Determination of Dimenhydrinate and Chlorpheniramine Maleate in Pharmaceutical Forms by new Gas Chromatography Method

Muhamad Esam Kaf Alghazal¹, Fadi Alrouh², Yaser Bitar¹, Saleh Trefi¹

¹Pharmaceutical Quality and Pharmaceutical Chemistry Department-Faculty of Pharmacy –

University of Aleppo-Syria

²Organic Chemistry Department–Faculty of Medicine- University of Al-Hawash –Syria

*Corresponding Author E-mail: salehtrefi@yahoo.com

ABSTRACT:

A specific and highly sensitive gas chromatographic method was developed for the simultaneous determination of chlorpheniramine maleate and dimenhydrinate using one drug as internal standard for the other one. The chromatographic conditions are aTRB-5-625 capillary column (60m × 0.25 mm i.d0.25μm film thickness) and nitrogen as a carrier gas at a flow rate of 1.0 mL min. The oven temperature was programmed at 230°C for 1 min, with a rise of 20°C min up to 250°C (held for 7 min). The injector and detector port temperatures were maintained at 290°C. Detection was carried out using flame ionization detector. Results of assay and recovery studies were statistically evaluated for its accuracy and precision using one drug as internal stander for the other one and vice versa. Objective: The objective of this study was to develop and validate a gas chromatographic method to separate and assay to antihistamine drugs, dimenhydrinate and chlorpheniramine maleate in pharmaceutical forms. This method was a practical additional choice in quality control laboratories.

KEYWORDS: chlorpheniramine maleate, dimenhydrinate, gas Chromatography.

INTRODUCTION:

Antihistamines are pharmaceutical agents which act by stimulating histamine action in the H1-receptors, thereby antagonizing most of the smooth muscles to alleviate or prevent the symptoms of hay fever and other allergies and put a stop to motion sickness, nausea, vomiting, and dizziness. In addition, since antihistamines may cause drowsiness as a side effect, some of them may be used as an opponent to insomnia. Some antihistamines are used in the handling of nervous and emotional conditions to help control anxiety and to relax patients before surgery¹.

Chlorpheniramine maleate (CPM) chemically, (RS)-3-(4-chlorophenyl)-3-(pyrid-2-yl) propyldimethylamine hydrogen maleate. It is an antihistamine drug that is widely used in pharmaceutical preparations for symptomatic relief of common cold and allergic diseases. It inhibits the effects of histamine on capillary permeability and bronchial smooth muscles. It is a first generation alkyl amine antihistamine used in the prevention of the symptoms of allergic conditions such as rhinitis and urticarial². Chlorpheniramine maleate is used for the treatment of allergic conditions including Conjunctivitis, rhinitis, urticarial and pruritic skin disorders, also used in formulations for treatment of coughs and cold³chlorpheniramine maleate (CPM) can be given as transdermal patch⁴. Dimenhydrinate (DMH) is the diphenhydramine salt of 8-chlorotheophylline. It is chemically known as a (1:1) compound of 8-chloro-3, 7-dihydro-1, 3-dimethyl-1H-purine-2, 6-dione with 2 (diphenylmethoxy)-N, N-dimethylethanamines. DMH is an antihistamine with antimuscarinic and significant sedative effects. It is mainly used as an antiemetic in the prevention and treatment of motion sickness. It is also used for the symptomatic treatment of nausea and vertigo caused by Meniere's disease and other vestibular disturbances⁵. The quantification of DMH and chlorpheniramine maleate in various drug Formulations and/or biological samples was addressed in several reports. Analytical methodology in these reports involved the use of spectrophotometry^6,7adsorptive stripping voltammetry⁸, capillary electrophoresis⁹, liquid chromatograph –tandem mass spectrometry (LC/MS/MS)¹⁰, high performance liquid chromatography(HPLC) with fluorescence detection¹¹, Equation Spectroscopic¹², HPLC with UV detection¹³and high performance thin layer chromatography (HPTLC)^14,15. Few GC/MS methods could be found in the scientific literature for the analysis of DMH merely for toxicological and forensic purposes^16,17.

