balance (Germany), shaking water bath (Taiwan) and furnace (Germany) were used through this study.

Selection of common solvent:

Main criteria for media selection was solubility and stability, i.e. Chlorpheniramine maleate (CPM) and Glyceryl Guaiacolate (GUA) should be soluble as well as stable for sufficient time in selected media. The media used in reported method was water – acetonitrile – methanol in a ratio of (80 – 10 – 10) respectively, which was selected as analytical media for present work.

Preparation of drug stock solutions (1000 mg/L):

A 0.1g from each standard drug was weighed and dissolved in (80% H₂O – 10% ACN – 10% MOH), transferred to a 100mL volumetric flask separately, it was then sonicated for 10 minutes and the final volume of both the solutions were made up to 100mL with the solvent to get stock solutions containing 1000μg/mL each of CPM and GUA in two different 100mL volumetric flasks. More diluted solutions were prepared by simple dilution of stock solution of drugs.

Diluent:

By using the stock solution of 100μg/mL, subsequent dilution was carried out by withdrawing different aliquots (0.1 – 10) mL from standard solution were transferred into a series of 10mL calibrated volumetric flasks and all were made up to the mark with mobile phase in order to prepare working standard solutions of different concentrations (1 – 100) μg/mL.

RESULTS AND DISCUSSION:

Determination wavelength of maximum absorbance:

By appropriate dilution of two standard drug solutions with (80% H₂O – 10% ACN – 10% MOH), solutions containing 100μg/mL of CPM and 100μg/mL of GUA were scanned separately in the range of (200‐ 400) nm to determine the wavelength of maximum absorption for both the drugs. CPM and GUA showed absorbance maximum at 261.4nm (λ₁) and 273nm (λ₂) respectively, as shown in Fig. 2.

Fig: 2: UV-VIS Spectra of CPM and GUA

Method A: Simultaneous Equation Method:

Two wavelengths selected for the method are 261.4nm and 273nm that are absorption maxima of CPM and GUA in (80% H₂O – 10% ACN – 10% MOH), respectively. The stock solutions of both the drugs were further diluted separately with (80% H₂O – 10% ACN – 10% MOH), to get a series of standard solutions of 1‐ 100μg/mL concentrations of CPM and 1‐ 100μg/mL concentrations of GUA. The absorbances were measured at the selected wavelengths for both the drugs at both wavelengths were determined as mean of Seventeen independent determinations. Concentrations in the sample were obtained by using following equations. 

        A1 ay – A2 ay1

Cx = ----------------------                                                                   Eq (i)

          ax1ay2 – ax2ay1

           A1 ax – a2 ax1

Cy = ----------------------                                                                   Eq(ii)

          ay1ax2 – ay2ax1

Where, A1 and A2 are absorbance’s of mixture at 261.4 nm and 273 nm respectively ax1 and ax2 are absorptivities of CPM at λ₁ and λ₂ respectively and ay1 and ay2 are absorptivities of GUA at λ₁ and λ₂ respectively. Cx and Cy are concentrations of CPM and GUA respectively.

Method B: Absorption Ratio Method (Q Method):

The solutions of CPM and GUA (10 µg /ml) were scanned in the range of 200 to 400 nm against (80% H₂O – 10% ACN – 10% MOH) as blank. For Q method, 270.4nm (isosbestic point) and 273nm (λ_max of Glyceryl Guaiacolate) were selected as wavelengths of measurements Fig. 3. Concentrations of CPM and GUA were determined using following equations²⁰.

Cx = (Qm ‐ Qy). A1 / (Qx ‐ Qy). ax1

and

Cy = (Qm ‐ Qx). A1 / (Qy ‐ Qx). ay1

Where

Qm = A2/ A1, Qx = ax2/ ax1, Qy = ay2/ ay1

Where, A1 and A2 are absorbance’s of mixture at 270.4 nm and 273 nm respectively ax1 and ax2 are absorptivities of CPM at λ₁ and λ₂ respectively and ay1 and ay2 are absorptivities of GUA at λ₁ and λ₂ respectively. Cx and Cy are concentrations of CPM and GUA respectively.

