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

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Codeine or methyl morphia. an alkaloid. was foremost isolated in 1832 from natural opium. It concentration ranges from 0. 2 % to 0. 8 % . Largely used for its analgetic. anti-tussive and anti-diarrheal capablenesss ( Tremlett. Anderson and Wolf. 2010 ) . Paracetamol besides known as Datril ( n-acetyl-p-aminophenol. APAP ) on the other manus. is a utile non- steroidal anti- inflammatory drug ( NSAID ) . It is normally used in the direction of hurting and febrility in a assortment of patients ( Kamberi. et Al. . 2004 ) .

Fig 1: Codeine [ NCBI. 2009 ] Fig 2: Acetaminophen [ NCBI. 2009 ] One of the technique involved in the extraction of codeine and paracetamol from its matrix. is the solvent extraction otherwise known as liquid – liquid extraction. This procedure entails the usage of two non-miscible liquids normally chloroform and H2O ; in fade outing the sample for two typical beds to organize after the mixture had been exhaustively shaken together ( Rubinson and Rubinson. 1998 ) . Separating the constituents of the infusion. is done through the usage of Thin Layer Chromatography. It is one of the criterion processs used in many forensic research lab when analysizing unknown drugs or mixtures ( Howlett and Steiner. 2011 ) . Separation of the mixtures occur based on the pH. mutual opposition of its constituents. dissolver and the thin bed stationary stage ( Howlett and Steiner. 2011 ) .

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Methods:
The finely divided sample was dissolved in 20ml of distilled H2O. This was so basified with NaOH solution to pH 12 utilizing litmus paper. The ensuing solution was subsequently filtered. 1. 0ml of trichloromethane was pipetted into the filtrate. After shaken and combined. two typical beds was observed. The bottom bed was extracted thrice utilizing a micro- pipette. On a thin chromatography home base. five musca volitanss were placed ( as shown in table 2 ) and the home base was developed utilizing chloroform/methanol. This was subsequently visualized with dragendorff’s reagent under the UV visible radiation. All detached constituents were observed. identified and recorded. Consequence:

Table of ascertained pH
SOLUTIONInitial pHFinal pH
Basified sample1012
Table 1
Table of Retention factor ( RF value )
Rf = Distance travelled by the substance ( centimeter ) Distance travelled by the dissolver ( centimeter ) SUBSTANCEDistance travelled by substance ( centimeter ) Distance travelled by Solvent ( centimeter ) Retention factor value ( Rf ) Chloroform extract3. 04. 00. 75




Codeine positive control3. 04. 00. 75
Paracetamol positive control4. 04. 01. 00
Chloroform ( negative control ) 3. 54. 00. 86
Diluted sample4. 04. 01. 00
Table 2
Diagram:




Fig 3: The Developed Chromatographic Plate.

Discussion:
Runing the trichloromethane infusions and diluted sample together with two positive controls and a negative control on a individual chromatographic home base at the same time. the keeping factor ( Rf ) of five different samples were determined. The RF value of the trichloromethane infusion ( 0. 75 ) tallied with that of the codeine positive control and that of diluted sample ( 1. 00 ) with the paracetamol positive control. This tentatively shows that. codeine and paracetamol were present in the sample. The solvent forepart ( i. vitamin E distance travelled by the assorted dissolvers ) is 4cm. this is rather close to the distances covered by all detached constituents ( between 3 – 3. 5cm ) . which makes the keeping factors. non a true representative of their existent values. It was subsequently discovered that. this is due to non leting the chromatographic home base to develop for a longer period of clip in the dissolver armored combat vehicle. The dissolver forepart besides dried up rapidly when the home base is taken out. . doing pulling a line at that point rather hard. Fortunately. this was overcome by the usage of visualising spray and UV lamp. Solvent extraction ( liquid-liquid ) . involved selective motion of constituents of a substance in mcg to gram measures between two non-miscible liquid stage ; its separation and selectivity is based on solubility differences and pH control severally ( Fifield and Kealey. 1995 ) .