This work describes a new simple, direct and selective capillary GC/FID method for the determination of chlorpheniramine maleate and dimenhydrinate.

Dimenhydrinate

Figure 1: Chemical structures of (A)chlorpheniramine maleate and(B) dimenhydrinate.

Materials:

Reference Standard of dimenhydrinate and chlorpheniramine maleate are given by Ibn-Alhaytham Industries, Aleppo-Syria. Chloroform (HPLC-grade). The tablet samples brand names are Beloramine (chlorpheniramine 4 mg produced by Mediotech Industries Aleppo, Syria), dimenhydrinate Barakat (dimenhydrinate 50 mg produced by Barkat pharmaceutical industries (Aleppo, Syria).

Apparatus:

GC-2010 gas chromatograph (GC) equipped with a split/split less injector and a flame ionization detector (FID) from SHIMADZU Technologies Inc. was used in this study. Nitrogen (ultrapure) was obtained from G 1010E nitrogen generator (Peak scientific) and used as carrier gas. 5μL gas-tight syringe (SGE Analytical science) were used.

Standard Solutions preparation:

For the standard solution in the first experiment, a precise quantity of dimenhydrinate was accurately weighed and transferred in to a volumetric flask, then dissolved in chloroform to a final volume10 ml. (20mg/ml). For the standard solution in the second experiment a precise quantity of chlorpheniramine maleate was accurately weighed and transferred in to a volumetric flask, then dissolved in chloroform to a final volume10 ml. (20mg/ml). This standard solution, was used for the preparation of all diluted solutions.

Internal standard solution preparation:

For internal standard solution, a precise quantity of chlorpheniramine maleate was accurately weighed and transferred in to a volumetric flask then dissolved in chloroform to a final volume of 10 ml (30mg/ml). A precise quantity of dimenhydrinate was accurately weighed and transferred in to a volumetric flask then dissolved in chloroform to a final volume of 10 ml (30mg/ml).

Linearity solutions:

In the first experiment chlorpheniramine maleate was considered as an internal standard (1 mL was added to each concentration) and dimenhydrinate was the studied substance (a series was prepared with increasing concentrations). In the second experiment, dimenhydrinate was considered as an internal standard (1 mL was added to each concentration) and chlorpheniramine maleate was the studied substance (a series was prepared with increasing concentrations). The concentration of the internal standard was 30 mg/ml and the concentration of the studied substance 20 mg/ml and final volume of each concentration 10 ml. A series of working standard drug solutions were prepared by diluting the standard solution with the chloroform. (1-2-4-6 mg/ml). Six replicates (2μL) of each working standard solution were injected immediately after preparation into the column and the peak area of the chromatograms was measured.

Sample preparation for tablets:

Twenty tablets of Beloramine (chlorpheniramine maleate 4 mg Mediotech Industries) were crushed and powdered, then a quantity of the powder containing 20 mg chlorpheniramine maleate was transferred into a 10 ml volumetric flask containing chloroform (C = 2 mg / ml), and (1 ml) internal standard of dimenhydrinate (30 mg/ml) was added. Twenty tablets of dimenhydrinate Barakat (dimenhydrinate 50 mg) were crushed and powdered, then a quantity of the powder containing 40 mg dimenhydrinate was transferred into a 10 ml volumetric flask containing chloroform (C = 4 mg / ml), and (1 ml) internal standard of chlorpheniramine maleate(30mg/ml) was added.

RESULTS AND DISCUSSION:

Chromatography:

A specific and highly sensitive capillary column gas chromatographic method was developed for the simultaneous determination of chlorpheniramine maleate and dimenhydrinate using on a TRB-5-625 capillary column (60m×0.25 mm i.d0.25 μm film thickness). In the first experiment six replicates (2μL) of each working standard solution were injected immediately after preparation into the detector at 290°C, in the mode at a split ratio of 0.5:10. Sharp and symmetrical peaks were achieved by programming the oven temperature at 230°C for 1min, then programmed to 250°C at a rate 5°C min and maintained for 7 min. The retention times of dimenhydrinate is 7.5 min and 9.3 min for chlorpheniramine maleate. The figure (2) show the chromatogram for first experiment, chlorpheniramine maleate was considered as an internal standard and dimenhydrinate was a studied substance.