Fig: 3: Absorbance spectra for the drugs and Q - isosbestic point

Analytical method validation:

Additional parameters to be evaluated when demonstrating accuracy and precision are part of the method development and optimization process, or are performed during the validation process when demonstrating acceptable method performance. These parameters include limits of detection and quantification, linearity of the method, range, recovery, robustness and selectivity²¹.

Linearity:

The linearity of measurement was evaluated by analysing different concentration of the standard solution of CPM and GUA. Absorbance of all solutions was measured at λ_max of each drug. For simultaneous equation method the Beer‐ Lambert’s concentration range was found to be for 1‐100μg/mL for CPM and 1‐100μg/mL GUA, as shown in Fig. 4. The calibration graph were obtained by plotting absorbance versus known concentrations, and R² (0.9996) for the both drugs, indicating that there is a strong correlation between the variation of concentration and response. Linearity was determined by the regression analysis as shown in Table 1.

Fig: 4: Linearity curve for CPM and GUA

Table 1: Statistical calculations.

Statistical factors	Value
	Chlorpheniramine Maleate	Glyceryl Guaiacolate
Linear equation	y= 0.0154 [X] + 0.0004	y = 0.0102[X] - 0.001
Slope (m)	0.0154	0.0102
Intercept	0.0004	0.001
Determination Coefficient R^2	0.9996	0.9996
Percentage linearity (R^2%)	99.96	99.96
Correlation Coefficient (r)	0.9998	0.9998
RSD	0.16	0.15
Linearity range μg/mL	1-100	1-100
Calculated (t) values t cal. =	193.461>> 2.110	193.461 >> 2.110

Limit of Detection (LOD) and Limit of Quantization (LOQ):

The LOD and LOQ of a Chlorpheniramine maleate (CPM) and Glyceryl Guaiacolate (GUA) by proposed methods were determined using calibration standards. LOD and LOQ were calculated by gradual dilution of lowest concentration and as 3.0 LOD respectively. The results obtained are shown in Table 3.

ACCURACY:

CPM and GUA were determined at three different selected concentrations (6, 28, 55) μg/mL. The obtained results were tabulated in Table 2, which indicated that the proposed method for the determination of three drugs is quite satisfactory in reality with respect to the procedure and parameters calculated. The Optical characteristics data and validation parameters were tabulated in Table 2.

Table 2: Recovery study of CPM and GUA for the two methods.

Method	Simultaneous estimation method
Drug	Taken	Found	% Recovery ± SD		%Error	R.S.D n =3
CPM	6	5.96	99.33	Mean = 99.79 S.D. =0.40	-0.67	0.18
	28	28.02	100.07		0.07	0.21
	55	54.98	99.96		-0.04	0.11
GUA	6	5.99	99.83	Mean = 99.91 S.D. = 0.12	-0.17	0.17
	28	27.96	99.86		-0.14	0.22
	55	55.03	100.05		0.05	0.15
Method	Cut cross point method (Q– isosbestic)
Drug	Taken	Found	% Recovery ± SD		%Error	R.S.D n =3
CPM	6	6.06	101	Mean = 100.30 S.D. = 0.61	1	0.15
	28	28.01	100.04		0.04	0.19
	55	54.93	99.87		-0.13	0.10
GUA	6	6.01	100.17	Mean = 100.08 S.D. = 0.13	0.17	0.13
	28	27.98	99.93		-0.07	0.21
	55	55.07	100.13		0.13	0.10

