Thin Layer Chromatography Lab Report

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CHROMATOGRAPHIC METHODS: After successful extraction of phospholipids from their source analysis can be performed for the detection of specific phospholipids. This section will discuss chromatographic methods used for the analysis of phospholipids. All systems of chromatography consist of a stationary and mobile phase. A monster placed on a stationary phase, i.e., a solid or a liquid, and the mobile phase, a gas or a liquid, is allowed by modifying the system. The components of the sample will be separated on the basis of their ranging physical and chemical properties, imparting different affinities for the two phases. Thin layer chromatography (TLC) was the first chromatographic method for assessing phospholipids, and is commonly used today. …show more content…

Most of the systems vary to include ratios of chloroform, methanol and water. triethylamine, ethanol, hexane, and isopropanol are also common solvents in the mobile phase. Phospholipids migrate to the stationary phase a certain distance on the basis of the composition and affinity for the mobile phase. Identification is based on the delay (Rf), wherein the ratio of the distance moved by the analyte (i.e., the phospholipids) from the origin of the distance moved by the flowing solvent from the origin. Each of the analyte will have its own Rf value under certain circumstances. The separation of the phospholipid classes can be improved by two-dimensional chromatography. This technique requires developing the TLC plate in a direction, then dried, and developed in a solvent mixture at a 90 ° the first development (Singh and Jiang, …show more content…

One progress on TLC called high performance TLC (HPTLC; Sherma and Jain, 2000).HPTLC makes use of gel qualities that are finer, so that thinner plates and smaller. This allows faster separation times and better separation efficiency. HPTLC has improved reduced resolution and detection limits, so that the to walk two dimensions. To phospholipids visible on the TLC plates are used detection reagents. spots corresponding phospholipids may be carbonized by the addition of phosphomolybdic acid, sulfuric acid or copper sulfate in phosphoric acid, and then heating of the sample. The result charred spots can be scraped off the plate and measured by densitometry. In order to visualize the spots without charring, solutions such as rhodamine 6G or berberine can be used for UV-detection. iodine vapor is commonly used; but it does not work well with lipids which do not contain double bonds (Hoving,

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