Linearity:

The linearity of an analytical procedure is its ability (within a given range) to obtain test results which are directly proportional to the concentration (amount) of analyte in the sample¹⁹. The linearity between the concentration and the relative area was analyzed. The linear equation obtained by the least squares method was y = 0.07519x - 0.384 for dimenhydrinate and y= 0.07394x - 0.3657 for chlorpheniramine maleate as shown in (Fig. 4, 5). The resulting correlation coefficients allowing estimating the quality of the curves were R² = 0.9986 and R² = 0.9991respectively, these values of R² greater than 0.99 indicates a satisfactory linearity. The linearity was demonstrated in the concentration range (1-6mg/mL).

Accuracy:

The accuracy of an analytical procedure: expresses the closeness of agreement between the value which is accepted either as a conventional true value or an accepted reference value and the value found. For the quantitative approaches, at least nine determinations across the specified range should be obtained¹⁹.

Table 2: Relative standard deviation of the six determinations of solutions of dimenhydrinate and chlorpheniramine

N	dimenhydrinate (6mg/ml)	chlorpheniramine maleate (6mg/ml)
1	6.077	6.059
2	6.074	6.057
3	6.065	6.065
4	6.063	6.056
5	6.067	6.057
6	6.062	6.059
Mean (±) SD	6.068±0.006	6.058±0.003
RSD%	0.101	0.0536

Accuracy was determined from three concentrations for dimenhydrinate and chlorpheniramine maleate, with six replicates. Results are reported in (Table1). For dimenhydrinate, the results indicated that the individual recovery ranged from 99.91% to 101.14%.

The recovery of dimenhydrinate by the proposed method was acceptable, as the mean recovery value was 99.39% in the range 95.0-105.0% and RSD was 2.07%, less than 5%. For chlorpheniramine maleate accuracy, the same approach was used and the individual recovery ranged from 99.55% to 100.99%. Considering the same criteria, the recovery of chlorpheniramine maleate by the proposed method was also satisfactory, with a mean recovery value of 99.45% and a RSD of 1.60%.

Precision:

The precision of an analytical procedure: expresses the closeness of agreement between a series of measurements obtained from multiple sampling of the same homogeneous sample under the prescribed conditions¹⁹. The solution (6 mg/ml) was injected six times (N = 6). The precision data are shown in (Table 2). The results show coefficient of variation (RSD) under the limit of 5%. RSD of 0.101% and 0.0536 % were calculated for dimenhydrinate and chlorpheniramine respectively.These results indicated that the intermediate precision of this method was acceptable for dimenhydrinate and chlorpheniramine

Application to tablet assays for quality control:

Finally, we applied our method to assay commercial tablets purchased in Syria. One formulation was analyzed for each drug. It was observed that, all the formulations tested had concentrations within the specification of the USP Pharmacopeia, which recommended that tablets should contain not less than 90% and not more than 110% of the labelled amount of the active pharmaceutical²⁰. Fuad A.R. et al. result of assaying chlorpheniramine tablets was 98.1+- 0.40²¹. Akwasi Ac. et al. result of assaying chlorpheniramine tablets was 100.90 +- 2.10²².Tarek BE. Et al result of assaying dimenhydrinate tablets was 98.85 ± 1.84⁵. The data of commercial tablets contents are reported in (Table 3). These results showed that our results are similar to other research results and the proposed method is suitable for the analysis of commercial formulations and allow a quick and easy quality control of dimenhydrinate and chlorpheniramine maleate content.

