1.paper chromatography

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Evaluation seminar on Comparison of TLC,GC,HPLC and it’s applications Presented by Veeresh.G.Bellatti, I M. Pharma, ( Pharmacognosy) Under the guidance of Mr.Manzoor Ahmed Dept. of Pharma. Analysis

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1. Comparison Between TLC and HPLC TLC HPLC

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Sl. Thin Layer Chromatography (TLC) High Performance Liq. Chromatograpy (HPLC) 1 It is an off line process. It is an on line process. 2 Sample and standards are handled simultaneously. Better analytical precision. No need for internal standard. Required for precision. Accuracy of analysis. Internal standard. 3 TLC Plates are Economic. Columns are expensive. 4 Sample preparation is simple. It is critical step , laborious and time consuming. 1. Comparison Between TLC and HPLC

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Sl Thin Layer Chromatography (TLC) High Performance Liq. Chromatograpy (HPLC) 5 Solvents of analytical grade are required. HPLC grade is required. 6 Choice of solvent for dissolving sample is not critical. Solvent should be removed before developing. Compatible with HPLC Column and Mobile Phase. 7 No prior treatment is required for solvents. Degassing and /or Filtrations is required. 8 Mobile phase consumption is low. Mobile phase consumption is high.

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Sl. Thin Layer Chromatography (TLC) High Performance Liq. Chromatograpy (HPLC) 9 As fresh mobile phase & stationary phase are used, No interference of previous analysis is possible Columns are reusable. 10 Corrosive mobile phase can be used. Corrosive mobile phase can’t be used. 11 The resolution is inferior. The resolution is superior. 12 Substance, sensitive to light and oxygen will create problems. It will create least problems.

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Sl. Thin Layer Chromatography (TLC) High Performance Liq. Chromatograpy (HPLC) 13 It can’t be fully automated but programmed. Fully automated. 14 Visual detection possible as it is open system. Not possible. 15 It is a simple to learn & operate. It is skilled & well trained personnel. 16 Strongly retained materials resulted in compact & sharpest spots. Least retained (first emerging) peaks are sharp. 17 Separational efficiency is 5000 N Separational efficiency is 10000 to 15000 N

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2. Comparision between HPLC and Gas Chromatography HPLC Gas Chromatography

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2. Comparision between HPLC and Gas Chromatography Sl High Performance Liq. Chromatograpy (HPLC) Gas Chromatograpy (GC) 1 It can use non- volatile & thermally unstable compounds. It can’t use. 2 It is complicated & expensive equipment. It is simple & inexpensive equipment. 3 It’s overall analysis is not that fast. It’s quite rapid. 4 In this, no universal detector available. In this, FID (Flame ionisation detector) universal detector available.

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. Sl High Performance Liq. Chromatograpy (HPLC) Gas Chromatograpy (GC) 5 It can use for inorganic ions. It can’t possible. 6 It have low degree of sensitivity. It have high degree of sensitivity. 7 It required skilled & well trained personnel. Fairly suitable for routine analysis. Not requires skilled personnel. 8 Tailing is less frequently occurring. In this, compound with polar or ionisable function groups causes tailing.

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Applications of Thin Layer Chromatography (TLC) : Separation of mixtures of drugs of chemical or biological origin, plant extracts etc. Separation of carbohydrates, vitamins, antibiotics, proteins, alkaloids and glycosides etc. Identification of drugs. Eg : Amino caproic acid, Amoxicillin tri hydrate Identification of related compounds in drugs. Eg: Allopurional. To detect the presence of foreign substance in drugs . Eg: carbimazole. To detect decomposition products in drugs. Eg:Diazepam

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Applications of Gas Chromatography : Qualitative analysis. Used in checking the purity of a compound. Used in presence of impurities. Quantitative analysis. a. Direct compression method. b. Calibration curve method. c. Internal standard method.

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Applications of HPLC Used to analyse multicomponent or determination of mixture of drugs . Used to isolate & identify drugs or metabolites in urine, plasma, serum etc can be carried out. Used to isolate & identify the mixture of components of natural or synthetic origin . Used for biopharmaceutical & pharmacokinetics studies. Used for stability studies . Used in purification of some compounds of natural or synthetic origin on preparative scale.

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Qualitative analysis. Used in checking the purity of a compound. Used in presence of impurities . Quantitative analysis. i. Direct compression method. ii. Calibration curve method. iii. Internal standard method .

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REFERENCES : HPLC- High performance liquid chromatography : -Quantitative analysis of pharmaceutical formulations –P.D. Sethi. Practical pharmaceutical chemistry – -4 th Edition, part- 2. A.D. Beckett & J.B. Stenlake. Instrumental methods of chemical analysis -R.B.K. Sharma. Pharmaceutical analysis – Dr. S. Ravi sankar.

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THANK YOU

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