Column chromatography

The concentration of CDCL and CSER is obtained by solving equation (1) and (2) CX = A1 / ax1 ………………..... (1) CY = [A2 – (ax2 × CX)] / ay2…...(2) Where,  A1 and A2are absorbance of mixture at λ1 and λ2respectively.  ax1 and ax2 are absorptivities of DCL at λ1 and λ2, respectively,  ay1 and ay2 are absorptivities of SER at λ1 and λ2, respectively,  CXandCYare concentrations of DCL and SER, respectively. Concentration of DCL and SER in the powder mixture is found by using equation (3) and (4)…

Paper Chromatography Paper Chromatography is an analytical method that is used to separate coloured chemicals substances, especially pigments. This is how paper chromatography is conducted: Materials Chromatography Paper (Can also be Coffee Filter) Large Beaker (max one litre) Paper Clips/Sellotape & Skewer Stick Water soluble black marker Water Procedure: Draw a ink dot with the marker on each one of the chromatography paper near the bottom end of the paper. Then stick or paper clip the…

Using Gel Filtration Chromatography Introduction: Chromatography techniques are one of the most useful methods use for the isolation or separation of macromolecule from a mixture. The most common used chromatography techniques are thin -layer chromatography (TLC), column chromatography, etc. The purpose of this lab exercise is using the gel filtration chromatography to separate different proteins from a mixture based on their size. Also, collecting elutes that run through the column, and…

Chromatography is a method of separation in which the components of a substance are separated and distributed between two phases, which are the mobile phases and stationary phase (1). There are multiple types of chromatograph, which includes paper chromatography, column chromatograph, retention chromatography, gas chromatography, liquid chromatography etc. Each technique has its own strengths and weaknesses, but all are useful in getting separations of different components (8). Since many…

Thin Layer chromatography was also use to separate the mixture of fluorene and fluorenone. Thin layer chromatography is a useful separation method when the components of a mixture are in question. Analyzing what the mixture consist of can be efficiently determine by TLC by calculating Rf and comparing these values. For the separation and isolation of a mixtures components, this method is extensive and not as efficient at column chromatography. This conclusion is represented in the data gathered.…

High performance liquid chromatography (HPLC) is a separation technique utilizing differences in distribution of compounds in two phases; called the stationary phase and the mobile phase. The stationary phase designates a thin layer created on the surface of fine particles and the mobile phase designates the liquid flowing over the particles. Under a certain dynamic condition, each component in a sample has a different distribution equilibrium depending upon the solubility in the phases and or…

According to Simon (2002) the silica gel is the most widely used absorbent and it is the stationary phase that is dominant in Thin layer chromatography, particles or molecules that are non-polar travel faster at the stationary phase and the molecules that are polar spend more time at the stationary phase and therefore this means that molecules that are non-polar travel larger distance on the TLC plate in comparison to molecules that are polar since the polar molecules stick to the polar…

The chromatography was performed on an ACQUITY TM UPLC system (Waters Corp., Milford, MA, USA). The UPLC system included quaternary solvent manager, a binary pump, degasser, autosampler with an injection loop of 10 μL and a column heater-cooler. The separation was performed on Acquity UPLC BEH TM C18 column (50 × 2.1 mm, i.d., 1.7 μm, Waters, USA) maintained at 40 °C. a mobile phase of acetonitrile containing 0.1% formic acid and aqueous 5 mM ammonium acetate buffer containing 0.1% formic acid…

The instrumentation of a HPLC apparatus is as shown in the diagram above. A HPLC apparatus is including a solvent reservoir, degasser, pump, injector, HPLC column, detector and data collection device. Solvent reservoir The solvent reservoir is usually a glassware or reagent bottle which contained the analytic sample. It is located at the upper part of the HPLC. It is also known as mobile phase reservoir since the sample will be moved. The flowing of the sample is depends on gravitational…

results are listed and taken into consideration to get an accurate comparison. 2. The type of column used is listed, this column contains the stationary phase and the mobile phase is pumped through the column under pressure. The dimensions of the column are listed as this will determine how much stationary phase is inside the column. As well as the size of the stationary phase particles, the amount in the column can determine how quickly the sample will pass through i.e. if there are a small…