CONCLUSION:

Newly GC-FID method was developed for determination of dimenhydrinate and chlorpheniramine maleate. The developed method has proved specific, precise and accurate for assaying the two drugs either individually or in mixtures. This method could be an additional analytical technique particularly in the quality control of raw materials, active pharmaceutical ingredients and pharmaceutical formulations. The results reported herein demonstrated that the quality of the analyzed formulations in sold in Syria was correct.

REFERENCES:

1. Sher NA, Siddiqui FA, Hasan NA, Shafi NI, Zubair AR, and Mirza AZ. Simultaneous determination of antihistamine antiallergic drugs, cetirizine, domperidone, chlorphenamine maleate, loratadine, meclizine and buclizine in pharmaceutical formulations, human serum and pharmacokinetics application, Royal society of chemistry.2014; 9(1):29:57.

2. Daravat BH, Reddy G.S., Kamarapu SK. Development and validation of RP-HPLC Method for simultaneous estimation of Chlorpheniramine maleate and Diethylcarbamazine Citrate in pharmaceutical dosage forms. Asian journal of Pharmaceutical and Clinical Research.2014; 7(3):98-102.

3. Ali A, Ahmed M, Mahmud T, Qadir MA, Nadeem K, Saleem A. Stability-indicating High-performance Liquid Chromatography Method for Simultaneous Determination of Aminophylline and Chlorpheniramine Maleate in Pharmaceutical Formulations. Indian journal for pharmaceutical sciences.2015; 77(5): 515–521.

4. Bagyalakshmi J., Nipu Sam P., George, Ravi T.K. Formulation and Evaluation of Chlorpheniramine Maleate from Methyl Cellulose Patches. Research J. Pharm. and Tech. 4(6): June 2011; Page 965-971.

5. Tarek BE, Karim AH, Randall Cl. Selective determination of dimenhydrinate in presence of six of its related substances and potential impurities using a direct GC/MS method. Journal of advanced research.2016; 7(1): 53–58.

6. Kar A., Aniuha G.I. Spectrophotometric determination of dimenhydrinate with reinecke salt. JPharm Sci. 1981; 70(6):690–691.

7. Mitic S.S., Miletic G.Z., Pavlovic A.N., Pecev E.T., Velimirovic D.S.: Quantitative estimation of dimenhydrinate in pharmaceuticals and human control serum using ligand-exchange reaction. Oxid Commun. 2012; 35(4):856–868.

8. Shubietah R.M., Abu Zuhri A.Z., Fogg A.G. Adsorptive stripping voltammetric determination of dimenhydrinate at a hanging mercury drop electrode. Microchim Acta. 1999; 130(3):165–171.

9. Ong C.P., Ng C.L., Lee H.K., Li S.F.Y. Determination of antihistamines in pharmaceuticals by capillary electrophoresis. J Chromatogr A. 1991; 588(1–2):335–339.

10. Tavares V., Macedo C.C., Montanhez L., Barros F.A.P., Meurer E.C., Campos D.R. Determination of dimenhydrinate in human plasma by liquid chromatography–electrospray tandem mass spectrometry: application to a relative bioavailability study. J Chromatogr B. 2007; 853(1–2):127–132.

11. Özkan C.K., Taşdemir U., Taş C., Savaşer A., Erol H., Özkan Y. Determination of Dimenhydrinate nasal delivery system in the blood by RP-LC. Chromatographia. 2013; 76(21–22):1521–1525.

12. Ladmeera, Hasumati Raj, Vineet Jain. Development and Validation of Analytical Method for Bromhexine Hydrochloride, Chlorpheniramine Maleate and Guaifenesin by Simultaneous Equation Spectroscopic Method. Asian J. Res. Pharm. Sci. 5(2): 2015; 76-82.

13. Barbas C., García A., Saavedra L., Castro M. Optimization and validation of a method for the determination of caffeine, 8-chlorotheophylline and diphenhydramine by isocratic high-performance liquid chromatography. Stress test for stability evaluation. J Chromatogr A. 2000; 870(1–2):97–103.

