Measurement of diazepam in tablets

Diazepam also known as Valium having the chemical formula (C16H13ClN2O) of Molecular Weight 284.75 gmol-1is a benzodiazepine derivative. Benzodiazepines are group of sedatives, muscle-relaxants and having anticonvulsant properties, and whose compounds are widely used therapeutically in the treatment of anxiety, alcoholism, insomnia , seizures , muscle spasms and a variety of other disorders. Its anticonvulsant properties aid in the treatment of seizures and its use as a sedative drug or tranquilizer reduces irratibility, moderates excitement and calms the nerve. At higher doses it is used as a hypnotic drug to induce sleep, producing drowsiness

In its pharmacological mechanism of action, diazepam enhances the effect of the neurotransmitter -Amino butyric acid (GABA) by binding to the benzodiazepine active site on the GABA receptor leading to central nervous system depression (Mohler, Fritschy and Rudolph, 2001). Diazepam is a derivative of 1, 4-benzodiazepin-2-one. It is Synthesize from 2-amino-5chlorobenzophenone which consists of the condensation of 2-amino-5chlorobenzophenone with ethyl ester of glycine hydrochloride. The amide nitrogen atom is methylated by dimethylsulfate, and this leads to the formation of diazepam. Diazepam is available in several commercial brands. There are several analytical methods that can be use to quantitatively determine diazepam in commercial pharmaceutical tablets which include, chromatography, titration, photometry and Molecular (UV/visible) Absorption Spectroscopy. The use of Molecular (UV/visible) Absorption Spectroscopy is prove to be a better method for the quantitative determination of diazepam in commercial pharmaceutical tablets. This method has been found to be a more rapid, accurate and gives reproducible results. In the spectrophotometric determination of diazepam in commercial pharmaceutical tablets, diazepam reacts with ninhydrin in an alkaline medium normally Sodium hydroxide ,at a pH range of 12-12.5 when heated for 60 seconds at 60 degrees Celsius giving a violet colour complex of maximum absorbance at wavelength of 530 nm. The

reaction obeys Beers law from 0.1 to 2mg/10 mL of diazepam at max value of 530 nm (Khan, Naz and Doger, 2012).

Methodology for the analysis of diazepam in tablets Prepare 100 cm3 of diazepam stock solution by weighing 20 Farcozepam 2 mg tablets and powder in an agate mortar. This amounts to 10 mg of diazepam and transferring in small amounts to 100 cm3 volumetric flask containing 50 cm3 of distilled water and stir to dissolve most of the solid particles. Make up the volume to 100 cm3 and add the rest of the powdered tablet into the flask. Continue stirring to dissolve the suspension. Allow the mixture to settle for 15 minutes. Filter the mixture through a filter funnel in a filter paper into a clean 100 cm3 volumetric flask. Discard the residue. Accurately prepare concentration of 0.5, 1,1.5,2 and 2.5 mg of diazepam solution by Pipetting 5 cm3 , 10 cm3, 15 cm3,20 cm3 and 25 cm3 aliquot of the Diazepam stock solution each in 100 cm3 conical flask. Add 0.5 cm3 of 1 M sulphuric acid (prepared from conc. H2SO4) measured in a 5 cm3 graduated cylinder follow by 0.5 cm3 of 2% Ninhydrin measured in a 5 cm3 graduated cylinder .Dilute the solutions to 100 cm3 with distilled water. Heat the samples at 60 degrees Celsius in water bath for 60 seconds. Note the violet colour solution. Determine the absorbances of the samples at 530 nm using the spectrophotometer, using distilled water as blank. For the construction of calibration curve for diazepam, plot the absorbance (y axis) against concentration of diazepam solution (x axis) and determine the best-fit least square line to calculate the concentration. This is use to determine the amount of diazepam found in Farcozepam 2 mg tablets.

Recommended Instrument: Spectrophotometer

Manufacturer: Perkin Elmer, Inc Model: Lambda 800 Cost: $US 8,892.00 The Lambda 800 provides accurate spectroscopic measurements. It is a premier, double-beam optical system providing accurate, stable and repeatable results. Its Optical system consist of Holographic grating monochromatic with 1440 lines/mm .Its Detector is a photomultiplier R6872 for high energy in whole UV/Vis range .The source is a pre-aligned tungsten-halogen and deuterium of Wavelength range: 185 to 900nm (UV/Vis resolution: < 0.05nm ). Wavelength accuracy is: 0.08 nm.Wavelength reproducibility is: < 0.020 nm. Its Standard deviation of 10 measurements is: < 0.005 nmUV/Vis. Size of Curvette holder is: 10 x 10mm curvettes.

Bibliography

CaeOnline.Com. http://www.caeonline.com/listing/product/144494/perkin-elmerlambda-800 (accessed October 20, 2013). Khan, W; Naz ,N; Doger ,N; Khan, N; Khan, A. N. Spectrophotometric Determination of Diazepam. J.Chem.Soc.Pak [Online] 2012, 34, pp 1-5. Mohler, H; Fritschy, J. M; Rudolph, U. A. New Benzodiazepine Pharmacology. The Journal of Pharmacology and Experimental Therapeutics [Online] 2001, 300, pp 1-3