Column chromatography

Methods Column chromatography was set up using a 60 ml syringe clamped to a ring stand. The syringe was plugged using a small amount of cotton. An Erlenmeyer flask was placed underneath the column to collect any excess material. Eleven 20 ml vials were labeled numbers 1 through 11. Silica gel was measured to 42 ml using a graduated cylinder and then moved into a 150 ml beaker. A slurry was created by adding 50 ml of hexane to the beaker containing the silica gel. The slurry was stirred for about one minute. This was the stationary phase for the column chromatography. Pour the slurry into the column that is plugged with cotton. Hexane was added to the beaker to move all the slurry into the column. More hexane was added to the column until the…

Introduction Separation, a process by which the distinguishing between substances through physical or chemical means in a mixture may be carried out. A widely employed technique for this purpose is called chromatography. 1 There are many different types of chromatography, including gas-liquid, paper, column and so on. 1 Chromatography differentiates by allowing the differences in polarity, vapor pressure, solubility and so on of different substances to be observed. 1 In our endeavor, we will be…

ABSTRACT: In this experiment, we determined the relative polarity of substances using the principles of chromatography. Three substances were tested using a thin layer chromatography. The column chromatography tested a mixture of two substance which were separated as a result of the chromatography. The results for both the thin layer and column chromatography supported the hypothesis. Therefore, chromatography is an effective tool that can be used to determine relative polarities. INTRODUCTION:…

Initially, the protein extract solution was subject to ion exchange column chromatography using SP Sepharose Fast Flow (GE Healthcare, Sweden). The column (with 5 ml of stationary phase) was equilibrated with five column volumes of sodium acetate/acetic acid 10mM pH5 and 10 ml of protein extract containing bacteriocin were injected into this. After washing the column with sodium acetate/acetic acid 10 mM pH5 (Buffer A), elution was performed using a linear gradient from sodium acetate/acetic…

瀀瀀ract The purpose of Isolation of Bixin from Annatto Seeds using Column Chromatography was to separate and purify macroscopic amounts of pure bixin compound from annatto seed ex-tract.1 A mixture of bixin, methylbixin, and norbixin were extracted from annatto seeds. Bixin was then isolated from the mixture through the use of the column chromatography technique. The isolated bixin was characterized through Thin Layer Chromatography (TLC) analysis, which displayed an Rf value of 0.317, which…

compounds and their formulations (Sasidharan et al., 2011). According to the World Health Organization (WHO), nearly 20,000 medicinal plants exist in 91 countries. The premier steps to exploit the biologically active compound from plant resources are extraction, pharmacological screening, isolation and characterization of bioactive compound, toxicological evaluation and clinical evaluation. The aim of the present study was to characterize the bioactive compounds from ethyl acetate fraction of…

A solution of butadiene and meso-tetraphenylporphyrin (100 mg, 0.162 mmol) dissolved in dichloromethane (100 ml, 1.57 mol) in a water-jacketed flask was irradiated with compact fluorescent lamps. The solution was then concentrated in vacuo and the product subjected to column chromatography to yield the product, aldehyde, and unreacted starting butadiene which could then be recycled. 1-Phenylbutadiene…

Introduction: The main purpose of this experiment was to take the Gas Chromatography/ Mass Spectrum part of the “Analysis of US bills for cocaine using gas chromatography / mass spectrometry” lab and improve upon the gas chromatographic results. This would be done by finding a way to separate the peaks of cocaine and doxepin. The lab done previously showed the use of two different methods to obtain samples of cocaine and doxepin for spiked dollar bills. The two extraction methods were…

Raju Chandra et al; A reliable and reproducible reversed-phase high performance liquid chromatography (RP-HPLC) was developed for the quantitative determination of quetiapine fumarate from marketed bulk tablets. The active ingredient of quetiapine fumarate separation achieved with C18 column using the methanol water mobile phase in the ratio of 30:70 (v/v). The active ingredient of the drug content quantify with UV detector at 359 nm. The retention time of quetiapine fumarate is 5.27 min. A good…

Protein X has a pI of 5.9, Protein Y has a pI of 7.6, and Protein Z has a pI of 8.5. For each of the methods below, describe how you would go about purfying Protein Y from Proteins X and Z (provide a pH that you would use and why.) Anion exchange chromatography: In anion exchange chromatography negatively charged ions will bind to the column. This will leave positively charged ions flushed through the column. By picking a pH of 6.5, protein X will flush through the column while protein Y and Z…