This was observed when trichloromethane was added to the basified filtrate. After vigorous shaking and settling down. chloroform being more dense. composed the bottom bed. with the aqueous stage up. Liquid-liquid extraction frequently involved high volume of organic dissolvers and hapless declaration of mixtures of organic stuffs ( Fifield and Kealey. 1995 ) . Thin Layer Chromatography is normally employed in the qualitative analysis of mixtures of non-volatile compounds like pharmaceuticals ( Skoog. et Al. . 2000 ) . Tender loving care can besides be used to corroborate the individuality of an unknown sample ( Lewis and Evans. 2011 ) . Dissolution of the codeine and paracetamol tablet in distilled H2O without weighing. shows that. TLC was ne’er designed for semi- quantitative analysis. This is due to troubles in reproducibly using aliquots of the mixture to the home base and so retrieving all of the detached constituents from the home base ( Skoog. et Al. . 2000 ) . Decision: Using the Rf values obtained in the tabular array 2 above and the ocular index reaction with the substances under the UV visible radiation. codeine was extracted to a high grade during the solvent extraction. tentatively identified by TLC ( due to its positive control holding the same Rf values with the trichloromethane infusion ( 0. 75 ) and both were the lone 1 that were seen under the UV visible radiation ) while paracetamol was extracted to a low grade ( due to its positive control holding the same Rf with the diluted sample ) . Multiple compounds can portion the same keeping factor ( Rf ) or bring forth similar chromophores when sprayed with sensing reagents ( Howlett and Steiner. 2011 ) . The survey by Lewis and Evans ( 2011 ) shows that if a topographic point from an unknown substance is developed on a TLC home base together with a topographic point from a substance that is suspected to be the unknown. and the two substance are found to hold the same Rf value. they are likely the same substance.

Future SUGGESTIONS AND RECOMMENDATIONS: Due to the restriction that is associated with utilizing TLC to precisely place a given sample. minimal criterions for drug testing and coverage in the forensic community are recommended by the Scientific Working Group for the Analysis of seized drugs ( SWGDRUG ) ( Howlett and Steiner. 2011 ) . In order for a drug designation to be confirmed to SWGDRUG specification. extra trials must includes. Infrared spectrometry and GC-MS ( Howlett and Steiner. 2011 ) .

Mentions:

Fifield. F. W. and Kealey. D. 1995. Principles and Practice of Analytical chemical science. ( 4th erectile dysfunction ) Glasgow. Blackie Academic and professional. Howlett. S. E. and Steiner. R. R. 2011. Validation of Thin Layer Chromatography with AccuTOF-DART™ Detection for Forensic Drug Analysis* . Forensic Sciences [ e-journal ] 56 ( 5 ) . pp. 1261–1267. Available through: Anglia Ruskin University Library website hypertext transfer protocol: //libweb. Anglia. Ac. uk [ Accessed on 11 March 2014 ] . Kamberi. M. . Riley. C. M. . Huang. C. C. and Xiaoyan. M. 2004. A validated. sensitive HPLC method for the finding of hint drosss in acetaminophen drug substance. Pharmaceutical and Biomedical Analysis [ e-journal ] 34 ( 1 ) . pp. 123–128. Available through: Anglia Ruskin University Library website hypertext transfer protocol: //libweb. Anglia. Ac. uk [ Accessed on 18 March 2014 ] . Lewis. R. and Evans. W. 2011. Chemistry. 4th erectile dysfunction. Hampshire. Palgrave Macmillan. NCBI. 2009. National Library of Medicine. [ on-line ] Available at: hypertext transfer protocol: //www. ncbi. nlm. National Institutes of Health. gov/pccompound [ Accessed 7 April. 2014 ] . Rubinson. J. F. and Rubinson. K. A. 1998. Contemporary chemical analysis. Upper Saddle River. NJ. Prentice Hall. Skoog. D. . West. D. . Holler. F. and Crouch. S. 2000. Analytic Chemistry- An debut. ( 7th erectile dysfunction ) . Boca raton. Thomson Learning Inc. Tremlett. M. . Anderson. B. J. and Wolf. A. 2010. Pro–con argument: is codeine a drug that still has a utile function in paediatric pattern? Pediatric Anesthesia [ e-journal ] 20 ( 2 ) . pp. 183–194. Available through: Anglia Ruskin University website hypertext transfer protocol: //libweb. Anglia. Ac. uk [ Accessed on 29 March 2014 ] .

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