Asian Journal of Research in Chemistry

Cijo John ¹, C. Chainulu ¹, Priyankar Ghosh ¹, Seema Srivastava ¹, Sudhirkumar Shukla ², S. Satyanarayana ³

Affiliations
1 Central Forensic Science Laboratory, Govt. of India, Hyderabad, 500 013, IN
2 Central Forensic Science Laboratory, Govt. of India, Chandigarh, 160 036, IN
3 Department of Chemistry, Osmania University, Hyderabad, India- 500 007, IN

Abstract

A new simple, precise, rapid and selective High-Performance Thin-Layer Chromatographic (HPTLC) method has been developed for the simultaneous determination of amitriptyline (AMTRL), atenolol (ATL) and propranolol (PRPL) using benzene: methanol: acetone: ammonia solution (25% NH3) (5:3:2:0.2; v:v) as the mobile phase and Merck HPTLC plates (0.2 mm thickness) pre coated with 60F254 silica gel on aluminium sheet as the stationary phase. Detection was carried out densitometrically using a UV detector at 254 nm. The retention factors (Rf value) of AMTRL, ATL and PRPL were 0.80, 0.37 and 0.69 respectively. The developed method was fully validated as per the guidelines given by the International conference on Harmonisation (ICH) for precision, recovery, accuracy, robustness, ruggedness and specificity and determination of the limits of detection and quantification. The amount of AMTR, PRPL and ATL present in marketed tablets was quantified and estimated using the developed method which was found to be in close agreement with the labelled content. The linear regression analysis for calibration plots showed good linear relationship in the concentration range of 0.1ug/spot to 10ug/spot with respect to peak area.

Keywords

High Performance Thin Layer Chromatography, Amitriptyline, Atenolol, Propranolol, Simultaneous Determination.

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Cijo John ¹, Mohd. Idris ¹, P. Sudhakar ¹, Priyankar Ghosh ¹, K. M. Varshney ¹, S. K. Shukla ²

Affiliations
1 Central Forensic Science Laboratory, Govt. of India, Hyderabad - 500 013, IN
2 Central Forensic Science Laboratory, Govt. of India, Chandigarh - 160 036, IN

Abstract

Clonazepam, a nitrobenzodiazepine and a scheduled drug is one of the highly abused drugs in recent times. An analytical methodology was developed for the detection and quantitation of clonazepam in chocolate sample. The method involves the extraction of the chocolate sample by solvent extraction with chloroform under basic conditions (pH: 8-9). The extract was used for the GC-MS analysis in SIM and full scan modes for the detection of the analyte present. For quantification, the extract was re-dissolved in the mobile phase, Acetonitrile: 40mM potassium di hydrogen-phosphate (6:4,v/v), pH adjusted to 3.9. The separation was achieved on a Phenomenex Gemini C18 HPLC reversed-phase column (150mmx4.6mmx5μ) with the mobile phase and diode array detection with UV range 200-370 nm. The calibration plot for the determination of clonazepam is based on linear regression analysis (y=51.405x + 84.924; r²=0.9989). The limit of detection of clonazepam was estimated as 0.1μg/ml with a signal-to-noise ratio of 3:1 and limit of quantitation of clonazepam was estimated as 0.35μg/ml with a signal-to-noise ratio of 10:1. The clonazepam detected in the chocolate was found to be 0.0763μg/mg of the chocolate. The precision studies given a % RSD range of 0.28-3.9 % with respect to peak area and 0.51-1.03 % with respect to retention time. The method is robust, specific and reliable for the qualitative and quantitative analysis of clonazepam in chocolate and the law enforcement laboratories can utilise the method for routine analysis.

Keywords

Clonazepam, Drug Abuse, Chocolate, GC-MS, HPLC.

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