Estimation of Etizolam and Propranolol in Combined Dosage Form by Spectroscopic Method

ESTIMATION OF ETIZOLAM AND PROPRANOLOL IN COMBINED DOSAGE FORM BY SPECTROSCOPIC METHOD

MASTER OF PHARMACY DISSERTATION PROTOCOL

SUBMITTED TO THE

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES KARNATAKA, BANGALORE.

BY

DEVI SWAPNA P V

Under The Guidance Of

Ms. PARAMITA DAS

DEPARTMENT OF PHARMACEUTICAL ANALYSIS

SRINIVAS COLLEGE OF PHARMACY, MANGALORE – 574143.

2012 – 2014

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA.

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

NAME OF THE INSTITUTION

COURSE OF STUDY AND SUBJECT

TITLE OF TOPIC

Propranolol

6.3 REVIEW OF LITERATURE

• Shivarkar N A(2)et al. have reported UV spectrophotometric method development & validation for simultaneous estimation of Flunarizine dihydrochloride & Propranolol hydrochloride in bulk drug & capsule dosage form by simultaneous and Q-analysis/absorption ratio method.The % recovery of Flunarizine dihydrochloride & Propranolol hydrochloride was found out to be 98.49 – 101.06 % (Method I) and 99.82 – 100.86% (Method II).The proposed method was validated as per ICH guidelines.

• Tulja R G(3) et al. have reported RP-HPLC method for simultaneous estimation of propranolol hydrochloride and alprazolam in bulk and formulation. Chromatographic separation of the drugs was achieved on a Waters C18 column (250 ×4.6 mm, i.d. 5 ì) using acetonitrile: water (adjusted to pH 2.3 with ortho phosphoric acid) in the ratio of 60:40 v/v as mobile phase. Flow rate was 1.0 mL/min and the detection wavelength was 214 nm. The two drugs were satisfactorily resolved with retention time values 1.737 min and 3.810 min for propranolol and alprazolam, respectively. The method was validated in terms of accuracy, precision, linearity, and limit of detection, limit of quantitation, robustness, and ruggedness as per ICH guidelines.
• The Merk Index(4), an Encyclopedia of chemicals, drugs and biologicals. Fourteenth ed. NJ; 2006
• Casacchia M(5) et al, have reported Etizolam in the treatment of generalized anxiety disorder. A double-blind study versus placebo.
• Inoue H(6) et al. have reported the Screening and determination of benzodiazepines in whole blood using solid-phase extraction and gas chromatography/mass spectrometry.
• Tanaka E(7)et al. have reported Simultaneous determination of twelve benzodiazepines in human serum using a new reversed-phase chromatographic column on a 2-microns porous microspherical silica gel.
• Soo V A(8) et al. have reported the quantitative screening for hypnotic-sedative drugs in which we use capillary gas chromatography with a nitrogen-phosphorus detector (GC/NPD) as the primary method and capillary gas chromatography-mass spectrometry (GC-MS) for confirmation. GC retention times of the acid-extracted underivatized drugs were stable (CVs less than 1%), and the detector response varied linearly over a 20-fold concentration range with a mean correlation coefficient for 11 drugs of 0.989.
• Doshi A K(9) et al. developed a spectrophotometric method for simultaneous estimation of Propranolol hydrochloride and Flunarizine dihydrochloride in combined dosage form.Linearity was observed in range of 24-64 μg/ml and 6-16 μg/ml for Propranolol hydrochloride and Flunarizine dihydrochloride respectively. Recovery studies confirmed the accuracy of proposed method and results were validated as per ICH guidelines.
• Jończyk A(10) et al. developed Spectrophotometric and HPLC methods for determination of hydrochlorothiazide, triamterene and propranolol hydrochloride. Both methods were appropriate for the determination of three compounds in pharmaceutical preparations containing their mixtures. Both the elaborated methods for the determination of the studied compounds give comparable results and can successfully be applied to the assay in their mixtures occurring in the composition of pharmaceutical preparations.

• Bhavar G(11) et al. reported HPTLC method for estimation of propranolol hydrochloride from bulk drug and tablet formulations. The separation was achieved on TLC plates using appropriate solvent system. The spots so developed were densitometrically scanned at 290 nm. The linearity of the method was found to be within the concentration range of 200-2000 ng/spot. The validation parameters, tested in accordance with the requirements of ICH guidelines, prove the suitability of this method. The method was successively applied for determination of drug in tablets, wherein, no interference from tablet excipients was observed.
• Salle E D(12) et al. has reported a sensitive gas chromatographic method for the determination of propranolol in human plasma.
• Prashanth K V(13) et al. reported spectrophotometric methods for the determination of propranolol hydrochloride (PPH) as base form (PPL) in bulk drug, and in tablets and capsules. The methods were based on the molecular charge-transfer complexation of propranolol base (PPL) with either 2,4,6-trinitrophenol (picric acid; PA) or 2,4-dinitrophenol (DNP). The yellow colored radical anions formed on dissociation, are quantitated at 425 nm (PA method) or 415 nm (DNP method). The assay conditions were optimized. Beer's law is obeyed in the concentration ranges 2.4-42.0 μg ml-1 in PA method and 9.0-126.0 μg ml- 1in DNP method, with respective molar absorptivity values of 4.97 × 103 and 1.66 × 103 l mol-1 cm-1.
• Krishnla R(14) et al. report a liquid-chromatographic method for assay of total and free propranolol and its metabolite, propranolol glycol, in plasma. The mobile phase consists of equal parts of methanol and a solution of potassium 50 mmol/L phosphate and 2.5 mmol/L 1-pentanesulfonic acid, pH 5.0. Detection was by fluorimetry, with excitation at 296 nm and emission at 338 nm.
• Palak P(15) et al. developed a high-performance thin layer chromatographic method. The method employed HPTLC aluminum plates precoated with silica gel 60F as the stationary phase while the solvent system was toluene:methanol: ethyl acetate: acetic acid (7 : 1.5 : 1.5 : 0.1 v/v/v/v). The Rf value was observed to be 0.07 ±0.02 and 0.67±0.02 for propranolol hydrochloride and flunarizine di hydrochloride. The densitometric analysis was carried out in absorbance mode at 240 nm.
• Patel V B(16) et al. have reported spectrophotometric method for the simultaneous determination of Escitalopram oxalate(ESC) and Etizolam(ETI) in combined tablet dosage form. The method is based on the simultaneous equations for analysis of both the drugs using 0.01 N HCl as solvent. The method was linear in the concentration range of 4-40µg/ml for ESC and 1-32 µg/ml for ETI with a correlation coefficient of 0.9999 and 0.9998 for respective drugs. The percent recovery was found in the range of 98.39-101.13% and 98.07-101.83 % for ESC and ETI, respectively. The proposed method was successfully applied for quantitative determination of Escitalopram oxalate and Etizolam in combined tablet dosage form for routine analysis.