Table 3: Optical characteristics data and validation parameters

Parameters	Values for CPM			Values for GUA
Absorption maxima (λ max)	261.4 nm			273 nm
Beer’s law limit (μg/ml)	1-100			1-100
Regression equation	y= 0.0154 [X] + 0.0004			y = 0.0102[X] - 0.001
Correlation coefficient (R2)	0.9996			0.9996
Molar absorptivity	λ1	λ2	λQ	λ1	λ2	λQ
	4264	1675	2672	1080	2034	1907
A(1%,1cm)	153.5			102.6
Accuracy (%Recovery ± SD)	Method A	Method B		Method A	Method B
	99.79 ± 0.40,	100.30 ± 0.61		99.91±0.12	100.08 ± 0.13
LOD (μg/ml)	0.1			0.1
LOQ (μg/ml)	0.33			0.33
Precision	Method A	Method B		Method A	Method B
Intraday*(Analyst 1)	100.05 ± 0.24,	99.96 ± 0.31		99.78 ± 0.15,	100.01± 0.34
Interday*(Analyst 2)	100.43 ± 0.33,	99.69 ± 0.25		100.22± 0.38,	99.98 ± 0.16

Quantitative assessment of drugs in tablets:

Different types of pharmaceutical formulations of drugs have been analyzed as described under recommended procedure, a good accuracy and precision were obtained as shown in Table 4. The absorbances of mixture solutions have concentration (5 and 0.1) μg/mL for GUA and CPM respectively, were measured at (261.4 and 273) nm for the simultaneous methods and at (270.4 and 273) nm for Q - method. The concentration of CPM and GUA present in the sample solution was calculated by using the equation generated for the two methods. Values were substituted in the respective formula to obtain concentrations. Obtained results were confirmed the reality, applicability and validity of the proposed method for the determination of CPM and GUA in pharmaceutical formulations. The results indicate that the recovery percentages for applying methods are with an acceptable range of (99.79 – 100.43%) for the quantity of drugs in tablets was accepted within the normal percentage according to official method.

Table 4: Estimated quantity of drugs in different formulated samples

Simultaneous estimation					Method
R.S.D n=3	%Mean amount Found	Mean amount found mg/ 5 ml.	Amount taken mg/ 5 ml.	Drug Type	Drugs name
0.13	99.71	0.997	1	CPM	SOOLAN
0.08	100.13	50.065	50	GUA
0.15	99.94	0.999	1	CPM	DECOPECT
0.21	99.99	49.995	50	GUA
0.10	100.07	1.001	1	CPM	TUSSILET
0.11	99.83	49.915	50	GUA
Cut cross point method (Q– isosbestic)					Method
R.S.D n=3	%Mean amount Found	Mean amount found mg/ 5 ml.	Amount taken mg/ 5 ml.	Drug Type	Drugs name
0.20	100.41	1.004	1	CPM	SOOLAN
0.13	100.16	50.08	50	GUA
0.13	99.92	0.999	1	CPM	DECOPECT
0.16	99.95	49.975	50	GUA
0.12	100.17	1.002	1	CPM	TUSSILET
0.10	99.87	49.935	50	GUA

CONCLUSION:

The most striking feature of this novel method is its simplicity, rapidity and economy, UV spectrophotometric method for the quantitative determination of CPM and GUA in standard and pharmaceutical formulated mixture samples simultaneously without any separation method. The new method can be employed for routine analysis in quality control drugs analysis. The described methods give accurate and precise results for the determination of CPM and GUA with recovery percentages range (99.00 – 100.43) for the drugs.

ACKNOWLEDGMENT:

The authors would like to express his gratitude to the University of Diyala, College of Education for Pure Science, Department of Chemistry for providing lab and research facilities to complete this work.

AUTHORS' CONTRIBUTION:

All the authors have contributed equally

CONFLICT OF INTERESTS:

Declared none.

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Received on 25.12.2019           Modified on 02.04.2020

Accepted on 01.05.2020         © RJPT All right reserved

Research J. Pharm. and Tech. 2021; 14(5):2608-2612.

DOI: 10.52711/0974-360X.2021.00459