14. DiGregorio D., Westgate E., Sherma J. Analysis of the active ingredient dimenhydrinate in motion sickness tablets by high-performance thin-layer chromatography with ultraviolet absorption densitometry. Acta Chromatogr. 2000; 10:190–194

15. El-Kafrawy D.S., Belal T.S. Validated HPTLC method for the simultaneous determination of cinnarizine and dimenhydrinate in their combined dosage form. J Assoc Arab Univ Basic Appl Sci. 2014

16. Farrell M., Heinrichs M., Tilelli J.A. Response of life threatening dimenhydrinate intoxication to sodium bicarbonate administration. J Toxicol Clin Toxicol. 1991; 29(4):527–535.

17. Kyle P.B., Spencer J.L., Purser C.M., Eddleman K.C., Hume A.S. Suspected pediatric ingestions: effectiveness of immunoassay screens vs. gas chromatography/mass spectroscopy in the detection of drugs and chemicals. J Toxicol Clin Toxicol. 2003; 41(7):919–925.

18. United Stated Pharmacopoeia USP 38/National Formulary NF33; 2015.

19. http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1__Guideline.pdf [Last accessed on 10 Dec 2017].

20. United Stated Pharmacopoeia USP 38/National Formulary NF33; 2015.

21. Fuad A.R., Normal-phase LC method for simultaneous analysis of pseudophedrine hydrochloride, dextromethorphan hydrobromide, chlorpheniramine maleate, and paracetamol in tablet formulations. Saudi pharmaceutical journal.2010; 18(2): 103–106

22. Akwasi Ac., Wilfred O. G., Godfred D., Joseph A., and Sylvester Ad., Validated RP-HPLC method for simultaneous determination and quantification of chlorpheniramine maleate, paracetamol and caffeine in tablet formulation. Springerplus. 2016; 5: 625.

23. Shasho HA, Sakur A.A., Trefi SA, Separation and assay of four antihistamine drugs Diphenhydramine, Chlorpheniramine, Cyproheptadine and Fexofenadine in Pharmaceutical Forms by a Single HPLC Method. International Journal of Pharmacy and Pharmaceutical Sciences, 10(4):53 · April 2018.

24. JY Nehete, VV Shewale, VN Deshmukh, MR Narkhede. Gas Chromatographic Ethanol Determination in Bhunimbadi Kwath. Research J. Pharm. and Tech.2 (4): Oct.-Dec. 2009; Page 876-87.

25. Shoeb Alahmad, Mhd. Amer Almardini, Mahziayahia. Validated HS-GC-FID Method for Determination of Residual Ethanol in Solid Dosage Form. Research J. Pharm. and Tech. 7(2): Feb. 2014; Page 184-187.

26. Atul Bajaj, Cijo John, Joydip Choudhury, Rohitashva Mani Tripathi. Development and Validation of Gas Chromatography/Mass Spectrometry Method for the Simultaneous Determination of Some Preservatives in Personal Care, Cosmetics and Pharmaceutical Formulations. Research J. Science and Tech. 2017; 9(3): 308-312.

27. Agilandeswari Devarajan, MK Mohan Maruga Raja. Identification and Analysis of Chemical Constituents of Rasam by Gas Chromatography–Mass Spectrometry (GC-MS). Research J. Pharm. and Tech 2017; 10(12): 4183-4187.

Received on 11.03.2019 Modified on 16.04.2019

Research J. Pharm. and Tech. 2019; 12(6): 2851 -2856.

DOI: 10.5958/0974-360X.2019.00480.3

	Dimenhydrinate		Chlorpheniramine maleate
Mean concentration level1 (2mg/ml) %	99.91	Mean concentration level1 (2mg/ml) %	99.55
Mean concentration level2 (4mg/ml) %	97.12	Mean concentration level2(4mg/ml) %	97.80
Mean concentration level3 (6mg/ml) %	101.14	Mean concentration level3 (6mg/ml) %	100.99
Mean recovery % (±) SD	99.39 ± 2.05	Mean recovery % (±) SD	99.45 ± 1.60
%RSD	2.07	%RSD	1.60