• Sanghavi N M(17) et al. worked out a simple colorimetric method for the determination of propranolol hydrochloride. The method involves nitration of the drug with a mixture of potassium nitrate and sulphuric acid. The coloured nitro-derivative has an absorption maxima at 360 nm. The working concentration range is 10–50 μg/ml. The method is applicable to analysis of pharmaceutical preparations
• Suryakant B(18) et al. developed an isocratic reversed phase HPLC method has been developed for the simultaneous determination of escitalopram oxalate and etizolam on a HiQ-silC18HS (250 x 4.6 mm) column using a mobile phase consisting of methanol:phosphate buffer pH-5 (70:30,v/v) at a flow rate of 1 mL/min and the detection was carried out at 254 nm. Retention times of escitalopram oxalate and etizolam were found to be 4.85 (± 0.31) min and 7.65 (± 0.56) min respectively
• Tulja R G(19) et al. developed and validated RP-HPLC method for the simultaneous determination of propranolol hydrochloride and diazepam in bulk and in pharmaceutical formulations. The separation was carried out on Waters C18 (250 ×4.6 mm i.d, 5 μ) column using acetonitrile: 0.4 % potassium dihydrogen ortho phosphate (adjusted to pH 3.52 with ortho phosphoric acid) in the ratio of 60:40 v/v as eluent. The flow rate was 1 ml/min and effluent was detected at 229 nm. The retention time of propranolol hydrochloride and diazepam were 2.330 and 6.663 min. respectively. The linear dynamic range was 2-24 μg/ml and 0.25- 3.0 μg/ml for propranolol hydrochloride and diazepam, respectively. Percentage recoveries for propranolol hydrochloride and diazepam were 100.03 and 99.72 %, respectively.
• Jayant B D(20) et al. have reported three methods for the for quantitative estimation of etizolam in bulk and in tablet dosage form by UV spectroscopy, HPTLC and RP-HPLC. In the first method, etizolam was estimated by measuring absorbance at λmax of 249 nm in 0.01 N HCl. In the second method, etizolam was estimated by HPTLC method using precoated plate of silica gel 60F254 and Toluene: Chloroform: Methanol (4.8:4.0:1.2 v/v/v) as mobile phase.. In the third method, etizolam was estimated by simple and specific RP-HPLC method using the Kromasil column at ambient temperature using a mobile phase consisting of water: acetonitrile (50:50% v/v) and flow rate was 1.0 ml/min. Quantitation was achieved with UV detection at 241 nm based on peak area with linear calibration curves at concentration ranges 4–32 µg/ml etizolam.

• To determine λ max in different solvent .
• To develop simple, selective, rapid, precise and economical analytical method for the estimation of Etizolam and Propranolol.

1. Procurement of the drug and marketed formulations.
2. Determination of wavelength of maximum absorbance and intensity of absorption in different solvents for analytical studies.
3. Develop analytical method for Etizolam and Propranolol by UV-Visible spectrophotometer using estimation method.

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3. Tulja R G., Gowri S D, Kadgapathi P, Satyanarayana B. A Validated RP HPLC Method for Simultaneous Determination of Propranolol hydrochloride and Alprazolam in Bulk and in Pharmaceutical formulations.Journal of Pharmacy Research. 2011; 4: 296--360.
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3. Jończyk A, Nowakowska Z. Determination of hydrochlorothiazide, triamterene and propranolol hydrochloride by the spectrophotometric method and highperformance liquid chromatography (HPLC).Acta Pol Pharm. 2001; 58: 339-44.
4. Bhavar G, Chatpalliwar V A. Quantitative Analysis of Propranolol Hydrochloride by High Performance Thin Layer Chromatography.Indian J Pharm Sci.2008;70: 395–8.
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1. Jayant B D, Vinay B P, Badmanaban R, Shital M P, Chhaganbahi N.Development and validation of UV-spectroscopic, HPTLC and RPHPLC methods for the estimation of etizolam in bulk and in tablet dosage form. Inventi Rapid: Pharm Analysis & Quality Assurance.2012, Article ID-" Inventi:ppaqa/433/12.