PRACTICAL REPORT ON THE ISOLATION AND IDENTIFICATION OF CODEINE AND PARACETAMOL Essay

Codeine or methyl morphine, an alkaloid, was first isolated in 1832 from raw opium. It concentration ranges from 0.2% to 0.8%. Mostly consumptiond for its analgesic, anti-tussive and anti-diarrheal capabilities (Tremlett, Anderson and Wolf, 2010). Paracetamol also known as acetaminophen (n-acetyl-p-aminophenol, APAP) on the other hand, is a utile non- steroidal anti- insurgent drug (NSAID). It is commonly used in the management of pain and fever in a variety of patients (Kamberi, et al., 2004). bod 1 CodeineNCBI, 2009 Fig 2 AcetaminophenNCBI, 2009 One of the technique involved in the declivity of codeine and paracetamol from its matrix, is the final result extraction otherwise known as liquid liquid extraction. This process entails the use of two immiscible liquids usu all in ally chloroform and body of water in dissolve the sample for two distinctive layers to form after the mixture had been good jolted together (Rubinson and Rubinson, 1998). Separating the components of th e extract, is done through the use of Thin layer Chromatography. It is one of the standard procedures used in many forensic laboratory when analysizing unknown drugs or mixtures (Howlett and Steiner, 2011). Separation of the mixtures occur based on the pH, mansion of its components, solvent and the thin layer stationary phase (Howlett and Steiner, 2011).METHODSThe finely change integrity sample was dissolved in 20ml of distilled water. This was then basified with NaOH solution to pH 12 using litmus paper. The resulting solution was afterwards filtered. 1.0ml of chloroform was pipetted into the filtrate. After shaken and combined, two distinctive layers was discover. The bottom layer was extracted thrice using a micro- pipette. On a thin chromatography scale, fivesome spots were placed ( as shown in table 2) and the domicile was developed using chloroform/methanol. This was later visualized with dragendorffs reagent under the UV light. All separated components were observed, determine and recorded. RESULTS tabularise of observed pHSOLUTIONInitial pHFinal pHBasified sample1012TABLE 1Table of Retention factor (RF value)Rf = Distance traveled by the import (cm) Distance travelled by the solvent (cm) SUBSTANCEDistance travelled by depicted object (cm)Distance travelled by event (cm)Retention factor value (Rf) Chloroform extract3.04.00.75Codeine positive guard3.04.00.75Paracetamol positive control4.04.01.00Chloroform (negative control) 3.54.00.86Diluted sample4.04.01.00TABLE 2DIAGRAMFig 3 The Developed Chromatographic Plate.DISCUSSIONRunning the chloroform extracts and dilute sample together with two positive controls and a negative control on a single chromatographic case simultaneously, the retention factor(Rf) of five different samples were determined. The RF value of the chloroform extract(0.75) tallied with that of the codeine positive control and that of cut sample(1.00) with the paracetamol positive control. This tentatively shows that, codein e and paracetamol were present in the sample. The solvent front(i.e distance travelled by the mixed solvents) is 4cm, this is quite close to the distances covered by all separated components(between 3 3.5cm), which makes the retention factors, not a true exemplar of their actual values. It was later discovered that, this is due to not al low gearing thechromatographic plate to develop for a longer period of time in the solvent tank. The solvent front also dried up quickly when the plate is taken out., making drawing a line at that bloom quite difficult. Fortunately, this was overcome by the use of visualizing spray and UV lamp. Solvent extraction(liquid-liquid), involved selective movement of components of a vegetable marrow in mcg to gram quantities between two immiscible liquid phase its detachment and selectivity is based on solubility differences and pH control respectively (Fifield and Kealey, 1995).This was observed when chloroform was added to the basified filtrate. Aft er vigorous shaking and settling down, chloroform be more dense, composed the bottom layer, with the aqueous phase up. Liquid-liquid extraction often involved high volume of organic fertilizer solvents and poor resolution of mixtures of organic materials (Fifield and Kealey, 1995). Thin Layer Chromatography is usually employed in the qualitative analytic thinking of mixtures of non-volatile compounds like pharmaceuticals (Skoog, et al., 2000). tender loving care can also be used to keep up the identity of an unknown sample ( Lewis and Evans, 2011). Dissolution of the codeine and paracetamol tablet in distilled water without weighing, shows that, TLC was never designed for semi- quantitative analysis. This is due to difficulties in reproducibly applying aliquots of the mixture to the plate and then recovering all of the separated components from the plate (Skoog, et al., 2000). CONCLUSION Using the Rf values obtained in the table 2 above and the visual indicator reaction with th e totals under the UV light, codeine was extracted to a high degree during the solvent extraction, tentatively identified by TLC (due to its positive control having the same Rf values with the chloroform extract(0.75) and both(prenominal) were the only one that were seen under the UV light) while paracetamol was extracted to a low degree (due to its positive control having the same Rf with the diluted sample). triple compounds can share the same retention factor(Rf) or produce akin(predicate) chromophores when sprayed with detection reagents (Howlett and Steiner, 2011). The study by Lewis and Evans( 2011) shows that if a spot from an unknown substance is developed on a TLC plate together with a spot from a substance that is suspected to be the unknown, and the two substance are found to have the same Rf value, they are in all probability the same substance.FUTURE SUGGESTIONS AND RECOMMENDATIONS Due to the limitation that is associated with using TLC to hardly identify a given sample, minimum standards for drug testing and reportage in the forensic community are recommended by the Scientific works Group for the analytic thinking of seized drugs (SWGDRUG) (Howlett and Steiner, 2011). In order for a drug credit to be confirmed to SWGDRUG specification, additional tests must includes, Infrared spectroscopy and GC-MS (Howlett and Steiner, 2011).REFERENCESFifield, F. W. and Kealey, D. 1995. Principles and utilize of Analytical chemistry. (4th ed) Glasgow, Blackie Academic and professional. Howlett, S. E. and Steiner, R. R. 2011. Validation of Thin Layer Chromatography with AccuTOF-DART Detection for rhetorical Drug Analysis*. Forensic Sciences e-journal 56 (5), pp. 12611267. Available through Anglia Ruskin University subroutine library website http//libweb.anglia.ac.uk Accessed on 11 borderland 2014. Kamberi, M., Riley, C. M., Huang, C. C. and Xiaoyan, M, 2004. A validated, sensitive HPLC method for the finding of trace impurities in acetaminophen drug substance. Pharmaceutical and Biomedical Analysis e-journal 34 (1), pp. 123128. Available through Anglia Ruskin University Library website http//libweb.anglia.ac.uk Accessed on 18 March 2014. Lewis, R. and Evans, W. 2011. Chemistry. 4th ed. Hampshire, Palgrave Macmillan. NCBI, 2009. National Library of Medicine. online Available at http//www.ncbi.nlm.nih.gov/pccompound Accessed 7 April, 2014. Rubinson, J. F. and Rubinson, K. A. 1998. present-day(a) chemical analysis. Upper Saddle River, NJ, Prentice Hall. Skoog, D., West, D., Holler, F. and Crouch, S. 2000. Analytical Chemistry- An introduction. (7th ed). Boca raton, Thomson Learning Inc. Tremlett, M., Anderson, B. J. and Wolf, A. 2010. professionalcon debate is codeine a drug that still has a useful role in pediatric practice? Pediatric Anesthesia e-journal 20 (2), pp. 183194. Available through Anglia Ruskin University website http//libweb.anglia.ac.uk Accessed on 29 March 2014.