Column chromatography

purified from rabbit sera by ammonium sulphate precipitation followed by negative ion exchange chromatography employing DEAE-cellulose. The antisera were decomplemented, IgG fraction precipitated by 40% (w/v) ammonium sulphate saturation in PB, pH 7.2 and the precipitate dialysed against the same buffer. The preparation thus obtained was dissolved in 1 mL of PB, pH 7.2 and loaded on to a DEAE-cellulose column equilibrated with PB. An inverse sigmoidal elution profile was obtained as shown in…

1.5 ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY (UPLC) Since from thirty years, High performance liquid Chromatography (HPLC) is well accepted technique used in laboratories worldwide. It is one of the analytical techniques used to identify, separate and quantify each analyte in a mixture. One of the primary drivers for the development of this technique has been the progression of the packing materials (particle size, packing technology, chemistry) used to impact the separation. The fundamental…

DISTILLATION The purpose of experiment 1, fractional distillation and gas chromatography, was to separate an unknown organic mixture into two liquids. The primary process of separation was fractional distillation, which utilizes the physical properties of the two unknown volatile liquids. Once the unknown mixture was successfully fractioned, gas chromatography was used to help determine the identity of the two unknown liquids. Fractional distillation and simple distillation both take advantage…

using affinity chromatography, LDH was further purified based upon its hydrophobicity. Using a ligand that had high affinity for enzymes with NAD+ cofactors, LDH was filtered from other enzymes that didn’t contain this property. Lastly, size exclusion chromatography was used to separate enzymes by their molecular weight. Knowing the molecular weight of LDH and using beads that had specific exclusion limits, the location of LDH was identified while the sample ran inside the column. Thereafter,…

In this experiment p-cresol was converted to n-propyl p-tolyl ether by using a modified form of the Williamson Ether method. The modified form used is called Phase Transfer Catalysis. The p-cresol was deprotonated by NaOH. The addition of tetrabutylammonium bromide provides the necessary help that the nucleophile needed to get into the organic phase in order to react with the electrophile. The reaction would have proceeded without this addition although it would have been a very slow reaction.…

This separation resulted in a HETP value of 8.114cm. Given the large HETP value, with normal value in the range of 3-7cm, this particular column was not very successful in separating toluene and cyclohexane. This could be seen by the low number of theoretical plates, 0.912. A theoretical plate is one cycle of evaporation and condensation in a distillation. In general, having more theoretical…

Separation of plasma proteins by molecular exclusion chromatography Sebastian Prisacariu 140013826 Introduction The components of blood is broken down into 3 parts. This breakdown is consisted of: Plasma which is a yellowish fluid that helps transport blood cells around the body and contains a small supply off proteins, platelets which are blood clotting agent, red blood cells which is a carry for oxygen and remover off carbon dioxide and white cells which is the bodies defence from infection…

The objective of Experiment one (both Week one and two) was the separation and examination of Ethanol from complex mixtures using solvent-solvent extraction and chromatography. Week one involved retrieving the concentration of Ethanol and the partition coefficient from standard solutions utilizing the calibration method. Week two was centered on the use of a standard addition method to find Ethanol concentrations in beer samples. Overall, the experiment addressed the issues associated with…

super-critical fluid chromatography[2,3], in urine by gas chromatography- mass spectrometry[4] and, simultaneously with its active metabolite in biological fluids, by HPLC[5- 10]. Amlodipine Besylate (ADB) )[FIG.1.B], chemically,…

mixture was quenched by adding water and extracted with methylene chloride. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and filtered. After removing the solvent, the crude product was purified by silica gel chromatography by using hexane/CH2Cl2 (4:1) as the eluent to afford the title compound as yellow solid (1.23 g, 76%). 1H NMR (400 MHz, CDCl3): δ 8.55 (s, 2H), 8.44 (s, 2H), 8.01 (dd, 2H), 7.76 (dd, 2H), 1.39 (s,…